Recovery process and prognosis of aphasic patients with left putaminal hemorrhage: relationship between hematoma type and language modalities.

To elucidate the precise recovery process and prognosis of language functions in aphasic patients with left putaminal hemorrhage, we investigated 48 aphasic patients classified into 4 groups according to the location and extent of hematoma. The hematoma extended to the corona radiata in all patients, extracapsular in type I (12 cases), to the anterior limb in type II (10 cases), to the posterior limb in type III (12 cases), and to both limbs in type IV (14 cases). The Standard Language Test for Aphasia was performed at 1 month, 3 months, and 6 months after the attack. The type II, III, and IV patients were divided into 2 groups, with and without ventricular rupture of the hemorrhage. At 3 and 6 months after the attack, the type I, II, and III patients showed significant improvement (P < .05) in all language modalities compared with the type IV patients. Most improvement in language modalities occurred in the first 3 months. The evaluation of patients with ventricular rupture after 6 months revealed poor recovery (P < .05) in oral commands, visual commands, confrontation naming, sentence repetition, narratives, verbal fluency, and writing in type II and III patients. In type IV patients, this evaluation showed poor recovery (P < .05) only in oral and written naming (kanji words). No significant difference in prognostic outcome was observed between the surgical treatment group and the nonsurgical treatment group. The classification of hemorrhage may be useful in predicting the outcome of aphasia with putaminal hemorrhage and in guiding clinicians in providing effective instructions to patients and their relatives.

Acute onset of intracranial subdural hemorrhage five days after spinal anesthesia for knee arthroscopic surgery: a case report.

INTRODUCTION: Spinal anesthesia is a widely used general purpose anesthesia. However, serious complications, such as intracranial subdural hemorrhage, can rarely occur. CASE PRESENTATION: We report the case of a 73-year-old Japanese woman who had acute onset of intracranial subdural hemorrhage five days after spinal anesthesia for knee arthroscopic surgery. CONCLUSION: This case highlights the need to pay attention to acute intracranial subdural hemorrhage as a complication after spinal anesthesia. If the headache persists even in a supine position or nausea occurs abruptly, computed tomography or magnetic resonance imaging of the brain should be conducted. An intracranial subdural hematoma may have a serious outcome and is an important differential diagnosis for headache after spinal anesthesia.

Laminin isoforms and their integrin receptors in glioma cell migration and invasiveness: Evidence for a role of alpha5-laminin(s) and alpha3beta1 integrin.

Glioma cell infiltration of brain tissue often occurs along the basement membrane (BM) of blood vessels. In the present study we have investigated the role of laminins, major structural components of BMs and strong promoters of cell migration. Immunohistochemical studies of glioma tumor tissue demonstrated expression of alpha2-, alpha3-, alpha4- and alpha5-, but not alpha1-, laminins by the tumor vasculature. In functional assays, alpha3 (Lm-332/laminin-5)- and alpha5 (Lm-511/laminin-10)-laminins strongly promoted migration of all glioma cell lines tested. alpha1-Laminin (Lm-111/laminin-1) displayed lower activity, whereas alpha2 (Lm-211/laminin-2)- and alpha4 (Lm-411/laminin-8)-laminins were practically inactive. Global integrin phenotyping identified alpha3beta1 as the most abundant integrin in all the glioma cell lines, and this laminin-binding integrin exclusively or largely mediate the cell migration. Moreover, pretreatment of U251 glioma cells with blocking antibodies to alpha3beta1 integrin followed by intracerebral injection into nude mice inhibited invasion of the tumor cells into the brain tissue. The cell lines secreted Lm-211, Lm-411 and Lm-511, at different ratios. The results indicate that glioma cells secrete alpha2-, alpha4- and alpha5-laminins and that alpha3- and alpha5-laminins, found in brain vasculature, selectively promote glioma cell migration. They identify alpha3beta1 as the predominant integrin and laminin receptor in glioma cells, and as a brain invasion-mediating integrin.

Growth potential of orbital cavernous hemangioma suggested by vascular endothelial growth factor and its receptor flk-1.

