Ham, J.S.;Jeong, S.G.;Lee, S.G.;Han, G.S.;Jang, A.;Yoo, Y.M.;Chae, H.S.;Kim, D.H.;Kim, H.J.;Lee, W.K.;Jo, C.
Asian-Australasian Journal of Animal Sciences
/
v.22
no.2
/
pp.289-295
/
2009
To develop a safer yogurt for immuno-compromised or allergy patients and to extend shelf-life, a plain yogurt was irradiated with doses of 0, 1, 3, 5, and 10 kGy using a gamma ray and the chemical and microbiological quality and allergenicity change were investigated. There was no difference in the content of protein, total solid, and amino acids of the plain yogurt by irradiation treatment and different storage temperatures (4, 20, and $35^{\circ}C$). The lactic acid bacterial counts of irradiated plain yogurt had approximately 3-decimal reduction at 3 kGy, and no viable cell at 10 kGy regardless of storage time and temperature. The binding ability of rabbit antiserum to milk proteins in irradiated plain yogurt showed that 10 kGy of irradiation produced significantly higher binding ability than other treatments. Sensory evaluation indicated that only appearance of the plain yogurt irradiated at 3 kGy or higher had a lower value than the non-irradiated control when stored at $20^{\circ}C$. Results suggest that irradiation of plain yogurt does not significantly affect the chemical and sensory quality of plain yogurt, but can extend the shelf-life, possibly reduce allergenicity, and provide a safer product.
The performance of an immuno-analytical system can be assessed in terms of its analytical sensitivity, i.e., the detection limit of an analyte, which is determined by the amount of analyte molecules bound to the capture antibody that has been immobilized onto a solid surface. To increase the number of the binding complexes, we have investigated a site-directed immobilization of an antibody that has the ability to resolve a current problem associated with a random arrangement of the insolubilized immunoglobulin. The binding molecules were chemically reduced to produce thiol groups that were limited at the hinge region, and then, the reduced products were coupled to biotin. This biotinylated antibody was bound to a streptavidin-coated surface via the streptavidin-biotin reaction. This method can control the orientation of the antibody molecules present on a solid surface and also can significantly reduce the possibility of steric hindrance in the antigen-antibody reactions. In a two-site immunoassay, the introduction of the site-directly immobilized antibody as the capture enhanced the sensitivity of analyte detection approximately 10 times compared to that of the antibody randomly coupled to biotin. Such a novel approach would offer a protocol of antibody immobilization in order for the possibility of constructing a high performance immunochip.
Clostridioides (Clostridium) difficile is a Gram (+), anaerobic, spore forming, rod shaped bacterium that can produce toxin. The objective of this study is to reveal the presence of C. difficile in meat products, to analyze the ribotype diversity by PCR and to evaluate the antibiotic susceptibility of isolated strains. The organism was isolated in 22 out of 319 (6.9%) examined meat product samples and 9 out of 22 (40.9%) isolates were identified as RT027 and all isolates had the ability of toxin production. In terms of antibiotic susceptibility, all isolates were susceptive to amoxicillin-clavulanic acid, tetracycline and vancomycin and 21 (95.4%) isolates to metronidazole. On the other hand, imipenem and cefotaxim resistance was observed in all. In conclusion, the results of this comprehensive study conducted in Turkey deduced the presence of C. difficile in different meat products. Therefore, these products can be evaluated as a potential contamination source of C. difficile from animals to humans especially for elders, youngsters, long terms wide spectrum antibiotic used and immuno-suppressed individuals.
To monitor the reproductive patterns of endangered common chimpanzee including annual reproductive cycle, amenorrhea, breeding season, and pregnancy diagnosis, Time-Resolved Fluorescence Immuno Assay (TR-FIA) was used to trace MRH (estradiol, progesterone, testosterone) and human chorionic gonadotropin (HCG). In result of this research, age was not the important factor in determining the reproduction capability in common chimpanzee; it was rather greatly influenced by the combination of various factors such as individual's fertilizing ability, presence of obstetrical diseases, the pattern of mating behavior, and the mental communication under the introduction of a mating partner. This research will play an important role in operating conservation project for common chimpanzees and can be extended also for shedding new light on understanding human menopause and obstetrical diseases.
We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.
