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Sample records for urine total protein

  1. Prevalence of isolated non-albumin proteinuria in the US population tested for both, urine total protein and urine albumin: An unexpected discovery.

    PubMed

    Katayev, Alexander; Zebelman, Arthur M; Sharp, Thomas M; Samantha Flynn; Bernstein, Richard K

    2017-04-01

    Isolated non-albumin proteinuria (NAP) is a condition when urine total protein concentrations are elevated without elevation of urine albumin. The prevalence of NAP in the US population tested for both, urine total protein and albumin was assessed in this study. The database of a US nationwide laboratory network was queried for test results when random urine albumin was ordered together with urine total protein and also when timed 24-hour urine albumin was ordered together with urine total protein. The total prevalence of NAP in the US population tested for both, urine total protein and albumin was calculated for patient groups having normal and low-normal urine albumin (random and timed) with elevated and severely increased urine total protein (random and timed). Also, the prevalence of NAP was calculated for patients with normal urine albumin to assess the probability of missing proteinuria if only urine albumin is measured. The prevalence of NAP in the random samples group was 10.1% (15.2% for females and 4.7% for males). Among patients with normal random albumin, there were 20.0% (27.3% of females and 10.7% of males) patients with NAP. The prevalence of NAP in the timed samples group was 24.6% (29.8% for females and 18.5% for males). Among patients with normal timed urine albumin, there were 36.2% (40.0% of females and 30.8% of males) patients with NAP. There was a strong positive association with female gender and NAP in most patients groups. Testing for only urine (micro)albumin can miss up to 40% of females and 30.8% of males with gross proteinuria. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  2. The Total Urine Protein-to-Creatinine Ratio Can Predict the Presence of Microalbuminuria

    PubMed Central

    Yamamoto, Kyoko; Yamamoto, Hiroyuki; Yoshida, Katsumi; Niwa, Koichiro; Nishi, Yutaro; Mizuno, Atsushi; Kuwabara, Masanari; Asano, Taku; Sakoda, Kunihiro; Niinuma, Hiroyuki; Nakahara, Fumiko; Takeda, Kyoko; Shindoh, Chiyohiko; Komatsu, Yasuhiro

    2014-01-01

    Background The Kidney Disease: Improving Global Outcomes chronic kidney disease (CKD) guidelines recommend that CKD be classified based on the etiology, glomerular filtration rate (GFR) and degree of albuminuria. The present study aimed to establish a method that predicts the presence of microalbuminuria by measuring the total urine protein-to-creatinine ratio (TPCR) in patients with cardiovascular disease (CVD) risk factors. Methods and Results We obtained urine samples from 1,033 patients who visited the cardiovascular clinic at St. Luke's International Hospital from February 2012 to August 2012. We measured the TPCR and the urine albumin-to-creatinine ratio (ACR) from random spot urine samples. We performed correlation, receiver operating characteristic (ROC) curve, sensitivity, and subgroup analyses. There was a strong positive correlation between the TPCR and ACR (R2 = 0.861, p<0.001). A ROC curve analysis for the TPCR revealed a sensitivity of 94.4%, a specificity of 86.1%, and an area under the curve of 0.903 for detecting microalbuminuria for a TPCR cut-off value of 84 mg/g of creatinine. The subgroup analysis indicated that the cut-off value could be used for patients with CVD risk factors. Conclusions These results suggest that the TPCR with an appropriate cut-off value could be used to screen for the presence of microalbuminuria in patients with CVD risk factors. This simple, inexpensive measurement has broader applications, leading to earlier intervention and public benefit. PMID:24614247

  3. Protein urine test

    MedlinePlus

    Urine protein; Albumin - urine; Urine albumin; Proteinuria; Albuminuria ... After you provide a urine sample, it is tested. The health care provider uses a dipstick made with a color-sensitive pad. The color change ...

  4. Normal values of urine total protein- and albumin-to-creatinine ratios in term newborns.

    PubMed

    El Hamel, Chahrazed; Chianea, Thierry; Thon, Séverine; Lepichoux, Anne; Yardin, Catherine; Guigonis, Vincent

    2017-01-01

    It is important to have an accurate assessment of urinary protein when glomerulopathy or kidney injury is suspected. Currently available normal values for the neonate population have limited value, in part because they are based on small populations and obsolete creatinine assays. We have performed a prospective study with the aim to update the normal upper values of the urinary total protein-to-creatinine and albumin-to-creatinine ratios in term newborns. Urine samples were collected from 277 healthy, full-term newborns within the first 48 hours (D0-1) and between 72 and 120 h of life (D3-4). Total protein, albumin, creatinine and osmolality were measured and the upper limit of normal (upper-limit) values determined. At D0-1 and D3-4, the upper-limit values for the total protein-to-creatinine ratio were 1431 and 1205 mg/g (162 and 136 g/mol) and those for the albumin-to-creatinine ratio were 746 and 301 mg/g (84 and 34 g/mol), respectively. The upper-limit values were significantly higher at D0-1 than at D3-4 only for the albumin-to-creatinine ratio. This study determined the upper limit of normal values for urinary total protein-to-creatinine and albumin-to-creatinine ratios in the largest population of newborns studied to date. These values can therefore be considered as the most clinically relevant data currently available for the detection and diagnosis of glomerular injury in daily clinical practice in this population.

