• Asian-Australasian Journal of Animal Sciences
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• v.32 no.5
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• pp.614-628
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• 2019

Objective: The inhibitory leukocyte immunoglobulin-like receptors (LILRBs) play an important role in innate immunity. The present study represents the first description of the cloning and structural and functional analysis of LILRB1 and LILRB3 isolated from two genetically disparate chicken lines. Methods: Chicken LILRB1-3 genes were identified by bioinformatics approach. Expression studies were performed by transfection, quantitative polymerase chain reaction. Signal transduction was analyzed by western blots, immunoprecipitation and flow cytometric. Cytokine levels were determined by enzyme-linked immunosorbent assay. Results: Amino acid homology and phylogenetic analyses showed that the homologies of LILRB1 and LILRB3 in the chicken line 6.3 to those proteins in the chicken line 7.2 ranged between 97%-99%, while homologies between chicken and mammal proteins ranged between 13%-19%, and 13%-69%, respectively. Our findings indicate that LILRB1 and LILRB3 subdivided into two groups based on the immunoreceptor tyrosine-based inhibitory motifs (ITIM) present in the transmembrane domain. Chicken line 6.3 has two ITIM motifs of the sequence LxYxxL and SxYxxV while line 7.2 has two ITIM motifs of the sequences LxYxxL and LxYxxV. These motifs bind to SHP-2 (protein tyrosine phosphatase, non-receptor type 11) that plays a regulatory role in immune functions. Moreover, our data indicate that LILRB1 and LILRB3 associated with and activated major histocompatibility complex (MHC) class I and ${\beta}2-microglobulin$ and induced the expression of transporters associated with antigen processing, which are essential for MHC class I antigen presentation. This suggests that LILRB1 and LILRB3 are transcriptional regulators, modulating the expression of components in the MHC class I pathway and thereby regulating immune responses. Furthermore, LILRB1 and LILRB3 activated Janus kinase2/tyrosine kinase 2 (JAK2/TYK2); signal transducer and activator of transcription1/3 (STAT1/3), and suppressor of cytokine signaling 1 genes expressed in Macrophage (HD11) cells, which induced Th1, Th2, and Th17 cytokines. Conclusion: These data indicate that LILRB1 and LILRB3 are innate immune receptors associated with SHP-2, MHC class I, ${\beta}2-microglobulin$, and they activate the Janus kinase/signal transducer and activator of transcription signaling pathway. Thus, our study provides novel insights into the regulation of immunity and immunopathology.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.187-195
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• 2022

Crucian carp, Carassius carassius (Weight 28.1±3.7 g, Length 10.0±1.0 cm) were challenged with Aeromonas hydrophila at 0, 2.0×10⁴, 2.0×10⁵, 2.0×10⁶, 2.0×10⁷ CFU/ml for 2 weeks. The lethal concentration 50 (LC₅₀) at 2 weeks of C. carassius challenged with A. hydrophila was 19.776×10⁵ CFU/ml. In hematological parameters, the hemoglobin and RBC counts were significantly decreased by A. hydrophila challenge, whereas there was no significant change in hematocrit. The inorganic plasma components such as magnesium and calcium were significantly decreased. In organic plasma components, glucose and cholesterol were significantly increased by A. hydrophila challenge, whereas total protein was significantly decreased. In enzymatic plasma components, ALP (Alkaline phosphatase) were significantly increased by A. hydrophila challenge. The results of this study suggest that the A. hydrophila challenge to C. carassius induced the significant physiological changes in the hematological parameters and plasma components as deadly pathogenic bacteria.

• The Korea Journal of Herbology
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• v.36 no.6
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• pp.27-37
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• 2021

Objectives : In the current study, we performed the 13-week repeated oral dose toxicity test and a 4-week recovery test of standardized Cornus officinalis Sieb. et Zucc. and Psoralea corylifolia L. 30 % ethanol extract (SCP) in Sprague-Dawley (SD) rats owing to aims for verifying no observed adverse effect level (NOAEL). Methods : The animal study was performed according to OECD guidelines for the testing of chemicals section 4 health effects test No.408 repeated dose 90-day oral toxicity study in rodents (03 October 2008). In the repeated dose toxicity study, SCP was orally administered to female and male rats at dose levels of 1,000, 2,000, and 4,000 mg/kg/day for 13-week. The control group and high dose (4,000 mg/kg/day) group were then monitored for 4 extra weeks to determine recovery time after the study period. 1) Results : Compared with the control group, there were no treatment-related adverse effects in clinical signs, body weight, hematology, serum biochemistry (Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, 𝛾-Glutamyl transpeptidase, Blood urea nitrogen, Creatinine, Glucose, Total cholesterol, Total protein, Creatine phosphokinase, Albumin, Total bilirubin, Triglyceride, Inorganic phosphorus, Albumin/Globulin ratio, Calcium ion, Sodium ion, Potassium ion, Chloride ion), necropsy findings and organ weight (Ovary, Adrenal gland, Pituitary, Thymus, Prostate, Testis, Epididymis, Spleen, Kidney, Heart, Lung, Brain, Liver) at any dose tested. Conclusions : Taken together, these results suggest that the NOAEL of SCP in both genders was considered as over 4,000 mg/kg. Results from this study provide scientific evidence for the safety of SCP.

