• Title/Summary/Keyword: Mucosal epithelial cells

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Cholera Toxin B Subunit-Porphyromonas gingivalis Fimbrial Antigen Fusion Protein Production in Transgenic Potato

  • Lee, Jin-Yong;Kim, Mi-Young;Jeong, Dong-Keun;Yang, Moon-Sik;Kim, Tae-Geum
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.268-274
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    • 2009
  • Porphyromonas gingivalis, the gram-negative anaerobic oral bacterium, initiates periodontal disease by binding to saliva-coated oral surface. The cholera toxin B subunit (CTB) genetically linked to FimA1 (1-200 aa) or FimA2 (201-337 aa) of the P. gingivalis fimbrial antigen were introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation method. The integration of CTB-FimA1 or CTB-FimA2 fusion genes were confirmed in the chromosome of transformed leaves by genomic DNA PCR amplification method. Synthesis and assembly of the CTB-FimA fusion proteins into oligomeric structures with pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding activities of CTB-FimA fusion proteins to intestinal epithelial cell membrane receptors were confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA showed that the expression levels of the CTB-FimA1 or CTB-FimA2 fusion proteins were 0.0019, 0.002% of the total soluble protein in transgenic tuber tissues, respectively The synthesis of CTB-FimA monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using edible plants for the production of enterocyte targeted fimbrial antigens that could elicit mucosal immune responses.

Techniques for laboratory diagnosis of cryptosporidiosis (크��토스포리디움증의 실험실적 진단법)

  • Noh, Jae-wuk;Kang, Du-weon;Jang, Du-hwan;Kang, Yung-bai
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.501-507
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    • 1991
  • Diagnosis of cryptosporidiosis is currently confirmed by the detection of the oocysts or endogenous stages in fecal or tissue samples. Various conventional staining methods and serodiagnostic techniques have been reported, but the latter has far been limited to a few laboratories. Cryptosporidium has recently been reported in mice and chiekens in Korea, but there has been no report on staining methods to the oocysts. The present study was performed by light and scanning electron microscopic observations, and discussed with staining properties of four conventional methods such as dichromate solution floatation method, Carbol fuchsin stain, Auramine-O stain and Giemsa stain method. Cryptosporidial oocysts were isolated from the laboratory mouse. In tissue sections of duodenum, jejunum, ileum, cecum and upper colon, numerous very small, basophilic bodies were observed on the border of mucosal epithelial cells. In scanning electron microscopic observations, a few of developmental stages of Cryptosporidium were seen. Two types of thick and thin-walled oocysts were recognized in the intestinal contents. Mean size of its were $5.19{\pm}0.23{\times}4.31{\pm}0.32{\mu}m$ and $5.14{\pm}0.25{\times}4.27{\pm}0.4{\mu}m$, respectively. Carbol fuchsin and Auramine-O stain methods are recommended as the satisfactory ones for the identification of Cryptosporidium oocysts. Giemsa stain was also recommended as available in the laboratory, because a few of developmental stage fo Cryptosporidium could be seen by it.

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Effects of Cryptospoyidium bnileyi infection on the bursa of Fabricius in chickens (닭에 있어서 닭와포자충 감염이 파브리시우스낭에 미치는 영향)

  • Lee, Jae-Gu;Kim, Hyeon-Cheol;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.35 no.3
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    • pp.181-188
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    • 1997
  • In order to clarify the effect of cryptosporidiosis on immune response, histopathological changes associated with experimentally occurring bursal cryptosporidiosis in chickens were chronologically observed as the first step. A total of 150 2-day-old chickens was each inoculated orally with a single dose of 5 × 105 Cryptospori,mum bailevi oocysts. The chickens showed a normal profile of oocyst shedding in droppings. The bursa indices throughout the experimental period indicated negligible reactions. Numerous cryptosporidia occurred in the microvillous border of bursal epithelium between days 4 and 16 postinoculation (PI). Appearance of the most mast cells was followed by a dramatic loss of the protozoa in the bursa of Fabricius (BF). The distribution of the coccidium coincided with heterophil infiltration in the epithelium and adjacent lamina propria. The histopathological lesion was marked diffuse chronic superficial purulent bursitis with heterophil infiltration in the epithelium and adjacent lamina proprla and mucosal epithelial hyperplasia. These results suggest that the bursitis may induce immunosuppressive effect.