  5. Urine Protein and Urine Protein to Creatinine Ratio

    MedlinePlus

    ... Less Common Questions Related Content On This Site Tests: Urinalysis ; Albumin ; Urine Albumin ; Protein Electrophoresis ; Total Protein , BUN , Creatinine , Creatinine Clearance , eGFR Conditions: Kidney Disease , Proteinuria , Pre-eclampsia , Diabetes , Hypertension , Multiple Myeloma , Urinary Tract Infection ...

  6. Pediatric reference intervals for random urine calcium, phosphorus and total protein.

    PubMed

    Slev, Patricia R; Bunker, Ashley M; Owen, William E; Roberts, William L

    2010-09-01

    The aim of this study was to establish age appropriate reference intervals for calcium (Ca), phosphorus (P) and total protein (UTP) in random urine samples. All analytes were measured using the Roche MODULAR P analyzer and normalized to creatinine (Cr). Our study cohort consisted of 674 boys and 728 girls between 7 and 17 years old (y.o.), which allowed us to determine the central 95% reference intervals with 90% confidence intervals by non-parametric analysis partitioned by both gender and 2-year age intervals for each analyte [i.e. boys in age group 7-9 years (7-9 boys); girls in age group 7-9 years (7-9 girls), etc.]. Results for the upper limits of the central 95% reference interval were: for Ca/Cr, 0.27 (16,17 y.o.) to 0.46 mg/mg (7-9 y.o.) for the girls and 0.26 (16,17 y.o.) to 0.43 mg/mg (7-9 y.o.) for the boys; for P/Cr, 0.85 (16,17 y.o.) to 1.44 mg/mg (7-9 y.o.) for the girls and 0.87 (16,17 y.o.) to 1.68 mg/mg (7-9 y.o.) for the boys; for UTP/Cr, 0.30 (7-9 y.o.) to 0.34 mg/mg (10-12 y.o.) for the girls and 0.19 (16,17, y.o.) to 0.26 mg/mg (13-15 y.o.) for the boys. Upper reference limits decreased with increasing age, and age was a statistically significant variable for all analytes. Eight separate age- and gender-specific reference intervals are proposed per analyte.

  7. Urine protein electrophoresis test

    MedlinePlus

    Urine protein electrophoresis; UPEP; Multiple myeloma - UPEP; Waldenström macroglobulinemia - UPEP; Amyloidosis - UPEP ... special paper and apply an electric current. The proteins move and form visible bands. These reveal the ...

  8. 24-hour urine protein

    MedlinePlus

    ... one urine sample (protein-to-creatinine ratio). Normal Results The normal value is less than 100 milligrams ... meaning of your specific test results. What Abnormal Results Mean Abnormal results may be due to: A ...

  9. Total protein

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003483.htm Total protein To use the sharing features on this page, please enable JavaScript. The total protein test measures the total amount of two classes ...

  10. Spot Urine Protein-to-Creatinine Ratio to Predict the Magnitude of 24-Hour Total Proteinuria in Preeclampsia of Varying Severity.

    PubMed

    Kucukgoz Gulec, Umran; Sucu, Mete; Ozgunen, Fatma Tuncay; Buyukkurt, Selim; Guzel, Ahmet Baris; Paydas, Saime

    2017-10-01

    The predictive value of spot urine protein-to-creatinine ratio (PCR) for estimating total 24-hour proteinuria in severe preeclampsia is unclear. This study aimed to assess the diagnostic accuracy of spot urine PCR for ascertaining the magnitude of proteinuria in women with preeclampsia of varying severity. A total of 205 patients with prediagnosed preeclampsia were included in this prospective cohort study. Patients were allocated into one of the three groups categorized by severity of disease, as follows: gestational hypertension, group 1 (n = 41); preeclampsia, group 2 (n = 88); and severe preeclampsia, group 3 (n = 76). We assessed the spot urine PCRs to determine significant proteinuria and the magnitude of proteinuria in these groups. The spot urine PCR was 0.53, with 81% sensitivity and 93% specificity to detect significant proteinuria. A significant correlation was found between PCR and 24-hour total proteinuria in group 1 (r = 0.473, P = 0.002). There were also significant correlations in group 2 (r = 0.814, P < 0.001) and group 3 (r = 0.912, P < 0.001). The established formula using spot urine PCR to estimate 24-hour total proteinuria in severe preeclampsia was Y = 832.02X + 378.74 mg (r 2  = 0.8304). Although 24-hour urine collection remains a merely reliable test to determine the degree of total proteinuria, our findings suggest that it is likely to assess the magnitude of proteinuria by the spot urine PCR, especially in severe preeclampsia. www.clinicaltrials.govNCT01623791. Copyright © 2017 The Society of Obstetricians and Gynaecologists of Canada/La Société des obstétriciens et gynécologues du Canada. Published by Elsevier Inc. All rights reserved.