• IMMUNE NETWORK
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• v.21 no.6
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• pp.43.1-43.14
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• 2021

Group 3 innate lymphoid cells (ILC3), which express IL-22 and IL-17A, has been introduced as one of pathologic cells in axial spondyloarthritis (axSpA). Dyslipidaemia should be managed in axSpA patients to reduce cardiovascular disease, and dyslipidaemia promotes inflammation. This study aimed to reveal the role of circulating ILC3 in axSpA and the impact of dyslipidaemia on axSpA pathogenesis. AxSpA patients with or without dyslipidaemia and healthy control were recruited. Peripheral blood samples were collected, and flow cytometry analysis of circulating ILC3 and CD4⁺ T cells was performed. The correlation between Ankylosing Spondylitis Disease Activity Score (ASDAS)-C-reactive protein (CRP) and circulating immune cells was evaluated. The effect of oxidized low-density lipoprotein cholesterol (oxLDL-C) on immune cell differentiation was confirmed. AxSpA human monocytes were cultured with with oxLDL-C, IL-22, or oxLDL-C plus IL-22 to evaluate osteoclastogenesis using tartrate-resistant acid phosphatase (TRAP) staining and real-time quantitative PCR of osteoclast-related gene expression. Total of 34 axSpA patients (13 with dyslipidaemia and 21 without) were included in the analysis. Circulating IL-22⁺ ILC3 and Th17 were significantly elevated in axSpA patients with dyslipidaemia (p=0.001 and p=0.034, respectively), and circulating IL-22⁺ ILC3 significantly correlated with ASDAS-CRP (Rho=0.4198 and p=0.0367). Stimulation with oxLDL-C significantly increased IL-22⁺ ILC3, NKp44^- ILC3, and Th17 cells, and these were reversed by CD36 blocking agent. IL-22 and oxLDL-C increased TRAP⁺ cells and osteoclast-related gene expression. This study suggested potential role of circulating IL-22⁺ ILC3 as biomarker in axSpA. Furthermore, dyslipidaemia augmented IL-22⁺ ILC3 differentiation, and oxLDL-C and IL-22 markedly increased osteoclastogenesis of axSpA.

• Korean Journal of Agricultural Science
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• v.25 no.2
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• pp.181-198
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• 1998

In order to determine the optimum pasteurization conditions by microwave heating(MWH) at $50^{\circ}C{\sim}70^{\circ}C$ for 30 minute compared with water bath heating(WBH) at $65^{\circ}C$ for 30minute during storage at $5^{\circ}C$, the chemical composition, microbiological changes and keeping quality were examined and the results were as follows: 1. The fat protein lactose, total solid contents of raw milk, at $50{\sim}70^{\circ}C$ for 30 min. in MWH and at 65 for $30^{\circ}C$ min. in WBH did not changed significantly during the storage at $5^{\circ}C$. 2. The pH and acidity for the raw milk untreated were 6.75 and 0.16%, and those of MWH heated and WBH milk wee 6.75~6.50 and 0.16%~0.19%, phosphatase test were negative at $61^{\circ}C$ for 20 min. at $62^{\circ}C$ for 15 min. at $63^{\circ}C$ for 10 min. at $64^{\circ}C$ for 5 min. at $65^{\circ}C$ for 5 min. in MWH and at $65^{\circ}C$ for 30 min. in WBH. 3. Whey protein content was $18.53mg/m{\ell}$ in raw milk untreated, however, those were decreased as the heating temperature increased. The proteolytic activity of treated milk by WBH(44%) was lower than that by MWH(94%). 4. Total bacteria counts were $2.8{\times}10^5CFU/m{\ell}$ in raw milk untreated, $2.8{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $2.4{\times}10^3CFU/m{\ell}$ at $70^{\circ}C$ for 30 min. in MWH and $3.0{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. in WBH. Because total bacteria count did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30 min. and $65^{\circ}C$ for 30 min. in WBH during the 10 days storaging, Also, total bacteria counts for treated milk were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 5. Coliform bacteria counts were $2.6{\times}10^3CFU/m{\ell}$ in raw milk untreated. There were not detected at $55^{\circ}C{\sim}70^{\circ}C$ for 30 min. in MWH and at $65^{\circ}C$ for 30 min. in WBH. Coliform bacteria counts were not detected after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 6. Thermoduric bacteria counts were $5.2{\times}10^4CFU/m{\ell}$ in raw milk untreated, $2.0{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $1.9{\times}10^3CFU/m{\ell}$ at $70^{\circ}C$ for 30min. in MWH and $2.2{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. in WBH. Because thermoduric bacteria counts did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30 min. and $65^{\circ}C$ for 30 min. in WBH during the 10days storaging. Also, thermoduric bacteria counts were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 7. Psychrotrophic bacteria counts were $2.8{\times}10^5CFU/m{\ell}$ in raw milk untreated, $2.0{\times}10^1CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $2.0{\times}10^1CFU/m{\ell}$ at $70^{\circ}C$ for 30 min. in MWH and $3.0{\times}10^1CFU/m{\ell}$ at $65^{\circ}C$for 30 min. in WBH. Because psychrotrophic bacteria counts did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30min. and $65^{\circ}C$ for 30 min. in WBH during the 10 days storaging. Also, psychrotrophic bacteria counts were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.8
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• pp.992-997
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• 2008

