2023年09月, Asian Federation of Laboratory Animal Science Assosiations, 第９回 AFLAS Congress 2023 トラベルアワード　　　　　　　　　　　　　　　
Analysis of storage and transport conditions of multiplex ICG sticks for diagnosis of infectious diseases in rats and mice
Noriko Tosa

2013年05月, 日本実験動物技術者協会, 第4回実験動物福祉奨励賞　　　　　　　　　　　　　　　
マウスの被毛異常または床敷湿潤が発生した後の環境エンリッチメントの効果
土佐 紀子

Simultaneous serodetection of major rat infectious pathogens by a multiplex immunochromatographic assay.　　　　　　　　　　　　　　　
Tosa N, Ishida T, Yoshimatsu K, Hayashimoto N, Shiokawa K, Takakura A, and Arikawa J
Exp. Anim., 70, 2, 161, 168, 2021年05月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）

Multiplex Immunochromatographic Assay for Serologic Diagnosis of Major Infectious Diseases in Laboratory Mice.
Noriko Tosa, Tomoko Ishida, Kumiko Yoshimatsu, Nobuhito Hayashimoto, Kanae Shiokawa, Akira Takakura, Jiro Arikawa
Journal of the American Association for Laboratory Animal Science : JAALAS, 58, 6, 790, 795, 2019年11月01日, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, Serologic monitoring of infectious diseases is important for microbial control in colonies of laboratory mice. Rapid and simple tests that do not require killing animals are valuable for this purpose. In this study, we developed a multiplex immunochromatographic assay (ICA) for detection of antibodies to mouse hepatitis virus (MHV), Sendai virus (also known as hemagglutinating virus of Japan [HVJ]), and Clostridium piliforme (The pathogen that causes Tyzzer disease), which are major infectious diseases in mice. For this assay, an ICA strip was put into a microtube containing 150 μL PBS and either 0.75 μL mouse serum or 1.5 μL whole blood. Binding antibodies were visualized by using protein A-conjugated colloidal gold. Under these conditions, multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. To evaluate the sensitivity and specificity of multiplex ICA, positive serum samples for each infectious disease were used. Sensitivities of the multiplex ICA test for MHV, HVJ, and C. piliforme were 100%, 100%, and 90%, respectively. No nonspecific reaction was observed in any of the 30 positive sera. In addition, 10 samples of uninfected sera did not show any bands except for the control line. These observations indicate high specificity of the multiplex ICA test. Moreover, the multiplex ICA could be applied to diluted blood. These results indicate that the multiplex ICA is appropriate for rapid, simple, and safe serologic testing of laboratory mice.

Comparison of immune response in mice sensitized to an animal allergen, Can f 1, and to a food allergen, ovalbumin
Noriko Tosa, Kumiko Yoshimatsu, Motoko Takahashi, Jiro Arikawa
Biomedical Research, 40, 1, 9, 15, 2019年, ［査読有り］, ［筆頭著者］, ［国内誌］
英語, 研究論文（学術雑誌）, Can f 1 belongs to the lipocalin superfamily and is considered to be an animal allergen. The immune response induced by Can f 1 in mice was compared with that induced by ovalbumin (OVA), a typical food allergen. Female BALB/c and C57BL/6 mice (6 weeks of age) were subcutaneously injected with Can f 1 or OVA with or without aluminum hydroxide (Alum) three times with intervals of two weeks. Serum levels of total IgE or antigen-specific IgE and production of IL13 and IFNγ from splenocytes were analyzed. Immunization with Can f 1 or OVA increased serum levels of both total IgE and antigen-specific IgE significantly irrespective of Alum. These results indicate that Can f 1 and OVA were able to induce allergic sensitization in mice. Splenocyte production of IL13 in mice immunized with Can f 1 or OVA with and without Alum were significantly increased after stimulation with each antigen. However, IL13 levels in the mice immunized with Can f 1 with Alum were significantly lower than those immunized without Alum. Increases in IFNγ levels after stimulation with Can f 1 or OVA were not remarkable. No influence of genetic backgrounds of BALB/c and C57BL/6 mice was found. Although Can f 1 induced Th2 type immune responses as was also the case for immunization with OVA, an inhibitory effect of Alum on induction of IL13 was observed only in mice immunized with Can f 1. These results suggest that the immune mechanism for allergic sensitization with Can f 1 is different from that with OVA.