  11. Diagnostic Accuracy of Urine Protein/Creatinine Ratio Is Influenced by Urine Concentration

    PubMed Central

    Yang, Chih-Yu; Chen, Fu-An; Chen, Chun-Fan; Liu, Wen-Sheng; Shih, Chia-Jen; Ou, Shuo-Ming; Yang, Wu-Chang; Lin, Chih-Ching; Yang, An-Hang

    2015-01-01

    Background The usage of urine protein/creatinine ratio to estimate daily urine protein excretion is prevalent, but relatively little attention has been paid to the influence of urine concentration and its impact on test accuracy. We took advantage of 24-hour urine collection to examine both urine protein/creatinine ratio (UPCR) and daily urine protein excretion, with the latter as the reference standard. Specific gravity from a concomitant urinalysis of the same urine sample was used to indicate the urine concentration. Methods During 2010 to 2014, there were 540 adequately collected 24h urine samples with protein concentration, creatinine concentration, total volume, and a concomitant urinalysis of the same sample. Variables associated with an accurate UPCR estimation were determined by multivariate linear regression analysis. Receiver operating characteristic (ROC) curves were generated to determine the discriminant cut-off values of urine creatinine concentration for predicting an accurate UPCR estimation in either dilute or concentrated urine samples. Results Our findings indicated that for dilute urine, as indicated by a low urine specific gravity, UPCR is more likely to overestimate the actual daily urine protein excretion. On the contrary, UPCR of concentrated urine is more likely to result in an underestimation. By ROC curve analysis, the best cut-off value of urine creatinine concentration for predicting overestimation by UPCR of dilute urine (specific gravity ≦ 1.005) was ≦ 38.8 mg/dL, whereas the best cut-off values of urine creatinine for predicting underestimation by UPCR of thick urine were ≧ 63.6 mg/dL (specific gravity ≧ 1.015), ≧ 62.1 mg/dL (specific gravity ≧ 1.020), ≧ 61.5 mg/dL (specific gravity ≧ 1.025), respectively. We also compared distribution patterns of urine creatinine concentration of 24h urine cohort with a concurrent spot urine cohort and found that the underestimation might be more profound in single voided samples

  12. Diagnostic Accuracy of Urine Protein/Creatinine Ratio Is Influenced by Urine Concentration.

    PubMed

    Yang, Chih-Yu; Chen, Fu-An; Chen, Chun-Fan; Liu, Wen-Sheng; Shih, Chia-Jen; Ou, Shuo-Ming; Yang, Wu-Chang; Lin, Chih-Ching; Yang, An-Hang

    2015-01-01

    The usage of urine protein/creatinine ratio to estimate daily urine protein excretion is prevalent, but relatively little attention has been paid to the influence of urine concentration and its impact on test accuracy. We took advantage of 24-hour urine collection to examine both urine protein/creatinine ratio (UPCR) and daily urine protein excretion, with the latter as the reference standard. Specific gravity from a concomitant urinalysis of the same urine sample was used to indicate the urine concentration. During 2010 to 2014, there were 540 adequately collected 24h urine samples with protein concentration, creatinine concentration, total volume, and a concomitant urinalysis of the same sample. Variables associated with an accurate UPCR estimation were determined by multivariate linear regression analysis. Receiver operating characteristic (ROC) curves were generated to determine the discriminant cut-off values of urine creatinine concentration for predicting an accurate UPCR estimation in either dilute or concentrated urine samples. Our findings indicated that for dilute urine, as indicated by a low urine specific gravity, UPCR is more likely to overestimate the actual daily urine protein excretion. On the contrary, UPCR of concentrated urine is more likely to result in an underestimation. By ROC curve analysis, the best cut-off value of urine creatinine concentration for predicting overestimation by UPCR of dilute urine (specific gravity ≦ 1.005) was ≦ 38.8 mg/dL, whereas the best cut-off values of urine creatinine for predicting underestimation by UPCR of thick urine were ≧ 63.6 mg/dL (specific gravity ≧ 1.015), ≧ 62.1 mg/dL (specific gravity ≧ 1.020), ≧ 61.5 mg/dL (specific gravity ≧ 1.025), respectively. We also compared distribution patterns of urine creatinine concentration of 24h urine cohort with a concurrent spot urine cohort and found that the underestimation might be more profound in single voided samples. The UPCR in samples with low

  13. Urine chemistry

    MedlinePlus

    ... rate 24-hour urine protein Acid loading test (pH) Adrenalin - urine test Amylase - urine Bilirubin - urine Calcium - urine Citric acid ... Urine dermatan sulfate Urine - hemoglobin Urine metanephrine Urine pH Urine specific gravity Vanillylmandelic acid (VMA)

  14. CSF total protein

    MedlinePlus

    CSF total protein is a test to determine the amount of protein in your spinal fluid, also called cerebrospinal fluid (CSF). ... The normal protein range varies from lab to lab, but is typically about 15 to 60 milligrams per deciliter (mg/dL) ...

  15. Correlation of Urine Biomarkers: Microalbuminuria and Spot Urine Protein among Diabetic Patients. Application of Spot Urine Protein in Diabetic Kidney Disease, Nephropathy, Proteinuria Estimation, Diagnosing and Monitoring.