Orbital cavernous hemangiomas (CHs) manifest as slowly developing symptoms indicative of slow growth. The present study investigated the involvement of angiogenic factors and their receptors in the growth of orbital CHs. Surgical specimens of orbital CHs were obtained from nine patients. Formalin-fixed, paraffin-embedded specimens were stained immunohistochemically using antibodies against Ki-67, CD31, alpha-smooth muscle actin (alpha-SMA), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and VEGF receptors (flt-1 and flk-1). CD31 was expressed in the single layer of endothelial cells lining the vascular cavity. The thick vascular walls were positive for alpha-SMA, indicating that the vascular walls were smooth muscle cells. Ki-67 antigen immunostaining was mostly positive in the vascular walls and the staining index ranged from 0% to 6.8% (mean +/- standard deviation, 2.7 +/- 1.9%). VEGF and bFGF immunostaining were positive in all specimens. Flt-1 immunostaining was negative in all specimens, but flk-1 immunostaining was positive in both endothelial cells and smooth muscle cells. These results suggest that both VEGF and its receptor flk-1 are important in the growth of orbital CH.

Participation of an abnormality in the transforming growth factor-beta signaling pathway in resistance of malignant glioma cells to growth inhibition induced by that factor.

OBJECT: Malignant glioma cells secrete and activate transforming growth factor-beta (TGFbeta) and are resistant to growth inhibition by that factor. Nevertheless, the mechanism underlying this effect remains poorly understood. In this study, the mechanism of the resistance to growth inhibition induced by TGFbeta was investigated. METHODS: The authors examined the expression of downstream components of the TGFbeta receptor, including Smad2, Smad3, Smad4, and Smad7, and the effect of TGFbeta1 treatment on the phosphorylation of Smad2 and the nuclear translocation of Smad2 and Smad3 by using 10 glioma cell lines and the A549 cell line, which is sensitive to TGFbeta-mediated growth inhibition. The expression of two transcriptional corepressor proteins, SnoN and Ski, and the effect of TGFbeta1 treatment on the expression of the SnoN protein and the cell cycle regulators p21, p15, cyclin-dependent kinase-4 (CDK4), and cyclin D1 were also examined. Expression of the Smad2 and Smad3 proteins was lower in the glioma cell lines than in the A549 cell line and in normal astrocytes. In particular, Smad3 expression was low or very low in nine of the 10 malignant glioma cell lines. Expression of Smad4 was low in four glioma cell lines, and expression of the Smad7 protein was similar when compared with protein expression in the A549 cell line and in normal astrocytes. The levels of Smad2 phosphorylation after TGFbeta1 treatment were lower in glioma cell lines than in the A549 cell line, except for one glioma cell line. Seven of the 10 glioma cell lines exhibited lower levels of nuclear translocation of Smad2 and Smad3, and two cell lines that expressed very low levels of Smad3 protein showed no nuclear translocation. All glioma cell lines expressed the SnoN protein and its expression was unaltered by treatment with TGFbeta1. Three glioma cell lines expressed high levels of the Ski protein. The expression of the p21(cip1), p15(INK4B), CDK4, and cyclin D1 proteins was not altered by TGFbeta1, treatment, except in one cell line that displayed a slight increase in p21 protein. Overall, the expression of the Smad2 and Smad3 proteins was low in the glioma cell lines, the phosphorylation and nuclear translocation of Smad2 and Smad3 were impaired, and the TGFbeta receptor signal did not affect the expression of the SnoN, p21, p15, cyclin D1, and CDK4 proteins. CONCLUSIONS: These results suggest that the ability to resist TGFbeta-mediated growth inhibition in malignant glioma cells is due to abnormalities in the TGFbeta signaling pathway.

Atypical meningioma with large cyst. Case report.

A 29-year-old male presented with loss of consciousness and generalized seizure, followed by right hemiparesis and speech disturbance. Computed tomography and magnetic resonance imaging showed a solid, enhanced tumor with a cyst in the left frontal area with surrounding edema and mild mass effect. The cyst wall was also enhanced. The preoperative diagnosis was cystic falx meningioma. The tumor was totally resected, but most of the cyst wall adhered tightly to the surrounding brain and could not be removed. Histological examination revealed atypical meningioma and tumor cells in the cyst wall. The patient received local radiotherapy to the residual cyst wall with a total dose of 50 Gy.

Reduced expression of the transporter associated with antigen processing 1 molecule in malignant glioma cells, and its restoration by interferon-gamma and -beta.

