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100 Advances in the Biology, Imaging and Therapies for Glioblastoma

In recent years there have been many publications in the area of immunohistochemistry in brain tumor pathology (Takei et al., 2007; Dunbar & Yachnis, 2010; Ikota et al, 2006). Extensive molecular studies have identified diagnostic and prognostic markers in gliomas (Labussiere et al., 2010). They can assist in diagnosis, provide prognostic information and potentially predict response to therapy (Rivera & Pelloski, 2010). As we mentioned above, we combined the clinical work by different experimental glioma models. Animal tumor sections were examined for tumor markers by routine haematoxylin and eosin staining and immunohistochemical analyses. Established animal models provide a basis for further experimental studies of genetic and protein expression fingerprints during human glioma tumorigenesis. Furthermore, in a present paper we reviewed clinical and experimental work in glioma patients operated at our department of neurosurgery. Immunohistochemical studies supplement conventional H&E histology. Immunohistochemically we evaluated the expression of possible biological markers in human gliomas, including proteolytic enzymes (cathepsins B and L), neural stem cell markers (nestin, musashi), marker for microglia (CD68) and others (e.g. kallikrein 6). Differently from other reports, we performed immunohistochemical staining for the panel of markers on the same group of patients. Increased expression of lysosomal cystein proteinases such as cathepsins B and L plays a functional role in tumor cell migration and metastasis (Lah et al., 2000). We found that Cat B expression was highly elevated in GBM compared to lower grade malignant tumors and benign tumors. Cat B was also highly expressed in the endothelial cells of about two third of GBM. The latter finding indicates that Cat B may be associated with the invasion of not only tumor but also endothelial cells in the process of angiogenesis. At the end of the nineties we first published the clinical study on prognostic impact of Cat B in tumors of CNS, revealing that survival time in all patients with weak total immunostaining score is significantly longer compared to survival of patients with strong staining. Intense Cat B staining of endothelial cells is also prognostic important in patients with glioblastoma indicating significantly shorter survival. Cat L is preferentially expressed in tumor cells, increasing with glioma progression, but is not significantly associated with new vasculature of glioblastoma. Nestin is expressed in tumor cells of primary gliomas to a greater extent than musashi. Nestin-positive tumor cells are localized more abundantly in the transition zone of the tumor. Nestin is expressed in the endothelial cells in both low- and high-grade tumors, whereas musashi is expressed only to a limited extent in endothelial cells in the high-grade tumors. The further research should confirm the hypothesis derived from our data, that is, that angiogenesis also may result predominantly from the bone marrow stem cells attracted to and differentiating onto blood vessels within the tumor. Nestin is shown to be a strong prognostic marker for glioma malignancy. Our study revealed that both microglia and tumor cells expressed CD68. Malignant astrocytoma cells were highly CD68 positive in accordance to previous report (Leenstra et al., 1995). We found that some authors recommended to identify macrophages on intraoperative consulatiton to distinguish a neoplastic process from a demyelinating (or other destructive noneoplastic) disorder using IHC staining for CD68 (Dunbar & Yachnis, 2010). We point out that the demonstration of macrophages within the astrocytomas by using macrophage-specific antibodies alone must be cautiously considered. We further conclude that specific immunostaining of CD68 in tumor cells can be used to predict the risk of overall death in patients with glioma.

**3. Conclusion** 


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**6** 

**Gray or White? – The Contribution** 

*1Division of Neurology, Department of Internal Medicine,* 

Ryuta Kinno1,2, Yoshihiro Muragaki3 and Kuniyoshi L. Sakai2

Symptoms of a glioma include not only headaches and seizures, but cognitive deficits including aphasia. One of the most important regions for aphasia is the anterior speech area, the damage of which causes Broca's aphasia, marked by effortful, distorted articulation, reduced speech output, and agrammatic syntax. These patients show relatively good comprehension of single words and simple sentences, but show trouble understanding sentences with more complex syntactic structures, such as passive sentences and sentences with object relative clauses (Schwartz et al., 1980; Caplan et al., 1985; Grodzinsky, 2000); this aspect of Broca's aphasia is called agrammatic comprehension (Goodglass & Menn, 1985; Menn and Obler, 1990; Pulvermüller, 1995). However, methodological problems have been raised (Badecker & Caramazza, 1985), and general processes of short-term memory or decision-making have been proposed to be disrupted in agrammatic comprehension (Just & Carpenter, 1992; Cupples & Inglis, 1993; Dick et al., 2001). Thus, for appropriately assessing a coginitive deficit, it is crucial to use an experimental task in which general cognitive

In our recent functional magnetic resonance imaging (fMRI) study with a picture-sentence matching task, we examined the effect of sentence structures strictly controlling general cognitive demands such as the memory load (Kinno et al., 2008), where a sentence was visually presented with a picture representing an action (Fig. 1; the same task and stimuli were used in the present study). The participants indicated whether or not the meaning of each sentence matched the action depicted by the corresponding picture. There were three main conditions with different sentence types: canonical / subject-initial active sentences (AS) (e.g., " ∆-ga ○-o hiiteru", " ∆ pulls ○"), noncanonical / subject-initial passive sentences (PS) (e.g., " ○-ga ∆-ni hikareru", " ○ is affected by ∆'s pulling it"; see Kinno et al. (2008) for ni direct passive form), and noncanonical / object-initial scrambled sentences (SS) (e.g., " ○-o ∆-ga hiireru", "as for , ○ pulling it"; this form is allowed not only in Japanese but in German, Finnish, and other languages). Under these conditions, each sentence had a

demands such as the memory load are stricly controlled.

**1. Introduction** 

*2Department of Basic Science, Graduate School of Arts and Sciences,* 

*3Department of Neurosurgery, Tokyo Women's Medical University* 

**of Gray Matter in a Glioma** 

*Showa University Northern Yokohama Hospital* 

**to Language Deficits** 

*The University of Tokyo* 

*Japan* 

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