    PubMed

    Aziz, Kamran M A

    2015-01-01

    Current study has invented a new method for utilizing spot urine protein among diabetic patients. There have been various efforts and strategies in research internationally to detect, diagnose and monitor nephropathy/DKD. Although 24-hour urine studies are gold standard, however, there exist some controversies about microalbuminuria and spot urine protein. The current study was designed to utilize spot urine protein among diabetic patients and to find its association with routine dipstick urine test for albumin, and microalbuminuria. The study demonstrated significant association of spot urine protein with urine dipstick albumin, and has demonstrated increasing spot urine protein with increasing albumin in urine (p-value < 0.0001). This study also demonstrated significantly higher levels of spot urine protein between the groups with nephropathy/DKD as compared to those without nephropathy/DKD (p-value < 0.0001). Similarly, spot urine protein and spot urine protein/creatinine were also significantly associated with microalbumin and microalbumin/creatinine in urine. Significant regression models for spot urine protein and microalbuminuria were also developed and proposed to detect and estimate microalbumin in urine while utilizing spot urine protein (< 0.0001). Synthesized regression equations and models can be used confidently to detect, rule out and monitor proteinuria and DKD. ROC curves were utilized to detect spot urine protein cutoff points for nephropathy and DKD with high specificity and sensitivity. Some important patents were also discussed in the paper regarding albuminuria/proteinuria detection and management. Current study has demonstrated and concluded, for the first time, that there exists a significant association of spot urine protein with routine dipstick albumin in urine and microalbuminuria. It is also essential to detect early, monitor and manage proteinuria, hypertension and dyslipidemia with good glycemic control to prevent diabetes

  16. The urine proteome for radiation biodosimetry: effect of total body vs. local kidney irradiation.

    PubMed

    Sharma, Mukut; Halligan, Brian D; Wakim, Bassam T; Savin, Virginia J; Cohen, Eric P; Moulder, John E

    2010-02-01

    Victims of nuclear accidents or radiological terrorism are likely to receive varying doses of ionizing radiation inhomogeneously distributed over the body. Early biomarkers may be useful in determining organ-specific doses due to total body irradiation (TBI) or partial body irradiation. The authors used liquid chromatography and mass spectrometry to compare the effect of TBI and local kidney irradiation (LKI) on the rat urine proteome using a single 10-Gy dose of x-rays. Both TBI and LKI altered the urinary protein profile within 24 h with noticeable differences in gene ontology categories. Some proteins, including fetuin-B, tissue kallikrein, beta-glucuronidase, vitamin D-dependent calcium binding protein and chondroitin sulfate proteoglycan NG2, were detected only in the TBI group. Some other proteins, including major urinary protein-1, RNA binding protein 19, neuron navigator, Dapper homolog 3, WD repeat and FYVE domain containing protein 3, sorting nexin-8, ankycorbin and aquaporin were detected only in the LKI group. Protease inhibitors and kidney proteins were more abundant (fraction of total scans) in the LKI group. Urine protein (Up) and creatinine (Uc) (Up/Uc) ratios and urinary albumin abundance decreased in both TBI and LKI groups. Several markers of acute kidney injury were not detectable in either irradiated group. Present data indicate that abundance and number of proteins may follow opposite trends. These novel findings demonstrate intriguing differences between TBI and LKI, and suggest that urine proteome may be useful in determining organ-specific changes caused by partial body irradiation.

  17. Polyphenols excreted in urine as biomarkers of total polyphenol intake.

    PubMed

    Medina-Remón, Alexander; Tresserra-Rimbau, Anna; Arranz, Sara; Estruch, Ramón; Lamuela-Raventos, Rosa M

    2012-11-01

    Nutritional biomarkers have several advantages in acquiring data for epidemiological and clinical studies over traditional dietary assessment tools, such as food frequency questionnaires. While food frequency questionnaires constitute a subjective methodology, biomarkers can provide a less biased and more accurate measure of specific nutritional intake. A precise estimation of polyphenol consumption requires blood or urine sample biomarkers, although their association is usually highly complex. This article reviews recent research on urinary polyphenols as potential biomarkers of polyphenol intake, focusing on clinical and epidemiological studies. We also report a potentially useful methodology to assess total polyphenols in urine samples, which allows a rapid, simultaneous determination of total phenols in a large number of samples. This methodology can be applied in studies evaluating the utility of urinary polyphenols as markers of polyphenol intake, bioavailability and accumulation in the body.

  18. Electrical conductivity and total dissolved solids in urine.

    PubMed

    Fazil Marickar, Y M

    2010-08-01

    The objective of this paper is to study the relevance of electrical conductivity (EC) and total dissolved solids (TDS) in early morning and random samples of urine of urinary stone patients; 2,000 urine samples were studied. The two parameters were correlated with the extent of various urinary concrements. The early morning urine (EMU) and random samples of the patients who attended the urinary stone clinic were analysed routinely. The pH, specific gravity, EC, TDS, redox potential, albumin, sugar and microscopic study of the urinary sediments including red blood cells (RBC), pus cells (PC), crystals, namely calcium oxalate monohydrate (COM), calcium oxalate dihydrate (COD), uric acid (UA), and phosphates and epithelial cells were assessed. The extent of RBC, PC, COM, COD, UA and phosphates was correlated with EC and TDS. The values of EC ranged from 1.1 to 33.9 mS, the mean value being 21.5 mS. TDS ranged from 3,028 to 18,480 ppm, the mean value being 7,012 ppm. The TDS levels corresponded with EC of urine. Both values were significantly higher (P < 0.05) in the EMU samples than the random samples. There was a statistically significant correlation between the level of abnormality in the urinary deposits (r = +0.27, P < 0.05). In samples, where the TDS were more than 12,000 ppm, there were more crystals than those samples containing TDS less than 12,000 ppm. However, there were certain urine samples, where the TDS were over 12,000, which did not contain any urinary crystals. It is concluded that the value of TDS has relevance in the process of stone formation.