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Excessive copper in feed not merely undermines animal health but affects food safety

  • Ma, Zicheng;Li, Yan;Han, Zifeng;Liu, Zhaohu;Wang, Hongyu;Meng, Fanliang;Liu, Sidang;Chen, Dawei;Liu, Mengda
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.31.1-31.12
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    • 2021
  • Background: Blackened intestines in slaughtered pigs have been commonly observed in China in recent years. However, no cause has been reported. Objectives: We attempted to determine whether the blackening of the pig intestine was related to an excess of copper (Cu) in their feed. Methods: In this study, we observed and collected porcine intestines in small- and large-scale pig slaughterhouses in Shandong province from May to October 2018. Twelve types of metal ions were detected in the black intestinal samples. Results: The Cu level in the intestine samples was mostly higher than the Chinese national limit for food. Further study showed that Cu supplementation in most commercial porcine feed also exceeded the national standard. An animal model (mouse) that could mimic the intestinal blackening in pigs was established. Compared to control mice, Cu accumulated in the liver and intestines of mice fed an excessive Cu level, confirming the excessive Cu in the feed may be considered the major cause of blackened porcine intestines. Microscopic examination revealed that black intestines had many particles containing Cu in the lamina propria of the intestinal mucosa, and the intestinal mucosal epithelial cells showed degeneration and necrosis. Conclusions: In conclusion, overuse of Cu in animal feed can lead to animal poisoning and Cu accumulation in animal products. Such overuse not only harms the health of livestock but can also affect public health.

Histochemical Studies of the Intestinal Mucosubstances in Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma, and Helirolenus dactylopterus (노래미, 쑤기미, 구실우럭 및 홍감펭 장관 점액질에 대한 조직화학적 연구)

  • Jeong, Gil-Nam;Jo, Un-Bock
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1669-1674
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    • 2007
  • The conventional histochemical stains were used to study the properties of mucosubstances of the intestine in four teleostean species, i. e., Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma, and Helicolenus dactylopterus, all belonging to the order Perciformes. The following methods were used; periodic acid Schiff (PAS), alcian blue (AB) pH at 2.5, AB pH at 1.0, AB pH at 2.5-PAS, and aldehyde fuchsin (AF) pH at 1.7-AB pH at 2.5 stain. The mucosal folds of intestines show differences in species and regions. Widely distributed in all portions studied, goblet cells situated between cylinderical epithelial cells are round or ovoid in shape. They were most densely distributed in distal intestine. In the middle and. distal intestine of Epinephelus chlorostigma and all regions of intestines of Agramus agramus, Inimicus japonicus, and Helicolenus dactylopterus, the presence of both acidic and neutral mucin was confirmed. The property of acidic mucosubstance was sialomucin. Neutral mucin was only encountered in the proximal intestine of Epinephelus chlorostigma. The amount and property of mucin showed differences in species and regions. In the distal intestine of Inimicus japonicus, the amount of acidic mucin is similar to that of neutral mucin. In all regions of intestine of Agramus agramus, proximal and middle intestine of lnimicus japonicus, middle and distal intestine of Epinephelus chlorostigma, and distal intestine of Helicolenus dactylopterus, acidic mucin occured more frequently than neutral mucin. The proximal and middle intestine of Helicolenus dactylopterus have more neutral mucin than acidic mucin.

Chronological observation of intestinal lesions of rots experimentally infected with Echinostoma hortense (흰쥐의 실험적 호르틴스극구흡충 감염에 있어서 장 병변에 대한 경시적 관찰)

  • 이순형;노태영
    • Parasites, Hosts and Diseases
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    • v.28 no.1
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    • pp.45-52
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    • 1990
  • Intestinal histopathological changes due to infection with Echinostcma hortense (Trematoda) were studied in rats after experimental infection with the metacercariae. The metacercariae were obtained from the tadpoles of Rana nigrcmaculata, a second intermediate host infected in the laboratory. Total 18 albino rats(Sprague-Dawley) were given 200 matacercariae each and sacrificed on the day 1, 3, 7, 11, 22 or 44 post-infection(PI) Segments of- the small intestine at 1, 3, 5, 8 and 30 cm posterior to the pylorus(PTP) were rejected and studied histopathologically. 1. The flukes were seen to have intruded into the intervillous space in the upper small intestine at early stages(1∼3 days PI), however, they were located mainly in the intestinal lumen at later stages(7∼44 days PI) . The flukes were sucking and destroying the epithelial layers of villi with their oral and ventral suckers. 2. Histopathological changes of the intestine were recognizable in as early as 1∼3 days after infection, and the changes became severer as the infection progressed. 3. The intestinal mucosa was histopathologically characterized by villous atrophy and crypt hyperplasia throughout the infection period. Major villous changes were blunting, fusion, severe destruction and loss of epithelial layers of villi. Villous/crypt(V/C) height ratio was remarkably reduced from 3 : 1 in controls to 1 : 1 in severely infected animals. In the stroma of villi, inaamma- tory cell infiltrations, vascular congestion, edema, and/or fibrosis were recognized. The goblet cells were increased in number after 11 days PI. It was revealed in the present study that the pathological changes in the intestine of rats infected with E. hortense were chieay confined to the mucosal layer of the upper small intestine, however, the changes were very severe accompanying remarkable destruction of villi and loss of mucosal integrity, and persistent until 44 days PI.

