Journal of the Korean Society of Food Science and Nutrition
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v.37
no.5
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pp.555-560
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2008
Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acid transporters, the system L amino acid transporters are the major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The L-type amino acid transporter 1 (LAT1), an isoform of system L amino acid transporter, is highly expressed in cancer cells to support their continuous growth and proliferation. 2-Aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) is a model compound for the study of amino acid transporter as a system L selective inhibitor. We have examined the effect and mechanism of BCH on cell growth suppression in HEp2 human head and neck squamous cell carcinoma. The BCH inhibited the L-leucine transport in a concentration-dependent manner with a $IC_{50}$ value of $51.2{\pm}3.8{\mu}M$ in HEp2 cells. The growth of HEp2 cells was inhibited by BCH in the timeand concentration-dependent manners. The formation of DNA ladder was not observed with BCH treatment in the cells. Furthermore, the proteolytic processing of caspase-3 and caspase-7 in the cells were not detected by BCH treatment. These results suggest that the BCH inhibits the growth of HEp2 human head and neck squamous cell carcinoma through the intracellular depletion of neutral amino acids for cell growth without apoptotic processing.
Chaiyabutr, N.;Komolvanich, S.;Preuksagorn, S.;Chanpongsang, S.
Asian-Australasian Journal of Animal Sciences
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v.13
no.3
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pp.334-347
/
2000
The present experiment was carried out to study the utilization of glucose in the mammary gland of crossbred Holstein cattle during feeding with different types of roughage. Sixteen first lactating crossbred Holstein cattle which comprised eight animals of two breed types, Holstein Friesian${\times}$Red Sindhi ($50{\times}50=50%$ HF) and Holstein Friesian${\times}$Red Sindhi ($87.5{\times}12.5=87.5%$ HF). They were divided into four groups of 4 animals each of the same breed. The utilization of glucose in the mammary gland was determined by measuring rates of glucose uptake and the incorporation of glucose into milk components in both groups of 50% HF and 87.5% HF animals feeding on either hay or urea treated rice straw. In early lactation, there were no significant differences of the total glucose entry rate and glucose carbon recycling among groups of crossbred animals feeding on either hay or urea treated rice straw. During lactation advance, the total glucose turnover rates and recycling of carbon glucose of crossbred HF animals feeding on urea treated rice straw were markedly higher than those of crossbred HF animals feeding on hay as roughage, whereas there were no significant changes for both groups of crossbred animals feeding on hay. The percentages and values of non-mammary glucose utilization showed an increase during lactation advance in the same group of both 50% HF and 87.5% HF animals. The percentage of glucose uptake for utilization in the synthesis of milk lactose by the mammary gland was approximately 62% for both groups of 87.5% HF and by approximately 55% for both groups of 50% HF animals feeding on either hay or urea treated rice straw. Intracellular glucose 6-phosphate metabolized via the pentose phosphate pathway accounted for the NADPH (reducing equivalent) of fatty acid synthesis in the mammary gland being higher in 87.5% HF animals during mid-lactation. A large proportion of metabolism of glucose via the Embden-Meyerhof pathway in the mammary gland was more apparent in both groups of 50% HF animals than those of 87.5 % HF animals during early and mid-lactation while it markedly increased for both groups of 87.5% HF animals during late lactation. It can be concluded that utilization of glucose in the mammary gland occurs in a different manner for 50% HF and 87.5% HF animals feeding on either hay or urea treated rice straw. The glucose utilization for biosynthetic pathways in the mammary gland of 50% HF animals is maintained in a similar pattern throughout the periods of lactation. A poorer lactation persistency in both groups of 87.5% HF animals occurs during lactation advance, which is related to a decrease in the lactose biosynthetic pathway.
