• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1485-1490
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• 2012

An electrochemical immunoassay based on osmium-hippuric acid (HA) conjugate films onto the electrode is presented for the detection of urinary HA. This is the first report on the use of the oxidative electropolymerization of 5-amino-1,10-phenanthroline (5-$NH_2$-phen) for immobilizing an antigen, osmium-conjugated HA. As a redox mediator, [Os(5-amino-1,10-phenanthroline)$_2$(4-aminomethylpyridine-HA)Cl]$^{+/2+}$ (Os-phen-HA) was successfully synthesized and electropolymerized onto the screen-printed carbon electrodes (SPCEs). The interaction between osmium-HA conjugate films and antibody-HA ($anti$-HA) was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The electrical signals were linearly proportional to urinary HA in the range of 0.1-5.0 mg/mL, which is sufficient for use as an immunosensor using a cutoff concentration of 2.0 mg/mL in urine samples. The proposed electrochemical immunoassay method can be extended to various applications for detecting a wide range of different small antigens in the health care area.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1483-1489
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• 2016

Most of amino acid (AA) digestibility values for feed ingredients are obtained using pigs cannulated in the distal ileum. The ileal-cannulated pig model uses pigs older than six weeks due to difficulties related to implanting the T-cannula in distal ileum of younger pigs and complications during the post-surgical recovery. However, to properly formulate the diet of weaned pigs, the nutritive value of feed ingredients should be determined with younger pigs. Thus, 25 weaned pigs were used to determine the apparent total tract digestibility (ATTD) of nutrients, energy, and apparent ileal digestibility (AID) and standardized ileal digestibility (SID) ileal AA digestibility of broken rice (BR), with or without multicarbohydrase (MC) and phytase (Phy) supplementation. Piglets were weaned at 23 d of age and individually housed in digestibility cages until 45 d of age. The trial consisted of 7 d of adaptation to the experimental diets and 3 d of excreta (feces and urine) collection. Ileal digesta was collected at slaughter (about 6 weeks of age). A completely randomized experimental design was used to determine the effects of MC and Phy. Reference diets (RD, 5% casein) was replaced by 30% of BR with or without MC, Phy, or MC+Phy. The RD was used to quantify endogenous AA losses. BR with Phy supplied had increased the ATTD of dry matter (p<0.05) and SID of histidine (p = 0.05), arginine, leucine, lysine, valine, alanine, and proline (p<0.05). BR with MC had been increased digestible energy and protein and SID for histidine (p<0.05). There was no interaction between Phy and MC on the BR nutrient digestibilities. Standardized amino acid digestibilities of BR, without enzymes, were lower than those values reported in the literature. The MC and Phy improved the digestibility of some nutrients and energy of BR in post-weaned piglet diets.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.472-477
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• 1996

Natural human epidermal growth factor (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C$_{18}$ HPLC system. Following characterization by Western blot analysis and double immu- nodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporatioin of 5-bromo-2'-deoxy uridine (BrdU), a non-radioactive alternative for [$^{3}$H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1 - 10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others. Based on the results from the HPLC analysis spiked with recombinant human epidermal growth factor (rhEGF) and amino acid sequencing, we concluded that the second fraction was nhEGF and the other three fractions were the derivatives of nhEGF. In addition, the proportion of nhEGF was approximately 46% is compared with that of the other three derivatives.

