Single-Component Pheromone Consisting of Bombykal in a Diurnal Hawk Moth, Neogurelca himachala sangaica.

Recent work has suggested that hawk moths share pheromone components but are sexually separated by qualitative and quantitative differences in their pheromone blends. During field assays on the sex pheromones of other species, a diurnal hawk moth, Neogurelca himachala sangaica (Lepidoptera: Sphingidae), was frequently captured, but the composition of the sex pheromone of this species was not known. Analysis of hexane extracts of the pheromone glands of calling female by gas chromatography (GC) using an electroantennographic detector (EAD) revealed two components that elicited EAD responses from male moth antennae. These components were identified by their mass spectra and retention indices on two GC columns as (10E,12Z)-10,12-hexadecadienal (E10,Z12-16:Ald) and a trace of its (10E,12E)-isomer (E10,E12-16:Ald) in 98:2 ratio. In field experiments, E10,Z12-16:Ald alone attracted male moths, and addition of E10,E12-16:Ald significantly reduced the attractiveness, even at the naturally-occurring ratio. Analysis of the data using a generalized linear mixed model showed that E10,Z12-16:Ald positively contributed to attractiveness, whereas E10,E12-16:Ald did so negatively, and it was concluded that the sex pheromone of N. himachala sangaica consists solely of E10,Z12-16:Ald, bombykal. The negative effect of E10,E12-16:Ald on attractiveness could promote the species-specificity of this single-component pheromone system.

Chondromodulin-1 directly suppresses growth of human cancer cells.

BACKGROUND: Chondromodulin-1 (ChM1), an endogenous anti-angiogenic factor expressed in cartilage, has been suggested to inhibit invasion of endothelial cells into cartilage. In addition, the ectopic administration of ChM1 has been reported to suppress tumorigenesis in vivo. However, it is unclear whether the anti-tumor effect is due to not only the anti-vascularization effect of ChM1, but also its direct action against oncocytes. In the present study, we sought to determine whether ChM1 has a direct action on tumor cells. METHODS: BrdU incorporation assay was performed on human umbilical vein endothelial cells (HUVECs), normal human dermal fibroblasts (NHDFs), HepG2 cells and HeLa cells in the presence or absence of recombinant human ChM1 (rhChM1). An adenovirus that expresses ChM1, Ad-ChM1, was established and applied to the tumor xenografted in vivo, and to in vitro tumor cells cultured on plates or in soft agar. Cell cycle-related proteins and the phosphorylation of Erk, Akt, and GSK3beta, the downstream molecules of the extracellular matrix-integrin signaling pathways, in HepG2 cells treated with or without Ad-ChM1 were detected by western blot analysis. Luciferase reporter assays of STAT, GAS, and ISRE, which participate in another cytokine signaling pathway, ware performed in HepG2, HeLa, and HUVEC cells. RESULTS: ChM1 suppressed BrdU incorporation in HUVECs and in HepG2 cells dose-dependently, but did not suppress BrdU incorporation in NHDFs and HeLa cells cultured on plates. In soft agar, however, ChM1 suppressed the growth of HeLa cells, as well as HepG2 cells. Western blot analyses demonstrated that ChM1 decreased the levels of cyclin D1, cyclin D3, and cdk6 and increased those of p21cip1 without affecting the phosphorylation levels of Erk, Akt, and GSK3beta in HepG2 cells. The luciferase reporter assay demonstrated that ChM1 suppressed the transcriptional activities of STAT and GAS but not of ISRE. CONCLUSION: ChM1 directly suppressed the proliferation of tumor cells in an anchorage-independent manner. However, ChM1 did not alter the phosphorylation of downstream molecules, at which the signaling pathways through growth factor and cytokine receptors converge with the anchorage-dependent pathway. Our results show that ChM1 has a direct anti-tumor effect; moreover, this effect occurs by inhibiting the STAT signaling pathway.

[Case of infected subdural hematoma diagnosed by diffusion-weighted imaging].