Variation in the bacterial conditions inside cages is correlated with intracage humidity and ammonia levels.　　　　　　　　　　　　　　　
Tosa N, Yoshimatsu K, Tadasuke Tsukiyama, Hatakeyama S, Arikawa J
Lab Animal and Environ, 21, 2, 87, 98, 2013年, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）

Environmental enrichment after the occurrence of wet bedding and the appearance of abnormal fur in mice is effective for solving those problems.
TOSA Noriko, YOSHIMATSU Kumiko, ARIKAWA Jiro
Lab Animal and Environ, 20, 1, 1, 8, 2012年, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）

Effect of environmental enrichment after the occurrence of wet bedding created by mice and abnormal fur in mice.　　　　　　　　　　　　　　　
Tosa N, Yoshimatsu K, Arikawa J
J Am Assoc Lab Anim Sci, 50, 5, 779, 780, 2011年09月, ［査読有り］, ［筆頭著者］

Rab7 is a critical mediator in vesicular transport of tyrosinase-related protein 1 in melanocytes
Tokimasa Hida, Hitoshi Sohma, Yasuo Kokai, Akinori Kawakami, Kuninori Hirosaki, Masae Okura, Noriko Tosa, Toshiharu Yamashita, Kowichi Jimbow
JOURNAL OF DERMATOLOGY, 38, 5, 432, 441, WILEY-BLACKWELL, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）, How melanosomal proteins such as enzymic proteins (tyrosinase and tyrosinase-related proteins, Tyrps) and structural protein (gp100) are transported from Golgi to melanosomal compartments is not yet fully understood. A number of small GTPases have been found to be associated with melanosomes and we have identified one of them, Rab7, a regulator of vesicular transport, organelle motility, phospholipid signaling and cytosolic degradative machinery, as being involved in the transport of Tyrp1 from Golgi to stage I melanosomes. This study further characterizes the role of Rab7 as a regulator of differential sorting of melanosomal proteins in this process. Murine melanocytes were transiently transfected with a plasmid encoding either wild-type (Rab7WT), constitutively active (Rab7Q67L) or dominant-negative (Rab7N125I and Rab7T22N) Rab7. Through immunocytostaining and confocal laser scanning microscopy, we quantitatively compared the bio-distribution of melanosomal proteins between Rab7WT-expressing cells and mutant Rab7-expressing cells. We also characterized their differential elimination from melanosomal compartments by Rab7 by utilizing a proteasome inhibitor, MG132. Our findings indicate that Rab7 plays an important role in differential sorting of tyrosinase, Tyrp1 and gp100 in early melanogenesis cascade, and that it is more specifically involved with Tyrp1 than tyrosinase and gp100 in the trafficking from Golgi to melanosomes and the specific exit from the degradative process.

The functional role of the glucocorticoid receptor (Gr) and Nur77 in thymocyte development　　　　　　　　　　　　　　　
Noriko Tosa, Takahiro Fukumoto, Tadaaki Miyazaki
Nuclear Receptors, 67, 86, Nova Science Publishers, Inc., 2011年03月
英語, 論文集(書籍)内論文, Nuclear receptors are involved in various aspects of intracellular signal transduction on a range of tissue and play an important role as regulators in numerous essential biological functions. In the thymus, these nuclear receptors also participate in positive or negative selection during T cell development. In particular, the glucocorticoid receptor (Gr) and Nur77 play central roles in apoptosis induction mediated by the T cell receptor (TCR) in mature thymocytes or glucocorticoids (GCs) in immature thymocytes, respectively. Recently, we demonstrated that death-associated protein 3 (DAP3) was critical for TCR-mediated induction of apoptosis downstream of Nur77 in immature thymocytes. The DAP3 is an evolutionarily conserved GTP binding protein that plays a number of roles in normal mitochondrial physiology and in apoptosis induced via the tumor necrosis factor (TNF) family of death receptors. This chapter reviews recent studies of the signal transduction mediated by Gr and Nur77 in thymocyte development, focusing on signaling molecules, such as DAP3, involved in the signaling pathways of Gr or Nur77. Briefly, discussion which have attracted attention are summarized as follows: 1) signaling molecules interacting with Gr or Nur77, 2) the functional role of Gr or Nur77 in subcellular localization, 3) the function of genes subject to induction by Gr or Nur77, 4) crosstalk and its physiological importance in the signal transduction mediated by Gr and Nur77. © 2011 by Nova Science Publishers, Inc. All rights reserved.

