Kim, Jang-Seoung;Chang, Ji-Hoon;Park, Eun-Jeong;Chung, Soo-Il;Yum, Jung-Sun
Journal of Microbiology and Biotechnology
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v.10
no.6
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pp.865-872
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2000
Helocobacter phylori is the major cause of gastritis, peptic ulcer, and a principal risk factor for gastric cancer. As the firs step towards a vaccine against H. pylori infection, Hy.pylori urease was expressed and purified as a recombinant apoenzyme (rUrease) in E. coli. In order to develop an effective immunization protocol using rUrease, the host immune responses were evaluated after the oral immunization of mice with rUrease preparations plus cholera toxin relative to various conditions, such as the physical nature of the antigen, the frequency of the booster immunization, the dose of the antigen, and the route of administration. The protective efficacy was assessed using a quantitative culture following an H. pylori SS1 challenge. It was demonstrated that rUrease, due to its particulated nature, was more superior than the UreB subunit as a vaccine antigen. The oral immunization of rUrease elicited significant systemic and secretory antibody responses, and activated predominantly Th2-type cellular responses. The bacterial colonization was significantly reduced (~100-fold) in those mice immunized with three or four weekly oran doses of rUrease plus cholera toxin (p<0.05), when compared to the non-immunized/challenged controls. The protection correlated well with the elicited secretory IgA level against rUrease, and these secretory antibody responses were highly dependent on the frequency of the booster immunization, yet unaffected by the dose of the antigen (25-200$\mu\textrm{g}$). These results demonstrate the remarkable potential of rUrease as a vaccine antigen, thereby strengthening the possibility of developing an H. pylori vaccine for humans.
The effect of an ethanolic extract of the plant Trianthema portulacastrum L. on the $CCI_4$-induced chronic hepatocellular damage of Swiss albino mice has been investigated. The normal mice received olive oil (0.2 ml/mouse) for five weeks. The $CCI_4$ control mice, on the other hand, received $CCI_4$ (0.05 ml/mouse) in olive oil for five weeks. The extract was administered at the dose of 100 mg/kg or 150 mg/kg for five weeks by gastric intubation in addition to $CCI_4$ treatment. The $CCI_4$ administraction alone caused hepatocellular necrosis, severe anemia, leucopaenia, lymphocytopaenia, neutrophilia, eosinophilia and haemoglobinaemia along with the alterations of plasma albumin and globulin. The administration of plant extract (at 100 or 150 mg/kg) restored the $CCI_4$-induced alterations of the haematological parameters to the normal level. The extract of T. portulacastrum elicited a marked protection against $CCI_4$-induced hepatotoxicity as indicated by the several haematological parameters, related indices of formed elements, and different fractions of plasma protein. We also observed the dose-dependent antihepatotoxic effect of the extraction on these mice. The 150 mg/kg of extract was found to be more effective in normalizing the toxic effects of $CCI_4$ on the above parameters of mice. These results suggest that the hepatoprotective effect of T. poltulacastrum could be caused by its critical involvement in modulating several factors associated with erythropoiesis, and the boosting of general immunity of the host.
This study was designed to find the most suitable method and wall material for microencapsulation of the Lactobacillus plantarum to maintain cell viability in different environmental conditions. To improve the stability of L. plantarum, we developed an encapsulation system of L. plantarum, using water-in-oil emulsion system. For the encapsulation of L. plantarum, corn starch and glyceryl monostearate were selected to form gel beads. Then 10% (w/v) of starch was gelatinized by autoclaving to transit gel state, and cooled down at $60^{\circ}C$ and mixed with L. plantarum to encapsulate it. The encapsulated L. plantarum was tested for the tolerance of acidic conditions at different temperatures to investigate the encapsulation ability. The study indicated that the survival rate of the microencapsulated cells in starch matrix was significantly higher than that of free cells in low pH conditions with relatively higher temperature. The results showed that corn starch as a wall material and glycerol monostearate as a gelling agent in encapsulation could play a role in the viability of lactic acid bacteria in extreme conditions. Using the current study, it would be possible to formulate a new water-in-oil system as applied in the protection of L. plantarum from the gastric conditions for the encapsulation system used in chicken feed industry.