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Protective Effect of Water Extract of Leonurus japonicus Houttuyn against HCl/EtOH-induced Gastric Mucosal Damage and Genotoxicity Evaluation using Micronucleus Test (익모초 물 추출물의 위염 예방효과 및 소핵시험을 이용한 유전독성 평가)

  • Nho, Jong Hyun;Lee, Hyun Joo;Jang, Ji Hun;Yang, Beo Dul;Woo, Kyeong Wan;Kim, A Hyeon;Seo, Jae Wan;Kim, Sun Young;Cho, Hyun Woo;Jung, Ho Kyung
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.282-289
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    • 2019
  • Gastritis is an inflammatory disease involving the stomach and is caused by several factors, including stress, non-steroidal anti-inflammatory drugs such as aspirin, liquor, and Helicobacter pylori. In Korea, Leonurus japonicus Houttuyn (LJW) has been used as traditional medicine for vaginal bleeding, hematuria, and bruise. Previous studies have reported that LJW exhibited hepatoprotective, cardioprotective, and anti-hyperlipidemic effect. However, the effect of the water extract of LJW on gastritis was not elucidated. Thus, we evaluated the anti-gastric effect and genotoxicity of LJW. LJW effectively prevented the degeneration of surface mucous cells and glandular epithelial cells and vascular congestion induced by HCl/EtOH. Micronucleus assay indicated that the rate of micronucleated polychromatic erythrocytes/polychromatic erythrocytes was not significantly different compared that of the control. Further experiments are required to determine the role of LJW in the gastric injury process such as cyclooxygenase signaling pathway and the secretion of mucus in the stomach.

Experimental activation of cryptosporidiosis in mice by immunosuppression (면역억제에 의한 마우스의 Cryptosporidium 발현 실험)

  • 채종일;신손문
    • Parasites, Hosts and Diseases
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    • v.28 no.1
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    • pp.31-38
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    • 1990
  • Cryptosporidium, a coccidian parasite first described by Tyzzer (1907) from a laboratory mouse, has become an important human enteric pathogen causing overwhelming diarrhea especially in immunocompromised patients such as AIDS. This parasite has been reported from over 20 countries and is recognized as a cosmopolitan species. In Korea, however, thEre has been no report on human as well as animal cryptosporidiosis. This study was performed so as to verify the presence of Cryptosporidium in Korea by activating the parasite from laboratory mice by immunosuppression. Total 65 conventionally.bred ICR mice including a control (5 mice) and 3 experimental groups (20 each) were used for this study. Group I was immunosuppressed with Prednisolone injection (1 mg IM, every other day) for 7 weeks. Group II (prednisolone injection and tetracycline administration) and Group III. (prednisolone injection and trimethoprim-sulfamethoxazole administration) were prepared to observe the effect of antibacterial agents on the activation of cryptosporidiosis. In fecal examinations of mice Cryptosporidium oocysts($4-6{\mu\textrm{m}}$ in size) were detected from 1 week after the start of immunosuppression and the mice began to die. In H-E stained tissue sections of the lower jejunum, numerous very small ($2~4{\;}{\mu\textrm{m}}$), dense, ovoid or spherical, slightly basophilic bodies were seen attached on the free border of mucosal epithelial cells. In scanning and transmission electron microscopic observations, these organisms were identified as various developmental stages of Cryptosperidium. The species is considered to be C. parvum. Cryptosporidiosis was activated not only in Group I but also in Group II and III, indicating no protective effects of the antibacterial agents used, although the mice in Group II and III lived longer than those in Group I. The present study confirmed that Cryptosporidium exists in laboratory mice bred in Korea, and predicts possible occurrence of human cryptosporidiosis in Korea.