An 80-year-old man who had undergone total gastrectomy and splenectomy for gastric cancer 13 years ago presented with headache, drowsiness, and high fever 1 month after a traffic accident. Brain CT scans revealed bilateral subdural fluid collections. Diffusion-weighted imaging (DWI) showed mixed high and low signal intensities in the left subdural fluid, and contrast-enhanced MR imaging revealed capsule enhancement of the left subdural fluid collection. The patient was diagnosed with left subdural empyema, and 2 burr-holes were drilled for drainage and irrigation. Operative findings revealed a neomembrane underneath the dura mater. Old hematoma and yellowish-white purulent fluid were present within the neomembrane. This confirmed the diagnosis of infected subdural hematoma (ISH). Abscess culture results were positive for Escherichia coli. The patient's symptoms resolved postoperatively with subsequent antibiotic therapy. However, 4 months after the operation, he suddenly died of severe sepsis and disseminated intravascular coagulation following cholecystitis, which was possibly associated with splenectomy. The clinical presentation, diagnosis, and treatment of an unusual case of ISH have been discussed. We emphasize that DWI and enhanced MR imaging may be useful for diagnosing ISH, and serial DWI evaluations may help in monitoring the therapeutic response in ISH.

Glucocorticoid inhibits bone regeneration after osteonecrosis of the femoral head in aged female rats.

Idiopathic osteonecrosis of the femoral head (ION) is a painful disease of the hip, the pathogenic mechanism of which is still unclear. The most common extraneous factor is steroid treatment. Steroids have inhibiting effects on bone formation and resorption. When bone regenerative treatments are indicated for ION patients who are exposed to steroids, we cannot ignore the effects of corticosteroid itself on bone healing. The aim of this study was to investigate the effects of glucocorticoid on bone regeneration after osteonecrosis of the femoral head in a rat model. Osteonecrosis was induced surgically on the left femoral heads of aged female rats (about 6 months old) on day 0. Methylprednisolone sodium succinate (MPSS) or normal saline was administrated at a dose of 100 mg/kg/day from day 7 to 11. Femoral heads were analyzed histologically. There were no pathological findings in the right femoral heads of the MPSS-treated and saline-treated rats, as control for the contralateral injury. The newly formed bone volume and the osteoclast number were significantly smaller in the MPSS-treated group. The normal bone marrow was regenerated in the saline-treated group, whereas most of the bone marrow space still contained fibroblast-like spindle-shaped cells in the MPSS-treated group on day 42. Alkaline phosphatase-positive cells were only seen in the areas around the regenerated bone marrow cavities. Thus, MPSS inhibits bone formation by suppressing osteoblast proliferation and resorption by suppressing the recruitment of osteoclast precursors. These findings may be useful when designing treatments for ION patients exposed to steroids.

High dose glucocorticoid hampers bone formation and resorption after bone marrow ablation in rat.

We analyzed the effect of glucocorticoid on bone regeneration after bone marrow ablation in tibiae of 8-week-old rats. Methylprednisolone sodium succinate (MPSS) was injected intramuscularly at a dose of 100 mg/kg/day for 3 days. Tibiae on days 1, 3, 5, 7, 10, 12, and 14 after ablation were subjected to tartrate-resistant acid phosphatase staining, immunohistochemistry, in situ hybridization, and transmission electron microscopy (TEM), and measurement of the volume of newly-formed bone and the osteoclast number. MPSS significantly decreased the newly-formed bone volume on day 7, and immature bone still remained on day 10 in the MPSS-treated group. The volume of this bone was significantly higher than that in the control group. However, there were no differences between the groups in the osteoclast number, the expression of mRNAs for osteoblast differentiation markers, and alkaline phosphatase and cathepsin K judged by immunohistochemistry. TEM findings showed no difference in the form of osteoblasts, whereas osteoclasts in the MPSS-treated group had less developed ruffled borders, compared to those in the control group. These results suggest that MPSS treatment affects neither the differentiation nor the shape of osteoblasts, and does not change the osteoclast number or the cathepsin K level. However, high dose MPSS inhibits both bone formation and resorption during bone regeneration after rat tibial bone marrow ablation, and inhibits ruffled border formation in osteoclasts. These data will be useful to develop bone regenerative therapies for bone diseases due to high dose steroid administration.