Dandelion(Taraxacum officinale), an oriental herbal medicine, has been shown to favorably affect choleretic, antioxidative and protection of gastric mucosa. The protective effects of common dandelion leaf and root extract were investigated by irradiation in Sprague-Dawley rats. Male SD rats, 6weeks, were orally injected with dandelion extract(100mg/kg) for 10days. Immediately after final injection, rats were whole body irradiated with 10Gy used irradiation facility(Elekta Linac, Sweden). At 24h-15days after irradiation, complete blood counted test and apoptosis in jejunal crypt cell. Stimulated recovery by the extract was observed in platelet but was not showed in the erythrocyte and leucocyte. The jejunal crypt cells were protected significantly(p<0.05) and the radiation-induced apoptosis was reduced(p<0.05). The survival rate were carried for 15days, the survival ratio was 15% and 85% for the control and experimental group. Observations intestinal inhibition of cell death, intestinal mucosa and tissue decreased systemic inflammation and vacuole. Base on these data, we propose that dandelion may be a useful radioprotector, especially since it is a relatively nontoxic natural product.
Journal of Physiology & Pathology in Korean Medicine
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v.24
no.6
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pp.996-1003
/
2010
This study was performed to investigate the effect of promoting gastrointestinal function and inhibiting of decreasing body temperature of ginger extract(Zingiber officinale) in rats. In order to elucidate the gastrointestinal function and inhibiting effect of body temperature of native ginger and improved ginger, water extracts of ginger were orally administrated into rats. The results are as follows: The gastrointestinal transit time was significantly decreased in native ginger(7.66hrs) and improved ginger(7.72hrs) extract administrated groups compare to control group(8.44hrs). The mean red faecal weight was increased in native ginger(30.6%) and improved ginger(31.1%) extract administrated groups compare to control group(24.9%) for 24hrs. Inhibiting effect of decreasing body temperature induced by serotonin was increased in native ginger($1.116^{\circ}C$) and improved ginger($1.416^{\circ}C$) extract administrated groups compare to positive control group($0.384^{\circ}C$) during 40 minutes. Gastrin and CGRP immunoreactive density was more strongly expressed in native ginger and improved ginger extract administrated groups compare to control group. Serotonin immunoreactive density was more weakly expressed in native ginger and improved ginger extract administrated groups compare to control group. These results suggest that ginger extracts may enhance physiological activity such as gastrointestinal motility, protection of mucosa and gastric acid secretion in gastrointestinal tracts, and inhibits decreasing body temperature
Objective: To determine the effect of gut pH and rumen microbial fermentation on glycerol encapsulated in alginate and alginate-chitosan polymers. Methods: Glycerol was encapsulated at 2.5%, 5%, 7.5%, or 10% (w/w) with sodium alginate (A) and alginate-chitosan (AC) polymers. Surface morphology and chemical modifications of the beads were evaluated using scanning electron microscopy and Fourier transform infrared (FTIR) spectra. Encapsulation efficiency was determined at the 5% glycerol inclusion level in two experiments. In experiment 1, 0.5 g of alginate-glycerol (AG) and alginate-chitosan glycerol (ACG) beads were incubated for 2 h at $39^{\circ}C$ in pH 2 buffer followed by 24 h in pH 8 buffer to simulate gastric and intestinal conditions, respectively. In experiment 2, 0.5 g of AG and ACG beads were incubated in pH 6 buffer at $39^{\circ}C$ for 8 h to simulate rumen conditions. All incubations were replicated four times. Free glycerol content was determined using a spectrophotometer and used to assess loading capacity and encapsulation efficiency. An in vitro experiment with mixed cultures of rumen microbes was conducted to determine effect of encapsulation on microbial fermentation. Data were analyzed according to a complete block design using the MIXED procedure of SAS (SAS Institute, Cary, NC, USA). Results: For AG and ACG, loading capacity and efficiency were 64.7%, 74.7%, 70.3%, and 78.1%, respectively. Based on the FTIR spectra and scanning electron microscopy, ACG treatment demonstrated more intense and stronger ionic bonds. At pH 6, 36.1% and 29.7% of glycerol was released from AG and ACG, respectively. At pH 2 minimal glycerol was released but pH 8 resulted in 95.7% and 93.9% of glycerol released from AG and ACG, respectively. In vitro microbial data show reduced (p<0.05) fermentation of encapsulated glycerol after 24 h of incubation. Conclusion: The AC polymer provided greater protection in acidic pH with a gradual release of intact glycerol when exposed to an alkaline pH.
New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.
