Tenascin-C regulates proliferation and migration of cultured astrocytes in a scratch wound assay.

Tenascin-C (TNC), an extracellular matrix glycoprotein, is involved in tissue morphogenesis like embryogenesis, wound healing or tumorigenesis. Astrocytes are known to play major roles in wound healing in the CNS. To elucidate the roles of TNC in wound closure by astrocytes, we have examined the morphological changes of cultured astrocytes in a scratch wound assay and measured the content of soluble TNC released into the medium. We have also localized the expression of TNC mRNA, TNC, glial fibrillary acidic protein (GFAP), vimentin and integrin beta1. After wounding, glial cells rapidly released the largest TNC isoform and proliferated in the border zones. Subsequently, they became polarized with unidirectional processes and finally migrated toward the denuded area. The proliferating border zone cells and pre-migratory cells intensely expressed TNC mRNA, TNC-, vimentin-, GFAP- and integrin beta1-like immunoreactivity, while the migratory cells showed generally reduced expression except the front. Exogenous TNC enhanced cell proliferation and migration, while functional blocking with anti-TNC or anti-integrin beta1 antibody reduced both of them. These results suggest that mechanical injury induces boundary astrocytes to produce and release TNC that promotes cell proliferation and migration via integrin beta1 in an autocrine/paracrine fashion.

Serial changes in signal intensities of the adjacent discs on T2-weighted sagittal images after surgical treatment of cervical spondylosis: anterior interbody fusion versus expansive laminoplasty.

BACKGROUND: There have been many reports about newly developed degenerative changes in the adjacent segments after anterior interbody fusion. It is a controversial issue whether the adjacent-segment disease in patients treated by anterior interbody fusion is the result of progressive cervical spondylosis at the adjacent levels or is caused by the arthrodesis. The aim of this study is to clarify the difference in postoperative effect on the adjacent segments between anterior interbody fusion and expansive laminoplasty. METHOD: This study included 14 patients who underwent pre- and postoperative MR images at 6 and 12 months. Seven patients underwent cervical interbody fusion and the other 7 patients underwent expansive laminoplasty. Disc degeneration was evaluated semiquantitatively by calculating the degenerative index (DI) that is a ratio of the intensity in the disc to that in the upper cervical cord. FINDINGS: In the anterior interbody fusion group, the adjacent disc intensities decreased within 12 months (F = 20.42; P < 0.01). The pre-operative mean DI was 0.59 +/- 0.16. The post-operative mean DIs were 0.56 +/- 0.16 at 6 months and 0.47 +/- 0.16 at 12 months. In the expansive laminoplasty group, the signal intensities of both the adjacent discs and the discs within the range of laminoplasty had no serial changes during the same period (F = 2.67; P = 0.09 and F = 0.15; P = 0.87 respectively). INTERPRETATION: Anterior interbody fusion had a significant influence on the adjacent discs even as soon as 12 months after surgery, but laminoplasty had no influence on them during the same period.

Radiographic and scintigraphic courses of union in cervical interbody fusion: hydroxyapatite grafts versus iliac bone autografts.

UNLABELLED: This study investigated the radiographic and scintigraphic courses of union in cervical interbody fusion using hydroxyapatite (HA) grafts or iliac bone autografts. METHODS: Twelve patients underwent both serial plain radiography and bone scintigraphy during the 12 mo after surgery. Serial plain radiographs were obtained every month until the end of the study period. Bone scintigrams with 99mTc-hydroxymethylene diphosphonate (HMDP) were obtained at 2 wk and at 1, 2, 3, and 6 mo. Uptake of 99mTc-HMDP in the graft was expressed as a ratio of the counts in the graft to those in the axis. RESULTS: In the HA graft group, the plain radiographs of all patients showed a radiolucent stripe that disappeared 7.3 +/- 1.5 (mean +/- SD) months after surgery. In the autograft group, a radiolucent stripe around the graft was not seen for any patient, and union was confirmed by follow-up radiographs within 6 mo after surgery. The serial changes in the 99mTc-HMDP uptake ratio showed no difference between the 2 groups. The 99mTc-HMDP uptake ratio peaked 1 mo after surgery and decreased rapidly to a plateau within 2 mo. CONCLUSION: In the HA graft group, despite the presence of a radiolucent stripe around the graft for more than 6 mo, the scintigraphic course of union was not different from that in the autograft group. The likelihood is that the presence of a radiolucent stripe around the HA graft in the early months after surgery is not always a sign of pseudoarthrosis.

