Tatsuki Oyoshi

Attenuation of inflammatory and neuropathic pain behaviors in mice through activation of free fatty acid receptor GPR40

BackgroundThe G-protein-coupled receptor 40 (GPR40) is suggested to function as a transmembrane receptor for medium- to long-chain free fatty acids and is implicated to play a role in free fatty acids-mediated enhancement of glucose-stimulated insulin secretion from pancreas. However, the functional role of GPR40 in nervous system including somatosensory pain signaling has not been fully examined yet.ResultsIntrathecal injection of GPR40 agonist (MEDICA16 or GW9508) dose-dependently reduced ipsilateral mechanical allodynia in CFA and SNL models and thermal hyperalgesia in carrageenan model. These anti-allodynic and anti-hyperalgesic effects were almost completely reversed by a GPR40 antagonist, GW1100. Immunohistochemical analysis revealed that GPR40 is expressed in spinal dorsal horn and dorsal root ganglion neurons, and immunoblot analysis showed that carrageenan or CFA inflammation or spinal nerve injury resulted in increased expression of GPR40 in these areas. Patch-clamp recordings from spinal cord slices exhibited that bath-application of either MEDICA16 or GW9508 significantly decreased the frequency of spontaneous excitatory postsynaptic currents in the substantia gelatinosa neurons of the three pain models.ConclusionsOur results indicate that GPR40 signaling pathway plays an important suppressive role in spinal nociceptive processing after inflammation or nerve injury, and that GPR40 agonists might serve as a new class of analgesics for treating inflammatory and neuropathic pain.

Immunoreactivity of Wnt5a, Fzd2, Fzd6, and Ryk in glioblastoma: evaluative methodology for DAB chromogenic immunostaining

The aim of this study was to determine the influence of Wnt5a and its receptors on the survival of glioblastoma patients and to determine reliable evaluation methods for immunohistochemistry. Diagnostic specimens from 41 histopathologically confirmed primary glioblastoma patients whose Gd-enhanced tumors had been totally removed were immunohistochemically stained for Wnt5a, Fzd2, Fzd6, and Ryk. The immunoreactivity was evaluated using the following methods: (A) grayscale optical density after color deconvolution, (B) percentage of stained cells, (C) density of stained cells, (D) staining amount (multiplication product of B and C), and (E) staining rank. The data sets of A to E were statistically evaluated by correlation matrix analysis and regression analysis. The influence of the expression of the markers on survival was analyzed using a proportional hazard model. The results of color deconvolution (A) were well correlated with the results of the staining rank (E). In the semiquantitative results (B, C, and D), the staining amount (D) tended to show a better correlation with results of color deconvolution (A). Among all data sets, color deconvolution (A) demonstrated the most preferable fit in a proportional hazard model, and the expression of Fzd2 and Fzd6 was associated with poor prognosis in glioblastoma patients.

TLR4, IL-6, IL-18, MyD88 and HMGB1 are highly expressed in intracranial inflammatory lesions and the IgG4/IgG ratio correlates with TLR4 and IL-6.

We determined distribution of plasma cells and IgG4/IgG index and factors associated with the index in intracranial inflammatory lesions. Specimens of nine patients were analyzed immunohistochemically using antibodies against CD45, CD68, CD3, CD4, CD8, CD20, CD138, lambda chain, kappa chain, IgG, IgG4, IL‐1α, IL‐6, IL‐18, toll‐like receptor (TLR) 2, TLR4, high‐mobility group box 1 (HMGB1), tumor necrosis factor‐alpha (TNF‐α), myeloid differentiation factor 88 (MyD88), and anaplastic lymphoma kinase (ALK). The relationship between all the factors was assessed using Spearmans rank correlation coefficient (ρ). Negative ALK staining was observed in all the patients. Plasma cells were detected in eight patients with varying degrees. The highest number of neutrophils, but no plasma cells, was observed in a patient with the shortest history of inflammation. IgG4/IgG index was independent of the number of plasma cells. The index was relatively highly correlated with IL‐6 (ρ = 0.7271) and TLR4 expression (ρ = 0.7246). IL‐6 expression was highly correlated with TLR4 expression (ρ = 0.8042). IL‐18 was maximally expressed in all the patients. TLR4 expression was strong, but TRL2 expression was weak. Positive HMGB1 staining was observed in all the patients, predominantly in the nuclei, but also in the cytoplasm in four patients. The cytoplasmic expression strongly correlated with IL‐1α expression (ρ = 0.9583). The cytoplasmic colocalization of HMGB1 and IL‐1α was histologically confirmed in cells with collapsing nuclei by the double‐staining method. The IgG4/IgG indexes varied case by case. IL‐6 and TLR4 expressions may influence IgG4/IgG index. The nuclei of cells with both IL‐1α and HMGB1 expressions in the cytoplasm collapse in the cell death stage. The cooperative high expression of TLR4, IL‐6, IL‐18, MyD88 and HMGB1 suggest their critical roles in the inflammation circuit.

