Academic literature on the topic 'Urine amino acids'
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Journal articles on the topic "Urine amino acids"
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Harada, Masashi, Sachise Karakawa, Hiroshi Miyano, and Kazutaka Shimbo. "Simultaneous Analysis of d,l-Amino Acids in Human Urine Using a Chirality-Switchable Biaryl Axial Tag and Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry." Symmetry 12, no. 6 (June 2, 2020): 913. http://dx.doi.org/10.3390/sym12060913.
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Although d,l-amino acids are symmetrical molecules, l-isomers are generally dominant in living organisms. However, it has been found that some d-amino acids also have biological functions. A new method for simultaneously analyzing d,l-amino acids in biological samples is required to allow unknown functions of d-amino acids to be investigated. d-Amino acids in urine are currently receiving increasing amounts of attention, particularly for screening for chronic kidney diseases. However, simultaneously analyzing d,l-amino acids in human urine is challenging because of interfering unknown compounds in urine. In this study, the axially chiral derivatizing agent (R)-4-nitrophenyl-N-[2-(diethylamino)-6,6-dimethyl-[1,1-biphenyl]-2-yl] carbamate hydrochloride was used to allow enantiomers of amino acids in human urine to be simultaneously determined by liquid chromatography electrospray ionization tandem mass spectrometry. The optimized method gave good linearities, precision results, and recoveries for 18 proteinogenic amino acids and their enantiomers and glycine. The chiral-switching method using (S)-4-nitrophenyl-N-[2-(diethylamino)-6, 6-dimethyl-[1,1-biphenyl]-2-yl]carbamate hydrochloride confirmed the expected concentrations of 32 of the 37 analytes. The method was successfully used to determine the concentrations of d-serine, d-alanine, d-asparagine, d-allothreonine, d-lysine, and the d-isomers of 10 other amino acids in five human volunteer urine samples.
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Nagata, Y., R. Konno, Y. Yasumura, and T. Akino. "Involvement of d-amino acid oxidase in elimination of free d-amino acids in mice." Biochemical Journal 257, no. 1 (January 1, 1989): 291–92. http://dx.doi.org/10.1042/bj2570291.
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The physiological role of D-amino acid oxidase was investigated by using mutant ddY/DAO- mice lacking the enzyme. Free D-amino acid concentrations in the mutant mice were significantly higher than those of control ddY/DAO+ mice in kidney, liver, lung, heart, brain, erythrocytes, serum and urine. The results suggest that the enzyme is involved in the catabolism of free D-amino acids in the body, and that free D-amino acids are also excreted into urine.
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Hortin, Glen L., and Bonnie Meilinger. "Cross-Reactivity of Amino Acids and Other Compounds in the Biuret Reaction: Interference with Urinary Peptide Measurements." Clinical Chemistry 51, no. 8 (August 1, 2005): 1411–19. http://dx.doi.org/10.1373/clinchem.2005.052019.
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Abstract Background: Biuret assays for total protein measurement are considered to react with all peptides longer than 2 residues. Some studies using biuret assays of urine suggest that small peptides generally are more abundant than proteins in urine, but it is not clear whether this is a problem of assay specificity. Methods: We analyzed the specificity and kinetics of a biuret reaction for solutions of amino acids, organic compounds, peptides, proteins, and ultrafiltered urine specimens and compared the results with standard clinical assays for protein measurement. Results: The biuret assay cross-reacted with several amino acids, dipeptides, and other organic compounds able to form 5- or 6-member ring chelation complexes with copper. Reactions with amino acids and dipeptides had higher absorbance maxima (blue color) than with larger peptides and proteins (purple). Compounds forming potential 4-, 7-, 8-, or 9-member ring complexes with copper had low reactivity. Amino acid amides, dipeptides, and longer peptides had substantial reactivity, except those containing proline. Proteins and polypeptides had similar biuret reactivities per peptide bond, but reaction kinetics were slower for proteins than peptides. Urine specimens ultrafiltered through 3-kDa–cutoff membranes had substantial biuret reactivity, but absorbance maxima were consistent with cross-reactive amino acids rather than peptides. Conclusions: Many compounds, including amino acids, amino acid derivatives, and dipeptides, cross-react in biuret assays. Our studies improve understanding of the specificity of endpoint and kinetic biuret assays widely used in clinical laboratories. Amino acids, urea, and creatinine contribute to overestimation of urinary peptide content by biuret assays.
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Leeuwenburgh, Christiaan, Polly A. Hansen, John O. Holloszy, and Jay W. Heinecke. "Oxidized amino acids in the urine of aging rats: potential markers for assessing oxidative stress in vivo." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 276, no. 1 (January 1, 1999): R128—R135. http://dx.doi.org/10.1152/ajpregu.1999.276.1.r128.
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Oxidative damage of proteins has been implicated in disease and aging. In vitro studies demonstrate that two unnatural amino acids, o,o′-dityrosine and o-tyrosine, are stable markers of protein oxidation. We have investigated the possibility that assaying these compounds in urine could provide a noninvasive way to determine levels of protein oxidation in vivo. Isotope dilution gas chromatography-mass spectrometry was used to quantify levels of o,o′-dityrosine and o-tyrosine in skeletal muscle and urine of aging rats subjected to two interventions: 1) dietary antioxidant supplementation and 2) exercise training. In both sedentary rats and exercise-trained rats, antioxidant therapy reduced levels of protein-bound o,o′-dityrosine in skeletal muscle. In contrast, antioxidant therapy or exercise training minimally affected o-tyrosine levels in this tissue. Levels of the oxidized amino acids in urine samples mirrored those of skeletal muscle proteins. Quantification of the levels of oxidized amino acids in urine may thus serve as a noninvasive measure of oxidative stress in vivo because they change in parallel with levels of protein-bound oxidized amino acids in skeletal muscle.
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Drotningsvik, Aslaug, Øivind Midttun, Linn Anja Vikøren, Adrian McCann, Per Magne Ueland, Gunnar Mellgren, and Oddrun Anita Gudbrandsen. "Urine and plasma concentrations of amino acids and plasma vitamin status differ, and are differently affected by salmon intake, in obese Zucker fa/fa rats with impaired kidney function and in Long-Evans rats with healthy kidneys." British Journal of Nutrition 122, no. 03 (August 2019): 262–73. http://dx.doi.org/10.1017/s0007114519001284.
