New lectins have been isolated and purified from mung bean (Phaseolus radiatus) through physiological saline extraction, ammonium sulfate salt fractionation and column chromatographies. Ion exchanger were eluted by linear salt gradient and then further purified through gel filtration. Thus obtained lectin named as MBL. The gene expressions of 5 cytokines (IL-1, IL-2, IL-6, $TNF-{\aphpa}$ and $IFN-{\gamma}$) from human peripheral blood mononuclear cells (PBMC) stimulated with MBL were investigated by using reverse transcription polymerase chain reaction (RT-PCR). PBMC ($1{\times}106$ cells/ml) isolated from healthy volunteers were stimulated with lectins (4 mg/ml) for various time intervals (1 to 96 hrs). After each of the various stimulated times, total RNA was isolated and assessed for different cytokines mRNA by RT-PCR. The mRNA encoding IL-1, IL-2 were detected continuously from 1 to 20 hrs, and IL-6 was detected up to 24 hrs. But the mRNA encoding $IFN-{\gamma}$ and $TNF-{\alpha}$ were detected to 8 hours only and showed short time response compared with other cytokines. The significant expression of all cytokines mRNA were observed at 4 hrs. These results suggested that MBL, as inducer of cytokines could elicit detectable cytokine mRNA from PBMC.
Ahn, Y.J.;Kim, M.J.;Kawamura, T.;Yamamoto, T.;Fujisawa, T.;Mitsuoka, T.
Journal of Ginseng Research
/
v.14
no.2
/
pp.253-264
/
1990
The growth responses of a variety of human intestinal bacteria to extracts of Panax ginseng and five other oriental medicinal Araliaceae were elraluattd in vitro and in vivo. The extracts enhanced the growth of Brifidobnnerilrm breve and B. longlim in media with or without carbon sources, suggesting that bifidus factors) might be involved in the phenomenon. This effect was most pronounced with water extract of P. ginseng, the growth of 27 bifidobacteria strains belonging to B adolescentis, B. longum, B. brim and B. infantis being greatly stimurated, whereas seven B. bifidum strains and other bacteria such as clostridia and Escherichin soli had little or no ability to utilise it for growth. Methanol extracts of p. ginseng were found to selectively inhibit growth of various clostridia including bifidobacteria. Paraputrificum, but this effect was not observed on other bacteria including bifidobacteria. The effect of ginseng extract intake (600 mg/day for two weeks) on the faecal microflora, pH, volatile fatty acids, ammonia, putrefactive products, and -glucuronidase, -glucosidase and nitroreductase activities, and on the blood components (triglyceride, total cholesterol and ammonia) were investigated using seven healthy human volunteers. The total concentration of faecal microflora including Bifidnkaderiifm app. during the period of ginseng extract intake %twas significantly unaffected from the preceding and subsequent control peroids. However, the frequency of occurrence of subjects having C. perfringens was significantly decreased. The faecal pH value was also significantly decreased, suggesting that the intake might increase the activity of Bifidobncterium spry. Other biochemical properties in faeces did not changed significantly. The levels of ammonia and triglycerid in blood were decreased with ginseng extract intake. These results may be an indication of at least one of the Pharmacological actions of p. ginseng as an adaptogen.
Mercury is a toxic and non-essential metal in the human body. Mercury is ubiquitously distributed in the environment, present in natural products, and exists extensively in items encountered in daily life. There are three forms of mercury, i.e., elemental (or metallic) mercury, inorganic mercury compounds, and organic mercury compounds. This review examines the toxicity of elemental mercury and inorganic mercury compounds. Inorganic mercury compounds are water soluble with a bioavailability of 7% to 15% after ingestion; they are also irritants and cause gastrointestinal symptoms. Upon entering the body, inorganic mercury compounds are accumulated mainly in the kidneys and produce kidney damage. In contrast, human exposure to elemental mercury is mainly by inhalation, followed by rapid absorption and distribution in all major organs. Elemental mercury from ingestion is poorly absorbed with a bioavailability of less than 0.01%. The primary target organs of elemental mercury are the brain and kidney. Elemental mercury is lipid soluble and can cross the blood-brain barrier, while inorganic mercury compounds are not lipid soluble, rendering them unable to cross the blood-brain barrier. Elemental mercury may also enter the brain from the nasal cavity through the olfactory pathway. The blood mercury is a useful biomarker after short-term and high-level exposure, whereas the urine mercury is the ideal biomarker for long-term exposure to both elemental and inorganic mercury, and also as a good indicator of body burden. This review discusses the common sources of mercury exposure, skin lightening products containing mercury and mercury release from dental amalgam filling, two issues that happen in daily life, bear significant public health importance, and yet undergo extensive debate on their safety.