Improvement in treatment results of glioblastoma over the last three decades and beneficial factors

Abstract Background. The purpose of this study is to elucidate the trend of glioblastoma outcome and scrutinize the factors contributing to better outcome over three decades. Methods. Survival time and the influencing factors were retrospectively analyzed in 223 newly diagnosed primary glioblastoma patients during 1980–2010. Appraised factors included age, sex, tumor site, year of surgery, extent of resections, use of surgery supporting system, Karnofsky Performance Status (KPS), chemotherapy, conventional external beam radiotherapy (EBRT), and CyberKnife stereotactic radiotherapy (CK-SRT) use. Results. The median survival time (MST) in all patients was 13.6 months. The MSTs for 4 periods were 9.8 (1980–1990), 13.7 (1991–2000), 12.9 (2001–2005), and 15.8 months (2006–2010), respectively (p = 0.0047). Total resection, subtotal resection, partial resection, and biopsy had MSTs of 31.8, 13.9, 11.4, and 7.0 months, respectively (p < 0.0001). Regarding chemotherapy, MSTs of the temozolomide base group and nimustine hydrochloride (ACNU) base group were 16.9 and 14.6 months, respectively, whereas the MST of patients without chemotherapy was only 9.8 months (p < 0.0001). The MSTs for 40-Gy EBRT plus CK-SRT and 60-Gy EBRT were 19.1 and 10.7 months, respectively (p < 0.0001). But in sub-selected patients, treated during 2001–2010, whose resection rate was total resection or subtotal resection, EBRT was completed and postoperative KPS was greater than or equal to 70, the MST with and without CK-SRT was 26.6 and 18.3 months, respectively (p = 0.1529). According to the Cox proportional hazards model, degree of resection, KPS, ACNU use, temozolomide use, bevacizumab use, EBRT dose, and CK-SRT use were good prognostic factors. Use of neuronavigation and use of intraoperative magnetic resonance imaging were related to higher resection rate, but not determined as prognostic factors. Conclusions. We observed a gradual improvement in glioblastoma outcome, presumably because of improvements in therapeutic modalities for surgery, anticancer agents, and radiation, but the efficacy of CK-SRT remains unclear.

A Novel Bilateral Approach for Suprasellar Arachnoid Cysts: A Case Report

The endoscopic method is used to treat suprasellar arachnoid cysts (SACs) but it is sometimes difficult to make sufficiently sized fenestrations. Creating a larger fenestration on the cyst wall is preferable to prevent closure of the stoma. In this paper, we report a novel endoscopic approach for SAC treatment in which we use bilateral burr holes to achieve a more extensive cyst fenestration. A 7-year-old girl was referred to our hospital because of incidentally detected hydrocephalus by computed tomography scans. Physical examination did not show any signs of intracranial hypertension, but a digital impression of her skull on X-ray implied chronic intracranial hypertension. Magnetic resonance imaging (MRI) revealed enlargement of both lateral ventricles and a cystic mass occupying the third ventricle. We performed cyst wall fenestration using a bilateral approach in which we created two burr holes to introduce a flexible endoscope and a rigid endoscope. The cyst wall was held by forceps with the flexible endoscope, and resection of the cyst wall was achieved by using a pair of scissors with the rigid endoscope. There were no postoperative complications, and MRI performed 1 year after treatment showed disappearance of the superior part of the cyst wall.