Preoperative MRI analysis of patients with idiopathic scoliosis: a prospective study.

STUDY DESIGN: A prospective trial of preoperative MRI study in patients with "idiopathic" scoliosis. OBJECTIVES: To investigate the prevalence of neural axis malformations and the clinical relevance of MRI in the evaluation of patients with idiopathic scoliosis undergoing surgical intervention. SUMMARY OF BACKGROUND DATA: With the development of MRI, neural axis abnormalities such as syringomyelia or Chiari malformations are increasingly being found in patients with "idiopathic" scoliosis. The risk of neurologic complications during correction of scoliosis without prior decompression surgery for syringomyelia has been documented; however, there have been no prospective studies for identifying the risk of neurologic complications as a result of scoliosis surgery in patients with asymptomatic neural axis malformations. METHODS: A total of 250 patients who were classified as having "idiopathic" scoliosis at first presentation and admitted for spinal surgery were evaluated. All patients were examined for neural axis abnormalities using MRI. The presence of neurologic symptoms and abnormal neurologic signs was also examined before and after surgical intervention. Neurologic complications during scoliosis surgery were reviewed in patients with neural axis abnormalities. RESULTS: There were 44 (18%) patients (13 males and 31 females) who had neural axis abnormalities on MRI, including syringomyelia with Chiari malformations in 22 patients, syringomyelia with tonsillar ectopia in 2, Chiari malformations in 13, tonsillar ectopia in 6, and low conus medullaris in 1. On clinical examination, 44 (18%) patients had abnormal neurologic signs and 26 (7%) patients complained of headache or back pain. There were significant differences between patients with and without neural axis abnormalities regarding the age at first visit, gender, curve pattern, sagittal profile of thoracic spine, presence of neurologic deficit, and complaint of pain. Only 12 of 44 patients needed neurosurgical treatment for foramen magnum decompression before correction of scoliosis. Neurologic status temporarily worsened in 3 patients, including 2 patients with neurosurgical treatment and 1 patient without neurosurgical treatment; however, there were no permanent neurologic complications as a result of scoliosis surgery. All patients without neurologic deficits or complaints of pain did not receive neurosurgical treatment, while they had no permanent neurologic complications. CONCLUSIONS: Foramen magnum decompression for neural axis malformations could prevent permanent neurologic complications during scoliosis surgery. There is little risk of neurologic complications in patients with "idiopathic" scoliosis whose neurologic status is normal, even if these patients have a neural axis malformation on MRI.

High-level expression of the coxsackievirus and adenovirus receptor messenger RNA in osteosarcoma, Ewing's sarcoma, and benign neurogenic tumors among musculoskeletal tumors.

