• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.11
• /
• pp.1653-1663
• /
• 2016

Meat quality is a complex trait influenced by many factors, including genetics, nutrition, feeding environment, animal handling, and their interactions. To elucidate relevant factors affecting pork quality associated with oxidative stress and muscle development, we analyzed protein expression in high quality longissimus dorsi muscles (HQLD) and low quality longissimus dorsi muscles (LQLD) from Duroc pigs by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis. Between HQLD (n = 20) and LQLD (n = 20) Duroc pigs, 24 differentially expressed proteins were identified by LC-MS/MS. A total of 10 and 14 proteins were highly expressed in HQLD and LQLD, respectively. The 24 proteins have putative functions in the following seven categories: catalytic activity (31%), ATPase activity (19%), oxidoreductase activity (13%), cytoskeletal protein binding (13%), actin binding (12%), calcium ion binding (6%), and structural constituent of muscle (6%). Silver-stained image analysis revealed significant differential expression of lactate dehydrogenase A (LDHA) between HQLD and LQLD Duroc pigs. LDHA was subjected to in vitro study of myogenesis under oxidative stress conditions and LDH activity assay to verification its role in oxidative stress. No significant difference of mRNA expression level of LDHA was found between normal and oxidative stress condition. However, LDH activity was significantly higher under oxidative stress condition than at normal condition using in vitro model of myogenesis. The highly expressed LDHA was positively correlated with LQLD. Moreover, LDHA activity increased by oxidative stress was reduced by antioxidant resveratrol. This paper emphasizes the importance of differential expression patterns of proteins and their interaction for the development of meat quality traits. Our proteome data provides valuable information on important factors which might aid in the regulation of muscle development and the improvement of meat quality in longissimus dorsi muscles of Duroc pigs under oxidative stress conditions.

• Journal of Food Hygiene and Safety
• /
• v.27 no.3
• /
• pp.317-324
• /
• 2012

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

• Journal of Life Science
• /
• v.28 no.11
• /
• pp.1347-1353
• /
• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
• /
• v.26 no.4
• /
• pp.356-368
• /
• 2021

To understand the temporal variation of phytoplankton communities in a coastal area, the biomass and diversity were weekly investigated in the inner bay of Yeong-do, Busan. In the study area, chlorophyll a concentration ranged from 0.43~7.58 mg m^-3 during the study, indicating the study area was in mesotrophic or eutrophic status. The fractions of chlorophyll a occupied by large phytoplankton (> 3 ㎛ diameter) exhibited an average of 80% of total chlorophyll a in this study. Among the large phytoplankton, while Bacillariophyta was the most dominant in spring and summer, Cryptophyceae prevailed in the fall and winter. On the contrary, in the picophytoplankton community less than 3 ㎛ in diameter, Mamiellophyceae was the most dominant in most seasons, Cryptophyceae was relatively high with an average of 17.7 ± 17.6% throughout the year, but seasonal variations were large. Dinophyceae rarely occupied a higher fraction up to 60.4% of the picophytoplankton community. By weekly monitoring at a coastal station for 13 months, it is suggested that phytoplankton communities in coastal waters could be changed on a short time scale. If data are steadily accumulated at the time-series monitoring site for a long time, these will provide important data for understanding the long-term dynamics of phytoplankton as well as the impact of climate and environmental changes.

• Journal of Life Science
• /
• v.32 no.11
• /
• pp.872-881
• /
• 2022

In this study, we optimized the composition of the indole-3-acetic acid (IAA) production medium using response surface methodology on Pantoea agglomerans SRCM 119864 isolated from soil. IAA-producing P. aglomerans SRCM 119864 was identified by 16S rRNA gene sequencing. There are 11 intermediate components known to affect IAA production, hence the effect of each component on IAA production was investigated using a Plackett-Burman design (PBD). Based on the PBD, sucrose, tryptone, and sodium chloride were selected as the main factors that enhanced the IAA production at optimal L-tryptophan concentration. The predicted maximum IAA production (64.34 mg/l) was obtained for a concentration of sucrose of 13.38 g/l, of tryptone of 18.34 g/l, of sodium chloride of 9.71 g/l, and of L-tryptophan of 6.25 g/l using a the hybrid design experimental model. In the experiment, the nutrient broth medium supplemented with 0.1% L-tryptophan as the basal medium produced 45.24 mg/l of IAA, whereas the optimized medium produced 65.40 mg/l of IAA, resulting in a 44.56% increase in efficiency. It was confirmed that the IAA production of the designed optimal composition medium was very similar to the predicted IAA production. The statistical significance and suitability of the experimental model were verified through analysis of variance (ANOVA). Therefore, in this study, we determined the optimal growth medium concentration for the maximum production of IAA, which can contribute to sustainable agriculture and increase crop yield.

