• Biomolecules & Therapeutics
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• v.27 no.4
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• pp.414-422
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• 2019

There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

• Animal Bioscience
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• v.37 no.6
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• pp.1053-1064
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• 2024

Objective: This study aims to investigate the interactive effect of a glycine equivalent (Gly_equi) and standardized ileal digestible threonine (SID Thr) levels in low crude protein diets on performance, blood biochemistry, pectoral muscular creatine content and oxidative stability of meat in broiler chickens from 21 to 42 days. Methods: A total of 1,500, twenty-one-day-old Cobb-Vantress male broiler chickens were distributed in a completely randomized 5×3 factorial arrangement of Gly_equi×SID Thr with five replicates of 20 birds each. Fifteen dietary treatments of 16.5% CP were formulated to contain five levels of total Gly_equi (1.16%, 1.26%, 1.36%, 1.46%, and 1.56%) and three levels of SID Thr (0.58%; 0.68% and 0.78%). Results: Interaction effects (p<0.05) of Gly_equi and SID Thr levels were observed for weight gain, carcass yield, pectoral muscular creatine content and serum uric acid. Higher levels of Gly_equi increased (p = 0.040) weight gain in 0.58% and 0.68% SID Thr diets compare to the 0.78% SID Thr diet. The SID Thr level at 0.68% improved (p = 0.040) feed conversion compared to other SID Thr diets. Levels of Gly_equi equal to or above 1.26% in diets with 0.78% SID Thr resulted in birds with higher (p = 0.033) pectoral muscular creatine content. The breast meat yield observed in the 0.68% SID Thr diet was higher (p = 0.05) compared to the 0.58% SID Thr diet. There was a quadratic effect of Gly_equi levels for pectoral pectoral muscular creatine content (p = 0.008), breast meat yield (p = 0.030), and serum total protein concentrations (p = 0.040), and the optimal levels were estimated to be 1.47%, 1.35%, and 1.40% Gly_equi, respectively. The lowest (p = 0.050) concentration of malondialdehyde in the breast meat was found in 0.68% SID Thr diets at 1.36% Gly_equi. Conclusion: The minimum dietary level of Gly_equi needed to improve performance in low crude protein diets is 1.26% with adequate SID Thr levels for broiler chickens.

• Korean Journal of Food Science and Technology
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• v.2 no.2
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• pp.8-16
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• 1970

Exploitation of leaf protein concentrates for human consumption is very important. Leaf protein concentrates can be easily prepared by mechanically mincing leaves material and press it for getting the juice. Crude protein can be separated from the juice by aging, adjusting the pH, or heating to $75-80^{\circ}C$ etc. This report deals with the extractability of total-N from 69 species of fresh leaves by mechanical process, and then compared the recovery of leaf protein concentrates from leaf extracts by treating with TCA, pH adjustment and heating. Results are summarized as follows. 1. In general, the greater the content of total-N of leaves the greater the percentage extraction. Extraction of the juice from leaves is needed at least two times. The simple equations are constituted between the total-N (T; %) and the first and second extractability ($E_1,\;E_2;\;%$) of the total-N of leaves, as follows: $E_1=0.8168T\;E_2=0.1830T$ 2. The optimum pH value for coagulating protein from extracts is considered to be 3.5 to 4.5. However, the products of leaf protein concentrate by the pH adjustment of extracts are generally dull in color with rich elasticity. 3. Recoveries of the leaf protein concentrate from extracts by treating methods were in the following order of TCA treatment> pH 4 treatment> pH 3 treatment> heat treatment. The yield of leaf protein concentrates decreased bout 10% with pH 4 treatment, 11.4% with pH 3 treatment, and 14.8% with heat treatment compared with the TCA treatment. 4. The heat treatment is the most benifitial method for the production of leaf protein concentrates with regard to properties of texture, color and yield of products and easiness of the treatment method.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.9
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• pp.1267-1278
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• 2009