Androgen receptors have a potential role in mediating the serotonin synthesis mechanism　　　　　　　　　　　　　　　
Takahiro Fukumoto, Noriko Tosa, Tadaaki Miyazaki
Nuclear Receptors, 113, 120, Nova Science Publishers, Inc., 2011年03月
英語, 論文集(書籍)内論文, Nuclear receptors are a class of proteins that have the ability to directly bind to DNA and regulate gene expression, and these receptors are classified as transcription factors. This report focuses on a new function of AR (androgen receptor). Androgen receptors (ARs) belong to the steroid receptor family and play an essential role in the generation and development of the prostate. Androgen receptors have similar conserved domains that are composed of an NTD (N-terminal domain), a DBD (DNA-binding domain), and an LBD (ligand-binding domain). The NTD works stabilize bound androgen and the AR-LBD mediates the interaction between AR and other proteins, which include Hsps (heat-shock proteins). In the absence of androgen, AR remains in the cytoplasm in an inactive form. After AR binds to androgens, activated AR can bind with other signal molecules and form functional complexes. Then, the complex translocates to the nucleus and regulates the gene expression for androgen regulated genes. Recently, some research has shown that AR can interact with DDC (L-dopa decarboxylase), a key molecule for serotonin (5-HT) synthesizing. Serotonin is a well known neurotransmitter but has been mentioned in the relationship with the generation of the prostate. Then, we introduce here that AR can regulate prostate cancer progression via the serotonin synthesis process. A suggested rewrite of the previous sentence, placed here to avoid ambiguity: This paper suggests that AR may play a role in regulating the progress of prostate cancer via the serotonin synthesis process. © 2011 by Nova Science Publishers, Inc. All rights reserved.

Critical function of death-associated protein 3 in T cell receptor-mediated apoptosis induction
Noriko Tosa, Atsushi Iwai, Taku Tanaka, Tomoka Kumagai, Takeshi Nitta, Satoko Chiba, Masahiro Maeda, Yousuke Takahama, Toshimitsu Uede, Tadaaki Miyazaki
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 395, 3, 356, 360, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2010年05月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Death-associated protein 3 (DAP3) is crucial for promoting apoptosis induced by various stimulations. This report demonstrates that DAP3 is also important for T cell receptor (TCR)-mediated apoptosis induction in immature thymocytes. Enforced expression of DAP3 accelerated the negative selection in developing thymocytes, using the reaggregate thymus organ culture system. In addition, expression of DAP3 accelerated TCR-mediated apoptosis induction in DO11.10 cells. We also demonstrated that DAP3 translocates into the nucleus during TCR-mediated apoptosis in a Nur77 dependent manner. It is concluded that DAP3 is critical for TCR-mediated induction of apoptosis at the downstream of Nur77. (C) 2010 Elsevier Inc. All rights reserved.

Functional role of death-associated protein 3 (DAP3) in anoikis
T Miyazaki, M Shen, D Fujikura, N Tosa, HR Kim, S Kon, T Uede, JC Reed
JOURNAL OF BIOLOGICAL CHEMISTRY, 279, 43, 44667, 44672, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2004年10月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Detachment of adherent epithelial cells from the extracellular matrix induces apoptosis, known as anoikis. Integrin stimulation protects cells from anoikis, but the responsible mechanisms are not well known. Here, we demonstrated that a pro-apoptotic GTP-binding protein, DAP3 (death-associated protein 3), is critical for induction of anoikis. Down-regulation of DAP3 expression by antisense oligonucleotides inhibited anoikis. Conversely, overexpression of DAP3 augmented cell death and caspase activation induced by cell detachment. Furthermore, the association of DAP3 with FADD and the activation of caspase-8 were induced by cell detachment. We also showed that DAP3 is phosphorylated by kinase Akt (PKB), and active Akt can nullify apoptosis induction by DAP3. Mutation of a consensus Akt phosphorylation site in DAP3 renders it resistant to suppression by active Akt in cells. Integrin ligation stimulates Akt activation and phosphorylation of DAP3 in intact cells, as well as suppresses the ability of DAP3 overexpression to augment anoikis. Involvement of DAP3 in anoikis signaling demonstrates a novel role for this GTP-binding protein in apoptosis induction caused by cell detachment.

