• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1271-1279
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• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• Journal of Food Hygiene and Safety
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• v.32 no.1
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• pp.75-81
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• 2017

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.445-450
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• 2016

Thermal-stable soluble proteins (TSSP) in livestock products has been recently reported. Therefore, the development of antibodies and immunoassay using a TSSP is useful because the presence of TSSP can be measured on processed food. In this study, the existence of TSSPs in pork fat and meat was confirmed and their antigenicity was investigated. The extracts from pork fat and meat by heating method were analyzed by SDS-PAGE with 5% stacking and 12% separating gels. The protein profiles from the raw pork fat and meat extracts (major band ranged 25 to 100 kDa) without cooking and heating treatments were significantly different compared to those from cooked and heated pork fat and meat extracts (several major bands > 100 kDa and < 30 kDa). This meant that non thermal-stable soluble proteins ranged from 25 to 100 kDa may be denaturated to insoluble proteins by cooking and heating treatments, and TSSPs were in pork fat and meat at kept their properties. The confirmed TSSPs were used as an immunogen to investigate their antigenicity. Eight mice (5 mice for pork fat and 3 mice for pork meat) were separately immunized with the TSSPs of pork fat and meat, and the anti-sera obtained from the immunized mice showed high titer values. Polyclonal antibodies against each target protein showed the specific reaction to pork fat and meat, individually. These indicated that TSSP could be used as an immunogen to produce antibodies such as monoclonal and polyclonal antibodies. In addition, antibodies specific to TSSP from pork fat and meat may be used as a bio-receptor in immunoassays for the identification of fraudulent adulteration with pork fat and meat in livestock products.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.53-61
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• 1996

This study was carried out to determine effectively the sex of rabbit embryos using H-Y antiserum. H-Y antiserum was obtained from inbred SD strain female rat which was immunized by injection of testis cell of inbred SD strain male rate into its spleen. The titer of antiserum was identified by sperm cytotoxicity test and culture of rabbit embryos with antiserum. The developed or undeveloped embryos were separated by exposure the embryos to the antiserum with H-Y antibody. Developed embryo were transferred to the recipients and sex of offspring were examined. 1. In the sperm cytotoxicity test, the rate of dead sperm showed no difference between two antisera from spleen and testis cell as antigens. It is confirmed that H-Y antibody in antiserum was absorbed by H-Y antigen in male rat spleen cells. 2. When rabbit morulae were exposed to antiserum and complement, the rate of embryos developed or arrested was 51 and 49% respectively and the rate was closely same as natural sex ratio of 50:50%. 3. When rabbit morulae were cultured for 12h in the medium containing antiserum produced by antigen of testis cell, the rate of embryos developed or arrested was 48 and 52% respectively and the rate was closely same as natural sex ratio of 50:50%. 4. Eighty rabbit embryos which were not affected by H-Y antiserum were transferred to four recipients. Two recipients were pregnant and born 13 pups among which 2 (14%) were male and 11 (86%) were female. In conclusion, existence of H-Y antibody in the serum from female rat immunized by injecting testis cell from newborn male rat to the spleen of the female rat was confirmed. When rabbitmorulae were exposed to H-Y antiserum and complement, about a half of embryos were developed to blastocysts. When the rabbit embryos not affected by H-Y antiserum were transferred, the rate of female offspring was 86%. Therefore, it was identified that most of embryos which were not affected by H-Y antiserum were female.

• Journal of Nutrition and Health
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• v.25 no.6
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• pp.468-475
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• 1992