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Effect of Cimetidine and Gastric Acidity on the Gastric Mucosal Retention of $^{99m}Tc-Pertechnetate$ in Rats (Cimetidine과 위산도 변화가 $^{99m}Tc-Pertechnetate$의 흰쥐 위벽 집적에 미치는 영향)

  • Kim, Sung-Hoon;Kim, Jong-Woo;Bahk, Yong-Whee
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.1
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    • pp.41-48
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    • 1989
  • $^{99m}Tc-Pertechnetate\;(TcO_4^-)$ is concentrated by the stomach after intravenous injection, allowing the detection of ectopic gastric mucosa. It has been used to develop a noninvasive test of gastric secretion. However the cellular site of concentration is still controversial, that is whether mucin-secreting epithelial cell or acid-secreting parietal cell. This study is planned to investigate the effects of cimetidine and gastric acidity on the retention of $TcO_4^-$ in the gastric wall of the rat. Also we further attempted to clarify the uptake and secreting cell of $TcO_4^-$ in the gastric mucosa. One hundred rats were divided into two groups, preliminary (40 rats) and main examination group (60 rats). Preliminary examination group was composed of fasting group (20 rats) for the detection of the time for reaching stable $TcO_4^-$ retention ratio in gastric wall and post-prandial group (20 rats) for the detection of the time for reaching the maximal gastric acidity. Main examination group was composed of fasting group (30 rats), which was subdivided into control group (10 rats), cimetidine group (10rats), $Mylanta^{(R)}$ group (10 rats) and post?prandial group (30 rats), which was subaivided into 90 min group (10 rats), 90 min cimetidine group (10 rats), and 120 min group (10 rats). Retention ratio (%) of $TcO_4$ in the gastric wall and the pH of the gastric contents were measured in the extracted stomach of the six groups. Gastric wall retention ratio of $TcO_4^-$ was calculated by the gastric wall radioactivity (cpm) divided by total gastric radioactivity (cpm) at 30 mins after intravenous injection of 0.4 mCi of $TcO_4^-$. The results were as follows: 1) The time required for reaching stable $TcO_4$ retention ratio and the lowest gastric PH were 30 min and 90 min, respectively. 2) In the fasting group, the gastric wall retention ratio of $TcO_4^-$ was significantly increased in the cimetidine group, compared with the control group (P < 0.01). However there was no significant difference between the control and $Mylanta^{(R)}$ group 3) The $TcO_4^-$ retention ratios of 90 min and 120 min groups were lower than that of the fasting control group (p < 0.05), either. After administration of cimetidine, the retention ratio was significantly increased in 90 min group (p < 0.01). 4) While $TcO_4^-$ retention ratio and gastric pH were well correlated in the post-prandial 120 min group (r=0.7112, p<0.05), in the post-prandial 90 min and 90 min cimetidine groups correlated poorly. However, there was no correlation in the three fasting groups at all. Referring the above results, we infer that $TcO_4^-$ is secreted into the gastric lumen by both parietal and non-parietal cells, with dominant non-parietal $TcO_4^-$ secretion in the fasting state and dominant parietal $TcO_4^-$ secretion in the stimulated state.

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Effects of Di-(2-ethylhexyl) phthalate (DEHP) on Ultrastructure of Rat Seminal Vesicle (Di-(2-ethylhexyl)phthalate (DEHP)가 흰쥐 저정낭의 미세구조에 미치는 영향)

  • Kil, Young-Chun;Kim, Wan-Jong;Shin, Kil-Sang
    • Applied Microscopy
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    • v.30 no.1
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    • pp.73-80
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    • 2000
  • Di-(2-ethylhexyl)phthalate (DEHP) has been known as one of endocrine disruptors. The present study was carried out to investigate the alterations of fine structure in rat seminal vesicle after oral intubation of DEHP in dosages of 1g/kg/day, 2g/kg/day or 3g/kg/day respectively in 0.5 ml of corn oil for If days. In rats treated with DEHP for 15 days, seminal vesicle exhibited extensive histological alterations compared to those observed in control groups. The size of the seminal vesicle and the mucosal folds decreased, but the lamina propria was considerably thickened. The ultrastructural changes of epithelial cells in seminal vesicle of rat treated with DEHP were characterized by the high nuclear/cytoplasmic ratio and the increased beterochromatin within irregular nuclear envelope. And also, the rough endoplasmic reticulum, Golgi complex and secretory vesicles were poorly developed. In conclusion, DEHP caused the ultrastructural and functional alterations of seminal vesicle in rats dose-dependently. It is suggested that these detrimental effects of DEHP on seminal vesicle are derived from the decrease level of testosterone.

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