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AbstractKidney function affects amino acid metabolism and vitamin status. The aims of the present study were to investigate urine and plasma concentrations of amino acids as well as plasma vitamin status in rats with impaired renal function (Zucker fa/fa rats) and in rats with normal kidney function (Long-Evans rats), and to explore the effects of salmon intake on these parameters and potential biomarkers of salmon intake in both rat strains. Male rats were fed diets with casein as sole protein source (control diet) or 25 % protein from baked salmon and 75 % casein for 4 weeks. Urine concentrations of markers of renal function and most amino acids and plasma concentrations of most vitamins were higher, and plasma concentrations of several amino acids including arginine, total glutathione and most tryptophan metabolites were lower in Zucker fa/fa rats compared with Long-Evans rats fed the control diet. Concentrations of kidney function markers were lower after salmon intake only in Zucker fa/fa rats. A trend towards lower urine concentrations of amino acids was seen in both rat strains fed the salmon diet, but this was more pronounced in Long-Evans rats and did not reflect the dietary amino acid content. Urine 1-methylhistidine, 3-methylhistidine, trimethylamineoxide and creatine concentrations, and plasma 1-methylhistidine and creatine concentrations were higher after salmon intake in both rat strains. To conclude, concentrations of amino acids in urine and plasma as well as vitamin status were different in Zucker fa/fa and Long-Evans rats, and the effects of salmon intake differed by rat strain for some of these parameters.
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Venta, Rafael. "Year-Long Validation Study and Reference Values for Urinary Amino Acids Using a Reversed-Phase HPLC Method." Clinical Chemistry 47, no. 3 (March 1, 2001): 575–83. http://dx.doi.org/10.1093/clinchem/47.3.575.
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Abstract Background: Reversed-phase HPLC (RP-HPLC) has become an alternative to ion-exchange chromatography for amino acid analysis in biological fluids. However, validation studies for its urine application are limited, and the corresponding reference values have not been reported extensively. We studied the long-term performance of a commercial HPLC method for urine amino acid analysis and established specific age-related reference values for urine amino acid excretion. Methods: Method performance was continuously assessed by recovery and precision studies with urine samples and controls, respectively. Healthy individuals were prospectively analyzed throughout a 5-year period. Excretion of individual amino acids, expressed as mmol/mol of creatinine, was included in six age-related groups for random urine samples (0–1 month, 1–12 months, 1–3 years, 3–8 years, 8–16 years, and >16 years) and in two groups for 24-h urine collections (8–16 years and >16 years). Results: Over a 1-year period, CVs for retention times were <0.5% and 3.3% for within- and between-run imprecision, respectively. For amino acid concentrations, within-run CVs were 2.9–17% and between-run CVs were 7.1–46% for the same period. Amino acid recoveries were 78–122%. Reference intervals for 35 amino acids were calculated and compared with the concentrations observed in patients diagnosed with specific pathologies. A few statistically significant differences were found between the reference intervals derived using random and 24-h urine collections. Conclusions: Long-term reliability of the RP-HPLC method for urine amino acid analysis has been demonstrated. Representative age-related reference intervals for the RP-HPLC method in both random urine and 24-h urine collections have been established, and their feasibility for diagnosis of aminoaciduria has been shown. These intervals could serve as a guide for laboratories changing to HPLC methods.
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Fan, Jing, Jing Hong, Jun-Duo Hu, and Jin-Lian Chen. "Ion Chromatography Based Urine Amino Acid Profiling Applied for Diagnosis of Gastric Cancer." Gastroenterology Research and Practice 2012 (2012): 1–8. http://dx.doi.org/10.1155/2012/474907.
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Aim. Amino acid metabolism in cancer patients differs from that in healthy people. In the study, we performed urine-free amino acid profile of gastric cancer at different stages and health subjects to explore potential biomarkers for diagnosing or screening gastric cancer.Methods. Forty three urine samples were collected from inpatients and healthy adults who were divided into 4 groups. Healthy adults were in group A (n=15), early gastric cancer inpatients in group B (n=7), and advanced gastric cancer inpatients in group C (n=16); in addition, two healthy adults and three advanced gastric cancer inpatients were in group D (n=5) to test models. We performed urine amino acids profile of each group by applying ion chromatography (IC) technique and analyzed urine amino acids according to chromatogram of amino acids standard solution. The data we obtained were processed with statistical analysis. A diagnostic model was constructed to discriminate gastric cancer from healthy individuals and another diagnostic model for clinical staging by principal component analysis. Differentiation performance was validated by the area under the curve (AUC) of receiver-operating characteristic (ROC) curves.Results. The urine-free amino acid profile of gastric cancer patients changed to a certain degree compared with that of healthy adults. Compared with healthy adult group, the levels of valine, isoleucine, and leucine increased (P<0.05), but the levels of histidine and methionine decreased (P<0.05), and aspartate decreased significantly (P<0.01). The urine amino acid profile was also different between early and advanced gastric cancer groups. Compared with early gastric cancer, the levels of isoleucine and valine decreased in advanced gastric cancer (P<0.05). A diagnosis model constructed for gastric cancer with AUC value of 0.936 tested by group D showed that 4 samples could coincide with it. Another diagnosis model for clinical staging with an AUC value of 0.902 tested by 3 advanced gastric cancer inpatients of group D showed that all could coincide with the model.Conclusions. The noticeable differences of urine-free amino acid profiles between gastric cancer patients and healthy adults indicate that such amino acids as valine, isoleucine, leucine, methionine, histidine and aspartate are important metabolites in cell multiplication and gene expression during tumor growth and metastatic process. The study suggests that urine-free amino acid profiling is of potential value for screening or diagnosing gastric cancer.
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Kohri, K., M. Takada, Y. Katoh, K. Kataoka, M. Iguchi, and T. Kurita. "Amino acids in urine and plasma of urolithiasis patients." International Urology and Nephrology 21, no. 1 (January 1989): 9–16. http://dx.doi.org/10.1007/bf02549896.
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Schneider, K., M. Neupert, G. Spitler, H. V. Henning, D. Matthaei, and F. Scheller. "Gas chromatography of amino acids in urine and haemofiltrate." Journal of Chromatography B: Biomedical Sciences and Applications 345 (January 1985): 19–31. http://dx.doi.org/10.1016/0378-4347(85)80131-6.
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Moodie, I. M., B. J. Hough, and D. Labadarios. "Determination of amino acids in urine by gas chromatography." Journal of High Resolution Chromatography 12, no. 7 (June 1989): 437–41. http://dx.doi.org/10.1002/jhrc.1240120703.