Functional role of DAP(death associated protein)3 in apoptosis induction
Miyazaki F, Shen M, Tosa N, Fujikura D, Kon S, Uede T, Reed JC
FASEB JOURNAL, 18, 4, A42, 2004年03月23日, ［査読有り］

Critical function of T cell death-associated gene 8 in glucocorticoid-induced thymocyte apoptosis
N Tosa, M Murakami, WY Jia, M Yokoyama, T Masunaga, C Iwabuchi, M Inobe, K Iwabuchi, T Miyazaki, K Onoe, M Iwata, T Uede
INTERNATIONAL IMMUNOLOGY, 15, 6, 741, 749, OXFORD UNIV PRESS, 2003年06月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Transcriptional expression of a gene or genes is absolutely required for induction of glucocorticold-induced thymocyte apoptosis. We have previously shown that expression of T cell death-associated gene 8 (TDAG8) is quickly induced exclusively in the thymus after dexamethasone (DEX) treatment. Here, we present data that TDAG8 expression is induced prior to induction of DEX-mediated apoptosis. In contrast, TDAG8 expression in thymocytes was not induced in the process of gamma-irradiation-mediated apoptosis. TDAG8 expression accelerated only DEX-induced, but not TCR-mediated or gamma-irradiation-induced, thymocyte apoptosis in transgenic mice overexpressing TDAG8. Interestingly, these effects were specifically detected in CD4(+)CD8(+) double-positive thymocytes. Moreover, activation of caspase-3, -8 and -9 was enhanced in thymocytes of TDAG8 transgenic mice after DEX stimulation. In conclusion, TDAG8 expression is involved in glucocorticold-induced signals to activate caspase-9, -8 and -3 for subsequent apoptosis induction in CD4(+)CD8(+) double-positive thymocytes.

Defective development of NK1.1+ T-cell antigen receptor + cells in zeta-associated protein 70 null mice with an accumulation of NK1.1+ CD3- NK-like cells in the thymus.
Iwabuchi K, Iwabuchi C, Tone S, Itoh D, Tosa N, Negishi I, Ogasawara K, Uede T, Onoe K
Blood, 91, 6, 1765, 1775, 2001年, ［査読有り］
英語, 研究論文（学術雑誌）

Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice
M Takiguchi, M Murakami, Nakagawa, I, MM Rashid, N Tosa, S Chikuma, A Hashimoto, T Uede
JOURNAL OF VETERINARY MEDICAL SCIENCE, 62, 1, 29, 36, JAPAN SOC VET SCI, 2000年01月, ［査読有り］
英語, 研究論文（学術雑誌）, MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3(+)CD4(-)CD8(-)B220(+) (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules. CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice.

Detection of various epitopes of murine osteopontin by monoclonal antibodies
H Hotta, S Kon, YU Katagiri, N Tosa, T Tsukamoto, AF Chambers, T Uede
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 257, 1, 6, 11, ACADEMIC PRESS INC, 1999年04月, ［査読有り］
英語, 研究論文（学術雑誌）, We immunized rats with recombinant murine osteopontin protein and obtained four monoclonal antibodies recognizing distinct epitopes of murine osteopontin. OPN1.2 recognized the amino-terminal half of OPN, while OPN2.2, OPN2.3, and OPN3.1 recognized the carboxy-terminal half of OPN. The epitope recognized by OPN2.2 was destroyed by further cleavage of the carboxy half of OPN. The epitope recognized by OPN2.3 was located in the amino-terminal end of the carboxy half of OPN, whereas that recognized by OPN3.1 was located in the carboxy-terminal end of the carboxy half of OPN. OPN1.2 and OPN2.2 recognized thrombin-cleaved osteopontin, whereas thrombin-cleaved osteopontin was not recognized by OPN2.3 and OPN3.1. Thus, these monoclonal antibodies will be useful in structure/function studies of the role of osteopontin in murine models of disease. (C) 1999 Academic Press.