Three beef steers fitted with permanent cannulae in the rumen and duodenum were used to determine the effects of protein supply from soyhulls (SH) and wheat bran (WB) on ruminal metabolism, blood metabolites, nitrogen metabolism, nutrient digestion and concentrations of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). In a 3${\times}$3 Latin square design, steers were offered rice straw and concentrates formulated either without (control) or with two brans to increase crude protein (CP) level (9 vs. 11% dietary DM for control and bran-based diets, respectively). The brans used were SH and WB that had similar CP contents but different ruminal CP degradability (52 vs. 80% CP for SH and WB, respectively) for evaluating the effects of protein degradability. Ruminal ammonia concentrations were higher for bran diets (p<0.01) than for the control, and for WB (p<0.001) compared to the SH diet. Similarly, microbial nitrogen and blood urea nitrogen were significantly increased (p<0.05) by bran and WB diets, respectively. Retained nitrogen tended (p<0.082) to be increased by SH compared with the WB diet. Intestinal and total tract CP digestion was enhanced by bran diets. In addition, bran diets tended (p<0.085) to increase intestinal starch digestion. Concentrations of SNAN fractions in RD and OD were higher (p<0.05) for bran diets than for the control, and for WB than for the SH diet. More rumendegraded protein supply resulting from a higher level and degradability of CP released from SH and WB enhanced ruminal microbial nitrogen synthesis and ruminal protein degradation. Thus, free amino acids, peptides and soluble proteins from microbial cells as well as degraded dietary protein may have contributed to increased SNAN concentrations in the rumen and, consequently, the omasum. These results indicate that protein supply from SH and WB, having a low level of protein (13 and 16%, respectively), could affect ruminal metabolism and nutrient digestion if inclusion level is relatively high (>20%).

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.8
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• pp.3581-3586
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• 2014

Aim: To investigate the effects of tetramethypyrazine (TMP) on proliferation and apoptosis of the human gastric carcinoma cell line 7901 and its possible mechanism of action. Methods: The viability of TMP-treated 7901 cells was measured with a 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT) and cell apoptosis was analyzed by flow cytometry. The distribution of cells in different phases of cell cycle after exposure of TMPs was analyzed with flow cytometry. To investigate the molecular mechanisms of TMP-mediated apoptosis, the expression of NF-${\kappa}Bp65$, cyclinD1 and p16 in SGC-7901 cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Results: TMP inhibited the proliferation of human gastric carcinoma cell line 7901 in dose and time dependent manners. Cell growth was suppressed by TMP at different concentrations (0.25, 0.5, 1.0, 2.0 mg/ml), the inhibition rate is 0.46%, 4.36%, 14.8%, 76.1% (48h) and 15.5%, 18.5%, 41.2%, 89.8% (72h) respectively. When the concentration of TMPs was 2.0mg/ml, G1-phase arrest in the SGC-7901 cells was significant based on the data for cell cycle distribution. RT-PCR demonstrated that NF-${\kappa}Bp65$ and cyclin D1 mRNA expression was significantly down-regulated in 7901 cells treated with 2.0 mg/ml TMP for 72h (p<0.05), while the p16 mRNA level was up-regulated (p<0.05). The protein expression of NF-${\kappa}Bp65$ and cyclin D1 decreased gradually with the increase in TMP concentration, compared with control cells (p<0.05), while expression of protein p16 was up-regulated (p<0.01). Conclusion: TMP exhibits significant anti-proliferative and pro-apoptotic effects on the human gastric carcinoma cell line SGC-7901. NF-${\kappa}Bp65$, cyclinD1 and p16 may also play important roles in the regulation mechanisms.

• Clinical and Experimental Pediatrics
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• v.56 no.4
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• pp.159-164
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• 2013

Purpose: Indoxyl sulfate and p-cresyl sulfate are important protein-bound uremic retention solutes whose levels can be partially reduced by renal replacement therapy. These solutes originate from intestinal bacterial protein fermentation and are associated with cardiovascular outcomes and chronic kidney disease progression. The aims of this study were to investigate the levels of indoxyl sulfate and p-cresyl sulfate as well as the effect of probiotics on reducing the levels of uremic toxins in pediatric patients on dialysis. Methods: We enrolled 20 pediatric patients undergoing chronic dialysis; 16 patients completed the study. The patients underwent a 12-week regimen of VSL#3, a high-concentration probiotic preparation, and the serum levels of indoxyl sulfate and p-cresyl sulfate were measured before treatment and at 4, 8, and 12 weeks after the regimen by using fluorescence liquid chromatography. To assess the normal range of indoxyl sulfate and p-cresyl sulfate we enrolled the 16 children with normal glomerular filtration rate who had visited an outpatient clinic for asymptomatic microscopic hematuria that had been detected by a school screening in August 2011. Results: The baseline serum levels of indoxyl sulfate and p-cresyl sulfate in the patients on chronic dialysis were significantly higher than those in the children with microscopic hematuria. The baseline serum levels of p-cresyl sulfate in the peritoneal dialysis group were significantly higher than those in the hemodialysis group. There were no significant changes in the levels of these uremic solutes after 12-week VSL#3 treatment in the patients on chronic dialysis. Conclusion: The levels of the uremic toxins p-cresyl sulfate and indoxyl sulfate are highly elevated in pediatric patients on dialysis, but there was no significant effect by probiotics on the reduction of uremic toxins in pediatric dialysis patients. Therefore, studies for other medical intervention to reduce uremic toxins are also necessary in pediatric patients on dialysis.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.12
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• pp.1752-1760
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• 2005

