Craniocervical junction abnormalities with atlantoaxial subluxation caused by ventral subluxation of C2 in a dog

Craniocervical junction abnormalities with atlantoaxial subluxation caused by ventral subluxation of C2 were diagnosed in a 6-month-old female Pomeranian with tetraplegia as a clinical sign. Lateral survey radiography of the neck with flexion revealed  atlantoaxial subluxation with ventral subluxation of C2. Computed tomography revealed absence of dens and atlanto-occipital overlapping. Magnetic resonance imaging showed compression of the spinal cord and indentation of caudal cerebellum. The diagnosis was Chiari-like malformation, atlantoaxial subluxation with ventral displacement of C2,  atlanto-occipital overlapping, and syringomyelia. The dog underwent foramen magnum decompression, dorsal laminectomy of C1, and ventral fixation of the atlantoaxial joint. Soon after the operation, voluntary movements of the legs were recovered. Finally, the dog could stand and walk without assistance. The dog had complicated malformations at the craniocervical junction but foramen magnum decompression and dorsal  laminectomy for Chiari-like malformation, and ventral fixation for atlantoaxial  subluxation resulted in an excellent clinical outcome.Keywords: Atlantoaxial subluxation, Atlanto-occipital overlapping, Chiari-like  malformation, Craniocervical junction abnormalities, Foramen magnum  decompression

Evaluation of immunohistochemical staining with PMab-38, an anti-dog podoplanin monoclonal antibody, in various canine tumor tissues

Podoplanin (PDPN) is a type I transmembrane sialoglycoprotein with O-glycosylation and a high content of sialic acid. PDPN has been reported to be expressed in various human tumors and promote tumor progression, epithelial-mesenchymal transition, and distant metastasis. PDPN is also expressed in cancer-associated fibroblasts (CAFs), which promote tumor growth. We have developed novel tumor specific anti-PDPN antibodies. PMab-38, which is an anti-dog PDPN monoclonal antibody, recognized PDPN expression in canine squamous cell carcinoma (SCC) and malignant melanoma. However, there has been no research into PMab-38 recognition of other types of tumors and systemic normal tissue. The objective of this study was to evaluate the staining positivity of PMab-38 by immunohistochemical staining of various paraffin-embedded canine tumor and systemic normal tissues. Immunohistochemical analysis revealed that PMab-38 positively stained tumor cells in 9/11 (82%) SCC and 9/11 (82%) pulmonary adenocarcinoma tissues. In the tumor stroma, large spindle-shaped mesenchymal cells, which were suspected to be CAFs, were stained by PMab-38 in almost all tumor types: 9/10 (90%) for anal sac adenocarcinoma and 11/12 (92%) for transitional cell carcinoma tissues. Almost all normal tissues were negatively stained with PMab-38 except part of the kidney glomerulus. Taken together, our findings provide evidence that various types of tumor cells were strongly stained by PMab-38, but most normal cells were not stained. These results indicate that PDPN recognized by PMab-38 might be a target for tumor antigen targeting therapy. Further studies are required to investigate the anti-tumor effect in various canine tumors by antibody therapy using PMab-38

Craniocervical junction abnormalities with atlantoaxial subluxation caused by ventral subluxation of C2 in a dog

Craniocervical junction abnormalities with atlantoaxial subluxation caused by ventral subluxation of C2 were diagnosed in a 6-month-old female Pomeranian with tetraplegia as a clinical sign. Lateral survey radiography of the neck with flexion revealed  atlantoaxial subluxation with ventral subluxation of C2. Computed tomography revealed absence of dens and atlanto-occipital overlapping. Magnetic resonance imaging showed compression of the spinal cord and indentation of caudal cerebellum. The diagnosis was Chiari-like malformation, atlantoaxial subluxation with ventral displacement of C2,  atlanto-occipital overlapping, and syringomyelia. The dog underwent foramen magnum decompression, dorsal laminectomy of C1, and ventral fixation of the atlantoaxial joint. Soon after the operation, voluntary movements of the legs were recovered. Finally, the dog could stand and walk without assistance. The dog had complicated malformations at the craniocervical junction but foramen magnum decompression and dorsal  laminectomy for Chiari-like malformation, and ventral fixation for atlantoaxial  subluxation resulted in an excellent clinical outcome.Keywords: Atlantoaxial subluxation, Atlanto-occipital overlapping, Chiari-like  malformation, Craniocervical junction abnormalities, Foramen magnum  decompression

Anti-tumor effects of the histone deacetylase inhibitor vorinostat on canine urothelial carcinoma cells.