Haptoglobin in Carnivora: a unique molecular structure in bear, cat and dog haptoglobins.
K Mominoki, N Nakagawa-Tosa, M Morimatsu, B Syuto, M Saito
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology, 110, 4, 785, 9, 1995年04月, ［国際誌］
英語, 研究論文（学術雑誌）, Haptoglobin (Hp), a hemoglobin-binding protein in plasma, consists of alpha and beta subunits and has a tetra-chain arrangement (beta-alpha-alpha-beta) connected by disulfide bridges in most mammals so far examined. Dog Hp has been reported to be unique compared with other Hps in respect that (1) the two alpha beta units are joined by a non-covalent interaction rather than a disulfide bridge and (2) the alpha chain has an oligosaccharide-binding sequence (Asn-X-Ser/Thr) and is glycosylated. To determine whether the unique structures of dog Hp are common in the Carnivora, we purified Hps from sera of bear and cat, and analyzed their subunit structure and partial amino acid sequences. The analyses by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, under both reducing and non-reducing conditions, revealed that bear and cat Hps have similar subunit arrangements to dog Hp, suggesting the absence of a disulfide bridge between two alpha chains. This was confirmed by amino acid sequence analysis of the alpha chains: that is, Cys15 participating in the inter-alpha chain disulfide bridge was replaced by Val in bear or Leu in cat and dog. Thus, the unique subunit arrangement of Hp reported in dog may be common in the Carnivora. In contrast to dog Hp, however, alpha chains of bear and cat Hps were found not to have the typical oligosaccharide binding sequence on their alpha chains and were not glycosylated.

HAPTOGLOBIN IN CARNIVORA - A UNIQUE MOLECULAR-STRUCTURE IN BEAR, CAT AND DOG HAPTOGLOBINS
K MOMINOKI, N NAKAGAWATOSA, M MORIMATSU, B SYUTO, M SAITO
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 110, 4, 785, 789, PERGAMON-ELSEVIER SCIENCE LTD, 1995年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Haptoglobin (Hp), a hemoglobin-binding protein in plasma, consists of alpha and beta subunits and has a tetra-chain arrangement (beta-alpha-alpha-beta) connected by disulfide bridges in most mammals so far examined, Dog Hp has been reported to be unique compared with other Hps in respect that (1) the two alpha beta units are joined by a non-covalent interaction rather than a disulfide bridge and (2) the alpha chain has an oligosaccharide-binding sequence (Asn-X-Ser/Thr) and is glycosylated, To determine whether the unique structures of dog Hp are common in the Carnivora, we purified Hps from sera of bear and cat, and analyzed their subunit structure and partial amino acid sequences, The analyses by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, under both reducing and non-reducing conditions, revealed that bear and cat Hps have similar subunit arrangements to dog Hp, suggesting the absence of a disulfide bridge between two a chains, This was confirmed by amino acid sequence analysis of the alpha chains: that is, Cys(15) participating in the inter-a chain disulfide bridge was replaced by Val in bear or Leu in cat and dog, Thus, the unique subunit arrangement of Hp reported in dog may be common in the Carnivora, In contrast to dog Hp, however, alpha chains of bear and cat Hps were found not to have the typical oligosaccharide binding sequence on their alpha chains and were not glycosylated.

STIMULATION OF HAPTOGLOBIN SYNTHESIS BY INTERLEUKIN-6 AND TUMOR-NECROSIS-FACTOR, BUT NOT BY INTERLEUKIN-1, IN BOVINE PRIMARY CULTURED-HEPATOCYTES
N NAKAGAWATOSA, M MORIMATSU, M KAWASAKI, H NAKATSUJI, B SYUTO, M SAITO
JOURNAL OF VETERINARY MEDICAL SCIENCE, 57, 2, 219, 223, JAPAN SOC VET SCI, 1995年04月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, The hepatic synthesis of acute phase proteins in ruminants has been suggested to be regulated by some mechanisms different from those in other species such as rodents and human. To explore possible regulatory factors unique to ruminants, we examined effects of interleukin (IL)-6, IL-1 and tumor necrosis factor (TNF), on haptoglobin (Hp) synthesis using a primary culture system of bovine hepatocytes. After bovine primary cultured hepatocytes were incubated in the presence of various concentrations of the cytokines, the synthesis and mRNA level of haptoglobin and albumin were measured by labeling with [S-35]-methionine and immunoprecipitation, and by Northern blot analysis, respectively. Hp synthesis was dose-dependently increased by recombinant human (rh) IL-6, and also by rhTNF-alpha, but to a less extent, while it was not affected by rhlL-1 beta. The stimulatory effect is mainly pretranslational, because mRNA level of Hp changed in parallel with protein synthesis. In contrast, albumin synthesis was suppressed by these three cytokines similarly. These results are inconsistent with the previously proposed view that TNF and IL-l overlap in their pathways leading to the transcriptional activation of many acute phase protein genes. In conclusion, there is a species-specific unique signaling system, especially for TNF, in transcriptional activation of bovine Hp gene.