Cirsium japonicum var. ussuriense is often used in treatment of human disease such as hemorrhage, blood congestion and inflammation. This study was accomplished to evaluate the antioxidant properties of the leaf (CLE) and flower (CFE) extracts of C. japonicum var. ussuriense to protect normal human red blood cells (RBC) and plasma samples against oxidative damage in vitro. CLE and CFE were prepared by extracting with hot water. In red blood cells and plasma, oxidative hemolysis and lipid peroxidation induced by the aqueous peroxyl radical generator [2,2'-azobis (2-amidinopropane) dihydrochloride, AAPH] were significantly suppressed by CLE or CFE in a dose-dependent manner at the same time. CLE and CFE also prevented the depletion of cytosolic antioxidant glutathione (GSH) in RBC. These results suggest that the leaves and flowers of C. japonicum var. ussuriense may have the antioxidant properties.
Objective: This research investigated the effects of human chorionic gonadotropin (HCG)-producing peripheral blood mononuclear cells (PBMCs) on the implantation rate and embryo attachment in mice. Methods: In this experimental study, a DNA fragment of the HCG gene was cloned into an expression vector, which was transfected into PBMCs. The concentration of the produced HCG was measured using enzyme-linked immunosorbent assay. Embryo attachment was investigated on the co-cultured endometrial cells and PBMCs in vitro. As an in vivo experiment, intrauterine administration of PBMCs was done in plaque-positive female mice. Studied mice were distributed into five groups: control, embryo implantation dysfunction (EID), EID with produced HCG, EID with PBMCs, and EID with HCG-producing PBMCs. Uterine horns were excised to characterize the number of implantation sites and pregnancy rate on day 7.5 post-coitum. During an implantation window, the mRNA expression of genes was evaluated using real-time polymerase chain reaction. Results: DNA fragments were cloned between the BamHI and EcoRI sites in the vector. About 465 pg/mL of HCG was produced in the transfected PBMCs. The attachment rate, pregnancy rate, and the number of implantation sites were substantially higher in the HCG-producing PBMCs group than in the other groups. Significantly elevated expression of the target genes was observed in the EID with HCG-producing PBMCs group. Conclusion: Alterations in gene expression following the intrauterine injection of HCG-producing PBMCs, could be considered a possible cause of increased embryo attachment rate, pregnancy rate, and the number of implantation sites.
The present study were performed to analysis the hematocrit and the red blood cells content into the blood plasma of the transgenic pigs harboring recombinent human erythropoietin gene (rhEPO). Mouse whey acidic protein (mWAP) linked to rhEPO gene was microinjected into pronuclei of porcine one-cell zygotes. After delivered of offspring, PCR analyses identified one mWAP-rhEPO transgenic founder offspring(F$_{0}$). The first generation of transgenic pig (F$_{0}$) harboring mWAP-hEPO appeared to be a male, and the second generation (F$_1$) pigs were made by natural mating of F$_{0}$ with domestic swine, and male and female transgenic pigs (F$_1$) were identified by PCR. The blood samples from transgenic and normal pigs were collected for 50 days during lactation and were counted the red blood cell (RBC) numbers and Hematocrit (HCT) content into the blood. The transgenic pigs expressing rhEPO in their blood gave rise to higher RBC numbers and HCT contents than control animals. rhEPO was secreted both in the blood and milk of genetically engineered pigs harboring rhEPO gene. Therefore, this study provides a model regarding the production of transgenic pig carrying hEPO transgene for biomedical research.earch.
