Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).
This experiment was conducted to investigate effects of dietary protease on immune responses of weaned pigs. Weaned pigs (n = 75; 7.06 ± 0.18 kg BW; 28 d old) were randomly assigned to 3 treatments (5 pigs/pen; 5 pens/treatment). Dietary treatments were positive control, a diet with required protein level (PC), negative control, a diet with lower protein level than PC (NC), and NC + 0.02% dietary protease (PRO). The dietary protease used in this experiment was a commercial product containing 75,000 protease units/g derived from Nocardiopsis prasina produced in Bacillus licheniformis. The dietary treatments did not contain any ingredients or additives that may provide antibacterial or physiological effects. Pigs were fed respective dietary treatments for 6 weeks. Blood was collected from randomly selected 2 pigs in each pen on d 1, 3, 7, and 14 after weaning. Measurements were number of white blood cells (WBC), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), and C-reactive protein (CRP). Pigs fed PRO had lower WBC on d 7 (14.84 vs 20.42 × 103/μL; p < 0.05) and TNF-α on d 7 (618 vs 889 pg/mL; p = 0.085) and 14 (437 vs 576 pg/mL; p = 0.069) than those fed NC, but there were no differences on WBC and TNF-α between PC and PRO. Pigs fed PRO had lower TGF-β1 on d 3 (630 vs. 1,588 and 1,396 pg/mL; p < 0.05) than those fed PC and NC. However, no differences were found on CRP among dietary treatments. In conclusion, addition of dietary protease reduced inflammatory immune responses of weaned pigs.
Nutritional concentrations by chemical analyses of mushroom fermented milk were protein 2.87%, fat 0.09%, carbohydrates 6.0%, dietary fiber 0.3%, lactose 2.01%, sucrose 1.23%, calcium 95.9 mg/100 g and iron 0.08 mg/100 g. The present study was undertaken to investigate the hypoglycemic effects of the equal volume of either water (streptozotocin (STZ)rontrol rats), mushrooms water-extract (STZ-extrart fed rats), mushroom fermented milk product (STZ-mushroom yogurt fed rats) or mushroom fermented milk supernatant (STZ-supernatant fed rats) (10%, v/w), in STZ-induced diabetic rats for 3 week period. The mushroom fermented milk given to the STZ-diabetic rats decreased the blood glucose significantly and increased the blood insulin, compared with the STZ-control rats. The supernatant and mushroom water extract also slightly retarded the development of hyperglycemia in the STZ-diabetic rats. Taken together the results, the mushroom yogurt may have a potential for the hypoglycemic effect in the STZ-diabetic rats.
Kim, Mi-Sun;Lee, Ye-Seul;Kim, Jong Sik;Shin, Woo-Chang;Sohn, Ho-Yong
Microbiology and Biotechnology Letters
/
v.42
no.3
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pp.258-266
/
2014
Sweet potato soju (SPS), a form of traditional distilled alcoholic liquor in Korea, is manufactured by the distillation of fermented broth under normal pressure, thus providing it for a uniquely smooth taste infused with the flavor of sweet potato. After distillation, the lees of SPS is produced as by-product and discarded. In this study, the ethanol and hot water extracts of lees of SPS, and their subsequent organic solvent fractions using hexane, ethylacetate (EA), butanol, and water residue were prepared in an effort at the efficient re-use of the lees of SPS. The ethanol extraction yield was 1.36-fold higher than that of the hot water extraction, and the EA fraction revealed the highest total polyphenol content among the solvent fractions. The various extracts and solvent fractions did not demonstrate hemolytic activity at up to 0.5 mg/ml concentrations against human red blood cells. In the bioactivity assay, only the EA fraction displayed a broad spectrum of anti-microbial activity against different pathogenic and food spoilage bacteria, and demonstrated significant anti-coagulation activity by inhibitions of thrombin, prothrombin and blood coagulation factors. Furthermore, only the EA fraction from the hot water extract of the lees of SPS showed anti-platelet aggregation activity, which is comparable to aspirin (a commercially available drug). Our results suggest that the EA fraction of the hot water extract prepared from the lees of SPS has a high potential as a novel resource for anti-microbial and anti-thrombosis agents.