Thresholds of ischemia salvageable with intravenous tissue plasminogen activator therapy: evaluation with cerebral blood flow single-photon emission computed tomographic measurements.

OBJECTIVE: This study investigated the cerebral blood flow (CBF) thresholds of ischemic cortices that were salvageable with intravenous tissue plasminogen activator (t-PA) infusion therapy. METHODS: We retrospectively reviewed data for 20 patients who were treated with intravenous low-dose (7.2 mg) native t-PA infusion therapy for distal embolic occlusions of middle cerebral artery divisions or branches, without early computed tomographic ischemic changes. All patients underwent pretreatment single-photon emission computed tomographic CBF measurements using (99m)Tc-N,N'-(1,2-ethylenediyl)bis-L-cysteine diethylester. Intravenous t-PA infusion was initiated within 6 hours (average, 3 h) after symptom onset for 14 patients and 6 to 14 hours (average, 8.8 h) after the last time the patient was noted to be in normal condition for the other 6 patients. Pretreatment single-photon emission computed tomographic and 3-month post-treatment computed tomographic scans were compared using computerized coregistration. Ischemic cortices in single-photon emission computed tomographic scans were divided into areas of reversible and irreversible ischemia. The degree of hypoperfusion was analyzed with an asymmetry index (AI). The AI was calculated as C(a)/C(b) x 100%, where C(a) represents the mean reconstructed counts for the ipsilateral ischemic area and C(b) represents the mean reconstructed counts for the corresponding contralateral area. RESULTS: Partial recanalization, with clinical improvement, at 60 minutes was confirmed by angiography for 14 of the 20 patients (70%). Seventeen of the 20 patients (85%) exhibited major neurological improvements (defined as decreases in National Institutes of Health Stroke Scale scores of > or =4 points) at 24 hours, suggesting that recanalization occurred within 24 hours for almost all patients. AIs for the 25 irreversible lesions ranged from 15.0 to 53.4% (37.3 +/- 11.6%), whereas AIs for the 38 reversible lesions ranged from 45.0 to 83.1% (69.3 +/- 8.6%). There was a significant difference in the AIs for these two groups (P < 0.0001). The ischemia in tissue with AIs of more than 53.4% was reversible. In contrast, ischemic tissue with AIs of less than 45.0% could not escape cerebral infarction with our treatment. The ischemia in tissue with AIs between 45.0 and 53.4% was reversible in some patients and irreversible in others. CONCLUSION: To save ischemic tissue with our intravenous t-PA infusion therapy, residual CBF should be at least 45% of the contralateral presumed normal CBF value. CBF thresholds for ischemia that would be surely salvageable with our intravenous t-PA infusion therapy might be approximately 50 to 55% of the contralateral presumed normal CBF values.

[An investigation of serum soluble interferon receptor levels in patients with renal cell carcinoma].

OBJECTIVE: Serum soluble interferon alpha/beta receptor (s-IFN-receptor) levels were determined in renal cell carcinoma (RCC) patients to study the clinical significance of the measurement. SUBJECTS AND METHODS: S-IFN-receptor levels were measured in RCC patients (n = 27) and healthy volunteers (n = 22) by enzyme immunoassay technique. RESULTS: Significantly higher serum s-IFN-receptor levels were observed in RCC patients compared with the healthy volunteers (p < 0.003). The high s-IFN receptor levels in the patients suggested seriousness and mal-prognosis of this disease. The 4-years survival rate of the higher level group (with the mean value of 2.7 +/- 1.7 ng/ml or more) was 53.3%, while the lower level group's rate was 78.7% (Statistical analysis result by Logrank (Mantel-Cox) test; p = 0.4289). CONCLUSION: Further study in more subjects is required to determine the feasibility of the s-IFN receptor levels as a prognosis marker, since correlation between the prognosis and s-IFN receptor level was not clarified by this study result.