• The Korean Journal of Pesticide Science
• /
• v.5 no.3
• /
• pp.1-11
• /
• 2001

New technologies will have a large impact on the discovery of new herbicide site of action. Genomics, combinatorial chemistry, and bioinformatics help take advantage of serendipity through tile sequencing of huge numbers of genes or the synthesis of large numbers of chemical compounds. There are approximately $10^{30}\;to\;10^{50}$ possible molecules in molecular space of which only a fraction have been synthesized. Combining this potential with having access to 50,000 plant genes in the future elevates tile probability of discovering flew herbicidal site of actions. If 0.1, 1.0 or 10% of total genes in a typical plant are valid for herbicide target, a plant with 50,000 genes would provide about 50, 500, and 5,000 targets, respectively. However, only 11 herbicide targets have been identified and commercialized. The successful design of novel herbicides depends on careful consideration of a number of factors including target enzyme selections and validations, inhibitor designs, and the metabolic fates. Biochemical information can be used to identify enzymes which produce lethal phenotypes. The identification of a lethal target site is an important step to this approach. An examination of the characteristics of known targets provides of crucial insight as to the definition of a lethal target. Recently, antisense RNA suppression of an enzyme translation has been used to determine the genes required for toxicity and offers a strategy for identifying lethal target sites. After the identification of a lethal target, detailed knowledge such as the enzyme kinetics and the protein structure may be used to design potent inhibitors. Various types of inhibitors may be designed for a given enzyme. Strategies for the selection of new enzyme targets giving the desired physiological response upon partial inhibition include identification of chemical leads, lethal mutants and the use of antisense technology. Enzyme inhibitors having agrochemical utility can be categorized into six major groups: ground-state analogues, group specific reagents, affinity labels, suicide substrates, reaction intermediate analogues, and extraneous site inhibitors. In this review, examples of each category, and their advantages and disadvantages, will be discussed. The target identification and construction of a potent inhibitor, in itself, may not lead to develop an effective herbicide. The desired in vivo activity, uptake and translocation, and metabolism of the inhibitor should be studied in detail to assess the full potential of the target. Strategies for delivery of the compound to the target enzyme and avoidance of premature detoxification may include a proherbicidal approach, especially when inhibitors are highly charged or when selective detoxification or activation can be exploited. Utilization of differences in detoxification or activation between weeds and crops may lead to enhance selectivity. Without a full appreciation of each of these facets of herbicide design, the chances for success with the target or enzyme-driven approach are reduced.

• Development and Reproduction
• /
• v.6 no.2
• /
• pp.117-122
• /
• 2002

Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX＋Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX＋Oil group and in OVX＋E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX＋Oil and OVX＋E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

• Radiation Oncology Journal
• /
• v.28 no.1
• /
• pp.32-38
• /
• 2010

Purpose: Although radiation-induced fibrosis is one of the common sequelae occurring after irradiation of skin and soft tissues, the treatment methods are not well standardized. This study aimed to establish the skin fibrosis mouse model by fractionated radiation for the further mechanism studies or testing the efficacy of therapeutic candidates. Materials and Methods: The right hind limbs of BALB/c mice received two fractions of 20 Gy using a therapeutic linear accelerator. Early skin damages were scored and tissue fibrosis was assessed by the measurement of a leg extension. Morphological changes were assessed by H&E staining and by Masson's Trichrome staining. TGF-${\beta}1$ expression from soft tissues was also detected by immunohistochemistry and PCR. Results: Two fractions of 20 Gy irradiation were demonstrated as being enough to induce early skin damage effects such as erythema, mild skin dryness, dry and wet desquamation within several weeks of radiation. After 13 weeks of irradiation, the average radiation-induced leg contraction was $11.1{\pm}6.2mm$. Morphologic changes in irradiated skin biopsies exhibited disorganized collagen and extracellular matrix fibers, as well as the accumulation of myofibroblasts compared to the non-irradiated skin. Moreover, TGF-${\beta}1$ expression in tissue was increased by radiation. Conclusion: These results show that two fractions of 20 Gy irradiation can induce skin fibrosis in BALB/c mice accompanied by other common characteristics of skin damages. This animal model can be a useful tool for studying skin fibrosis induced by radiation.

• Korean Journal of Microbiology
• /
• v.42 no.1
• /
• pp.40-46
• /
• 2006

Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

• Development and Reproduction
• /
• v.5 no.2
• /
• pp.175-180
• /
• 2001

The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.