ISOLATION AND PRIMARY CULTURE OF BOVINE HEPATOCYTES - ALBUMIN SYNTHESIS AND ADRENERGIC ACTIVATION OF GLYCOGENOLYSIS
N NAKAGAWATOSA, M MORIMATSU, K MOMINOKI, H NAKATSUJI, B SYUTO, M SAITO
JOURNAL OF VETERINARY MEDICAL SCIENCE, 56, 1, 125, 129, JAPAN SOC VET SCI, 1994年02月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, We describe a technique for isolation and primary culture of bovine hepatocytes, and their metabolic characterization. Hepatocytes were isolated from the caudate lobe of bovine liver by perfusion with 0.25 mM ethyleneglycol tetraacetic acid and 0.05% collagenase. The viability and yield of the cells were 70-92% and 0.1-3.6 x 10(7) cells/g liver, respectively. When the isolated hepatocytes were cultured in Williams' medium E, they began to spread in 3 hr and formed monolayers in 24 hr. These monolayers were retained for at least 6 days. To monitor the metabolic activities specific to liver, synthesis and secretion of albumin were measured by labeling with [S-35]-methionine and immunoprecipitation. This activity was low in isolated hepatocytes, but increased after culturing 1-3 days, and decreased again after 6 days. Glycogenolytic activity was also assessed by measuring glucose release to the medium by stimulation with epinephrine. The glycogenolytic response to epinephrine was also enhanced by culturing the hepatocytes 1-3 days, but was decreased after 6 days. Since the isolated bovine hepatocytes retained the liver-specific activities of albumin synthesis and glycogenolysis for several days in culture, these cells are useful for cellular and molecular studies on the functions of bovine liver.

PURIFICATION AND IDENTIFICATION OF A SERUM-PROTEIN INCREASED BY ANTHELMINTIC DRUGS FOR DIROFILARIA-IMMITIS IN DOGS
N TOSA, M MORIMATSU, M NAKAGAWA, F MIYOSHI, E UCHIDA, M NIIYAMA, B SYUTO, M SAITO
JOURNAL OF VETERINARY MEDICAL SCIENCE, 55, 1, 27, 31, JAPAN SOC VET SCI, 1993年02月, ［査読有り］, ［筆頭著者］
英語, 研究論文（学術雑誌）, Polyacrylamide gel electrophoretic analysis of canine serum protein has revealed that the administration of anthelmintics elicits an increase in a certain serum protein. This protein, named PT60, was partially purified by ammonium sulfate fractionation and preparative electrophoresis. The purified PT60 gave a single band with the molecular size of 53 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-reducing conditions. After reduction with 2-mercaptoethanol, two bands appeared at 35 kDa and 17 kDa, indicating that PT60 consists of two subunits which are linked with each other by disulfide bonds. PT60 had the capacity to bind to hemoglobin. In an immunodiffusion test, an antiserum against PT60 cross-reacted with canine haptoglobin (Hp). N-terminal amino acid sequences of two PT60 subunits were identical to those of alpha and beta subunits of canine Hp, respectively. Thus, PT60 was identified as Hp.

コモンマーモセットの飼育管理における試行錯誤 ～個性豊かなマーモセットたちと向き合って～　　　　　　　　　　　　　　　
土佐紀子, 日本実験動物技術者協会北海道支部会誌, 45, 2022年04月, ［招待有り］, ［筆頭著者］

コモンマーモセット飼育室の稼働 ～沢山の方たちに助けて頂いて～　　　　　　　　　　　　　　　
土佐紀子, 日本実験動物技術者協会北海道支部会誌, 44, 6, 10, 2021年04月, ［招待有り］, ［筆頭著者］
日本語, その他

プロポロリスコーティング床敷はケージ内常在菌の増殖とマウスアレルゲンの発生を抑制する
土佐紀子, 吉松組子, 築山忠維, 畠山鎮次, 有川二郎, 日本実験動物学会総会講演要旨集, 61st, 280, 2014年05月01日
日本語

DAP3(death associated protein 3)を介するアポトーシスシグナルの解明
宮崎忠昭, SHEN M, 藤倉大介, 土佐紀子, 今重之, 上出利光, REED J, 日本分子生物学会年会プログラム・講演要旨集, 27th, 2004年

