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Takeshi SHINOGI Tomoko SUZUKI Yoshihiro NARUSAKA Pyoyun PARK 《Journal of General Plant Pathology》2002,68(1):38-45
The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to
pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves
by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H2O2. A small amount of non-specific generation of H2O2 was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H2O2 at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only
in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated
susceptible leaves. In addition, generation of H2O2 at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the
H2O2 generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane
fragments from altered portions of the invaginated plasma membrane.
Received 21 September 2001/ Accepted in revised form 25 October 2001 相似文献
4.
Takeshi Shinogi Tomoko Suzuki Takayuki Kurihara Yoshihiro Narusaka Pyoyun Park 《Journal of General Plant Pathology》2003,69(1):7-16
Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection
of O2
−, diaminobenzidine (DAB) methods for microscopic detection of H2O2, and cerium chloride methods for ultrastructural detection of H2O2. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves
of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified
ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves.
These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation
in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently
induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated.
After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated
with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin
layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS
generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in
the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated
with the expression of susceptibility.
Received: June 3, 2002 / Accepted: July 31, 2002 相似文献
5.
Kasumi Sudo Mariko Ochiai Naoyuki Aihara Noriyuki Horiuchi Atsushi Yamamoto Sachiko Matsumoto Koji Oishi 《Journal of toxicologic pathology》2021,34(2):137
Batch safety tests (BSTs) of veterinary vaccines are conducted using small laboratory animals to assure the safety of vaccines according to several criteria, including clinical signs and change in body weight. Although the latter is used as an evaluation index in BSTs, there have been no reports on the internal changes that affect body weight during the test period. Therefore, we analyzed BST via pathological examination of the tested animals. Here, BSTs were performed for 176 batches using mice and 126 batches using of guinea pigs. Most of the gross findings could be classified into four lesion types (nodules, adhesions, ascites, no apparent signs), with only one vaccine inducing lesions that could not be classified into any of these four types. Histopathological examination revealed that the reactions caused by BST were pyogenic and/or granulomatous inflammation. Nodular or adhesive lesions comprised more severe pyogenic granulomatous inflammation than ascites or cases with no apparent gross lesions. These nodular or adhesive lesions were more frequently induced by vaccines that contained an adjuvant than by vaccines that did not contain an adjuvant. The cases with “exceptional” gross findings histologically presented severe necrosis of the hematopoietic system. Additional testing showed that these “exceptional” lesions were induced when a specific type of light liquid paraffin was injected along with other vaccine additives. Our results show that body weight loss and/or lesions during BST were induced by proinflammatory properties of the tested vaccines and that BST is a sensitive method for detecting unexpected effects of vaccine components. 相似文献
6.
Kanae SHIOKAWA Chandika D. GAMAGE Nobuo KOIZUMI Yoshihiro SAKODA Kenta SHIMIZU Yoshimi TSUDA Kumiko YOSHIMATSU Jiro ARIKAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(2):221-230
The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was
assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then,
truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared
between two recombinant expression hosts Escherichia coli and Pichia
pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris
(tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E.
coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with
E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a
Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results
from flaB gene PCR of kidney samples and WB with whole Leptospira cell.
Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained
from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an
adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and
sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32
expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of
background reaction. 相似文献
7.
Oonuma T Morimatsu M Ochiai K Iwanaga T Hashizume K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(3):279-284
Molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) is a nuclear IkappaB protein that is also known as interleukin-1-inducible nuclear ankyrin repeat protein and inhibitor of nuclear factor kappaBzeta (IkappaBzeta). We previously observed that MAIL-deficient mice were affected by atopic dermatitis-like skin lesions and demonstrated the importance of MAIL in the skin. In this study, we investigated MAIL expression in mouse keratinocytes. MAIL mRNA was constitutively expressed in the skin epidermis. MAIL expression was also confirmed in primary keratinocytes and the PAM212 keratinocyte cell line. The inhibitors of nuclear factor kappaB (NF-kappaB)-Bay11-7082 and the IkappaBalphaM supersuppressor-considerably downregulated MAIL expression in the keratinocytes. Immunoreactivity for NF-kappaB components was localized in the cytoplasm and nucleus of normal unstimulated keratinocytes. The expression level of MAIL in the skin did not change following lipopolysaccharide (LPS) administration to mice. Interestingly, in accordance with the in vivo findings, the MAIL expression level did not change following LPS stimulation even in primary keratinocytes; however, MAIL expression was strongly increased by interleukin-1 stimulation. These results collectively suggest that the constitutive expression of MAIL in keratinocytes is controlled, at least in part, by NF-kappaB and that there may be LPS-specific repressive mechanisms that inhibit MAIL induction. 相似文献
8.