This study was performed to investigate the antiulcer effects of Opuntia dillenii Haw. on the stomach ulcer induced by restraint and water-immersion stress in rats. For this experiment, 48 male Sprague-Dawley strain were used. The experimental groups were divided into four: a control (C) and 3 Opuntia dillenii Haw. treatment groups (E-1, E-2, E-3). Each dose of Opuntia dillenii Haw. was 30 mg/kg bw (E-1), 60 mgfKg bw (E-2) and 120 mg/kg bw (E-3). The rats were allocated to each group by 12 and observed for 4 weeks. The results were as following: 1. The stomach surface pH in each group showed no significant difference, but the values of Opuntia dillenii Haw. treatment groups were higher than the value of the control group. 2. The gastric wall mucus was increased in all Opuntia dillenii Haw. treatment groups compared with the control group. Especially in E-1 difference was higher (p<0.05) and in E-2 difference was significantly higher (p<0.01). 3. At shear rate 11.25, 45.0, 90.0, $225\;sec^{-1}$, whole blood viscosity and plasma viscosity were measured. Most of the values of Opuntia dillenii Haw. treatment groups were low compared with that of the control group. At shear rate 90.0, $225\;sec^{-1}$ the values of whole blood viscosity in E-1 were significantly low (p<0.05) and at shear rate 11.25, $45.0\;sec^{-1}$, more significant (p<0.01). At shear rate 11.25, 45.0, 90.0, $225\;sec^{-1}$ the values of whole blood viscosity in E-2 were significantly low (p<0.01). At shear rate $90.0\;sec^{-1}$ the value of plasma viscosity in E-1 was significantly low (p<0.05) and at shear rate 90.0, $225\;sec^{-1}$ the values of plasma viscosity in E-2 we resignificantly low (p<0.01). 4. Less severe ulcers were obsered in Opuntia dillenii Haw. treatment groups than in the control group. Especially E-1 groups tissues had only slight ulcers and necrosis of tissue was not observed in this group. From the results of this study, it can be concluded that the oral administratio-n of Opuntia dillenii Haw. results in protection of stomach ulcer by stimulating the secretion of gastric mucus and improving the gastric mucosal microcirculation.
Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.
Choi, Yong-Sung;Lee, Sun Ju;Yim, Hyeon Woo;Choe, Byung-in;Lee, Jae Won;Oh, Sang-cheul;Shin, Im Hee;Huh, Jung-Sik;Kwon, Ivo;Kim, Jin Seok;Yoo, Soyoung;Cho, Hyunin;Lee, Mi-Kyung;Shin, Hee-Young;Kim, Duck-An
The Journal of KAIRB
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v.1
no.1
/
pp.5-21
/
2019
Purpose: Institutional review board (IRB) classifies risks of clinical trials into less than minimal risk, minor increase over minimal risk, and more than minimal risk. Based on classification and evaluation for risk, IRB decides whether permitting consent exemption or asking additional protection for clinical research subject or not. The purpose of this study is to analyze how IRB members evaluate minimal risk by sending questionnaire survey with 12 predetermined scenarios. Methods: IRB members and researchers (pediatrician, gastroenterologist, neurologist, and neurosurgeon) in 11 different hospitals were asked to answer survey questions via email or online. We analyzed the differences of answers among several subgroups in each predetermined scenarios. Result: Responders were 212 personnel(110 researchers and 102 IRB members) from 11 centers. There were significant differences between IRB members and researchers in response such as blood sampling, skin prick test, one time catheterization in a girl, spinal tapping in child, non-enhance MRI in child, non-enhance MRI with chrolal hydrate in a child, spinal tapping without anesthesia in adult, bioequivalence test, gastric endoscopy, and non-enhance CT. significant differences between medical IRB members and non-medical members were also revealed in one time catheterization in a girl, spinal tapping in a child, non-enhance MRI in a child, bioequivalence test. Depending on researchers' department, they responded differently in several questionnaires as well. Conclusions: We have found that IRB members and researchers evaluate the risks differently. Researchers compared to IRB members, medical IRB members compared to non-medical members answered less than minimal risk in many cases. In assessing and evaluating the risks associated with the study, medical IRB members answered predetermined scenarios as less dangerous compared to non-medical IRB members. Difference among researchers where also revealed significantly. Researchers answered predetermined scenarios as less dangerous compare to other department researchers, especially in predetermined scenarios containing procedures they are familiar with.
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