Functional role of DAP3 (death associated protein 3) in anoikis
T Miyazaki, M Shen, N Tosa, S Kon, T Uede, JC Reed, FASEB JOURNAL, 17, 7, C316, C316, 2003年04月
FEDERATION AMER SOC EXP BIOL, 英語, 研究発表ペーパー・要旨（国際会議）

アポトーシス誘導におけるDAP3の機能
宮崎忠昭, CHEN M, 土佐紀子, 今重之, 上出利光, REED J, 日本分子生物学会年会プログラム・講演要旨集, 26th, 2003年

The functional analysis of DIG1 molecule, a putative G protein-coupled receptor, in dexamethasone-induced thymocyte apoptosis.
N Tosa, M Murakami, M Iwata, WY Jia, M Yokoyama, K Iwabuchi, M Inobe, T Uede, FASEB JOURNAL, 15, 4, A315, A315, 2001年03月
FEDERATION AMER SOC EXP BIOL, 英語, 研究発表ペーパー・要旨（国際会議）

胸腺内NK-T細胞の分化経路　　　　　　　　　　　　　　　
岩渕和也, 岩渕千雅子, 刀祢さおり, 土佐紀子, 上出利光, 小野江和則, 日本呼吸器外科学会雑誌, 15, (), 1, 2001年

ウシ血清ハプトグロビン濃度の測定法の確立と急性相におけるその変動
森松正美, 土佐紀子, 内貴正治, 斉藤昌之, 家畜生化学研究会報, 29, 61, 68, 1992年09月
日本語

『一般外科医のための外傷診療DCRトレーニングコース』開発における有用性の証明
科学研究費助成事業
2022年04月01日 - 2027年03月31日
村上 壮一, 土佐 紀子, 方波見 謙一, 倉島 庸, 平野 聡, 本間 宙, 七戸 俊明, 伊澤 祥光
日本学術振興会, 基盤研究(C), 北海道大学, 22K10408

実験動物主要感染症の迅速・簡便な個体別血清診断法：多項目イムノクロマト法の開発
科学研究費助成事業
2020年04月01日 - 2023年03月31日
土佐 紀子, 林元 展人, 石田 智子, 森松 組子
学術・科学技術の研究に供する動物の微生物モニタリングは、研究結果の信頼性の担保、および動物福祉の観点から必要不可欠である。近年、多系統少数個体維持の増加や個別換気型飼育機の普及等により、従来の微生物モニタリング方法、すなわち「おとり動物」の感染症の検出を指標とする間接的な方法では感染症を摘発することが困難となっている。この問題を解決するためには、動物を安楽死させることなく微量全血を用い、マウス、ラットの主要感染症を同時に且つ迅速・簡便に検出できる個体別血清診断法の確立が必要である。その方法として有望と考えられる「多項目イムノクロマト(ICG)法」の開発について申請者らは取り組んで来た。
本研究ではこれまでの成果を基に、１）多項目ICG法の検出条件が確定したマウス・ラットの感染症（マウス：マウス肝炎ウイルス感染症、センダイウイルス感染症、ティザー病。ラット：唾液腺涙腺炎ウイルス感染症、センダイウイルス感染症、腎症候性出血熱）の検出における本法の実用性を検証し、２）上記感染症に加え、主要感染症であるマイコプラズマ感染症（マウス、ラット）とティザー病（ラット）のICG法を確立することを目的としている。２年度（令和３年度）においては、以下の研究を実施した。
１．多項目ICGスティックの最適な保管条件・最長保管期間を明らかにする実験において、保管期間６ヶ月および１２ヶ月の多項目ICGスティックの各保管条件における検出・感度特異性への影響の結果を得た。
２．乾燥血液を用いた時のICG法の検出・感度特異性を明らかにするため、乾燥血液を作製するための条件を検討した。
３．多項目ICGスティックの輸送条件を確定するための実験設備の準備を行った。
４．Myco（マウス、ラット）とTyzzer（ラット）のICG法を確立するため、抗原の準備を行い、ICGメンブレンに塗布する抗原処理方法の実験に着手した。
日本学術振興会, 基盤研究(C), 北海道大学, 20K06456