  19. Total morphine stability in urine specimens stored under various conditions.

    PubMed

    Chang, B L; Huang, M K; Tsai, Y Y

    2000-09-01

    The stability of total morphine in urine stored under various conditions was studied using control and experimental specimens. Samples in the control group were prepared using drug-free urine spiked with morphine at three concentration levels (300, 1000, and 2500 ng/mL), each with the pH adjusted to 5.5, 6.5, and 7.5. Samples in the experimental group came from 20 alleged heroin addicts (provided by Taipei Municipal Psychiatric Hospital). Samples in both groups were divided into two categories--one with and one without the precipitate (formed at 0 degrees C) removed. Samples in each of these two categories were further divided into two sub-groups--one with and one without sodium azide (0.05%) added. Total morphine contents in these samples were first determined by gas chromatography-mass spectrometry prior to storage and at 6, 12, 18, and 24 months following storage at -20, 4, 25, and 35 degrees C. Effects of sample treatment (azide addition and precipitate removal), pH, and storage temperature and length were evaluated by examining the percentage of total morphine remaining at the four time intervals following the initial determination. Major findings were as follows: (1) total morphine decomposition was minimal when stored for 12 months at -20 degrees C, which is a common current practice; (2) samples with lower initial sample pH had slower total morphine decomposition rates; and (3) azide addition appeared to have no detectable effect, whereas precipitate removal appeared to marginally reduce the decomposition rate, especially for samples with lower pH.

  20. Kidney function and urine protein composition in healthy volunteers during space station fitness tests.

    PubMed

    Fomina, Elena V; Lisova, Natalia Iu; Kireev, Kirill S; Tiys, Evgeny S; Kononikhin, Alexey S; Larina, Irina M

    2015-05-01

    There is a close physiological connection between muscular activity and kidney function. During physical exercise (PE) the qualitative and quantitative composition of urine changes. This paper explores the influence of moderate PE on urine protein composition. The study of urine protein composition will help to make corrections to the existing methods of countermeasures. There were 10 healthy men who exercised on a treadmill similar to the one onboard the International Space Station. We analyzed their urinary proteome composition, potassium level, sodium level, and their level of osmotically active substances before and after PE. After moderate PE, a small increase in urine flow speed and a constant glomerular filtration rate were noted. The average-group index of total protein excretion within the urine was reliably increased. From the 148 proteins identified in the urine, 64 were associated with known tissue origin. We found that protein penetration into the urine had a positive correlation with their tissue expression. Selectivity of the glomerular barrier during PE decreased and high-molecular weight proteins penetrated through the glomerular barrier more easily after PE. Performance of moderate intensity physical exercise of short duration did not lead to an increase in the glomerular filtration rate nor did diuresis increase above the limits of baseline variability. However, the protein excretion rate increased after PE. We also observed that protein composition drift indicated a change in the set of biological processes in which a given protein participated, in some cases activating, in some cases inactivating them.

  1. Stone former urine proteome demonstrates a cationic shift in protein distribution compared to normal.

    PubMed

    Kolbach-Mandel, Ann M; Mandel, Neil S; Hoffmann, Brian R; Kleinman, Jack G; Wesson, Jeffrey A

    2017-08-01

    Many urine proteins are found in calcium oxalate stones, yet decades of research have failed to define the role of urine proteins in stone formation. This urine proteomic study compares the relative amounts of abundant urine proteins between idiopathic calcium oxalate stone forming and non-stone forming (normal) cohorts to identify differences that might correlate with disease. Random mid-morning urine samples were collected following informed consent from 25 stone formers and 14 normal individuals. Proteins were isolated from urine using ultrafiltration. Urine proteomes for each sample were characterized using label-free spectral counting mass spectrometry, so that urine protein relative abundances could be compared between the two populations. A total of 407 unique proteins were identified with the 38 predominant proteins accounting for >82% of all sample spectral counts. The most highly abundant proteins were equivalent in stone formers and normals, though significant differences were observed in a few moderate abundance proteins (immunoglobulins, transferrin, and epidermal growth factor), accounting for 13 and 10% of the spectral counts, respectively. These proteins contributed to a cationic shift in protein distribution in stone formers compared to normals (22% vs. 18%, p = 0.04). Our data showing only small differences in moderate abundance proteins suggest that no single protein controls stone formation. Observed increases in immunoglobulins and transferrin suggest increased inflammatory activity in stone formers, but cannot distinguish cause from effect in stone formation. The observed cationic shift in protein distribution would diminish protein charge stabilization, which could lead to protein aggregation and increased risk for crystal aggregation.

  2. THE URINE PROTEOME FOR RADIATION BIODOSIMETRY: EFFECT OF TOTAL BODY VERSUS LOCAL KIDNEY IRRADIATION

    PubMed Central

    Sharma, Mukut; Halligan, Brian D.; Wakim, Bassam T.; Savin, Virginia J.; Cohen, Eric P.; Moulder, John E.