In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with $0.001\;mg/mL{\sim}0.1\;mg/mL$ silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)-$1{\beta}$ was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-$1{\beta}$.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1164-1172
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• 2011

Two experiments were conducted to evaluate the efficacy of Trichoderma reesei derived phytase for pigs fed diets with fixed calcium to total phosphorus ratios (1.5:1). In Exp. 1, 280 weaned pigs (initial BW of $10.32{\pm}1.94$ kg) were allocated to one of five dietary treatments on the basis of weight and gender in a randomized complete block design. Treatments were the low phosphorus (0.6% Ca, 0.4% total P and 0.23% available P) diets supplemented with 0, 250, 1,000, or 2,000 FTU phytase/kg of diet and a positive control diet (PC; 0.85% Ca, 0.58% total P and 0.37% available P). The treatments were applied to seven pens with eight pigs per pen, half male and half female. In Exp. 2, six barrows fitted with ileal T-cannula (initial BW = $35.1{\pm}1.6$ kg) were assigned to three dietary treatments with a double $3{\times}3$ Latin square design. The dietary treatments were the low-phosphorus diet (0.53% Ca, 0.34% total P and 0.14% available P), the low phosphorus diet plus 1,000 FTU phytase/kg and a positive control diet (0.77% Ca, 0.50% total P and 0.30% available P). In Exp. 1, there were linear increases (p<0.01) in weight gain, phosphorus absorption, bone strength, calcium and phosphorus content of fat-free dried bone and plasma phosphorus concentrations with increasing dose rate of phytase. The performance of pigs fed the diets with 250, 1,000, or 2,000 FTU of phytase/kg did not differ from pigs fed the PC diet. Pigs fed diets with 1,000 or 2,000 FTU of phytase/kg did not differ from pigs fed the PC diet in bone characteristics. The apparent digestibility of dry matter, crude protein, ash and energy was not affected by dietary treatment. However, pigs fed the PC diet excreted more fecal phosphorus (g/d, p<0.01) and fecal phosphorus per BW gain (g/kg) than pigs fed the diets with phytase. Phytase linearly decreased (p<0.01) fecal phosphorus excreted per BW gain (g/kg), plasma calcium concentration as well as plasma and bone alkaline phosphatase activity. In Exp. 2, phytase supplementation in the low-P diet increased (p<0.05) the apparent ileal digestibility (AID) of Ca, P, leucine, lysine, phenylalanine, alanine and cysteine, tended to AID of crude protein, isoleucine, threonine, asparagine and serine. In conclusion, the novel phytase originated from Trichoderma reesei is effective in releasing Ca, P, and amino acids from corn soy based diet for pigs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.3
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• pp.466-470
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• 2000