OBJECT: It remains unclear whether malignant glioma cells can deliver tumor antigens efficiently to major histocompatibility complex (MHC) Class I molecules. To elucidate the mechanism of antigen presentation in malignant gliomas, the authors examined the expression of the transporter associated with antigen processing 1 (TAP1), which transports antigens to MHC Class I molecules, and low-molecular-mass polypeptide 2 (LMP2), which is a subunit of a proteasome. They also analyzed the effects of interferon (IFN)-gamma and IFN-beta on the expression of these molecules. METHODS: Five glioma cells expressed undetectable or very low levels of TAP1 protein and did not express TAP1 messenger (m)RNA. Normal brain tissue and glioma tissue specimens also showed undetectable levels of TAP1 protein and the same levels of LMP2 protein as lymphoblastoid B cells. Treatments of the tumor cells with IFN-gamma, or -beta enhanced the expression of both TAP1 protein and mRNA as well as the expression of MHC Class I molecules. The expression of LMP2 protein was not altered by the IFN treatments. The authors analyzed structural alterations in the TAP1 promoter region in eight malignant glioma cell lines. Single nucleotide polymorphism was found in 446 bp up-stream of the translation start site of the TAP1 gene, and a point mutation was found in 34 bp upstream of the TAP1 gene. CONCLUSIONS: Malignant glioma cells may be less immunogenic due to low levels of TAP1 expression. Upregulated expression of TAP1 and MHC Class I molecules by IFN-gamma and -beta may enhance antigen presentation in malignant glioma cells.

Expression of syndecans, a heparan sulfate proteoglycan, in malignant gliomas: participation of nuclear factor-kappaB in upregulation of syndecan-1 expression.

Invasion of tumor cells into the surrounding normal brain tissues is a prominent feature of malignant gliomas. Malignant glioma cells secrete thrombospondin-1 which participates in the motility of glioma cells and binds cell surface heparan sulfate proteoglycan. To clarify the invasion mechanism of tumor cells, expression of the syndecans (syndecan-1, -2, -3, and -4), a major cell surface heparan sulfate proteoglycan family, was analyzed in malignant gliomas. Involvement of nuclear factor-kappaB (NF-kappaB) on syndecan-1 expression was also investigated. Using reverse transcription-PCR, the authors analyzed the expression of syndecan-1, -2, -3, and -4 in 10 malignant glioma cell lines, 2 glioblastoma specimens, and 2 normal brain specimens. All malignant glioma cell lines and glioblastoma specimens expressed all types of syndecan mRNA, except in one glioma cell line that lacked syndecan-3 expression. On the other hand, normal brain specimens expressed syndecan-2, -3, and -4 mRNA, but did not syndecan-1 mRNA. Syndecan-1 protein was localized in the cell surface of all malignant glioma cell lines by flow cytometry. Various levels of active nuclear factor-kappa B (NF-kappaB) was detected in all malignant glioma cell lines using immunoblotting. The expression of active NF-kappaB and syndecan-1 increased in U251 glioma cells after tumor necrosis factor-alpha or interleukin-1beta treatment, which can activate NF-kappaB. The amplification of active NF-kappaB and syndecan-1 by tumor necrosis factor-alpha or interleukin-1beta was suppressed by an inhibitor of NF-kappaB activation (emodin). Emodin also downregulated the expression of syndecan-1 mRNA in U251 cells. These results indicate that malignant glioma cells express all types of syndecans and suggest that NF-kappaB participates in the upregulation of the syndecan-1 expression at the transcriptional level, and increased expression of syndecan-1 could associate with extracellular matrices including thrombospondin-1.

Establishment and partial characterization of five malignant glioma cell lines.

Five malignant glioma cell lines (YMG1, 2, 3, 4, and 5) were established from surgical specimens obtained from patients with glioblastoma or anaplastic astrocytoma, and these lines were partially characterized. Three glioma cell lines (YMG1, 3, and 5) were weakly positive for GFAP by Western blot analysis and two cell lines were negative. S-100 protein was positive in all glioma cell lines. The expression of p53, p16, p15, cyclin-dependent kinase 4 (CDK4), and EGF receptor (EGFR) proteins was examined by Western blotting. YMG1 and 2 cell lines showed accumulation of p53 protein and loss of p16 and p15 expression. YMG3 and 4 showed accumulation of p53 protein and expression of p16 and p15 proteins. YMG5 revealed weak expression of p53 protein, suggesting wild-type p53, and loss of p16 and p15 expression. All cell lines expressed various levels of CDK4 protein. YMG1, 2, and 3 showed higher EGFR protein expression and YMG4 and 5 showed lower EGFR expression compared to U251 glioblastoma cells, which express high levels of EGFR. Fluorescence in situ hybridization analysis for EGFR gene expression did not show any amplification in the glioma cell lines. Immunohistochemical studies revealed that the patterns of p53 and EGFR expressions in the original tumor tissues were mostly correlated with those in the malignant glioma cell lines. These results suggest that the characteristics of p53 and EGFR expression in the malignant glioma cell lines were passed over from the original tumor tissues. These newly established malignant glioma cell lines can be used for further analysis of the mechanisms of tumor growth and progression.