Glucose and protein in urine are among the important substances for urine analysis and have generally been measured based on a reagent strip test. In this study, these two substances were measured using mid-infrared absorption spectroscopy. Samples were prepared from a commercial synthetic urine product. Glucose and albumin were added as well as red blood cells, which are expected to create the most spectroscopic interference of any substance. Concentrations of these substances were varied independently. Optimal wavelength regions were determined from a partial least squares regression analysis (glucose 980 - 1150/cm, albumin 1400 - 1570/cm). Interference by other substances increased the differences between measured and predicted values. Albumin measurement in particular weres heavily influenced by the presence of glucose and red blood cells. Depending on the inference by other substances, measurement errors were 29.85${\sim}$45.19 mg/dl for a glucose level between 0 and 1000 mg/dl and 14.0${\sim}$93.11 mg/dl for an albumin level of 0 ${\sim}$ 500 mg/dl. Our study proposes an alternative to the chemical test-strip analysis, which shows only discrete concentration levels.
The contribution of endogenous transport systems to the blood-brain barrier (BBB) transport of basic and acidic drugs was studied by using a carotid injection technique in rats and an isolated bovine cerebrovascular disease state were compared between the normotensive rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) which have been well established as an animal model with pathogenic similarities to humans. Basic drugs such as eperisone, thiamine and scopolamine inhibited, in a concentration dependent manner the in vivo uptake of $[{^3}H]choline$ through BBB, whereas amino acids and acidic drugs such as salicylic acid and valproic acid did not inhibit the uptake. The uptake of $[^3H]choline$ by B-CAP increased with time and showed a remarkable temperature dependency. The uptake of $[^3H]choline$ by B-CAP showed the very similar inhibitory effects as observed in the in vivo brain uptake, and was competitively inhibited by a basic drug, eperisone. The in vivo BBB uptakes of $[^3H]acetic$ acid and $[^{14}C]salicylic$ acid were dependent on pH of the injectate and the concentration of drugs. Several acidic drugs such such as salicylic acid, benzoic acid and valproic acid inhibited the in vivo uptake of $[^3H]acetic$ acid, whereas amino acid, choline and a basic drug such as eperisone did not inhibit the uptake. The uptake of acetic acid by B-CAP was competitively inhibited by salicylic acid. The permeability surface area product (PS) through BBB for $[^3H]choline$ in SHRSP was significantly lower than that in WKY. The concentration of choline in the brain dialysate in SHRSP was about half of that in WKY, while no significant difference was observed in the plasma concentration of choline between SHRSP and WKY. No significant difference was observed in the transport of monocarboxylic acids, glucose and neutral amino acid through BBB between SHRSP and WKY. From these results, it was concluded that BBB transport system of choline contributes to the transport of basic drugs through BBB, that acidic drugs can be transported via a moncarboxylic acid BBB transport system and that the specific dysfuntion of the BBB choline transport in SHRSP was ascribed to the reduction of the maximum velocity of choline concentration in the brain interstitial fluids.
This study aims to verify for humans the suitability of the enzyme-fixed hydrogel used for the patch sensor of the blood sugar testing system without blood sampling, which utilizes reverse iontophoresis. Using acrylate monomers, hydrogel was synthesized to which a certain unit of enzyme is fixed. In order to analyze the material property of the synthesized hydrogel, a structural analysis was performed using FT-IR spectroscopy, while the DSC was used to verify the thermal stability. In addition, with the UV-Vis spectrophotometer, it was verified that the degree of active enzyme is at least 50% greater than the standard product. The SEM was used to verify secure fixation of the enzyme onto the surface. As a result, it was observed that the enzyme is successfully fixed to the surface. Since the hydrogel makes direct contact with a patient's skin, it is essential to evaluate the toxicity when making direct contact with the skin. For that purpose, various sets of tests were undertaken according to the ISO 10993-cytotoxicity, intracutaneous reactivity, skin irritation test and maximization sensitization. Consequently, it was successfully verified that the enzyme-fixed hydrogel have bioavailability.