    2009-01-01

    Victims of nuclear accidents or radiological terrorism are likely to receive varying doses of ionizing radiation inhomogeneously distributed over the body. Early biomarkers may be useful in determining organ-specific doses due to total body irradiation (TBI) or partial body irradiation. We used liquid chromatography and mass spectrometry to compare the effect of TBI and local kidney irradiation (LKI) on the rat urine proteome using a single 10 Gy dose of X-rays. Both TBI and LKI altered the urinary protein profile within 24 hours with noticeable differences in Gene Ontology categories. Some proteins including fetuin-B, tissue kallikrein, beta-glucuronidase, vitamin D-dependent calcium binding protein and chondroitin sulfate proteoglycan NG2 were detected only in the TBI group. Some other proteins including major urinary protein-1, RNA binding protein 19, neuron navigator, Dapper homolog 3, WD repeat and FYVE domain containing protein 3, sorting nexin-8, ankycorbin and aquaporin were detected only in the LKI group. Protease inhibitors and kidney proteins were more abundant (fraction of total scans) in the LKI group. Up/Uc ratio and urinary albumin abundance decreased in both TBI and LKI groups. Several markers of acute kidney injury were not detectable in either irradiated group. Present data indicate that abundance and number of proteins may follow opposite trends. These novel findings demonstrate intriguing differences between TBI and LKI, and suggest that urine proteome may be useful in determining organ-specific changes caused by partial body irradiation. PMID:20065682

  3. The Urine Preservative Acetic Acid Degrades Urine Protein: Implications for Urine Biorepositories and the AASK Cohort Study.

    PubMed

    Almaani, Salem; Hebert, Lee A; Rovin, Brad H; Birmingham, Daniel J

    2017-05-01

    Patients enrolled in the African American Study of Kidney Disease and Hypertension (AASK) Cohort Study who exhibited overt proteinuria have been reported to show high nonalbumin proteinuria (NAP), which is characteristic of a tubulopathy. To determine whether African American Study of Kidney Disease and Hypertension nephropathy (AASK-N) is a tubulopathy, we obtained urine samples of 37 patients with AASK-N, with 24-hour protein-to-creatinine ratios (milligrams per milligram) ranging from 0.2 to 1.0, from the National Institute of Diabetes and Digestive Kidney Diseases repository and tested for seven markers of tubular proteinuria. By protocol, each sample had been collected in acetic acid (0.5%; mean final concentration). Compared with samples from patients with lupus nephritis or healthy black controls, AASK-N samples had lower amounts of six markers. Four markers (albumin, β -2-microglobulin, cystatin C, and osteopontin) were undetectable in most AASK-N samples. Examination by SDS-PAGE followed by protein staining revealed protein profiles indicative of severe protein degradation in 34 of 37 AASK-N urine samples. Treatment of lupus nephritis urine samples with 0.5% acetic acid produced the same protein degradation profile as that of AASK-N urine. We conclude that the increased NAP in AASK-N is an artifact of acetic acid-mediated degradation of albumin. The AASK-N repository urine samples have been compromised by the acetic acid preservative. Copyright © 2017 by the American Society of Nephrology.

  4. The Urine Preservative Acetic Acid Degrades Urine Protein: Implications for Urine Biorepositories and the AASK Cohort Study

    PubMed Central

    Almaani, Salem; Hebert, Lee A.; Rovin, Brad H.

    2017-01-01

    Patients enrolled in the African American Study of Kidney Disease and Hypertension (AASK) Cohort Study who exhibited overt proteinuria have been reported to show high nonalbumin proteinuria (NAP), which is characteristic of a tubulopathy. To determine whether African American Study of Kidney Disease and Hypertension nephropathy (AASK-N) is a tubulopathy, we obtained urine samples of 37 patients with AASK-N, with 24-hour protein-to-creatinine ratios (milligrams per milligram) ranging from 0.2 to 1.0, from the National Institute of Diabetes and Digestive Kidney Diseases repository and tested for seven markers of tubular proteinuria. By protocol, each sample had been collected in acetic acid (0.5%; mean final concentration). Compared with samples from patients with lupus nephritis or healthy black controls, AASK-N samples had lower amounts of six markers. Four markers (albumin, β-2-microglobulin, cystatin C, and osteopontin) were undetectable in most AASK-N samples. Examination by SDS-PAGE followed by protein staining revealed protein profiles indicative of severe protein degradation in 34 of 37 AASK-N urine samples. Treatment of lupus nephritis urine samples with 0.5% acetic acid produced the same protein degradation profile as that of AASK-N urine. We conclude that the increased NAP in AASK-N is an artifact of acetic acid–mediated degradation of albumin. The AASK-N repository urine samples have been compromised by the acetic acid preservative. PMID:28104821

  5. Correlation of random urine protein creatinine (P-C) ratio with 24-hour urine protein and P-C ratio, based on physical activity: a pilot study.