Simultaneous up-regulation of urokinase-type plasminogen activator (uPA) and uPA receptor by hepatocyte growth factor/scatter factor in human glioma cells.

Several lines of evidence indicate that hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, c-Met, may play an important role in progression of human glioma. In this study, effects of HGF/SF on urokinase- type plasminogen activator (uPA)-mediated proteolysis network were examined in c-Met-positive human glioma cell lines. Treatment of the glioma cells with various concentrations of HGF/SF resulted in an enhanced secretion of uPA proteins accompanying increased transcription of uPA mRNA in a dose dependent fashion. The levels of uPA receptor (uPAR) mRNAs were also elevated simultaneously upon HGF/SF stimulation, and the cell-surface associated uPA activity was also elevated by the treatment. Since concomitant expression of HGF and its receptor c-Met are frequently observed in malignant gliomas, these results suggest that HGF/SF participates in invasive process of malignant glioma cells not only by its motility-stimulating activity but also through enhanced degradation of the extracellular matrix induced by autocrine activation of uPA proteolysis network.

Expression of heat shock proteins in developing and degenerating rat testes.

In the testis, several types of heat shock proteins (HSPs) have been identified and characterized, although the cellular basis of the HSPs remains elusive. In the present study, alterations in the cellular localization of HSPs, including HSP 25, 60, 70, and 90, were studied during the developing and degenerating periods in the rat testis using immunohistochemistry and Western blotting. HSP25 was expressed in neither germ cells nor somatic cells on all days examined. In contrast, HSP 60 was expressed in Leydig cells during neonatal and prepuberty periods, and only in spermatogonia and primary spermatocytes after puberty. HSPs 70 and 90 were expressed in germ cells, Sertoli cells, and Leydig cells during neonatal and early developing testes, and in spermatocytes and round spermatids after puberty. Besides, there was faint expression of HSP 90 protein in spermatogonia in this period. In the degenerative condition, all HSP proteins were markedly expressed in germ cells after surgery. It would appear that HSPs play roles in unique homeostasis in testes.

Magnetic resonance imaging and angiographic appearance of meningioma of the fourth ventricle--two case reports.

A 47-year-old female presented with a transitional type meningioma entirely confined to the fourth ventricle. The tumor was totally resected. A 67-year-old female had undergone resection of an intraventricular transitional type meningioma of the fourth ventricle 9 years previously. She presented with a new meningioma of an obviously different origin at the posterior rim of the foramen magnum. The new meningioma was totally resected and the histological diagnosis was atypical meningioma. The magnetic resonance (MR) imaging characteristics of these two intraventricular meningiomas of the fourth ventricle were isointense and slightly hypointense to gray matter on the T1-weighted images, and hyperintense to gray matter on the T2-weighted images, with intense and homogeneous enhancement with gadolinium. Angiography showed the two intraventricular meningiomas fed by branches of the posterior inferior cerebellar arteries or superior cerebellar arteries. The second meningioma with dural attachment was fed by the right occipital artery. Intraventricular meningiomas of the fourth ventricle are not supplied by meningeal branches from vertebral and external cerebral arteries. MR imaging is the most useful tool in preoperative diagnosis, but cerebral angiography should also be performed to confirm the feeding vessels and the correct diagnosis.

[Monoclonal antibody against an urinary protein secreted from the renal tubules--immunohistochemical analysis of renal cell carcinoma].