Park, Sung-Jin;Kang, Jun-Chul;Lee, Da-young;Cho, Ju-Hyun;Yoon, Min-Ho
Journal of Food Hygiene and Safety
/
v.35
no.3
/
pp.284-289
/
2020
This study was performed with hot water extract (LJF-W) and 70% ethanol extract (LJF-70E) of Litsea japonica fruit to investigate the immunostimulatory activity and bone health-promoting effect of L. japonica fruit. The production of pro-inflammatory mediator (nitric oxide) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) in murine macrophage RAW264.7 cells were estimated to examine the immunostimulatory activity of the fruit extracts. The immunostimulatory activity of LJF-W was higher than that of positive control (geinsenosides). However, there was no effect in LJF-70E. Futhermore, both LJF-W and LJF-70E appeared to stimulate the proliferation of MC3T3-E1 cells, proving the effect as a bone health agent. From this result it could be presumed that L. japonica fruit extracts not only stimulate the immune system, but also the ability to promote bone health.
The Pancreatic islet are the clusters of endocrine cells scattered through out the exocrine pancreas. Transplantation of a sufficient pancreatic islets can normalize blood glucose level so that may prevent devastating complications of type I diabetes(IDDM) and other side effects of the IDDM. Recently, there are several approaches to transplant sufficient pancreatic islet, and it was comprised in increase or regeneration of the endogenous $\beta$-cell mass from donor's pancreas, but relatively few studies have been devoted to the morphological characters of the isolated and 3 day cultured pancreatic islets. We investigated morphological pattern of intracellular structure of isolated and 3 day cultured pancreatic islets. The morphological characters of the pancreatic islets were observed by scanning electron microscope and transmission electron microscope, and insulin distribution of the each islets were observed by transmission electron microscope, and were labeled with insulin antibody. Intracellular structures including nuclei, mitochondria, RER, Golgi complex and many secretory granules were normally appeared in the isolated pancreatic islets which was extracted immediately dornor's pancreas, however, There is a significant morphological changes between the 3 day cultured pancreatic islets and isolated islets. 3 day cultured pancreatic islet's $\beta$-cells had normal nuclei but increased cytoplasm mass and RER and developed Golgi complex. Insulin secretory granules were decreased in numbers rather than isolated pancreatic islet. In this study, the pattern of intracellular structure variation was examined during pancreatic islet culture. Most distinct features are variation of the insulin secretory granules, and developed RER, and dilated golgi complex. Therefore, we suggested that the various change of the morphological characters on cultured pancreatic islets were responsible for the function(biosynthesis and secretion of insulin) and growth. These results were also cultured islets have greater ability to recover and maintain normoglycemia than isolated islet transplantation.
Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.
Objective: This experimental study was performed to examine the in vitro and in viva anti-inflammatory and anti-allergic effects of Mori Cortex. Methods: Water extract of Mori Cortex was studied to its ability to stimulate or inhibit macrophage 264.7 cells to produce inflammatory and allergic mediators. Cytokines such as $IL-1{\beta}$, IL-6, IL-10 and $TNF-{\alpha}$ were measured by immunochemical assay. In vitro, the macrophages 264.7 were classified into four groups. One group was a normal group. The other group was a (-) control group stimulated with LPS. And the third group was a (+) control group pretreated for 1 hour with hydrocortisone. And the fourth group was a sample group pretreated for 1 hour with Mori Cortex. After pretreatment, macrophage were incubated with lipopolysaccharide(LPS) $100\;ng/m{\ell}$ for 12 hour and media collected and $IL-1{\beta}$, IL-6, IL-10 and $TNF-{\alpha}$ concentrations in supernatants were measured each by Enzyme linked immuno-soubent assay. Mori Cortex were used $50\;{\mu}g/m{\ell},\;100\;{\mu}g/m{\ell},\;250\;{\mu}g/m{\ell},\;500\;{\mu}g/m{\ell},\;and\;1,000\;{\mu}g/m{\ell}$. Hydrocortisones were used $10^{-8}M,\;10^{-7}M,\;10^{-6}M,\;10^{-5}M\;and\;10^{-4}M$. In vivo, the SD rats were classified into three groups. One group was a normal group injected with normal saline into the abdominal cavity. The other was a control group prescribed to compound 48/80 after normal saline injection. And the third was a sample group prescribed to compound 40/80 after Mori Cortex injection. Then, the release of histamine, IL-6 and $TNF-{\alpha}$ were measured. Results : In vitro, Man Cortex significantly increased the release of $IL-1{\beta}\;and\;TNF-{\alpha}$ by LPS-stimulated macrophage 264.7 cells. And it significantly decreased the release of IL-10. In IL-6, Mori Cortex of low concentration significantly decreased the release of IL-6, but that of high concentration acted in reverse. In vivo, Man Cortex didn't show significant inhibitory effects on the release of histamine and IL-6 in comparison with that of the control group. But it significantly increased the release of $TNF-{\alpha}$ in comparison with that of the control group.
Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.
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