Objectives: The purpose of this study was to investigate the food, nutrient intake, and diet quality of postmenopausal women at high risk of osteoporosis (OP) and cardiovascular disease (CVD) compared with those of control subjects. Methods: A total of 1,131 post-menopausal women aged over 45 years, who took the 2010-2011 Korean National Health and Nutrition Examination Survey (KNHANES), were included for analysis. These participants were classified into the following groups: the OP group, with a risk of OP (n=135); the CVD group, with a risk of CVD (n=373); the OP+CVD group, with a risk of OP and CVD concurrently (n=218); and the control group (n=405) according to bone mineral density (BMD) and CVD risk. Anthropometric measurements, blood profiles, dietary intake, and dietary quality indices were measured and compared among the four groups. Results: Waist circumference, total body fat percentage, blood pressure, fasting plasma glucose, total cholesterol, triglyceride, and LDL-cholesterol were higher, and HDL-cholesterol and BMD were lower in the OP+CVD group than in the control group. In the food frequency questionnaire, the OP+CVD group had significantly higher frequencies of grain (except for multi-grain) and lower frequencies of fruit and dairy product. The frequency of consumption of red meat, processed meat, and carbonated beverages was higher in OP+CVD group. In nutrient density analysis, proteins and vitamin $B_2$ levels were significantly lower in the OP+CVD group than in the control group. The nutritional quality index (INQ) values of calcium were in the order of 0.63, 0.58, 0.56, and 0.55 in each group, and it was urgent to improve the dietary intake for calcium in postmenopausal women. In addition, vitamin $B_2$ was inadequately consumed by all groups. Conclusions: These results suggest that it is necessary to increase the intake of vitamin $B_2$ and calcium and decrease the frequency of intake of red meat, processed meat, and carbonated beverages in postmenopausal women with the risk of OP and CVD.
A feeding trial was conducted to investigate the effects of dietary fish meal replacement by a blend of lysine cell mass, corn protein concentrate and poultry by-product meal on the growth and blood chemistry of the starry flounder Platichthys stellatus. The fish meal replacer (FMR) was prepared to have the same level of protein as fish meal (FM). With a commercial diet as a positive control, five experimental diets (basal, FM42, FM32, FM22 and FM12) were formulated to contain 52% protein and 10% lipid. The dietary FM levels decreased from 52% (basal) to 42, 32, 22 and 12% with concomitant increase in the FMR to 10, 20, 30, 40 and 50%, respectively. Juvenile starry flounder with an average body weight of 177.3 g were randomly distributed in each (30 fish/tank) of 18 plastic tanks ($139{\times}99{\times}54cm$). After a 45-day feeding trial, the survival rate ranged from 95.6% (FM22) to 100% (control and FM42), while the weight gain of the fish groups varied from 49.7 to 58.4 g. The results clearly revealed that starry flounder can grow well on a diet containing low FM (12%) with a high level of FMR (50%) without any adverse effects.
Jo, Joo-hyun;Im, Ji-sung;Kim, Jong-gyu;Park, Jung-hyun;Choi, Hag-soon;Hwang, Geu-won;Song, Yung-sun
Journal of Korean Medicine Rehabilitation
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v.31
no.1
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pp.33-46
/
2021
Objectives The aim of this study is to evaluate anti-inflammatory and anti-arthritic effects of Sogyunghwalhyel-tang-gamibang (SGHHTGB) in cell and animal models and also to suggest one of putative mechanisms underlying its anti-arthritic effects. Methods Enzyme-linked immunosorbent assay was applied to measure the concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in culture medium and blood serum and nitric oxide (NO) was assayed by Griess reagent. The expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. Results In a cell model using RAW264.7 macrophages stimulated with the endotoxin lipopolysaccharide (LPS), the drug, at its non-cytotoxic concentrations, inhibited the production of the pro-inflammatory cytokine TNF-α, IL-1β and IL-6. In addition, it suppressed the expression of the inflammatory enzyme iNOS and COX-2, and reduced the synthesis of the enzyme product NO (as stable nitrite) and PGE2 in activated macrophages. Meanwhile, in an animal model using rheumatic arthritis (RA) mice induced with injection of type II collagen antibody (CAb) and LPS, the drug improved clinical symptom of arthritis and reduced paw thickness and inflammatory cell infiltration. In blood of RA mice, the drug reduced serum levels of TNF-α, IL-1β, IL-6, nitrite, and PGE2, all inflammatory mediators produced by activated macrophages. Conclusions SGHHTGB may ameliorate CAb and LPS-induced RA in mice, presumably by inactivating macrophages that are capable of initiating joint inflammation by producing pro-inflammatory cytokines and expressing inflammatory enzymes.
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