地域における重症外傷患者の救命率改善を目指した新規DCSトレーニングコースの開発
科学研究費助成事業
2016年04月01日 - 2020年03月31日
村上 壮一, 澤村 淳, 大滝 純司, 土佐 紀子, 倉島 庸, 平野 聡, 本間 宙, 七戸 俊明, 伊澤 祥光
全国の外傷外科医117名に協力を依頼、Delphi法を用いたアンケート調査によりCronbachα=0.94を以て35項目の一般外科医が修得すべき外傷診療技能を選定した。この項目に対する経験および習熟度を北海道内の全一般外科医730名に対しアンケート調査を行い444名の有効回答を分析、脾臓摘出術、横隔膜損傷修復術、腹腔内ガーゼパッキング、肝損傷ガーゼパッキング、肝損傷部縫合術、一時的閉腹法、緊急開腹術、FASTの8項目をトレーニングすべき項目として抽出した。これによりシミュレーター、生体ブタ、献体、e-learningを用いた一般外科医に対する新規外傷診療トレーニングコースが完成した。
日本学術振興会, 基盤研究(C), 北海道大学, 16K11392

環境要因によるＴリンパ球レパトア形成と実験動物アレルギー発症との関連性の解明
科学研究費助成事業
2015年04月01日 - 2018年03月31日
土佐 紀子
本研究から、Tリンパ球レパトア形成が確立する期間である胎仔期および幼若期のマウスにおける抗原暴露が実験動物アレルギーの誘導を抑制したことから、胎仔期および幼若期の環境要因が実験動物アレルギー発症に影響を与え、その影響はレパトア形成と関係する可能性が示唆された。さらに、動物アレルゲンであるCan f 1 感作マウスにおいてアルミニウム塩アジュバントによりCan f 1特異的IgEとIL-13のレベルが抑制され、この現象は食物アレルゲンである卵白アルブミンでは認められなかったことから、実験動物アレルギーの誘導機構は食物アレルギーとは異なることが示唆された。
日本学術振興会, 基盤研究(C), 北海道大学, 15K07776

実験動物由来人獣共通感染症および主要感染症モニタリング用多項目イムノクロマト法
科学研究費助成事業
2015年04月01日 - 2018年03月31日
有川 二郎, 土佐 紀子, 高倉 彰
実験用ラットの感染症（センダイウイルス感染症、ティザー病、マイコプラズマ感染症、唾液腺涙腺炎ウイルス感染症、腎症候性出血熱）、及び、実験用マウスの感染症（マウス肝炎、センダイウイルス感染症、ティザー病、マイコプラズマ感染症）について、上記感染症病原体の菌体抽出抗原もしくは固定ウイルス粒子とProtein A標識金コロイド粒子を用い、IgG検出イムノクロマト法テストストリップを作成した。ラットのマイコプラズマ感染症とティザー病を除き、免疫血清や動物血清を用いた解析で、十分な感度と特異性を示し、血清診断法として有効な試験法であった。また本法は、希釈全血液も使用可能で、簡便かつ迅速な試験法であった。
日本学術振興会, 基盤研究(C), 北海道大学, 15K07717

ステロイド依存性アポトーシスにおけるDIG-1遺伝子の機能解析
科学研究費助成事業
1998年 - 2000年
上出 利光, 土佐 紀子, 岩田 誠
1.正常マウスの胸腺細胞および末梢リンパ球では発現していないが、デキサメサゾン(DEX)で刺激すると30分以内に主にCD4+CD8+胸腺細胞に発現する遺伝子としてDIG-1遺伝子を同定した。
2.DIG-1遺伝子は7回膜貫通型蛋白をコードしており、G蛋白結合受容体に属する分子である。
3.DIG-1遺伝子をlckプロモーターの支配下においた遺伝子導入マウス(以下DIG-1 TG)の3系統を樹立した。
4.DIG-1 mRNAの発現が高いB系統、中等度のC系統、低いE系統である。対照群に比して、BとC系統のTGでは、胸腺重量が有意に低下していた。B、C、Eの全ての系統において100μgのDEX腹腔内投与により、対照群に比し有意な胸腺細胞の重量低下とアポトーシス細胞の増加を認めた。アポトーシスを示す細胞は主に胸腺皮質に存在し、CD4+CD8+胸腺細胞であった。胸腺に比べて少数ではあるがTGではDEX投与後脾臓でもアポトーシスを示す細胞が増加した。
5.アポトーシス関連分子であるBcl-2とBAX蛋白の発現をDEX投与前後で検討したが、DEX投与前後で有意な変化はなく、対照群と比較しても有意差は認めなかった。
日本学術振興会, 基盤研究(B), 北海道大学, 10470053