Full textDissertations / Theses on the topic "Urine amino acids"
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James, M. D. "Analyses of amino acids in human urine." Thesis, Swansea University, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.637390.
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Analyses of amino levels in human urine were studied in order to evaluate their potential use in tests which could be used for the early detection of cancer. Three suitable reagents for derivatisation were studied and it was found that of the three reagents considered for use in this study, {dansyl chloride, dabsyl chloride and OPA}, dansyl chloride proved the most suitable as it was the most sensitive, enabled accurate reproducible separation of a greater number of amino acids than the other reagents and was the easiest of the three with which to work. A control group of 27 healthy individuals, were provided with five sample bottles in order for them to provide a number of separate samples taken throughout the day. The data that were obtained from these sample were used to establish the amino acid levels in the control group of healthy individuals. The results also demonstrate that the daily variation in amino acid levels in a healthy individual was not responsible for the difference between individuals. In the Discussion and Summary it can be seen that the pattern of concentrations of individual amino acids obtained is different for each form of cancer. Each pattern adds further evidence for the presence of a particular form of tumour. The final conclusion is that the lower average range of the amino acid alanine in cancer patients is statistically significant, it is therefore possible to use the depressed average level of alanine along with raised average level of isoleucine in data obtained from a number of samples taken from any individual patient over a period of 2-3 days in comparison with the average levels in the control group of healthy volunteers to assess a patient for the presence of any form of cancerous tumour within that individual.
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Vitor, Aline de Paula. "Determinação de aminoácidos por eletroforese capilar com detecção UV/vis para o estudo do perfil metabólico urinário do refluxo vésico-ureteral." Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/46/46136/tde-26042013-110017/.
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Uma avaliação da concentração dos aminoácidos primários em amostras de urina de crianças com refluxo vésico-ureteral (VUR) em busca de caminhos para o diagnóstico não invasivo desta doença. Dois métodos analíticos por eletroforese capilar com detecção UV/vis foram desenvolvidos para a quantificação dos analitos. No método 1 empregou-se a detecção UV/vis direta em 200 e 214 nm com as condições eletroforéticas eletrólito tampão fosfato 90 mmol L-1 pH 2,1; tensão de +15 kV; injeção de 7 s a 0,5 psi; capilar de 75 µm de diâmetro interno; 40,2 cm de comprimento total e 30,0 cm de comprimento efetivo. No método 2, fez-se uso da detecção indireta em 254 nm, com as condições eletroforéticas eletrólito tampão TEA 20 mmol L-1 e DNB 10 mmol L-1 pH 10,84; modificador de fluxo DDAB a 4 mmol L-1; tensão de -15 kV; injeção de 7 s a 0,5 psi; capilar de 75 µm de diâmetro interno; 50,2 cm de comprimento total e 40,0 cm de comprimento efetivo. O método 1 apresentou parâmetros de validação linearidade, precisão intra-dia e inter-dia, seletividade, robustez e recuperação satisfatórios. A quantificação de creatinina, fenilalanina (Phe), histidina (His), triptofano (Trp), tirosina (Tyr) nas amostras de urina foi possível pelo método 1, porém inviável para quantificação de arginina (Arg). O método 2 apresentou valores de robustez e recuperação satisfatórios para os aminoácidos alanina (Ala), aspartato (Asp), glutamato (Glu) e glicina (Gly) satisfatórios, mas a quantificação dos mesmos na maioria das amostras de urina diluída não foi possível por estarem em nível de concentração abaixo da detecção ou quantificação. Para avaliar a potencialidade dos resultados como ferramenta no diagnóstico do VUR, os aminoácidos His, Phe, Trp e Tyr, quantificados em todas as amostras, foram empregados como variáveis na classificação das amostras em dois grupos distintos (1) grupo de crianças saudáveis e (2) grupo de crianças diagnosticadas com VUR. A classificação realizada pelo método de análise de componente principal (PCA) apresentou valores estatísticos satisfatórios e poder de predição: R2 (capacidade de ajuste) e Q2 (capacidade de predição) foram 0.9993 e 0.65, respectivamente com os dois componentes principais (PC1 e PC2). A separação total com valor de Q2 desejável (acima de 0,8) poderia ser alcançada com uma quantidade maior de informação, sendo neste caso, número maior de aminoácidos quantificados. Assim, este trabalho abre caminho para estudos mais aprofundados na investigação da concentração dos aminoácidos primários em pacientes com VUR, objetivando o desenvolvimento de um potencial biomarcador para VUR.An assessment of the concentration of primary amino acids in urine samples from children with vesicoureteral reflux (VUR) using capillary electrophoresis separation with UV/vis detection has been proposed to help establishing a means for non invasive diagnosis of the disease. Two analytical methods were developed. Method 1 used direct UV/vis detection at 200 and 214 nm, 90 mmol L-1 phosphate buffer at pH 2.1, high voltage separation at +15 kV, injection of 0.5 psi during 7 s, and a fused-silica capillary of 75 µm inner diameter, 40.2 cm total length, and 30.0 cm effective length. Method 2 used indirect UV/vis detection at 254 nm, TEA at 20 mmol L-1 and DNB at 10 mmol L-1 electrolyte at pH 10.84, 4 mmol L-1 DDAB as flow modifier, separation voltage at -15 kV, injection of 0,5 psi during 7 s, fused-silica capillary of 75 µm inner diameter, 50.2 cm total length, and 40,0 cm effective length. Method 1 presented satisfactory results for linearity, intra-day and inter-day precision, selectivity, robustness, and recovery. By method 1 it was possible to quantify creatinine, phenylalanine (Phe), histidine (His), tryptophan (Trp), tyrosine (Tyr) but not arginine (Arg) in the urine samples under investigation. Method 2 presented satisfactory robustness and recovery for alanine (Ala), aspartate (Asp), glutamate (Glu) and glycine (Gly), but the contents of these metabolites in the urine samples were not established because they lay below the limits of detection and quantitation. To assess the potentiality of the results as diagnostic tool for VUR condition, the concentrations of the amino acids His, Phe, Trp and Tyr, quantified in all samples, were used as variables in a classification procedure where samples were divided in two distinct groups: (a) a group of healthy children and (b) a group of children diagnosed with VUR. The classification by principal component analysis (PCA) showed a partial separation with good statistics and prediction power: R2 (goodness of fit) and Q2 (goodness of prediction) were 0.9993 and 0.65, respectively with two components analysis (PC1 and PC2). Values of Q2 greater than 0.8 are usually desired and it could be provided if more information was available, such as a greater number of amino acids being quantified. Thus, this research opens the way for further investigative studies of amino acids concentration in patients with primary VUR, aimed at developing a potential biomarker for VUR.