PURPOSE: The sensitivity of human tumor tissues to infection with recombinant adenoviruses correlates with the expression of the coxsackievirus and adenovirus receptor (CAR). CAR has been shown to function as the primary receptor for adenoviruses and to play a critical role in adenovirus entry into host cells. It is important for clinical gene therapy to determine the expression level of CAR in tumor tissues. EXPERIMENTAL DESIGN: We analyzed the expression of CAR mRNA in 154 musculoskeletal tumor tissues from 154 patients and 10 normal mesenchymal tissues from 3 patients using reverse transcription-PCR and real-time quantitative PCR. An adenovirus infection assay was performed in two cell lines that were established from CAR-positive osteosarcoma tissue and CAR-negative malignant fibrous histiocytoma tissue. RESULTS: Ninety-nine of 154 tumors were detected as CAR positive by reverse transcription-PCR. We found that the expression levels of CAR mRNA varied markedly between different tumors as determined by real-time quantitative PCR. CAR mRNA was expressed at high levels in osteosarcoma, Ewing's sarcoma, neurofibroma, and schwannoma; at intermediate levels in exostosis, giant cell tumor, liposarcoma, synovial sarcoma, malignant peripheral nerve sheath tumor, and hemangioma; and at low levels in alveolar soft part sarcoma and desmoid. Whereas the osteosarcoma cell line that expressed a high level of CAR mRNA, like its parent tumor, had a high efficiency of adenovirus infection, the malignant fibrous histiocytoma cell line with almost undetectable expression of CAR mRNA, like its parent tumor, had a low efficiency of infection. CONCLUSIONS: Our data showed the great variations in CAR mRNA expression among human musculoskeletal tumors and mesenchymal tissues and implicated the potential usefulness of adenoviral vectors in gene therapy for osteosarcoma, Ewing's sarcoma, neurofibroma, and schwannoma. Efficient transduction with adenovirus for gene therapy could be realized in appropriate, sensitive tumor types.

Automatic measurement of polyethylene modification in metal-backed artificial hip joints using three-dimensional CT.

A new method has been developed for automatic measurement of polyethylene linear modification using three-dimensional CT in total hip arthroplasty (THA) and bipolar hemiarthroplasty (BHP). We obtained a three-dimensional digital image of the metal components by widening the maximum window width, adjusting the proper cutoff threshold level, and removing the metal artifact. The centric coordinates of both the metal-backed cup and the femoral head were calculated from this image. Modification was defined as a change in distance between those two points from their original interval. Phantom studies of the accuracy and reproducibility of the method indicated that the average error ranged from 0.02 to 0.12 mm and the standard deviation ranged from 0.01 to 0.05 mm. Clinical in vivo measurement was performed without error of computer software on 19 hips in which modification of highly cross-linked polyethylene components was significantly large.

Chondromodulin-I expression in rat articular cartilage.

The localization and expression of chondromodulin-I (ChM-I), an angiogenesis inhibitor, in the rat articular cartilage during maturation from 2 to 10 weeks of age were examined by immunohistochemistry, Western blot analysis and ribonuclease protection assay, and the results were compared with those in the epiphyseal cartilage. ChM-I was found to be diffusely immunostained in the inter-territorial space of the cartilage matrix from the intermediate to the deep layers at the immature stage. As the articular cartilage matured, the immunoreactivity was localized around the hypertrophic chondrocytes in the deep layer and the immunoreactivity became weak after maturation. In contrast, the ChM-I immunoreactivity was intense in the epiphyseal cartilage at all ages examined. ChM-I was detected by Western blotting as a broad band or occasionally as a cluster of multiple bands (approximately 25 kDa) in both the articular and the epiphyseal cartilage. The intensity of the bands decreased gradually with age in the articular cartilage, but was unchanged in the epiphyseal cartilage at all ages. Ribonuclease protection assay revealed that ChM-I mRNA also decreased gradually with age in the articular cartilage in parallel with the maturation of the articular cartilage, while no decrease in ChM-I mRNA was found in the epiphyseal cartilage. The expression of ChM-I mRNA in the articular cartilage was less than that in the epiphyseal cartilage at all ages. The decrease in amount of ChM-I in the mature articular cartilage suggests that ChM-I plays a more important role in the maintenance of avascularity in the immature articular cartilage than in the mature one. The avascular condition may be preserved by angiogenic inhibitors or mechanisms other than ChM-I in the mature articular cartilage.

Idiopathic scoliosis as a presenting sign of familial neurologic abnormalities.