Min, Eun Young;Kim, Kwang Il;Cho, Mi Young;Jung, Sung-Hee;Han, Hyun-Ja
Journal of Marine Life Science
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v.4
no.1
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pp.1-13
/
2019
Fucoidan is a physiologically functional ingredient extracted from seaweed brown algae, which is a sulfated polysaccharide containing fucose as a main molecule backbone. Fucoidan has a variety of immune-modulating or -stimulating effects, including promoting antigen uptake and enhancing anti-bacterial, anti-viral and anti-tumor effects. In addition, recent studies have suggested the possibility of use of fucoidan as a vaccine adjuvant in the field of human vaccine. Use of fucoidan as supplementary feeds have already been studied, but the development of fucoidan as an adjuvant of fish vaccine is still premature. However, the intracellular uptake of fucoidan differs depending on the molecular weight of fucoidan, and there is a limit to the study on specific immune response including the production of antibodies to fish caused by an artificial infection of pathogen. Although the safety of fucoidan has been demonstrated in animal cells, there is a need to confirm the safety of fucoidan in fish. Therefore, active research in this field is needed to use fucoidan as a vaccine adjuvant. This study discussed the effects of fucoidan on immune stimulation, humoraland cellular- immunity including humans and animals. The prospect of fucoidan as a vaccine adjuvant in fisheries also reviewed.
Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that 2. tenezla first penetrate into the mucosal intraepithelial Iymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope labelled uracil ($^3H-uracil$) . Third, the E. tenella sporozoites viability was assayed after preincubation of them with thicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (I) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schisogonic cycle of E. tenella in 3~4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merogoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48~60 hours, and decreased thereafter. The uptake amount of $^3H-uracil$ depended not only upon the inoculum sixte of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.
The search for tumor-avid agents for use in nuclear medicine imaging or therapy is a field of ongoing importance. Metallothionein (MT) is an intracellular protein that binds many metals with isotopes having imaging or radiotherapeutic potential. The purpose of the study was to determine whether uptake of radioisotopes that bind to MT is increased in tumor. We measured the uptake of Cd-109 and Ga-67 in tumor and normal tissues of sarcoma-bearing mice. Tumors were grown subcutaneously in female Balb/C mice from cultured Balb/3T3 cells transformed by the Moloney murine sarcoma virus (MMSV). When the tumors reached about 1 cm in diameter, mice were injected subcutaneously with Cd-109 and Ga-67. Eighteen and seventy-two hours later, the mice were sacrified. Organs and tissues were removed, weighed, and activity per mg tissue determined by gamma well-counting. Uptake of Cd-109 by MMSV tumors exceeded that by normal tissues examined, with the exception of liver and kidney (the organs known to be richest in MT). The tumor-to-tissue ratios of uptake for Cd-109 were far greater than those for Ga-67 for many normal tissues of great importance in terms of background activity (bone, intestine, fat, muscle, and blood). We concluded that metals that bind to MT may be useful for oncologic imaging or rediotherapy of cancer.
Hop, Huynh Tan;Arayan, Lauren Togonon;Reyes, Alisha Wehdnesday Bernardo;Huy, Tran Xuan Ngoc;Baek, Eun Jin;Min, WonGi;Lee, Hu Jang;Lee, Chun Hee;Rhee, Man Hee;Kim, Suk
Journal of Microbiology and Biotechnology
/
v.27
no.10
/
pp.1885-1891
/
2017
In this study, we evaluated the inhibitory effect of a rice bran mixture extract (RBE) on Brucella abortus pathogenesis in professional (RAW 264.7) and nonprofessional (HeLa) phagocytes. We fermented the rice bran mixture and then extracted it with 50% ethanol followed by gas chromatography-mass spectrometry to identify the components in RBE. Our results clearly showed that RBE caused a significant reduction in the adherence of B. abortus in both cell lines. Furthermore, analysis of phagocytic signaling proteins by western blot assay revealed that RBE pretreatment resulted in inhibition of phosphorylation of JNK, ERK, and p38, leading to decline of internalization compared with the controls. Additionally, the intensity of F-actin observed by fluorescence microscopy and FACS was remarkably reduced in RBE-pretreated cells compared with control cells. However, the intracellular replication of B. abortus within phagocytes was not affected by RBE. Taken together, these findings suggest that the phagocytic receptor blocking and suppressive effects of RBE on the MAPK-linked phagocytic signaling pathway could negatively affect the invasion of B. abortus into phagocytes.