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Harnevik, Lotta. "Molecular genetic studies on cystinuria." Doctoral thesis, Linköping : Univ, 2007. http://www.bibl.liu.se/liupubl/disp/disp2007/med1034s.pdf.
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McGregor, Neil Roland. "An investigation of the association between toxin producing staphylococcus, biochemical changes and jaw muscle pain." University of Sydney. Prosthetic Dentistry, 2000. http://hdl.handle.net/2123/369.
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Objectives: To assess the expression of the symptoms of jaw muscle pain and its association with alterations in biochemistry, other symptoms and the carriage of staphylococci. Methods: Three different study populations were assessed. The first was selected and examined by the author and consisted of 43 pain and 41 age and sex matched controls. The second was a study of CFS patients who were blinded to the author and the author subsequently examined the associations between jaw muscle symptom reporting and the standardised biochemistry measures. The third study was also blinded to the author but included an investigation of staphylococci and certain cytokine and biochemistry measures. Results: The three studies clearly establish an association between the carriage of toxicogenic coagulase negative staphylococci and the expression of jaw muscle pain in both males and females. These associations were homogeneous and were found whether the patients were selected on the basis of having jaw muscle pain or selected from within a population of patients selected on the basis of having Chronic Fatigue Syndrome. The studies associated the changes with variations in biochemistry and these were in turn associated with symptom expression within the jaw muscle pain patients. These biochemical alterations included the dysregulation of immune cell counts, cytokines, electrolyte and protein metabolism. These symptoms and biochemical changes were associated with pain severity and illness duration and staphylococcal toxin production. From the data a model was developed which shows the mechanisms involved in the development of chronic pain in the jaw muscles. Conclusions: The carriage of toxicogenic coagulase-negative staphylococci were found to be associated with the expression of jaw muscle pain and the alterations in biochemistry associated with these symptoms.
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McGregor, Neil Roland. "An investigation of the association between toxin producing staphylococcus, biochemical changes and jaw muscle pain." Thesis, The University of Sydney, 1999. http://hdl.handle.net/2123/369.
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Objectives: To assess the expression of the symptoms of jaw muscle pain and its association with alterations in biochemistry, other symptoms and the carriage of staphylococci. Methods: Three different study populations were assessed. The first was selected and examined by the author and consisted of 43 pain and 41 age and sex matched controls. The second was a study of CFS patients who were blinded to the author and the author subsequently examined the associations between jaw muscle symptom reporting and the standardised biochemistry measures. The third study was also blinded to the author but included an investigation of staphylococci and certain cytokine and biochemistry measures. Results: The three studies clearly establish an association between the carriage of toxicogenic coagulase negative staphylococci and the expression of jaw muscle pain in both males and females. These associations were homogeneous and were found whether the patients were selected on the basis of having jaw muscle pain or selected from within a population of patients selected on the basis of having Chronic Fatigue Syndrome. The studies associated the changes with variations in biochemistry and these were in turn associated with symptom expression within the jaw muscle pain patients. These biochemical alterations included the dysregulation of immune cell counts, cytokines, electrolyte and protein metabolism. These symptoms and biochemical changes were associated with pain severity and illness duration and staphylococcal toxin production. From the data a model was developed which shows the mechanisms involved in the development of chronic pain in the jaw muscles. Conclusions: The carriage of toxicogenic coagulase-negative staphylococci were found to be associated with the expression of jaw muscle pain and the alterations in biochemistry associated with these symptoms.
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Rodrigues, Karina Trevisan. "Investigação do refluxo vésico-ureteral por abordagens metabolômicas alvo e global em urina utilizando como plataformas analíticas CE-MS, CESI-MS, RPLC-MS e HILIC-MS." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/46/46136/tde-14122017-133339/.
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O refluxo vésico-ureteral (RVU) é uma das condições urológicas comumente diagnosticada entre crianças. Altos graus dessa condição podem causar cicatrização renal, insuficiência renal e hipertensão arterial. A uretrocistografia miccional é o método mais comumente utilizado para o diagnóstico, no entanto, esse procedimento envolve sedação, cateterismo vesical e expõe a criança a uma quantidade significativa de radiação. A investigação metabolômica pode fornecer novos entendimentos sobre a doença e visa a descoberta de metabólitos específicos associados a ela. Assim, existe um potencial considerável para a implementação de perfil metabólico em análises clínicas. Dessa forma, buscou-se estabelecer uma alternativa não invasiva para identificar crianças com o RVU através da metabolômica. Para a investigação metabolômica alvo um método por eletroforese capilar acoplada ao espectrômetro de massas (CE-MS) com analisador to tipo ion trap foi desenvolvido e validado para a determinação de 27 aminoácidos em urina. Os parâmetros experimentais relacionados às configurações da interface CE-MS, eletrólito (BGE) e espectrômetro de de massas (MS) foram otimizados, proporcionando uma boa separação dos 27 aminoácidos, incluindo os isômeros L-leucina, L-isoleucina e L-alloisoleucina, em menos de 30 min. O líquido auxiliar (SHL) foi composto de 0,5% (v/v) ácido fórmico em 60% (v/v) água/metanol à uma vazão de 5 µL min-1. O BGE consistiu de 0,80 mol L-1 de ácido fórmico e 15% (v/v) de metanol. Um procedimento de stacking por pH foi implementado para aumentar a detectabilidade (uma solução de NH4OH a 12,5% (v/v) foi injetada a 0,5 psi/9 s antes das amostras). O método foi validado de acordo com os protocolos FDA e ICH, exibindo parâmetros aceitáveis. A quantificação bem sucedida dos aminoácidos em amostras de urina de um estudo piloto do RVU foi alcançada. A avaliação estatística dos resultados mostrou que alguns dos aminoácidos avaliados podem carregar informações que possibilitam discriminar as amostras de urina entre os grupos teste e controle. Para a análise metabolômica global urinária, métodos por RPLC-MS e HILIC-MS foram otimizados. Cinco