In this experiment, the effects of cage density (CD) and probiotic supplementation (PS) on laying performance, metabolic profile, and egg quality in peak-producing hens were evaluated. After blocking according to the cage location, Lohman layers (n = 180, 46 wks of age) were allocated randomly to two levels of CD (540 vs. 360 $cm^2$/hen) and three levels of PS (0, 0.15, and 0.30%). Probiotic contained Enterococcus faecium culture (10${\times}$10$^9$ cfu/g). Egg production (EP) and feed consumption (FC) were measured daily; egg weight (EW) was measured bi-weekly; BW was measured before and after the experiment; and blood samples were obtained at the end of the experiment. The data were analyzed using two-way ANOVA. Increasing CD decreased FC (125.0 vs. 120.8 g/d, p<0.0001) and FCR (1.93 vs. 1.87, p<0.0001) and did not alter EP, EW, and BW. Increasing level of PS linearly decreased FC (p<0.02) and FCR (p<0.006). Averages were 123.9, 123.2, and 121.6 g/d for FC and 1.91, 1.92, and 1.86 for FCR in hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. Hens placed in high-density cages had greater serum corticosterone concentration than hens placed in normal-density cages (12.8 vs. 11.3 $\mu$g/dL, p<0.04); CD did not affect concentrations of other metabolites. Increasing level of PS linearly increased serum glucose, albumin, and creatine concentrations and quadratically increased total protein, globulin, Ca, and P concentrations. Average concentrations (mg/dL) were 260, 297, and 305 for glucose; 6.28, 8.09, and 7.58 for total protein; 1.98, 2.48, and 2.38 for albumin; 4.30, 5.62, and 5.19 for globulin; 0.40, 0.52, and 0.54 for creatine; 16.0, 16.5, and 16.3 for Ca; and 6.27, 8.14, and 7.17 for P in hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. There was no effect of CD on egg quality. Increasing level of PS linearly improved yolk color (YC) and quadratically increased albumen index (AI) and Haugh unit (HU). The mean values were 9.67, 9.75, and 10.58 for YC; 8.94, 6.93, and 8.72% for AI; and 85.6, 74.9, and 82.9 for HU for hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. There was also CD by PS effect on FC, EP, and serum glucose, total protein, albumin, globulin, creatine, Ca and P concentrations. In conclusion, increased CD partially depressed laying performance and caused stress. Probiotic supplementation improved laying performance and metabolic profile. It also partially alleviated the adverse effects of stress resulting from increased caging density.

• Journal of Nutrition and Health
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• v.19 no.1
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• pp.16-22
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• 1986

This study was designed to investigate the nutritional effect of mugowrt powder supple mentation to rice diets with different protein levels. Thirty female albino rats weighing 39-43g were adopted for the feeding trial for 4 weeks. The different 5 kind of experimental diets were performed . Control diet was commercially available forage for rats, experimental diet (I-C) highly milled rice, (Ⅰ) highly milled rice 95% and mugwort powder 5%, (Ⅱ-C)highly milled rice 95% and milk casein 5%, and (Ⅱ) highly milled rice 90%, milk casein 5 % and mugwort powder 5%. Growth rate was remarkably high in the dietary group fed on highly milled rice supplemented with 5% mugwort powder (protein 8%) (P<0.05), but it showed the tendency to be rather low in the group fed on highly milled rice supplemented with 5% casein and 5% mugwort powder (protein 12%). Food efficiency as well as protein efficiency appeared similar to the growth rate. Hematodcrit level demonstrated the same tendency as growth rate, but hemoglobin content was observed to increase by diets supplemented with increasing amount of mugwort regardless of protein level. Each nutrient intake was increased by adding mugwort powder to diets after a week's feeding, but it was increased by 8% protein diet, and decreased by 12% protein diet in 3 weeks as well as 4 weeks after feeding . The absorption rate of carbohydrate and protein decreased by feeding mugwort supplemented diets regardless of protein level and feeding period, and that of lipid increased with 12% protein diet.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.5
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• pp.373-384
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• 2001