The current study was undertaken to determine the prolinged effects of fasting-refecding on the lipoprotein lipase (LPL) activity and lipogenesis of adipose tissus in male Sprague-Dawley rals and to investigate the effects of various degrecs of food restriction during refeeding on the LPL activity and lipogenesis. The control group (n=5) was fed ad libitum and killed in the fed state at the beginning of the experiment. All rats except conteol group were fasted for 2 days (n=50). Five rats were killed at the end of fasting and others (n=45) were refed either ad libitum (ad libitum group) or mildly restricted (20% food restricted group), or esverely restricted diet (40% food restricted group). Rats were killed on the day of 7th, 14th, and 21st of refeeding. Lipogenesis was determined by the amount of glucose converted to the total lipid. Body weight and epididymal adipose tissue weight returned to control states by 5 days in ad libitym group and by 14 days in 20% food restricted group. As expected, in 40% food restriction during refeeding weight and epididymal adipose tissue weight did not rcturn to control states until day 21. On day 21 after refeeding, the serum total cholesterol concentration of ad libitum group was significantly (p<0.05) higher than that of control group. The serum HDL-C concentration of 40% food restricted group during refeeding was significantly (p<0.05) higher than that of control group. However, there were no significant dif-ferences in serum HDL-C/total cholesterol (TC) ratio and trigluceride concentration among the groups. Fasting for 2 days decreased lipogenesis and LPL activity (p<0.05). On day 21 after refecding, the lipogencsis of ad libitum group was significantly (p<0.05) lower than that of control group. The lipogenesis of 40% food restricted group during refeeding was significantly (p<0.05) higher than that of conrtol group. Ad libitum group and 20% food restricted group during refecding allowed heparin-releasable (HR) LPL or total extractable (TE) LPL activity to return to control states. 40% food restricted group during refeeding delayed the return of HR-LOL or TE-LPL activity to return to control states until day 21 of refeeding. These results suggest that food restriction during refeeding can partially or completely prevent the overshoot of LPL activity, and this may influence the rate of lipid accumulation in adipose tissue during refeeding.

• Journal of Life Science
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• v.30 no.11
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• pp.983-989
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• 2020

The balance between bone-resorbing osteoclasts and bone-forming osteoblasts is key to bone health. An imbalance between osteoclasts and osteoblasts leads to various bone-related disorders, such as osteoporosis, osteomalacia, and osteopetrosis. However, the bone-resorption inhibitor drugs that are currently used may cause side effects. Natural substances have recently received much attention as therapeutic drugs for the treatment of bone health. This study was designed to determine the effect of Tenebrio molitor larvae ethanol extract (TME) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. To measure the effect of TME on osteoclast differentiation, RAW264.7 cells were treated with RANKL with or without TME for 5 days. The tartrate-resistant acid phosphatase (TRAP) activity was significantly inhibited by treatment of TME without cytotoxicity up to 2 mg/ml. In addition, TME effectively suppressed expression of osteoclast differentiation-related marker genes and proteins such as TRAP, NFATc1, and c-Src. TME also significantly inhibited the p38 mitogen-activated protein kinase (MAPK) signaling pathway without affecting ERK and JNK signaling in RANKL-induced RAW264.7 cells. Consequently, we conclude that TME suppresses osteoclast differentiation by inhibiting RANKL-induced osteoclastogenic genes expression through the p38 MAPK signaling pathways. These results suggest that TME and its bioactive components are potential therapeutics for bone-related diseases such as osteoporosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1551-1558
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• 2009

The management of skeletal health in the twenties is the number one priority for preventing fracture or osteoporosis occurrence in later life cycle. Therefore, the factors influencing bone mineral density were examined by anthropometric measurements, food intakes, bone mineral density and biochemical nutritional indexes in blood among college students. Among the students who received health examinations from May to December, 2007, 532 male and 507 female students who agreed to participate in the study were selected. Nutritional intakes were estimated with 3-day 24-hour recall method. Bone mineral density was measured by ultrasonic attenuation passing through right calcaneus bone and expressed as broadband ultrasound attenuation and t-score was calculated by WHO criteria. Red blood cell count, hemoglobin, hematocrit values were measured by whole blood analysis, and alkaline phosphatase, serum calcium and serum iron were used as indexes for biochemical nutritional status. Data analysis was conducted using SPSS 14.0 program, and protecting and risk factors on bone health status were analyzed by logistic regression analysis between normal bone health group (t-score$\underline{\geq}$ -1.0) and osteopenia group (t-score<-1.0). The results showed that more people belong to the normal bone health group probably because this study was conducted among those with their peak bone density. Biochemical nutritional status and nutrition intakes of both groups for normal and osteopenia did not show statistically significant difference except MCHC, animal protein and animal iron intakes in female. According to the results of logistic regression analyses, dietary intakes of animal protein, animal iron and zinc showed protecting effects against osteopenia. Therefore, dietary nutritional intakes of micronutrients, especially iron and zinc are important for bone health of young people.