colunas com diferentes propriedades foram investigadas para RPLC e quatro colunas para HILIC; adicionalmente, foram investigados a influência dos aditivos e pH da fase móvel. As condições ótimas foram determinadas avaliando o formato de pico, a relação sinal-ruído, o tempo de retenção, o número de molecular features detectados e sua distribuição durante o gradiente de eluição. A melhor condição obtida para RPLC utiliza a coluna CSH C18 e fase móvel composta por 0,1% (v/v) ácido fórmico em água (A) e 0,1% (v/v) ácido fórmico em acetonitrila (B). Para HILIC, o melhor desempenho foi obtido com a coluna zwitteriônica ZIC-HILIC e fase móvel composta por 10 mmol L-1 acetato de amônio pH 6,8 (B) e 95% (v/v) acetonitrila e 5% 200 mmol L-1 acetato de amônio pH 6,8 (A). As amostras de urina dos grupos controle e teste foram submetidas à análise metabolômica global por RPLC-MS usando o método otimizado e por CESI-MS. Os resultados indicaram que diversas rotas metabólicas podem ter sido alteradas pelo RVU. Alteração dos níveis de carnitinas e acilcarnitinas, aminoácidos e derivados, purinas e outros foi observada. Ainda, a presença de acilcarnitinas na urina podem indicar danos mitocondriais e a diminuição de triptofano e aumento do ácido quinurênico indicam uma alteração no metabolismo do triptofano.Vesicoureteral reflux (VUR) is one of the most commonly urologic conditions diagnosed among children. A high degree of this condition can cause kidney scarring, kidney failure and high blood pressure. Voiding cystourethrography is the standard method for diagnosis; however, this procedure involves sedation, bladder catheterization and exposes the child to a significant amount of radiation. Metabolomics has provided new insights about the disease and aims to discover specific metabolites associated with it. Thus, there is a considerable potential for the implementation of metabolic profile in clinical analyses. Thus, we attempted to establish a noninvasive alternative to identify children with VUR through metabolomics approach. For target metabolomics, a CE-MS method was developed and validated for the separation and quantitative analysis of 27 amino acids in urine. Experimental parameters related to the CE-MS interface (based on co-axial sheath liquid, SHL), background electrolyte (BGE) and mass spectrometer (MS) settings were optimized providing a good separation of 27 amino acids, including the isomers L-leucine, L-isoleucine and L-alloisoleucine, in less than 30 min. The SHL was composed of 0.50% (v/v) formic acid in 60% (v/v) methanol-water delivered at a flow rate of 5 µL min-1. The BGE consisted of 0.80 mol L-1 formic acid and 15% (v/v) methanol. A pH stacking procedure was implemented to enhance sensitivity (a 12.5% (v/v) NH4OH solution was injected at 0.5 psi/9 s prior to samples). The proposed method was thoroughly validated according to FDA and ICH protocols exhibiting acceptable parameters. A successful quantification of amino acids in urine samples from the VUR cohort was achieved. The statistical evaluation of the results showed that some of the amino acids may carry information for the discrimination of the urine samples between the test and control groups. For untargeted metabolomics analysis, methods by RPLC-MS and HILIC-MS were optimized. Five columns with different properties were investigated for RPLC and four columns for HILIC; additionally, the influence of additives and pH of the mobile phase were investigated. The optimum conditions were determined assessing the peak shape, signal-to-noise ratio, retention time, number of molecular features detected and their distribution during the elution gradient. The best condition obtained for RPLC uses CSH C18 column and mobile phase composed by 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B). For HILIC, the best performance was obtained with the zwitterionic ZIC-HILIC column and mobile phase composed by 10 mmol L-1 ammonium acetate pH 6.8 (B) and 95% (v/v) acetonitrile and 5% (v/v) 200 mmol L-1 ammonium acetate pH 6.8 (A). Urine samples from the control and test groups were submitted to global metabolomics analysis by RPLC-MS using the optimized method and by CESI-MS. The results indicated that several metabolic pathways may have been altered by VUR. Changes of carnitine and acylcarnitine levels, amino acids and derivatives, purines and others was observed. Furthermore, the presence of acylcarnitines in the urine may indicate mitochondrial damage and the decrease of tryptophan and increase of the kynurenic acid indicate a change in the metabolism of tryptophan.
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Huang, Yi-chiuan, and 黃奕銓. "1. An improved assay for simultaneous analysis of 3-nitrotyrosine and 3-bromotyrosine in human urine by stable isotope dilution capillary liquid chromatography nanospray ionization tandem mass spectrometry2. Analysis of Thiol-Containing Amino Acids in Hu." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/75274478314817320035.
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碩士國立中正大學
化學所
98
Post-translational modification of proteins could affect protein functions. Nitration of protein tyrosine (Tyr), forming 3-nitrotyrosine (3NT), is implicated in cardiovascular diseases, cancer, and inflammatory diseases. At physiological concentrations of bromide, hypobromous acid can be a major oxidant produced by eosinophil peroxidase, leading to bromination of Tyr forming 3-bromotyrosine (3BT), which is considered a biomarker of cancer, allergic inflammatory disorders, asthma and other respiratory diseases. In this study, we have developed an isotope dilution capillary liquid chromatography nanospray ionization tandem mass spectrometry (capLC-NSI/MS/MS) assay for simultaneous analysis of protein-bound 3NT and 3BT in human urine. The detection limits were 0.2 pg (881 amol) for 3NT (S/N = 8) and 0.5 pg (1.92 fmol) for 3BT (S/N = 11) injected on-column, which were 50- and 10-fold lower for 3NT and 3BT, respectively, compared to the previous report ( Toxicol. Lett. 181, 31–39). Urinary protein was hydrolyzed by acid and purified by only one reversed phase solid-phase extraction (SPE) column to enrich Tyr, 3NT and 3BT. The fraction containing enriched 3NT and 3BT was analyzed by capLC-NSI/MS/MS. The high content Tyr in urinary proteins was quantified by high-performance liquid chromatography with fluorescence detection. Both 3NT and 3BT were detected and quantified in 0.1 mL of human urine samples. The use of a single SPE column in sample preparation simplifies the assay procedures. The high sensitivity and specificity of this capLC-NSI/MS/MS method render it a valuable tool in measurement of 3NT and 3BT in the human urinary protein as promising noninvasive biomarkers for protein tyrosine nitration and bromination in vivo.
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Yang, Shu-shu, and 楊琇琇. "1. Stability of adducts derived from 2’-deoxyoxanosine with glutathione and side chains of amino acids2. Simultaneous analysis of aldehydes in human urine by GC/MS." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/74118347615345976069.