Cellular proliferation is an intricately regulated process mediated by the coordinated interactions of critical growth control genes. Two of these factors in mammalian cells are the p53 and mdm-2 genes. A protein product of the mem-2 oncogene has been recently shown to associate with the protein encoded by the tumor suppressor gene p53. The p53 tumor suppressor protein is stabilized in response to DNA damage and other stress signals and causes the cell to undergo growth arrest or apoptosis, thus preventing the establishment of mutations in future cellular generations. Mutation or loss of p53 is a very common event in tumor progression. It occurs in about 50% of all tumors analysed including of colon, lung, breast and liver. The cellular mdm-2 gene, which has potential transforming activity that can be activated by overexpression, is amplified in a significant percentage of human sarcoma and in other mammalian tumors. Proteins encoded by the mdm-2 gene are able to bind to the p53 protein and, when overexpressed, can inhibit p53's transcriptional activation function, thus mdm-2 can act as a negative regulator of p53 function. Experimental study was performed to observe the relationship between p53 gene mutation and mdm-2 protein expression and apply the results to the clinical activity. 36 golden syrian hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek(control side) was treated with mineral oil as the same manner to the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were examined for histology and immunohistochemistry observation, and were completely dissected by microdissection and DNA from both tissue were isolated by proteins K/phenol/chloroform extraction. Segments of the hamster p53 exons 5, 6, 7 and 8 were amplified by PCR using the oligonucleotide primers, and then confirmational change was observed by SSCP respectively. The results were as follows : 1. Dysplasia at 6 weeks, carcinoma in situ at 8 weeks and invasive carcinoma from 10 weeks could be observed in experimental groups. 2. p53 mutations were detected in 10 of the 36(28%) and the exons 6(6 of the 10 : 60%) was the most hot spot area among the highy conserved region(exons 5, 6, 7 & 8). 3. Immunohistochemical study confirmed 22 of the 36(61%) of p53 expression involving 10 of p53 mutations. 4. mdm-2 expression of was showed in 3 of the 36(8%) involving 1 of the 22 of p53 expression and 2 of the 14 of p53 non-expression. From the above results, mutation of p53 gene or expression of p53 protein may have the influence of the DMBA induced carcinoma of hamster buccal pouch but the expression of mdm-2 protein may not have relationship with tumorigenesis.

• Korean Journal of Community Nutrition
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• v.21 no.4
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• pp.366-377
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• 2016

Objectives: The purpose of the study was to compare intake of energy nutrients, physical characteristics, and the prevalence of metabolic syndrome according to protein intake group. Methods: Subjects were 827 men aged 40-65 years. The results presented were based on data from the 2012-2013 National Health and Nutrition Examination Survey and analyzed using SPSS. The odds ratio (OR) of metabolic syndrome was assessed according to the protein intake group and intake pattern of protein-rich foods. Results: The mean of protein intake was $73.96{\pm}0.71g$. According to level of protein intake, four groups (deficient, normal, excess 1, excess 2) were created and their percentages were 8.3%, 39.6%, 37.1%, and 15.0% respectively. The mean of daily energy intake was $2,312.33{\pm}24.08kcal$. It was higher in excess group 2 than in the deficiency group (p < 0.001). Moreover, the intake of all energy nutrients increased significantly with protein intake group (p < 0.001). The main contribution to daily protein included mixed grains ($10.96{\pm}0.32g$), milled rice ($7.14{\pm}0.30g$), chicken ($3.50{\pm}0.21g$), and grilled pork belly ($3.04{\pm}0.16g$). With regard to physical characteristics, and blood pressure and blood test results, only body mass index increased significantly according to protein intake groups (p < 0.05). The prevalence of metabolic syndrome in subjects was 38.5%, and there was no significant correlation with protein intake group. The OR of metabolic syndrome increased with protein intake, and was higher 4.452 times in excess group 2 than in the normal group (p < 0.05). Conversely, the OR of metabolic syndrome according to the frequency of protein-rich food intake did not show a significant correlation. Conclusions: The results of this study can be used as significant supporting data to establish guidelines for protein intake in middle-aged men.