Knowledge of avian host responses to brucellosis is critical to understanding how birds resist this infection; however, this mechanism is not well established. On the other hand, temperature has a major involvement in the physiology of living organisms, and cell death induced by heat is attributed to protein denaturation. This study demonstrates the direct bactericidal effect of a high temperature ($41^{\circ}C$) on Brucella abortus that resulted in the gradual reduction of intracellular bacteria and inhibited bacterial growth within avian macrophage HD11 in an increasing period of time. On the other hand, this study also revealed that high temperature does not affect the rate of bacterial uptake, as confirmed by the bacterial adherence assay. No significant difference was observed in the expression of target genes between infected and uninfected cells for both temperatures. This study suggests the susceptibility of B. abortus to bacterial death under a high temperature with an increased period of incubation, leading to suppression of bacterial growth.
International Journal of Industrial Entomology and Biomaterials
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v.33
no.2
/
pp.85-91
/
2016
Exosomes are homogenous vesicles of 40-100 nm diameter produced endogenously. Exosomes are generated by inward budding into multi-vesicular bodies (MVB) and then released to extracellular space. Exosomes contain various nucleic acid and protein cargoes from their cells of origin and this endosomal cellular molecules are used for intracellular communication and for both promotion and suppression of immune responses. Recently, they are also considered as delivery vehicle for therapeutic proteins due to their characteristics of stability in body fluids and ability for target uptake. Also, they show less immune reactivity because the isolated exosome harboring therapeutic proteins can be from the same host. White-spotted flower chafer, Protaetia brevitarsis is one of the major insect commercially reared in Korea. There are bacterial and fungal pathogens causing diseases in the beetle, and these diseases incur economic loss to the larva-rearing farms. Due to their endosomal cargoes, exosomes are good candidates in use of disease diagnosis. In this study, we isolated insect exosome from the hemolymph of P. brevitarsis, and verified it by analysis of the exosome-specific surface proteins and RNA.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.2
/
pp.281-286
/
2015
In this study, free radical scavenging activities (ABTS, DPPH, NBT, TBARS, and ORAC) and anti-obesity potential were evaluated using 50% ethanol extract from fermented Curcuma longa L. (FCE50). FCE50 showed free radical scavenging activities and anti-oxidant potential. Lipid accumulation and intracellular TG content were significantly reduced by 25.8% and 28.6%, respectively, by $250{\mu}g/mL$ of FCE50 compared to adipocytes. Glucose uptake was significantly reduced by 12.0%. FCE50 significantly reduced mRNA expression of acetyl-CoA carboxylase in 3T3-L1 cells. These results indicate that the anti-adipogenic effect of FCE50 might be due to its radical scavenging activity and anti-oxidant potential.
In single atrial cells isolated from the rabbit the properties of inward current of Na-Ca exchange were investigated using the whole cell voltage clamp technique. The current was recorded during repolarization following brief 2 ms depolarizing pulse to +40 mV from a holding potential of * 70 mV. Followings are the results obtained: 1. When stimulated every 30 seconds, the inward currents were activated and reached peak values 6-12 ms after the beginning of depolarizing pulse. The mean current amplitude was 342 pA/cell. 2. The current decayed spontaneously from the peak activation and the time course of the relaxation showed two different phases fast and slow phase. The time constants were 10-18 ms and 60-140 ms, respectively. 3. The recovery of inward current was tested by paired pulse of various intervals. The peak current recovered exponentially with time constant of 140 ms and 1 p M isoprenaline accelerated the recovery process. 4. Relaxation time course was also affected by pulse interval and time constant of the fast phase was reduced almost linearly according to the decrease of pulse interval between 30 sec and 1 sec. 5. The peak activation was increased in magnitude by long prepulse stimulation, 5 p M Bay K, 1 p M isoprenaline or internal and external application of c-AMP. 6. The relaxation time constant of the fast phase was prolonged by 5 p M Bay K or c-AMP, and shortened by isoprenaline. However the time course of the slow relaxation phase was not so much changed. From the above results, it could be concluded that increase of the calcium current by Bay K or c-AMP results in the potentiation and prolongation of intracellular calcium transient, and the facilitation of Ca uptake by SR might be a mechanism of shortening the time constant of current relaxation by short interval stimulation or isoprenaline.
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