• Journal of Nutrition and Health
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• v.6 no.3
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• pp.9-15
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• 1973

Some effect on the protein metabolism in growing albino rats by feeding on the rice mixed with wheat or barley have been studied. The species of wheat and barley used in this experiment were either 80% polished or nonpolished wheat, barley and naked barley. The growing rats to be examined were fed on 30% wheat or barley mixed with rice diets for 8 weeks. The total nitrogen, creatinine, amino acid nitrogen and urea-nitrogen contents in the liver and the creatinine and urea-nitrogen contents in the urine have been measured. The results obtained are summarized as follows: 1. The total nitrogen contents in the liver and the serum were no remarkable difference by feeding on each mixed diet, compared with the rice diet group. 2. The creatinine contents in the liver of the unpolished wheat and barley mixed diet groups were the similar to that of the rice diet group, but these were higher by feeding on the polished wheat and barley mixed with rice diets. 3. The amino acid nitrogen contents in the liver of the polished naked barley mixed with rice diet groups were the similar to that of the rice diet group, but these were higher by feeding on the other mixed diets than the rice diet. 4. The urea-nitrogen contents in the serum of the polished wheat and naked barley mixed with rice diet groups were higher than that of the rice diet group, but these were significantly lower by feeding on the polished barley mixed with rice diets than the others. 5. The creatinine and the urea-nitrogen contents in the urine of the original wheat and barley mixed with rice diet groups were higher than that of the polished wheat and barley mixed with rice diet groups. In the view of the above results, it could be seen that the protein metabolism was remarkable change according to polish of the wheat and barley.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1273-1273
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• 2001

The efficiency of the luminal fermentation process influences overall efficiency of luminal production, animal health and reproduction. Ruminant production systems have a significant impact on the global environment, as well. Animal wastes contribute to pollution of the environment as ammonia volatilized to the air and nitrate leached to ground water. Microbial protein synthesis in the rumen satisfies a large proportion of the protein requirements of animals. Quantifying the microbial synthesis is possible by using markers for lumen bacteria and protozoa such as nucleic acids, purine bases, some specific amino acids, or by isotopic $^{15}N,^{32}P,\;and\;^{35}S$ labelled feeds. All those methods require cannulated animals, they are time-consuming and some methods are very expensive as well. Many attempts have been made to find an alternative method for indirect measurement of microbial synthesis in intact animals. The present investigations aimed to assess possibilities of NIRS for prediction of purine nitrogen excretion and ruminal microbial nitrogen synthesis by NIR spectra of urine. Urine samples were collected from 12 growing sheep,6 of them male, and 6- female. The sheep were included in feeding experiment. The ration consisted of sorghum silage and protein supplements -70:30 on dry matter basis. The protein supplements were chosen to differ in protein degradability. The urine samples were collected daily in a vessel containing $60m{\ell}$ 10% sulphuric acid to reduce pH below 3 and diluted with tap water to 4 liters. Samples were stored in plastic bottles and frozen at $-20^{\circ}C$ until chemical and NIRS analysis. The urine samples were analyzed for purine derivates - allantoin, uric acid, xantine and hypoxantine content. Microbial nitrogen synthesis in the lumen was calculated according to Chen and Gomes, 1995. Transmittance urine spectra with sample thickness 1mm were obtained by NIR System 6500 spectrophotometer in the spectral range 1100-2500nm. The calibration was performed using ISI software and PLS regression, respectively. The following statistical results of NIRS calibration for prediction of purine derivatives and microbial protein synthesis were obtained.(Table Omitted). The result of estimation of purine nitrogen excretion and microbial protein synthesis by NIR spectra of urine showed accuracy, adequate for rapid evaluation of microbial protein synthesis for a large number of animals and different diets. The results indicate that the advantages of the NIRS technology can be extended into animal physiological studies. The fast and low cost NIRS analyses could be used with no significant loss of accuracy when microbial protein synthesis in the lumen and the microbial protein flow in the duodenum are to be assessed by NIRS.