Efficacy of bevacizumab therapy for unresectable malignant glioma: A retrospective analysis

Bevacizumab (BEV), an inhibitor of vascular endothelial growth factor A, has been used for primary and recurrent malignant gliomas in Japan since June, 2013. Previous randomized controlled studies demonstrated that BEV prolonged the progression-free survival, but not the overall survival (OS) of patients with newly diagnosed glioblastoma. The aim of the present study was to elucidate the effect of BEV on the OS of patients with unresectable malignant gliomas. Of the 440 cases of malignant glioma initially treated in our institute between 2000 and 2015, 88 were not suitable for maximal resection due to patient age, physical condition, tumor location and extent, or the patients wishes. Based on the biopsy results, the pathological diagnosis was glioblastoma, anaplastic astrocytoma and anaplastic oligodendroglioma in 60, 19 and 9 patients, respectively. Kaplan-Meier and log-rank analyses were performed to investigate the effect of BEV on OS. OS was longer in the BEV group (n=24) compared with that in the non-BEV group [n=64; median survival time (MST), 566 vs. 243 days, respectively; hazard ratio (HR)=0.413; 95% confidence interval (CI): 0.216-0.787; P=0.003]. In the 41 patients who received temozolomide (TMZ) and radiotherapy and the 31 patients with glioblastoma who received TMZ and radiotherapy, OS was longer in the BEV group compared with that in the non-BEV group (MST, 568 vs. 334 days, HR=0.404, 95% CI: 0.175-0.933, P=0.016; and MST, 566 vs. 160 days, HR=0.253, 95% CI: 0.099-0.646, P=0.001, respectively). In the Cox hazard model analysis of 41 patients who underwent TMZ-based chemoradiotherapy after biopsy, the use of BEV was the strongest independent beneficial factor associated with prolonged OS (HR=0.101; P=0.0002). Our retrospective survey suggested that BEV prolongs the OS of patients with unresectable malignant gliomas. However, these results must be verified by a well-designed prospective randomized controlled trial.

Calcifying pseudoneoplasm of the neuraxis in direct continuity with a low-grade glioma: A case report and review of the literature: CAPNON associated with low-grade glioma

Calcifying pseudoneoplasms of the neuraxis (CAPNON) are presumed to be a non‐neoplastic reactive pathology, based on the frequent finding of granulomatous inflammation. To our knowledge, there are few reports of CAPNON in association with a neoplasm. Here, we report the case of a 62‐year‐old man presenting with headache, which was caused by CAPNON in the left cingulate gyrus. CT scan revealed a calcified mass exhibiting gradual growth and increasing peritumoral edema. MRI showed an intra‐axial hypointense mass on T1‐ and T2‐weighted images. Development of a peri‐lesional hyperintense lesion on T2‐weighted images suggested local edema or tumoral invasion. Gadolinium‐enhanced T1‐weighted images revealed mild peripheral enhancement of the calcified nodule. L‐methyl‐11C methionine‐positron emission tomography revealed the uptake of tracer in the calcified nodule. The calcified mass and its enveloping brain tissue were removed using a parietal craniotomy. The calcified tissue was surrounded by spindle‐shaped cells positive for GFAP and nestin. The MIB‐1 labeling index of spindle cells was around 10% (i.e. a hot spot). Fourteen months after surgery, gadolinium‐enhanced MRI evidenced growth of a tiny residual lesion. Therefore, this report illustrates a potential case of CAPNON arising from low‐grade glial neoplasm.

Concurrent occurrence of an inherited 16p13.11 microduplication and a de novo 19p13.3 microdeletion involving MAP2K2 in a patient with developmental delay, distinctive facial features, and lambdoid synostosis

A female patient presented with developmental delay, distinctive facial features, and congenital anomalies, including a heart defect and premature lambdoid synostosis. The patient showed a paternally inherited 16p13.11 microduplication and a de novo 19p13.3 microdeletion involving the mitogen-activated protein kinase kinase 2 gene (MAP2K2), in which mutations cause the cardio-facio-cutaneous (CFC) syndrome. Reports of patients with overlapping 19p13.3 microdeletions of this region describe similar clinical manifestations including distinctive facial features: prominent forehead, horizontal/down-slanting palpebral fissures, long midface, pointed chin/angular jaw, sparse eyebrows, and underdeveloped cheekbones. Some of these findings overlapped to that of the patients with 16p13.11 microduplications and CFC syndrome. Although craniosynostosis was occasionally observed in patients with dominant-negative mutations in RAS/MAP kinase signaling genes (RASopathies) related to CFC syndrome, it was also reported in two patients with 16p13.11 microduplications. Genetic contributions of both chromosomal aberrations were discussed.