Shin JH Sakoda Y Kim JH Ochiai K Umemura T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(11):1167-1169
Rabbits were immunized with inactivated influenza virus via the subarachnoidal (SA) or subcutaneous (SC) route, and the antibody titers in cerebrospinal fluid (CSF) and serum were assayed. There were no nervous signs or morphological lesions related to SA immunization. In the SC group, the antibody titer was elevated in serum, but not elevated in CSF. In the SA group, the antibody titer was significantly elevated in serum and even in CSF, and their antibody titers were much greater than in the SC group. The present results suggest that intrathecal immunization is more effective than SC immunization at inducing a protective immune response against the transneural spread of viruses. 相似文献
9.
Ochiai N Fujimura M Motoyama T Ichiishi A Usami R Horikoshi K Yamaguchi I 《Pest management science》2001,57(5):437-442
Osmotic-sensitive (os-1) mutant alleles in Neurospora crassa exhibit resistance to dicarboximides, aromatic hydrocarbons and phenylpyrroles. We have previously reported that the os-1 mutants can be classified into two groups based on their resistance to fungicides and osmotic stress: type I, which are highly resistant to iprodione and fludioxonil but moderately sensitive to osmotic stress, and type II, which are highly sensitive to osmotic stress but moderately resistant to fungicides. To explain the mechanism of resistance to these fungicides, we cloned and sequenced the mutant os-1 genes that encode putative osmo-sensing histidine kinase. Within the os-1 gene product (Os1p), the type I strains, NM233t and Y256M209, carried a stop codon at amino acid position 308 and a frameshift at amino acid position 294, respectively. These mutation sites were located on the upstream of histidine kinase and the response regulator domains of Os1p, strongly suggesting that type I strains are null mutants. The null mutants, NM233t and Y256M209, were highly resistant to iprodione and fludioxonil; thus Os1p is essential for these fungicides to express their antifungal activity. The amino acid changes in Os1p, 625Pro from Leu, 578Val from Ala, and 580Arg from Gly were found in the type II strains, M16, M155-1 and P5990, respectively. Os1p is novel in having six tandem repeats of 90 amino acids in the N terminal. Each amino acid change of the type II strains was located on the fifth unit of six tandem repeats. Type II strains with single amino acid changes were more sensitive to osmotic stress than the null mutants (type I), indicating that the amino acid repeats of Os1p were responsible for an important function in osmo-regulation. 相似文献
10.
Yoshiaki Nakagawa Yoshihiro Soya Katsumi Nakai Nobuhiro Oikawa Keiichiro Nishimura Tamio Ueno Toshio Fujita Norio Kurihara 《Pest management science》1995,43(4):339-345
The ability to stimulate N-acetylglucosamine (GluNAc) incorporation in-vitro of a number of N-tert-butyl-N,N′-dibenzoylhydrazines having various substituents on both phenyl rings was measured in cultured integument excised from the rice stem borer (Chilo suppressalis Walker). The relationship between in-vitro and larvicidal potency was approximately linear. The substituent effects on variations in the potency were similar between in-vitro and larvicidal activities. An inhibitor of oxidative detoxication, piperonyl butoxide, had no synergistic effects on the in-vitro potency. The ability of some dibenzoylhydrazines to inhibit GluNAc incorporation at exposure periods longer than the optimum for stimulation was also measured in a similar cultured integument system. The relationship between the inhibitory and stimulatory potency indices was linear, indicating that the larvicidal activity of dibenzoylhydrazines is closely related to its ability to stimulate as well as to inhibit GluNAc incorporation into the larval cuticle. 相似文献