Ketamine is a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist and a short-acting general anaesthetic agent for human and veterinary use. We previously reported that treatment with ketamine impairs oxidative burst activity of canine peripheral blood leukocytes. In this study, the effect of ketamine on phagocytic capacity of canine peripheral blood leukocytes was examined in vitro. Phagocytic capacity was analyzed by using a flow cytometry. Ketamine directly decreased the phagocytic capacity of peripheral blood polymorphonuclear cells (PMN) and monocytes but not total peripheral blood mononuclear cells (PBMC). In addition, the phagocytic capacity of PMN and monocytes was inhibited by the ketamine-treated PBMC but not PMN culture supernatant. These results suggest that ketamine has a direct inhibitory effect on the phagocytic capacity of canine peripheral blood phagocytes and involves the production of soluble factor(s) from canine PBMC, which may suppress the phagocytic capacity.
Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.
Lim, Seung Yeop;Heo, Hyun;Kim, Sang Ho;Won, Byeong Hee
Journal of the Ergonomics Society of Korea
/
v.32
no.4
/
pp.345-353
/
2013
Objective: This research measured physiological and psychological effects of Vibroacoustic stimulation(VA) to scapular and sacrum of supine position on the mattress. Background: When vibroacoustic stimulation applies to human body, it has a positive influence on physiological and psychological effects by stimulating the organs, tissues and cells of whole body. Method: This experiment was conducted to 10 normal males in two conditions: no stimulation and vibroacoustic stimulation. No stimulation experiment was executed as a supine position for 30 minutes without any vibrational stimulus, while vibroacoustic stimulation was transmitted by the vibrational speaker, which uses 40Hz frequency. Subjects had a laser Doppler flowmeter probe in scapular, sacrum, and also had 8 channel electroencephalogram(EEG) measurement sensor in the scalp. Blood pressure and skin temperature were measured in two conditions with an underlying posture for 30 minutes. Additionally, blood flow rate and EEG were measured before and after for two minutes on two conditions. Results: According to the vibroacoustic stimulation, blood flow rate and skin temperature were increased, while blood pressure was decreased. When using vibroacoustic stimulation compared to no stimulation, blood flow rate went approximately two times higher, and skin temperature also higher 3~4 times. Furthermore, the relative alpha power of brain wave was significantly increased when we applied to vibroacoustic stimulation. Conclusion: This experiment tested the VAT embedded in mattress in two conditions. According to this experiment, VAT decreases blood pressure, improves not only a physiological effect on blood flow rate as well as skin temperature, but also psychological functions by increasing relative alpha power. Application: The results of the publishing trend analysis might help physiological and psychological effects of vibroacoustic stimulation.
Background: This study was aimed to differentiate two forms of CTLA-4 (CD152) in activated peripheral blood lymphocyte and clarify the mechanism how cytoplasmic form of this molecule is targeted to cell surface. Methods: For this purpose we generated 2 different anti-human CD152 peptide antibodies and 5 different N'-terminal deletion mutant CTLA4Ig fusion proteins and carried out a series of Western blot and ELISA analyses. Antipeptide antibodies made in this study were anti-CTLA4pB and anti-CTLA4pN. The former recognized a region on extracellular single V-like domain and the latter recognized N'-terminal sequence of leader domain of human CD152. Results: In Western blot, the former antibody recognized recombinant human CTLA4Ig fusion protein as an antigen. And this recognition was completely blocked by preincubating antipeptide antibody with the peptide used for the antibody generation at the peptide concentration of 200 ug/ml. These antibodies were recognized human CD152 as a cytoplasmic sequestered- and a membrane bound- forms in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocyte (PBL). These two forms of CD152 were further differentiated by using anti-CTLA4pN and anti-CTLA4pB antibodies such that former recognized cytosolic form only while latter recognized both cytoplasmic- and membraneforms of this molecule. Furthermore, in a transfection expression study of 5 different N'-terminal deletion mutant CTLA4Ig, mutated proteins were secreted out from transfected cell surface only when more than 6 amino acids from N'-terminal were deleted. Conclusion: Our results implies that cytosolic form of CTLA-4 has leader sequence while membrane form of this molecule does not. And also suggested is that at least N'-terminal 6 amino acid residues of human CTLA-4 are required for regulation of targeting this molecule from cytosolic- to membrane- area of activated human peripheral blood T lymphocyte.
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