    PubMed

    Sadjadi, Seyed-Ali; Jaipaul, Navin

    2010-09-07

    Quantification of proteinuria is usually predicated upon 24-hour urine collection. Multiple factors influence urine collection and the rate of protein and creatinine excretion. Urine collection is often incomplete, and therefore creatinine and protein excretion rates are underestimated. A random urine protein-creatinine (P-C) ratio has been shown over the years to be a reliable alternative to the 24-hour collection for detection and follow up of proteinuria. However, urine protein excretion may be influenced by physical activity. We studied 48 patients with proteinuria and varying levels of physical activity to determine the correlation between the measures of urine protein excretion. The correlation coefficient (r) between 24-hour urine total protein and random urine P-C ratio was 0.75 (P < 0.01) in the overall study population, but varied according to the level of proteinuria and physical activity in a stratified analysis: r = 0.99 (P < 0.001) and r = 0.95 (P < 0.01) in bedridden patients; r = 0.44 (P = not significant [NS]) and r = 0.54 (P = NS) in semiactive patients; and r = 0.44 (P = NS) and r = 0.58 (P < 0.05) in active patients with nephrotic- (>3500 mg/day) and non-nephrotic (<3500 mg/day) range proteinuria, respectively. The correlation appeared to be stronger between random urine and 24-hour urine P-C ratio for the overall study population (r = 0.84; P < 0.001), and when stratified according to the level of proteinuria and physical activity: r = 0.99 (P < 0.001) and r = 0.92 (P < 0.01) in bedridden patients; r = 0.61 (P = NS) and r = 0.54 (P = NS) in semiactive patients; and r = 0.64 (P < 0.02) and r = 0.52 (P < 0.05) in active patients with nephrotic and non-nephrotic range proteinuria, respectively. We conclude that the random urine P-C ratio is a reliable and practical way of estimating and following proteinuria, but its precision and accuracy may be affected by the level of patient physical activity.

  6. Correlation of random urine protein creatinine (P-C) ratio with 24-hour urine protein and P-C ratio, based on physical activity: a pilot study

    PubMed Central

    Sadjadi, Seyed-Ali; Jaipaul, Navin

    2010-01-01

    Quantification of proteinuria is usually predicated upon 24-hour urine collection. Multiple factors influence urine collection and the rate of protein and creatinine excretion. Urine collection is often incomplete, and therefore creatinine and protein excretion rates are underestimated. A random urine protein-creatinine (P-C) ratio has been shown over the years to be a reliable alternative to the 24-hour collection for detection and follow up of proteinuria. However, urine protein excretion may be influenced by physical activity. We studied 48 patients with proteinuria and varying levels of physical activity to determine the correlation between the measures of urine protein excretion. The correlation coefficient (r) between 24-hour urine total protein and random urine P-C ratio was 0.75 (P < 0.01) in the overall study population, but varied according to the level of proteinuria and physical activity in a stratified analysis: r = 0.99 (P < 0.001) and r = 0.95 (P < 0.01) in bedridden patients; r = 0.44 (P = not significant [NS]) and r = 0.54 (P = NS) in semiactive patients; and r = 0.44 (P = NS) and r = 0.58 (P < 0.05) in active patients with nephrotic- (>3500 mg/day) and non-nephrotic (<3500 mg/day) range proteinuria, respectively. The correlation appeared to be stronger between random urine and 24-hour urine P-C ratio for the overall study population (r = 0.84; P < 0.001), and when stratified according to the level of proteinuria and physical activity: r = 0.99 (P < 0.001) and r = 0.92 (P < 0.01) in bedridden patients; r = 0.61 (P = NS) and r = 0.54 (P = NS) in semiactive patients; and r = 0.64 (P < 0.02) and r = 0.52 (P < 0.05) in active patients with nephrotic and non-nephrotic range proteinuria, respectively. We conclude that the random urine P-C ratio is a reliable and practical way of estimating and following proteinuria, but its precision and accuracy may be affected by the level of patient physical activity. PMID:20856681

  7. Optimizing Urine Processing Protocols for Protein and Metabolite Detection.

    PubMed

    Siddiqui, Nazema Y; DuBois, Laura G; St John-Williams, Lisa; Will, Thompson J; Grenier, Carole; Burke, Emily; Fraser, Matthew O; Amundsen, Cindy L; Murphy, Susan K

    In urine, factors such as timing of voids, and duration at room temperature (RT) may affect the quality of recovered protein and metabolite data. Additives may aid with detection, but can add more complexity in sample collection or analysis. We aimed to identify the optimal urine processing protocol for clinically-obtained urine samples that allows for the highest protein and metabolite yields with minimal degradation. Healthy women provided multiple urine samples during the same day. Women collected their first morning (1 st AM) void and another "random void". Random voids were aliquotted with: 1) no additive; 2) boric acid (BA); 3) protease inhibitor (PI); or 4) both BA + PI. Of these aliquots, some were immediately stored at 4°C, and some were left at RT for 4 hours. Proteins and individual metabolites were quantified, normalized to creatinine concentrations, and compared across processing conditions. Sample pools corresponding to each processing condition were analyzed using mass spectrometry to assess protein degradation. Ten Caucasian women between 35-65 years of age provided paired 1 st morning and random voided urine samples. Normalized protein concentrations were slightly higher in 1 st AM compared to random "spot" voids. The addition of BA did not significantly change proteins, while PI significantly improved normalized protein concentrations, regardless of whether samples were immediately cooled or left at RT for 4 hours. In pooled samples, there were minimal differences in protein degradation under the various conditions we tested. In metabolite analyses, there were significant differences in individual amino acids based on the timing of the void. For comparative translational research using urine, information about void timing should be collected and standardized. For urine samples processed in the same day, BA does not appear to be necessary while the addition of PI enhances protein yields, regardless of 4°C or RT storage temperature.