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碩士國立中正大學
化學所
96
Excessive production of nitric oxide (NO) during cellular inflammation is known to damage DNA, and it plays an important role in cancers related to chronic diseases and inflammation. 2’-Deoxyoxanosine (dO) is a DNA lesion originated from reaction between 2’-deoxyguanosine (dGuo) and NO or nitrous acid (HNO2). However the O-acylisourea ring structure of dO is susceptible to reaction with nucleophilies forming ring-opened adducts. In this study, we react dO with the nucleophilic side chains of GSH, NAc-Tyr and NAc-Lys, forming dO-S-GSH, dO-NAc-Tyr and dO-NAc-Lys. The half lives of dO-S-GSH, dO-NAc-Tyr and dO-NAc-Lys at pH 7 and 8 at 37 °C are 11.3 and 2.22, 5.53 days and 1.36, 91.3, and 102 days, respectively. Thus, the stabilities of cross-linked products by reacting dO-DNA with amino acid in the protein follow the order of amide＞thiol ester＞phenol ester. The poor stability of thiol ester bond and phenol ester bond at pH 8 explains why modification on Cys and Tyr in the cross-links of dO-DNA with lysozyme were not detected. Lipid peroxidation is a cellular process implicated in the development of several diseases, including cancers. Aldehydes derived from lipid peroxidation react with biomolecules, such as DNA and proteins. A highly sensitive and specific assay has been developed for simultaneous quantification of aldehydes derived from lipid peroxidation, including acrolein, crotonaldehyde, hexanal, glyoxal, methylglyoxal, malondialdehyde and 2,4-decadienal in human urine. The urine sample was added tridecanal as internal standard and reacted with methanolic O-2,3,4,5,6-pentafluorobenzylhydroxylamine (PFBHA). The reaction mixture was extracted with hexane, followed by purification with a silica solid phase extraction (SPE) column. The PFB oxime derivatives eluted from the SPE column were analysed by gas chromatography-negative chemical ionization mass spectrometry (GC-NICI/MS) under the selective ion monitoring (SIM) mode of the specific fragment ions M-, [M–20]-, [M–50]- or [M–PFB]- for quantification in 0.4 and 0.5 mL of urine sample. Analysis of lipid peroxidation-derived aldehydes in human urine provides a noninvasive measure of in vivo exposure to reactive aldehydes capable of damaging biomolecules.
Books on the topic "Urine amino acids"
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Parker, James N., and Philip M. Parker. Maple syrup urine disease: A bibliography and dictionary for physicians, patients, and genome researchers [to Internet references]. San Diego, CA: ICON Health Publications, 2007.
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Rabier, Daniel. Amino Acids. Oxford University Press, 2016. http://dx.doi.org/10.1093/med/9780199972135.003.0083.
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Amino acids present in the different biological fluids belong to two groups: the protein group, with the 21 classical amino acids constituting the backbone of the protein, and the nonprotein group, appearing in different metabolic pathways as intermediate metabolites. It is important to know and to be able to recognize the latter, as they are the markers of many inherited metabolic diseases. Three kinds of pathways must be considered: the catabolic pathways, the synthesis pathways, and the transport pathways. A disorder on a catabolic pathway induces an increase of all metabolites upstream and so an increase of the starting amino acid in all fluids. Any disorder on the synthetic pathway of a particular amino acid will induce a decrease of this amino acid in all fluids. When a transporter is located on a plasma membrane, its deficiency will result in normal or low concentration in plasma concomitant to a high excretion in urine.
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Prunty, Helen, Jamie L. Fraser, Charles P. Venditti, and Robin H. Lachmann. Branched Chain Amino Acids. Oxford University Press, 2016. http://dx.doi.org/10.1093/med/9780199972135.003.0016.
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This chapter describes the four most common disorders affecting the degradation of branched chain amino acids: maple syrup urine disease, methylmalonic acidemia, propionic acidemia and isovaleric acidemia. These conditions most commonly present with encephalopathy in the newborn period, although cases with later onset have also been described. Although adult patients are less prone to acute metabolic decompensations, they do develop a number of long-term complications, both neurological and visceral. Management shares features with other disorders of protein metabolism and centers on a low-protein diet and the use of disease-specific amino acid supplements.
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Daudon, Michel, and Paul Jungers. Cystine stones. Edited by Mark E. De Broe. Oxford University Press, 2018. http://dx.doi.org/10.1093/med/9780199592548.003.0203_update_001.
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Cystinuria, an autosomal recessive disease (estimated at 1:7000 births worldwide), results from the defective reabsorption of cystine and dibasic amino acids (also ornithine, arginine, lysine, COAL) by epithelial cells of renal proximal tubules, leading to an abnormally high urinary excretion of these amino acids. Due to the poor solubility of cystine at the usual urine pH, formation of cystine crystals and stones ensues. Incidence of homozygotes is estimated at 1 in 7000 births worldwide, but is lower in European countries and much higher in populations with frequent consanguinity. Cystine stones represent 1–2% of all stones in adults and 5–8% in paediatric patients, with an equal distribution between males and females.Cystinuria is caused by inactivating mutations in the gene SLC3A1 or SLC7A9, both encoding proteins contributing to the function of the heterodimeric transport system of cystine.Cystine nephrolithiasis may present in infants, most frequently in adolescents or young adults, sometimes later. Cystine calculi are weakly radio-opaque. Stone analysis using infrared spectroscopy (or X-ray diffraction) allows immediate and accurate diagnosis. Urinary amino acid chromatography quantifies urinary cystine excretion, needed to define the therapeutic strategy.Urological treatment of cystine stones currently uses extracorporeal stone wave lithotripsy or flexible ureterorenoscopy with Holmium laser, that is, minimally invasive techniques. However, as cystine stones are highly recurrent, preventive therapy is essential.Medical treatment combines reduced methionine and sodium intake, to lower cystine excretion; hyperdiuresis (> 3 L/day) to reduce cystine concentration; and active alkalinization preferably using potassium citrate (40–80 mEq/day) to increase cystine solubility by rising urine pH up to 7.5–8. If these measures are insufficient to prevent recurrent stone formation, a thiol derivative (D-penicillamine or tiopronin), which converts cystine into a more soluble disulphide, should be added. Close monitoring and adherence of the patient to the therapeutic programme are needed to ensure life-long compliance, the key for successful prevention in the long term.
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Bockenhauer, Detlef, and Robert Kleta. Approach to the patient with renal Fanconi syndrome, glycosuria, or aminoaciduria. Edited by Robert Unwin. Oxford University Press, 2018. http://dx.doi.org/10.1093/med/9780199592548.003.0041_update_001.