Solitary plasmacytoma of the skull: immunohistochemical study of angiogenic factors and syndecan-1--two case reports.

Two cases of solitary plasmacytomas of the skull are presented, and some biological aspects of the tumor examined. A 75-year-old woman presented with a tumor in the right parietal region. The serum level of immunoglobulin G (IgG) was high and a urine test for Bence Jones protein was negative. A reddish vascular mass was totally removed at surgery. The serum level of IgG was within normal limits after the operation. Postoperative radiotherapy was not performed. A 58-year-old woman presented with a tumor in the occipital region. Serum levels of Igs were within normal limits. A urine test for Bence Jones protein was positive for Ig kappa chain. Bone marrow aspiration revealed no evidence of systemic myelomatosis. The tumor mass was totally removed at surgery and she received local radiation therapy (total 50 Gy). Three months after the surgery, Bence Jones protein (kappa chain) was detected in both the urine and serum and bone scintigraphy showed a weak hot spot in the iliac bone, suggesting development to multiple myeloma. Immunohistochemical studies showed that most tumor cells were positive for vascular endothelial growth factor and syndecan-1, and some tumor cells were strongly positive for basic fibroblast growth factor in both cases. The Ki-67 staining indices were 11.3% and 15.6%. Tumor tissues were negative for p53. These results suggest that solitary plasmacytoma of the skull expresses the angiogenic factors, vascular endothelial growth factor, and basic fibroblast growth factor, in accordance with the high vascularity of the tumors, and syndecan-1 may be an immunohistochemical marker of solitary plasmacytoma of the skull.

Trastuzumab (Herceptin) enhances class I-restricted antigen presentation recognized by HER-2/neu-specific T cytotoxic lymphocytes.

PURPOSE: Numerous examples from animal models and clinical trials showed that HER-2-derived peptides are naturally processed as a CTL epitope and can be recognized by tumor-specific CTLs in several tumors with HER-2 overexpression. The humanized anti-HER-2 monoclonal antibody, Herceptin, has been designed to specifically antagonize the HER-2 function by directing against the extracellular domain of the HER-2 protein. One of the actions of Herceptin includes the internalization and degradation of HER-2, which might increase the amount of HER-2-derived peptides available for loading to MHC class I. EXPERIMENTAL DESIGN: In the present study, we investigated how Herceptin treatment of HER-2-overexpressing targets affects lysis by HER-2-specific CTLs. RESULTS: We showed that Herceptin sensitized HER-2-overexpressing tumors to lysis by HLA-A2-restricted or HLA-A24-restricted CTLs, without any effect of the expression of MHC class I, costimulatory molecules, adhesion molecules, or TAP-1 on the targets. Furthermore, the enhancement of cytolytic activity with Herceptin was inhibited by addition of a specific proteasome inhibitor, lactacystin. CONCLUSIONS: These results suggested that Herceptin treatment might enhance the class I-restricted presentation of endogenous HER-2 antigen via the proteasome step, resulting in higher susceptibility of HER-2-overexpressing tumors to lysis by the HER-2-specific CTLs.

Overexpression of heparin-binding growth-associated molecule in malignant glioma cells.

The highly invasive and angiogenic characteristics of malignant gliomas depend on the production of growth factors and angiogenic factors. Heparin-binding growth-associated molecule (HB-GAM) is a secreted growth factor that is mitogenic for endothelial cells. To examine the expression profile of HB-GAM in malignant glioma cells, messenger ribonucleic acid (mRNA) expression was analyzed in 10 malignant glioma cell lines, two glioblastoma tissue specimens, and two normal brain tissue specimens by the reverse transcription-polymerase chain reaction. HB-GAM mRNA was expressed in all specimens including normal brain tissue specimens. Western blot analysis revealed that HB-GAM protein contents in glioma cell lines and glioblastoma tissues were 1.8 to 6.3 times higher than those in normal brain tissues. The effect of neutralizing anti-platelet-derived growth factor (PDGF) antibody was also examined on the production of HB-GAM in malignant glioma cells, since malignant glioma cells secrete PDGF that upregulates HB-GAM expression. Treatment of U251 and T98G glioblastoma cells with the anti-PDGF antibody did not affect the HB-GAM production. These results suggest that HB-GAM is overexpressed in malignant glioma cells and is involved in tumor growth.

Quantification of thrombospondin-1 secretion and expression of alphavbeta3 and alpha3beta1 integrins and syndecan-1 as cell-surface receptors for thrombospondin-1 in malignant glioma cells.