Canine urothelial carcinoma (cUC) is the most common tumor of the lower urinary tract in dogs. Although chemotherapy and radical surgery have improved the overall survival, most dogs with cUC succumb to metastasis or recurrence. Therefore, the development of an effective systematic therapy is warranted. In this study, a comprehensive drug screening test using a cUC cell line was performed and the anti-tumor effect of a histone deacetylase (HDAC) inhibitor was evaluated. Comprehensive drug screening was performed on cUC cells. Based on this screening, the anti-proliferation effect of vorinostat, an HDAC inhibitor clinically applied in humans, was evaluated using several cUC cell lines in sulforhodamine B and flow cytometry assays. Western blot analysis was also performed to evaluate the degree of acetylation of histone H3 as well as the expression and phosphorylation of cell cycle-related molecules. The anti-tumor effect of vorinostat in vivo was evaluated using a xenograft model. Finally, immunohistochemistry was performed on acetyl-histone H3 in cUC and the relationship between the degree of acetylation and prognosis was examined using Kaplan-Meier survival analysis. Drug screening revealed that HDAC inhibitors consistently inhibited the growth of cUC cells. Vorinostat inhibited the growth of 6 cUC cell lines in a dose-dependent manner and induced G0/G1 cell cycle arrest. Western blot analysis showed that vorinostat mediated the acetylation of histone H3, the dephosphorylation of p-Rb, and the upregulation of p21 upon exposure to vorinostat. Furthermore, inhibition of tumor growth was observed in the xenograft model. In clinical cUC cases, neoplastic urothelium showed significant deacetylation of histones compared to the normal control, where lower histone acetylation levels were associated with a poor prognosis. In conclusion, the therapeutic potential of vorinostat was demonstrated in cUC. Histone deacetylation may be related to cUC tumor progression

Tomohiro Nishikado, Oral History (2nd, 1): Development of “Sky Fighter”

平成28年度メディア芸術連携促進事業 連携共同事業「ゲーム産業生成におけるイノベーションの分野横断的なオーラル・ヒストリー事業」47 p

A novel model for treatment of hypertrophic pachymeningitis

Objective Immunoglobulin (Ig)G4-related disease is a major cause of hypertrophic pachymeningitis (HP), presenting as a progressive thickening of the dura mater. HP lacks an animal model to determine its underlying mechanisms. We developed a suitable animal model for the treatment of HP. Methods We longitudinally evaluated dura in mice with a mutation (Y136F) in the linker for activation of T cells (LAT), which induced type 2 T helper (Th2) cell proliferation and IgG1 (IgG4 human equivalent) overexpression. Mice were therapeutically administered daily oral irbesartan from 3 to 6 weeks of age. Human IgG4-related, anti-neutrophil cytoplasmic antibody-related, and idiopathic HP dura were also immunohistochemically examined. Results LATY136F mice showing dural gadolinium enhancement on magnetic resonance imaging had massive infiltration of B220(+) B cells, IgG1(+) cells, CD138(+) plasma cells, CD3(+) T cells, F4/80(+) macrophages, and polymorphonuclear leukocytes in the dura at 3 weeks of age, followed by marked fibrotic thickening. In dural lesions, transforming growth factor (TGF)-beta 1 was produced preferentially in B cells and macrophages while TGF-beta receptor I (TGF-beta RI) was markedly upregulated on fibroblasts. Quantitative western blotting revealed significant upregulation of TGF-beta 1, TGF-beta RI, and phosphorylated SMAD2/SMAD3 in dura of LATY136F mice aged 13 weeks. A similar upregulation of TGF-beta RI, SMAD2/SMAD3, and phosphorylated SMAD2/SMAD3 was present in autopsied dura of all three types of human HP. Irbesartan abolished dural inflammatory cell infiltration and fibrotic thickening in all treated LATY136F mice with reduced TGF-beta 1 and nonphosphorylated and phosphorylated SMAD2/SMAD3. Interpretation TGF-beta 1/SMAD2/SMAD3 pathway is critical in HP and is a potential novel therapeutic target