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Up to 80% of filtered salt and water is returned back into the circulation in the proximal tubule. Several solutes, such as phosphate, glucose, low-molecular weight proteins, and amino acids are exclusively reabsorbed in this segment, so their appearance in urine is a sign of proximal tubular dysfunction. An entire orchestra of specialized apical and basolateral transporters, as well as paracellular molecules, mediate this reabsorption. Defects in proximal tubular function can be isolated (e.g. isolated renal glycosuria, aminoacidurias, or hypophosphataemic rickets) or generalized. In the latter case it is called the Fanconi–Debre–de Toni syndrome, based on the initial clinical descriptions. However, in clinical practice it is usually referred to as just the ‘renal Fanconi syndrome’. Severity of proximal tubular dysfunction can vary, and may coexist with some degree of loss of glomerular filtration capacity. Causes include a wide range of insults to proximal tubular cells, including a number of genetic conditions, drugs and poisons.
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Lachmann, Robin, and Elaine Murphy. Aminoacidopathies, urea cycle disorders, and organic acidurias. Edited by Patrick Davey and David Sprigings. Oxford University Press, 2018. http://dx.doi.org/10.1093/med/9780199568741.003.0180.
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Aminoacidopathies are caused by deficiencies in enzymes involved in amino acid metabolism and are often characterized by the accumulation of a toxic amino acid. The two diseases most likely to be encountered in adult medicine are phenylketonuria, which is caused by a deficiency of phenylalanine hydroxylase, and maple syrup urine disease (MSUD), which is due to a branched-chain amino acid decarboxylase deficiency. High levels of phenylalanine progressively damage the developing brain, leading to severe learning difficulties. The high levels of leucine which accumulate in MSUD produce an acute encephalopathy which, if not treated, can rapidly become fatal.
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Cassiman, David, and Wouter Meersseman. Tyrosinemia Type I. Oxford University Press, 2016. http://dx.doi.org/10.1093/med/9780199972135.003.0013.
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Tyrosinemia type 1 (HT-1) is a rare metabolic disorder affecting degradation pathways of the amino acid tyrosine. HT-1 presents with liver, kidney and/or bone disease and can cause acute porphyria attacks. Biochemical diagnosis is made by measuring raised plasma tyrosine and detection of succinylacetone in urine. Long-term management with diet and nitisinone leads to excellent short term results, but since long term effects are largely unknown, life-long treatment and follow-up for liver malignancy, bone disease and kidney disease seem necessary. HT-1 is treatable by liver transplantation.
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Riazi, Roya. Branched chain amino acid metabolism: Requirements in healthy adult men and patients with maple syrup urine disease. 2003.
Find full textBook chapters on the topic "Urine amino acids"
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Carpenter, Kevin. "Branched Chain Amino Acids and Maple Syrup Urine Disease." In Branched Chain Amino Acids in Clinical Nutrition, 145–56. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-1923-9_12.
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Lou, Marjorie F., and Paul B. Hamilton. "Separation and Quantitation of Peptides and Amino Acids in Normal Human Urine." In Methods of Biochemical Analysis, 203–71. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2006. http://dx.doi.org/10.1002/9780470110454.ch3.
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Martini, C., M. Bruno, M. Petrarulo, D. Cosseddu, and F. Linari. "Sulfur Amino Acids, Thiol Drugs, and Related Mixed Disulfides from Urine Samples of Cystine Stone Patients." In Urolithiasis, 567–69. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4899-0873-5_174.
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Ferreira, Carlos R., and Kristina Cusmano-Ozog. "Spurious Elevation of Multiple Urine Amino Acids by Ion-Exchange Chromatography in Patients with Prolidase Deficiency." In JIMD Reports, 45–49. Berlin, Heidelberg: Springer Berlin Heidelberg, 2016. http://dx.doi.org/10.1007/8904_2016_552.
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van Gennip, Albert H., Sandra Busch, Eline G. Scholten, Lida E. Stroomer, and Nico G. Abeling. "Simple Method for the Quantitative Analysis of Dihydropyrimidines and N-Carbamyl-ß-Amino Acids in Urine." In Advances in Experimental Medicine and Biology, 15–19. New York, NY: Springer US, 1991. http://dx.doi.org/10.1007/978-1-4615-7703-4_4.
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Giordano, Giuseppe, Antonina Gucciardi, Paola Pirillo, and Mauro Naturale. "Quantification of Underivatized Amino Acids on Dry Blood Spot, Plasma, and Urine by HPLC-ESI-MS/MS." In Methods in Molecular Biology, 153–72. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9639-1_13.
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Giordano, Giuseppe, Iole Maria Di Gangi, Antonina Gucciardi, and Mauro Naturale. "Quantification of Underivatised Amino Acids on Dry Blood Spot, Plasma, and Urine by HPLC–ESI–MS/MS." In Methods in Molecular Biology, 219–42. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-445-2_18.
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Leah, J. M., T. Palmer, M. Griffin, C. J. Wingad, A. Briddon, and V. G. Oberholzer. "Urine Amino Acid Analysis by HPLC in the Investigation of Inborn Errors of Metabolism." In Practical Developments in Inherited Metabolic Disease: DNA Analysis, Phenylketonuria and Screening for Congenital Adrenal Hyperplasia, 250–53. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4131-1_39.
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Inoue, Gosuke, and Kenji Toba. "The Effect of Amino Acid Infusion on Partial Pressure of Oxygen in Pelvic Urine of the Rat." In Oxygen Transport to Tissue X, 609–13. New York, NY: Springer US, 1988. http://dx.doi.org/10.1007/978-1-4615-9510-6_75.
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Kratzin, H. D., A. I. Pick, and N. Hilschmann. "Complete Amino-Acid Sequence of a Kappa Light Chain Fragment Isolated from the Urine of Amyloidosis Patient MAL." In Amyloid and Amyloidosis 1990, 165–68. Dordrecht: Springer Netherlands, 1991. http://dx.doi.org/10.1007/978-94-011-3284-8_41.
Full textConference papers on the topic "Urine amino acids"
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Rabbani, Naila, Alberto de a Fuente, and Paul Thornalley. "Risk prediction of early decline in renal function in diabetic kidney disease with algorithm including fractional excretion of glycated amino acids." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2021. http://dx.doi.org/10.29117/quarfe.2021.0096.