Malignant glioma cells secrete thrombospondin-1 (TSP-1) which participates in the motility of glioma cells, and binds to cell surface alphavbeta3 and alpha3beta1 integrins, and syndecan-1. This study evaluated the amount of TSP-1 secretion from malignant glioma cells, and the expression of alphavbeta3 and alpha3beta1 integrins, and syndecan-1. The amounts of TSP-1 in the supernatants from 10 malignant glioma cell lines and eight non-glioma malignant tumor cell lines were measured by enzyme-linked immunosorbent assay. Expression of alphavbeta3 and alpha3beta1 integrins, and syndecan-1 were examined by flow cytometry. The amounts of TSP-1 secreted by malignant glioma cells were 43 to 2431 ng/l x 10(6) cells/24 h (mean +/- SD = 626 +/- 792). Seven of 10 glioma cell lines secreted more than 100 ng of TSP-1 and three of these cell lines secreted more than 1 microg. Seven of eight non-glioma cell lines secreted less than 100 ng of TSP-1. All glioma cell lines expressed alpha3beta1 integrin and syndecan-1, and seven of 10 glioma cell lines expressed alphavbeta3 integrin. Treatment of the glioma cell lines with TGF-beta2 did not change the expression of alphavbeta3 integrin. These results suggest that malignant glioma cells secrete high levels of TSP-1, which may be important in the migration of glioma cells via interactions with alphavbeta3 and alpha3beta1 integrins, and syndecan-1.

Cell density regulates thrombospondin-1 production in malignant glioma cells.

Thrombospondin-1 (TSP-1) is a multifunctional matrix protein implicated in cancer cell adhesion, migration, and invasion, inhibition of angiogenesis, and activation of latent transforming growth factor-beta (TGF-beta). The effect of cell density was investigated on the production of TSP-1, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) by two glioblastoma cell lines. The effect of TGF-beta was also examined. The amount of intracellular TSP-1 protein decreased significantly as the cell density increased in cultures of both T98G and A172 cells. The amount of intracellular TSP-1 was highest in sparse tumor cell cultures and lowest in densely confluent tumor cell cultures. The maximum reduction of TSP-1 protein production was 56.8% and 44.6% in T98G and A172 cells, respectively. The cell density did not affect the production of bFGF or VEGF. TGF-beta2 treatment did not affect the production of TSP-1, bFGF, or VEGF proteins. Treatment with excess TGF-beta2 resulted in a slight but significant decrease (22%; P < 0.02) of TGF-beta2 production by A172 cells, but not by T98G cells. The present results indicate that the production of TSP-1 protein is regulated by cell density of glioblastoma cells, while that of angiogenic factors is not affected by tumor cell density. This suggests that high tumor cell density may tilt the angiogenic balance in favor of angiogenesis.

Technical considerations of transsphenoidal removal of fibrous pituitary adenomas and evaluation of collagen content and subtype in the adenomas.

Pituitary adenomas are usually soft, but 5-13.5% are fibrous adenomas which are difficult to remove. Magnetic resonance (MR) imaging and operative findings were evaluated in eight patients (12.1%) with fibrous pituitary adenoma among 66 patients. Tumor specimens were examined histologically and immunohistochemically for collagen content and subtypes. Seven patients had clinically inactive non-functioning pituitary adenomas and one patient growth hormone-secreting adenoma. All patients underwent transsphenoidal surgery. Four cases were giant adenomas with suprasellar extension of more than 2 cm. T1- and T2-weighted MR imaging showed the tumors as nearly isointense to the surrounding brain, except in one case where the tumor was high intense on T2-weighted MR imaging. All tumors required piecemeal resection using a micro-dissector and tumor forceps. Four tumors of maximum size more than 3 cm needed a second operation. The interface between the thinned normal pituitary gland and fibrous adenoma was intended to identify at the anterior-superior portion in recent four cases, which was helpful to remove the tumors and preserve pituitary functions. Histological examination revealed prominent deposition of collagen in the perivascular area. The percentage of collagen content in fibrous adenomas was more than 5% and significantly higher than that in soft adenomas and normal pituitary glands. Immunohistochemical examination showed positive staining for collagen types I and III in the fibrous adenomas, but only for type V collagen in the normal pituitary glands. Large fibrous adenomas can be resected by transsphenoidal surgery which may require two-stage operations. Identification of the interface between the normal pituitary gland and adenoma is helpful to remove fibrous adenomas and to preserve pituitary functions. We propose that firm adenomas containing more than 5% collagen are "fibrous" adenomas.