This study was carried out to observe the effects of dietaryn-6, n-3 fatty acids and vitamin A levels on humoral immunity in rat. Sixty eight male Sprague-Dawley rats were fed 6 different experimental diets for 6 weeks. The diets were composed of 10% of either corn oil or fish oil with various levels of vitamin A ; deficient(12450 IU/kg diet) adequate(4000IU/kg diet) and excess(400,000 IU/kg diet) The weight of spleen from the excess vitamin A-fish oil group showed the lowest value of all the groups when spleen weight was expressed/100g body weight. The number of PFC to SRBC was not affected by dietary at type and vitamin A levels. Hemagglutination titers were significantly lower in fish oil groups compared to corn oil groups and the values of vitamin A deficient groups were lower than the ones of adequate and excess vitamin A groups. IgM contents is serum were significantly lower in fish oil groups than in corn oil groups. IgG contents were higher in fish oil groups than in corn oil groups and the highest levels was recorded in excess vitamin A-fish oil group which showed the smallest speen size. Light microscopical examination showed that spleen tissues of fish oil groups were well developed than those of the corn oil groups and vitamin A deficient and excessive groups showed poor development than the adequate groups. Therefore it is suggested that adequate amounts of vitamin A consumption is necessary for healthy individuals and fish oil intake along with excess vitamin A should be avoided in order to maintain immune function properly.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.22
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• pp.9655-9660
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• 2014

Background: Nigella Sativa (NS) is an herb from the Ranunculaceae family that exhibits numerous medicinal properties and has been used as important constituent of many complementary and alternative medicines (CAMs). The ability of NS to kill cancer cells such as PC3, HeLa and hepatoma cells is well established. However, our understanding of the mode of death caused by NS remains nebulous. The objective of this study was to gain further insight into the mode and mechanism of death caused by NS in breast cancer MCF-7 cells. Materials and Methods: Human breast cancer cells (MCF-7) were treated with a methanolic extract of NS, and a dose- and time-dependent study was performed. The $IC_{50}$ was calculated using a Cell Titer $Blue^{(R)}$ viability assay assay, and evidence for DNA fragmentation was obtained by fluorescence microscopy TUNEL assay. Gene expression was also profiled for a number of apoptosis-related genes (Caspase-3, -8, -9 and p53 genes) through qPCR. Results: The $IC_{50}$ of MCF-7 cells was $62.8{\mu}L/mL$. When MCF-7 cells were exposed to $50{\mu}L/mL$ and $100{\mu}L/mL$ NS for 24h, 48h and 72h, microscopic examination (TUNEL assay) revealed a dose- and time-dependent increase in apoptosis. Similarly, the expression of the Caspase-3, -8, -9 and p53 genes increased significantly according to the dose and time. Conclusions: NS induced apoptosis in MCF-7 cells through both the p53 and caspase pathways. NS could potentially represent an alternative source of medicine for breast cancer therapy.

• Journal of Korean Neurosurgical Society
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• v.63 no.5
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• pp.579-589
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• 2020

Objective : No optimum genetic rat Huntington model both neuropathological using an adeno-associated virus (AAV-2) vector vector has been reported to date. We investigated whether direct infection of an AAV2 encoding a fragment of mutant huntingtin (AV2-82Q) into the rat striatum was useful for optimizing the Huntington rat model. Methods : We prepared ten unilateral models by injecting AAV2-82Q into the right striatum, as well as ten bilateral models. In each group, five rats were assigned to either the 2×10¹² genome copies (GC)/mL of AAV2-82Q (×1, low dose) or 2×10¹³ GC/mL of AAV2-82Q (×10, high dose) injection model. Ten unilateral and ten bilateral models injected with AAV-empty were also prepared as control groups. We performed cylinder and stepping tests 2, 4, 6, and 8 weeks after injection, tested EM48 positive mutant huntingtin aggregates. Results : The high dose of unilateral and bilateral AAV2-82Q model showed a greater decrease in performance on the stepping and cylinder tests. We also observed more prominent EM48-positive mutant huntingtin aggregates in the medium spiny neurons of the high dose of AAV2-82Q injected group. Conclusion : Based on the results from the present study, high dose of AAV2-82Q is the optimum titer for establishing a Huntington rat model. Delivery of high dose of human AAV2-82Q resulted in the manifestation of Huntington behaviors and optimum expression of the huntingtin protein in vivo.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.229-235
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• 2004

To characterize the recent outbreaks of canine distemper (CD) in Korea, we carried out epidemiological investigations by clinical observations, serum neutralizing (SN) antibody titer determination and RT-PCR on the 315 dogs which were clinically suspected as canine distemper virus (CDV) infection. One hundred and sixty two of 315 dogs were infected with CDV. Breed or gender did not seem to have effects on the prevalence of CD. The major part of dogs were in young age from 6 weeks to 18 weeks of age, and were not vaccinated or incompletely vaccinated. Clinical signs of dogs with CD were multi systemic and extremely variable. Dogs died from CD had significantly more ocular signs and neurologic signs than those of dogs survived (p<0.05). The SN titers against CDV of 157 (96.9%) dogs were under 1:16, which is less than protective level. One possible explanation for recent outbreaks of CD in Korea might be low antibody titers against CDV because of vaccination failure. Therefore, to reduce the impact of virulent infection in the dog population, dogs should be vaccinated adequately and prophylactic measures should include isolation of young dogs from the dog population until vaccination can be expected to provide protection.