• Bulletin of the Korean Chemical Society
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• v.34 no.12
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• pp.3817-3824
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• 2013

Various separation modes in HPLC, such as anion exchange (AE), reversed-phase (RP), and affinity (AF) chromatography were examined for the separation of selenium species in human blood serum and urine. While RP- and AE-HPLC were mainly used for the separation of small molecular selenium species, double column AF-HPLC achieved the separation of selenoproteins in blood serum efficiently. Further, the effluent of AF-HPLC was enzymatically hydrolyzed and then analyzed with RP HPLC for selenoamino acid study. The versatility of the hybrid technique makes the in-depth study of selenium species possible. For quantification, post column isotope dilution (ID) with $^{78}Se$ spike was performed. ORC ICP/MS (octapole reaction cell inductively coupled plasma/mass spectrometry) was used with 4 mL $min^{-1}$ Hydrogen as reaction gas. In urine sample, inorganic selenium and SeCys were identified. In blood serum, selenoproteins GPx, SelP and SeAlb were detected and quantified. The concentration for GPx, SelP and SeAlb was $22.8{\pm}3.4\;ng\;g^{-1}$, $45.2{\pm}1.7\;ng\;g^{-1}$, and $16.1{\pm}2.2\;ng\;g^{-1}$, respectively when $^{80}Se/^{78}Se$ was used. The sum of these selenoproteins ($84.1{\pm}4.4\;ng\;g^{-1}$) agrees well with the total selenium concentration measured with the ID method of $87.0{\pm}3.0\;ng\;g^{-1}$. Enzymatic hydrolysis of each selenium proteins revealed that SeCys is the major amino acid for all three proteins and SeMet is contained in SeAlb only.

• Journal of Practical Agriculture & Fisheries Research
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• v.8 no.1
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• pp.43-51
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• 2006

An experiment was conducted to examine how the response of dairy cows to a change from twice to thrice-daily milking is affected by deficiencies in the dietary supplies of three amino acids, His, Met and Lys. The amino acid deficiencies were obtained by feeding the cows a diet of grass silage and a cereal-based supplement containing feather meal as the sole protein supplement. Taken overall, the results show that when cows were given the feather meal diet, even though dietary ME was in considerable excess, a deficiency of specific amino acids prevented any increase in milk yield in response to increasing the frequency of milking from twice to thrice daily. The results of half-udder milking showed that when cows consumed the diet deficient in amino acids, milking one half of the gland more frequently reduced the secretion of protein and lactose by the control gland. Neither MBF nor the ratio of 3-MH/creatinine in urine was affected by thrice-daily milking. The present results go a stage further in showing that, against a background of insufficient dietary amino acids, the stimulus of thrice-daily milking is not sufficient to induce a measurable change in the partition of amino acid use between body and udder.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.4 no.2
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• pp.249-255
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• 2001

Methylmalonic acidemia is a rare congenital autosomal recessive metabolic disease. It is caused by blocking in the pathways of isoleucine, valine, threonine, methionine, cholesterol and odd-chain fatty acids to succinyl CoA, resulting in the increase of L-methylmalonyl CoA and methylmalonic acid. In most cases, there are symptoms such as recurrent vomitings, lethargy and laboratory abnormalities including metabolic acidosis and hyperammonemia from the neonatal period. We had a 6-month-old infant with methylmalonyl acidemia who presented with recurrent vomiting episodes since 3 months of age, failure to thrive and developmental delay. The laboratory findings showed hyperammoninemia and ketotic metabolic acidosis. Plasma amino acid analysis showed nonspecific finding. Urine organic acid ananysis by gas chromatography and mass spectrometry detected large amount of methylmalonic acid excreted in the urine. We restrained the supply of protein in the amount of 1~1.5 g/kg of body weight a day using leucine, isoleucine and valine-r-estrained milk and administered vitamine $B_{12}$, in the amount of 1mg per day. During the follow-up in the outpatient clinic, He could control his head and showed increased muscle strength.