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Background and aims: Diabetic kidney disease occurs in ca. 40% patients with diabetes. Approximately 1 in 5 patients with type 1 diabetes mellitus (T1DM) and 1 in 3 patients with type 2 diabetes mellitus (T2DM) develop early decline in renal function (EDRF), requiring renal dialysis after 5 - 20 years. Currently, at the time of normoalbuminuria or new onset microalbuminuria (incipient diabetic nephropathy), it is uncertain which patients are at risk of EDRF. With Joslin Kidney Study investigators, we found patients with T1DM who later developed EDRF (Decliners) have higher fractional excretion (FE) of 6 glycated amino acids - fructosyl-lysine and 5 advanced glycation endproducts (AGEs), compared to patients with stable renal function (Non-decliners). However, FE of any single glycated amino acid could not classify Decliners or Non-decliners. The aim of this study was to apply artificial intelligence machine learning to develop diagnostic algorithms to classify Decliners and Non-decliners by optimum combination of levels of glycated and oxidized amino acids in plasma and urine, related FEs and conventional clinical chemistry variables. Materials and methods: Patients with T1DM with stable renal function (n = 63) and EDRF (n = 22) were recruited for this study. Data on levels of 14 glycated and oxidized amino acids in plasma, urine, related FEs, glycated hemoglobin A1C, log(urinary albumin creatinine ratio, ACR), age, gender and duration of diabetes at the time of new onset microalbuminuria were included as features in algorithm development. Algorithms were trained and tested on 90%/10% data split, repeated 1000 times, using the Extreme Gradient Boosting method. Results: The algorithm gave an optimal classification of Decliners and Non-decliners. Optimum with features: A1C, log[ACR], FE(Nꞷ-carboxymethylarginine, CMA), FE(glyoxal-derived hydroimidazolone, G-H1) and plasma concentration of Nε-carboxymethyl-lysine (CML) free adduct; For The diagnostic performance for risk prediction of future EDRF was (mean ± SD): sensitivity 74 ± 9%, specificity 91 ± 45 and accuracy 87 ± 4%. The positive likelihood ratio LR+ was 11.0, indicating that this method gives strong, often conclusive evidence of future EDRF in patients with T1DM. In contrast, algorithms with A1C and logACR only as features gave LR+ 2.6, providing small evidence of risk of future EDRF. Conclusion: With conclude that FEs of glycated amino acids are novel risk predictors of EDRF, likely linked to reporting of early-decline of cationic amino acid transporter function in the renal tubular epithelium. Genetic polymorphism of these amino acid transporters has been linked to rapid decline in renal function in genome-wide association studies. Measurement of only 3 glycated amino acids, CMA, G-H1 and CML, produced an algorithm with optimum risk prediction of EDRF. With further validation, including in patients with T2DM and with chronic kidney disease without diabetes, this method may markedly improve clinical risk prediction of EDRF.
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Delani, F., M. Tagliaferri, D. Macconi, C. Lupini, N. Perico, and G. Rumuzzi. "PLATELET ACTIVATING FACTOR (PAF) AS A MEDIATOR OF PROTEINURIA IN ISOLATED PERFUSED KIDNEY (IPK)." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643485.
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PAF amplifies tissue damage in glomerulonephritis and can promote proteinuria stimulating platelet and neutrophil cationic protein release. We used IPK to establish whether PAF directly causes proteinuria. Kidneys were isolated from male Sprague-Dawley rats and perfused at constant pressure (100 mmHg) in a closed circuit with a Krebs-Henseleit buffer containing glucose urea creatinine, BSA (1%), Ficoll 70 (3.5%) and amino acids. After 25 min stabilization period, a basal 10 min clearance period was followed by PAF (1.8 nM f.c. n = 6) or vehicle (n = 5) injection into the renal artery. As seen in the figure PAF but not vehicle significantly (p<0.01) increased urine protein excretion. No significant changes in GFR (as creatinine clearance) were observed after PAF or vehicle injection (Basal: 0.786 ± 0.075 PAF: 0.658 ± 0.070. Basal 0.653 ± 0.081, vehicle 0.639 ± 0.074 ml/min/g kidney). The data indicate that PAF may directly increase glomerular permeability to proteins.
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Heeb, M. J., F. Espana, M. Geiger, D. Collen, D. C. Stump, and J. H. Griffin. "IMMUNOLOGICAL SIMILARITIES BETWEEN PLASMA AND URINARY PROTEIN C INHIBITORS (PCIs) AND URINARY UROKINASE INHIBITOR (UKI)." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643816.
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Plasma PCIs have similar MW (∼ 57K), amino acid composition, and heparin dependence (Suzuki et al 1983, JBC 258:163) as urinary UKI (Stump et al 1986, JBC 261:12759). Urinary PCI of ∼ 50K MW has a similar heparin dependence and urokinase (UK) competes with activated protein C (APC) for this PCI (Geiger et al 1986, Circ. 74:11-234). For comparison, three forms of PCI, one from urine and two from plasma, were purified, and each exhibited heparin-dependent UK and APC inhibitory activity and formed heparin-dependent complexes with APC. The APC-PCI complexes were visible on immunoblots (nondenaturing gels) developed using: A) monoclonal anti-UKI + 125I-antimouse IgG; B) polyclonal anti-plasma PCI + 125I-plasma PCI; and C) monoclonal anti-protein C + 125I-protein C. The three forms of purified PCI were detected by methods A and B. Two new bands of APC-inhibitor complexes were seen upon incubation of plasma with APC in the presence of heparin, and the same pattern was visualized by methods A, B, and C. In the absence of heparin, only one APC-inhibitor band was visualized by methods A and B, but two bands were visualized by method C. Plasma immunodepleted of UKI by monoclonal anti-UKI-Sepharose showed no detectable antigen or complexes with APC as visualized by methods A and B. However, the UKI-depleted plasma contained components which formed a reduced amount of complexes with APC as visualized with protein C antibodies, i.e. method C. Heparin stimulates tenfold the PCI activity of normal plasma. Based on amidolytic assays of APC using S-2366, the UKI-depleted plasma was very deficient (< 15%) in heparin-dependent PCI activity, whereas the weak heparin-independent PCI activity was slightly reduced. This indicates that the majority of heparin-dependent PCI activity of plasma is immunologically.related to UKI. These studies suggest that the two slightly different forms of plasma PCI, the urinary UKI, and the urinary PCI are very similar if not identical proteins and that plasma may contain a minor heparin-independent PCI which is not immunologically related to these proteins.