BACKGROUND: Monoclonal antibodies (mABs) against urinary proteins originated from renal tubules were obtained in order to find a possible tumor marker for renal cell carcinoma. METHODS: Urine samples were collected from healthy adult males. After removal of Thamm-Horsfall protein (THP) by sodium chloride precipitation, supernatant urine was ultrafiltered, thus, THP-free urinary proteins were obtained. Then, proteins of plasma origin were removed by circulating the urinary proteins through an affinity column which contained immobilized rabbit anti-human whole serum immunoglobulins. BALB/c mice were immunized with the THP- and plasma protein-free urinary proteins. The spleen cells were fused with mouse myeloma cells. Hybridomas were screened by indirect immunofluorescence of fresh frozen kidney tissue sections using culture supernatants. RESULTS: Five hybridomas producing mAbs which reacted with renal tubular segments were established. One mAb reacted strongly with distal renal tubules. Using this mAb, formalin fixed and paraffin embedded sections of 39 renal cell carcinomas were stained by immunoperoxidase method. Twenty-six of the 39 cancer tissues (66.7%) were stained positive. According to the histological classification, positive staining rate of clear cell subtype, and mixed subtype was 75% (6/8), 80% (8/10) and 66.7% (6/9), respectively. According to the tumor grade, positive staining rate of G1, G2, G3 was 66.7% (6/9), 71.4% (15/21) and 55.5% (5/9), respectively. CONCLUSION: These findings indicate that most of renal cell carcinomas were originated from common precursor cells for both proximal and distal tubules, because normal proximal tubular cells were not stained by the mAb.

Identification of a novel protein (VBP-1) binding to the von Hippel-Lindau (VHL) tumor suppressor gene product.

The tumor suppressor VHL gene product (pVHL), recently reported to bind to elongins B and C, is thought to regulate transcription elongation. To establish whether the VHL gene may have other functions, we here searched for additional cellular protein(s) that might bind to pVHL using a two-hybrid system and identified seven independent clones, including elongin C, but not elongin B. Three clones (unknown, imidopeptidase, and unknown) presumably bind to the N-terminal nonconserved region, whereas the four other clones [elongin C, the HIV tat-binding protein, the actin-binding protein Filamin (ABP280), and the HIBBJ46 (named VBP-1)] were found to bind to the wild-type pVHL but not to a C-terminal 156-amino acid deletion mutant. Interestingly, the HIV tat-binding protein and Filamin could bind to C-terminal 26-amino acid deleted pVHL, but elongin C and VBP-1 failed to do so. Thus, elongin C and VBP-1 require the C-terminal end of pVHL for binding. It was also established that epitope-tagged pVHL strongly forms complexes with VBP-1 in vivo using immunoprecipitation Western blotting analysis. VBP-1 was widely expressed in various cell lines tested, in which VHL mRNA can be detected. When the VBP-1 protein was solely expressed, it located to the cytoplasm and did not localize to the nucleus. However, when coexpressed with VHL, it can translocate to the nucleus. These results indicate that VBP-1 can form a complex with VHL protein in vivo and hence VHL affects the intracellular localization of VBP-1 protein.

Mobile sediment of CaCO3 grains inside the cyst of an intraventricular teratoma. Case report.

The case of a 40-year-old woman with a mature cystic teratoma in the lateral ventricle is reported. An unusual mobile sediment of radiopaque material in the cyst was identified as grains of calcium carbonate. It was considered to have precipitated from the cystic fluid under specific chemical conditions.

Rhinocerebral mucormycosis--case report.

A rare case of rhinocerebral mucormycosis occurred in a 74-year-old diabetic male with gradually progressive right visual loss and total ophthalmoplegia. Computed tomography and magnetic resonance imaging revealed an invasive right orbital apex mass, destroying the medial wall of the orbit and extending into the right cavernous sinus and right middle fossa. Laboratory data demonstrated no signs of inflammation. A carcinomatous lesion originating in the paranasal sinuses and extending into the intracranial space was diagnosed. The mass was totally removed through a subfrontal approach to confirm the histological diagnosis and decompress the optic nerve. The histological diagnosis was mucormycosis. Despite aggressive medical therapy, dissemination resulted in mucor pleuritis and mucor encephalitis or meningitis. He died of septic shock and acute renal failure.