STUDY DESIGN: Familial cases of "idiopathic" scoliosis associated with neurologic abnormalities are reported with a review of the literature. OBJECTIVE: To investigate the prevalence of neurologic abnormalities such as syringomyelia, Chiari 1 malformation, and tonsillar ectopia in patients with genetically determined "idiopathic" scoliosis. SUMMARY OF BACKGROUND DATA: Idiopathic scoliosis is widely considered to be a genetic disorder of unknown etiology. Magnetic resonance imaging (MRI) studies have shown that several cases of "idiopathic" scoliosis show neurologic abnormalities including syringomyelia and Chiari 1 malformation. Recently, several familial cases of either syringomyelia or Chiari malformation were reported, and it is suspected that genetic factors may influence the development of the craniovertebral malformation. It was hypothesized that some cases of "idiopathic" scoliosis include a craniovertebral malformation that is genetically determined. METHODS: This study, using clinical examinations and MRI, investigated 71 patients with scoliosis and a family history of "idiopathic" scoliosis in third-degree relatives for the presence of neurologic abnormalities. If neurologic abnormalities were confirmed with MRI, the relatives affected with scoliosis were also examined. RESULTS: Nine (13%) patients showed neurologic abnormalities on MRI. Magnetic resonance imaging showed syringomyelia with Chiari 1 malformation in four patients, Chiari 1 malformation in three patients, and tonsillar ectopia in two patients. Among the relatives of these patients, 4 of 15 individuals affected with scoliosis also showed neurologic abnormalities on MRI. CONCLUSIONS: It is suggested that familial neurologic abnormalities may have a wide range of expression, and that some patients with "idiopathic" scoliosis present with genetically determined craniovertebral malformations such as syringomyelia, Chiari 1 malformation, and tonsillar ectopia.

Association between estrogen receptor gene polymorphisms and curve severity of idiopathic scoliosis.

STUDY DESIGN: Analysis of the estrogen receptor gene of girls with idiopathic scoliosis. OBJECTIVES: To determine whether estrogen receptor gene polymorphisms correlate with curve severity of adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Studies suggest that idiopathic scoliosis is a familial condition and that curve progression is related to genetically determined factors, such as skeletal and sexual growth. METHODS: A total of 304 girls with idiopathic scoliosis were followed until growth maturation. Height, arm span, menarcheal age, and age at growth maturation were recorded, and curve severity was measured using Cobb's method. The estrogen receptor gene, which contains polymorphic PvuII and XbaI sites, was amplified from lymphocyte deoxyribonucleic acid by polymerase chain reaction. RESULTS: The mean maximum Cobb measurements for patients with genotypes XX and Xx were greater than for those with genotype xx (P = 0.002). The risk of curve progression, defined as progression of >5 degrees from initial evaluation, was higher with genotype Xx than with xx (P = 0.03). Patients with genotypes XX and Xx had a significantly higher risk for operative treatment than those with genotype xx (21.4%, 24.7% vs. 7.6%, P< 0.001). Growth examination around the time of the growth spurt revealed that the XbaI site polymorphism was also related to the age of growth maturation. The frequency of patients with growth maturation at >or=16 years was higher for genotypes XX and Xx than for genotype xx (33.3%, 29.9% vs. 16.8%, P= 0.013). CONCLUSION: Our results suggest that the XbaI site polymorphism is associated with curve severity. DNA analysis may predict curve progression.

Prediction of curve progression in idiopathic scoliosis from gene polymorphic analysis.

Three hundred and four girls with adolescent idiopathic scoliosis were investigated to determine if DNA polymorphisms in the vitamin D receptor (VDR), estrogen receptor (BR), and CYP17 gene were related to curve progression of idiopathic scoliosis. The results suggested that XbaJ site polymorphism in the ER gene was associated with curve progression. The Cobb's curve angle with genotype XX and Xx was statistically greater than that with genotype xx. The curve progression risk (approximately 5 degrees) was higher for genotype XX and Xx than for genotype xx. Furthermore, patients with genotype XX and Xx had a higher risk of receiving operative treatment than those with genotype xx. In conclusion, DNA analysis may predict curve progression, although other polymorphisms were not associated with curve severity.