• Korean Journal of Veterinary Service
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• v.24 no.4
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• pp.379-382
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• 2001

To confirm the effect of food and water deprivation prior to Newcastle disease(ND) virus vaccination, three hundred chicks were divided into five groups with three replications. ND vaccine were sprayed to at 1 -day old chicks at commercial hatchery. Secondary and third vaccination was conducted at 2-week old and 24-day old chicks by LaSota strain. Control was conventionally vaccinated without withdrawing the food and water before or after vaccination. In group 2(G2) and 3(G3), LaSota strain was vaccinated to chicks before and after fasting the food and water for 3 and 2 hours, respectively. Group 4(G4) has the same fasting time of group 2, but supplemented the skim milk in vaccin dilution water. In group 5(G5), skim milk was added into group 3. Weight gain, feed intake and feed conversion were weekly measured for 5 weeks. Blood was collected from wing vein at 24 and 35 days of age. Each serum antibody level were measured by hemagglutination inhibition(HI) test. The average weight gain, feed intake, feed conversion of all group were not significantly different. Weight gain of each groups was 1910.30(control), 1875.28(G2), 1952.12(G4) and 1896.05(G5), respectively. Feed intake of all group was recorded at 3160.67(control), 3167.07(G2), 3189.48(G3), 3157.85(G4) and 3178.16(G5), respectively. The feed conversion of each groups was 1.655(control), 1.688(G2), 1.633(G3), 1.699(G4) and 1.676(G5), respectively. The HI titer of G4 was $ 5.50{\Pm}$1.40 and significantly higher than the other groups (p<0.05)(control : $4.36{\Pm}$1.87 , G2 : $5.18{\Pm}$2.14, G3 : $4.51{\Pm}$2.19, G : $5.28{\Pm}$1.58 at 35 days old. The results of this experiment indicated that two or three hours of fasting time before or after vaccination would be able to show the higher antibody level against ND virus.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.347-356
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• 2008

The purpose of this study was to evaluate the efficacy and cross-protection of serovar 12 against serovar 4 or 5 of H parasuis with M+$Parapac^{(R)}$. A total of 141 piglets from 2(A and B) farms were used and divided into experimental group and control group in each farm. Farm A has been detected H parasuis serovar 12, whereas farm B has been detected H parasuis serovar 4 or 5 from post-weaned pigs with PMWS. The piglets were vaccinated intramuscularly with 2.0ml of M+$Parapac^{(R)}$ in experimental group or normal saline in control group at 1 week of age. A same booster dose was given at 3 weeks of age. In order to value the antibody titer to H parasuis using by tube agglutination test, blood samples were collected from piglets at the aged of 1 week, 6 and 14 weeks. In experimental group and control group, the average antibody titers were $32.5{\pm}21.0,\;114.5{\pm}34.0,\;98.1{\pm}55.4$ and $32.9{\pm}18.6,\;25.8{\pm}36.9,\; 746.7{\pm}1,215.8$ at the aged of 1 week, 6 and 14 weeks, respectively. The cumulative clinical sign indexes by standard guideline of Schering-Plough Animal Health Corp were 486 and 1,069, respectively. The average daily gains and feed conversion rates were $0.553{\pm}0.016kg$ and $0.492{\pm}0.004kg$, and 1.99 and 2.24, respectively. The average gross lesion scores were $1.0{\pm}0.8$ and $1.9{\pm}0.6$, respectively. According to these results, the M+$Parapac^{(R)}$ containing H parasuis serovar 12 may be induce circulating antibodies that cross-react with serovar 4 or 5 and have a protection of PMWS with H parasuis.