• Clinical and Experimental Pediatrics
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• v.53 no.3
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• pp.329-334
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• 2010

Purpose : The neonatal screening test for homocystinuria primarily measures methionine by using a dried blood specimen. We investigated the incidence and clinical manifestations of homocystinuria, isolated hypermethioninemia, and transient hypermethioninemia among patients with hypermethioninemia on a neonatal screening test. Methods : We performed a retrospective study of 58 patients transferred to Shoonchunhyang Hospital because of hypermethioninemia on a neonatal screening test between January 1996 and August 2009. We analyzed the level of amino acid from plasma and urine, as well as blood homocysteine.Results : Almost half of the 58 patients were identified as normal. Whereas only 3 (5.1%) patients were identified as having homocystinuria, about 20.7% (12 cases) of the patients had isolated hypermethioninemia. The ages of these two groups at initial detection of hypermethioninemia on plasma amino acid analysis were $50.0{\pm}22.5$ days and $34.9{\pm}13.5$ days, respectively. Both groups were put on diets, and they showed a normal developmental course as a result of early diagnosis and treatment. Conclusion : Hypermethioninemia without homocystinuria, referred to as isolated hypermethioninemia, was also detected. Thus, the impact of hypermethioninemia on a neonatal screening test should be carefully evaluated through analysis of amino acid levels from blood and urine, and we need to detect and treat an early stage of isolated hypermethioninemia as well as homocystinuria.

• Journal of Nutrition and Health
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• v.7 no.2
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• pp.37-51
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• 1974

This Study was carried out to observe the effect of nutritional condition on the change of protein metabolism in the animal body by feeding on imbalanced protein diet. A total 242 growing male albino rats, weighing $115{\sim}120$ gm, were used for the experimental animals. The rats were fed on the standard diet(st), protein flee diet(pf) and imbalanced protein diet(ib) for twelve weeks respectively. Hemoglobin, packed cell volume in blood, and total nitrogen, amino acid nitrogen, urea-nitrogen, creatinine, transaminases(GPT, GOT) in liver and serum, and total nitrogen in small intestine, and total nitrogen, urea-nitrogen In small intestine, and total nitrogen, urea-nitrogen, creatinine, urea-nitrogen/creatinine ratio in urine were measured. The results obtained are as follows; 1. The gained body weight were lower in pf group and ib group than those of st group. The gained body weight fed for 12 weeks, were 80% lower in pf group than those of st group, and the body weight of pf group for $50{\sim}75$ days feeding were $40{\sim}60%$ decreased, compared with the stating weight, and then all of them died. 2. The change of the brain, liver, kidney, spleen and small intestine by feeding on imbalanced diet for 12 weeks were no remarkable difference with the starting weight, but those of protein free diet group were half or more decrease and those were significantly lower in spleen and small intestine especially than the other organ 3. The contents of hemoglobin in pf group for 8 weeks feeding, and the packed cell volume in pf group for 8 weeks feeding and in ib group for 12 weeks feeding were decreased. but those of the other feeding group were almost same value. 4. The total nitrogen in the liver, small intestine and serum of each diet group were no remarkable difference respectively. The contents of amino acid nitrogen in pf group for 2 and 6 weeks feeding were increased. 5. On transaminases: a) The cycle of increase and decrease of GPT activities were come periodically and the interval of cycle were fast in the early stage of feeding and slow there-after. b) The GPT activities were decreased gradually in pf group after feeding and those were increased in ib group for 6 weeks feeding but decreased there-after. The frequency of cycle were more GPT than GOT and specially those of GPT in early stage of feeding were two or three times while GOT was one. c) The interval of increase and decrease in GOT and amino acid nitrogen cycle were similar tendency. 6. The contents of total nitrogen, creatinine and urea-nitrogen of pf group in urine were decreased very sharply from sharting feeding to one week but increased dully from six weeks to eight weeks feeding. The contents of urea-nitrogen of ib group were increased dully by feeding on ten weeks but decreased by feeding on twelve weeks. From the above results, it is concluded that the trend of the metabolic change is maintained equally by homeostatic mechanism using the endogenous protein source during a certain period by imbalanced protein diet feeding. The homeostatic mechanism is come peridically, very fast in early stage of feeing and than slow there-after.