Contiguous gene deletion neighboring TWIST1 identified in a patient with Saethre-Chotzen syndrome associated with neurodevelopmental delay: Possible contribution of HDAC9: Saethre-Chotzen syndrome

Saethre‐Chotzen syndrome (SCS) is an autosomal dominant craniosynostotic disorder characterized by coronal synostosis, facial asymmetry, ptosis, and limb abnormalities.

Giant radiation-induced cavernous haemangioma before reduced-intensity bone marrow transplantation for acute lymphoblastic leukaemia

Although performed less frequently, cranial irradiation is used to treat leukaemia and lymphoma and prevent tumour infiltration to the central nervous system (CNS). One of the late complications of cranial irradiation is radiation-induced cavernous haemangioma (RICH), which develops cumulatively over up to 10 years after cranial irradiation in ~40% patients [1–3]. Rather than radiation-induced secondary neoplasm, RICH is regarded as a vascular disorder caused by radiation-induced damage to small intracranial vessels [4]. Although asymptomatic, RICH is incidentally detected during imaging tests in most cases. Asymptomatic patients are only observed and not treated even after diagnosis [2, 5, 6]. Few cases of RICH developing after haematopoietic stem cell transplantation (HSCT) have been reported; however, there are no reports in which RICH was observed prior to transplantation nor is there any known method to manage RICH that develops during HSCT [6]. Here, we report the case of a 10-year-old boy with second relapse of acute lymphoblastic leukaemia (ALL) with a giant 30-mm RICH. Prior to conditioning, the patient developed haemorrhage and hydrocephalus that required sub-occipital craniotomy to excise the haemangioma. Reduced-intensity conditioning was performed ~1 month after craniotomy, followed by allogeneic bone marrow transplantation from an unrelated donor. The patient was a 10-year-old boy with healthy parents and no siblings. At age 2 years, he was diagnosed with B cell precursor ALL (initial exam findings of white blood cell count: 37,500/μL; chromosome G-banding: 46,XY,t(5;22) (q35;11.2), der(9;15)(q10;q10); CNS lesions: negative). He was started on multi-agent systemic chemotherapy (KYCCSG-ALL02 [ref. 7]) and achieved remission. At age 5 year, he experienced an isolated CNS relapse during maintenance therapy; thus, remission-induction therapy was restarted using the ALL-REZ BFM 2002 protocol [8], resulting in a second remission. Thereafter, at age 6.5 years, the patient received radiation therapy to the whole brain (18 Gy/10 Fr) and whole spine (15 Gy/10 Fr). At age 8 years, the planned treatment was ended with maintained remission. At age 10 years, routine examination revealed a significant increase in white blood cell count at 44,600/μL; the patient was diagnosed with second relapse isolated to the bone marrow. Chromosome G-banding demonstrated a karyotype of 46,XY,t(5;9;22)(q35;q34;q11.2), and fluorescence in situ hybridisation testing revealed a bcr-abl fusion signal, indicating that the patient had Philadelphia chromosome-positive ALL. Cranial magnetic resonance imaging (MRI) prior to starting the treatment revealed a large RICH measuring 11 mm in diameter (Fig. 1a). The patients were started on systemic chemotherapy (EsPhALL regimen [ref. 9]) with adjuvant imatinib. At the end of early intensification therapy at age 10.9 years, the levels of minor bcr-abl chimeric mRNA were below the detection threshold, and the patient was referred to our hospital for allogeneic HSCT. On admission, physical findings included mild anaemia; lymphadenopathy or hepatosplenomegaly were not observed. The patient did not exhibit any abnormal neurological findings. He was administered one course of intensification therapy and scheduled for HSCT from an unrelated donor with a conditioning regimen including * Takuro Nishikawa adu44150@ams.odn.ne.jp