  8. OSMOTIC DIURESIS AND ITS EFFECT ON TOTAL ELECTROLYTE DISTRIBUTION IN PLASMA AND URINE OF THE AGLOMERULAR TELEOST, LOPHIUS AMERICANUS

    PubMed Central

    Forster, Roy P.; Berglund, Fredrik

    1956-01-01

    Quantitative evaluations have been made of the chief anions and cations in plasma, urine, and pericardial fluid taken both from freshly captured goosefish and from those undergoing "laboratory diuresis." Measurements included: Na, K, Ca, Mg, Cl, SO4, PO4, protein, HCO3, NH3, pH, titratable acidity, freezing point depression, creatine, trimethylamine oxide, and plasma volume. The total patterns of electrolyte distribution in these body fluids are presented. The morphologically undifferentiated aglomerular tubule acts as a barrier to the free diffusion of monovalent electrolytes, while transporting actively the divalent ions, especially Mg. Urine taken from freshly captured fish is hypotonic to plasma, low in electrolyte, and as much as 50 per cent of its total osmolarity is accounted for by nitrogenous components. Of these creatine is transported most actively by the renal tubule cells. With the onset of diuresis immediately after capture, plasma osmolarity slowly rises and urine suddenly becomes isotonic with plasma as chloride floods into the urine. The active movement of Mg continues during diuresis and urine/plasma concentration ratios of 100 or more are sustained for days while the animals are kept in the laboratory. Na follows chloride and never reaches 50 per cent of plasma values, and K never appears in urine in more than mere traces. Electrolytes in this system are viewed as not being in true equilibrium but rather as constituting a biological steady state with the distribution across renal cells being maintained against passive diffusion by the expenditure of cellular energy. PMID:13286453

  9. Comparison of urine specific gravity values from total-solids refractometry and reagent strip method.

    PubMed

    Chatasingh, S; Tapaneya-Olarn, W

    1989-01-01

    The comparison of specific gravity values of 561 urine samples from TS meter and reagent strip was made. The data were divided into two groups: group 1-less than 2+ protein contained urine samples and group 2--equal or more than 2+ protein contained urine samples. The results revealed that the specific gravity values from both methods in both groups were statistically different (p less than 0.01) but they were correlated at r = 0.84 (p less than 0.001) and r = 0.73 (p less than 0.001) in group 1 and group 2, respectively. It was concluded that the reagent strip is suitable for use as a screening test but it should not be considered when precise measurement is necessary.

  10. Estimation of daily protein intake based on spot urine urea nitrogen concentration in chronic kidney disease patients.

    PubMed

    Kanno, Hiroko; Kanda, Eiichiro; Sato, Asako; Sakamoto, Kaori; Kanno, Yoshihiko

    2016-04-01

    Determination of daily protein intake in the management of chronic kidney disease (CKD) requires precision. Inaccuracies in recording dietary intake occur, and estimation from total urea excretion presents hurdles owing to the difficulty of collecting whole urine for 24 h. Spot urine has been used for measuring daily sodium intake and urinary protein excretion. In this cross-sectional study, we investigated whether urea nitrogen (UN) concentration in spot urine can be used to predict daily protein intake instead of the 24-h urine collection in 193 Japanese CKD patients (Stages G1-G5). After patient randomization into 2 datasets for the development and validation of models, bootstrapping was used to develop protein intake estimation models. The parameters for the candidate multivariate regression models were male gender, age, body mass index (BMI), diabetes mellitus, dyslipidemia, proteinuria, estimated glomerular filtration rate, serum albumin level, spot urinary UN and creatinine level, and spot urinary UN/creatinine levels. The final model contained BMI and spot urinary UN level. The final model was selected because of the higher correlation between the predicted and measured protein intakes r = 0.558 (95 % confidence interval 0.400, 0.683), and the smaller distribution of the difference between the measured and predicted protein intakes than those of the other models. The results suggest that UN concentration in spot urine may be used to estimate daily protein intake and that a prediction formula would be useful for nutritional control in CKD patients.

  11. Inflammatory and fibrotic proteins proteomically identified as key protein constituents in urine and stone matrix of patients with kidney calculi.

    PubMed

    Boonla, Chanchai; Tosukhowong, Piyaratana; Spittau, Björn; Schlosser, Andreas; Pimratana, Chaowat; Krieglstein, Kerstin

    2014-02-15

    To uncover whether urinary proteins are incorporated into stones, the proteomic profiles of kidney stones and urine collected from the same patients have to be explored. We employed 1D-PAGE and nanoHPLC-ESI-MS/MS to analyze the proteomes of kidney stone matrix (n=16), nephrolithiatic urine (n=14) and healthy urine (n=3). We identified 62, 66 and 22 proteins in stone matrix, nephrolithiatic urine and healthy urine, respectively. Inflammation- and fibrosis-associated proteins were frequently detected in the stone matrix and nephrolithiatic urine. Eighteen proteins were exclusively found in the stone matrix and nephrolithiatic urine, considered as candidate biomarkers for kidney stone formation. S100A8 and fibronectin, representatives of inflammation and fibrosis, respectively, were up-regulated in nephrolithiasis renal tissues. S100A8 was strongly expressed in infiltrated leukocytes. Fibronectin was over-expressed in renal tubular cells. S100A8 and fibronectin were immunologically confirmed to exist in nephrolithiatic urine and stone matrix, but in healthy urine they were undetectable. Conclusion, both kidney stones and urine obtained from the same patients greatly contained inflammatory and fibrotic proteins. S100A8 and fibronectin were up-regulated in stone-baring kidneys and nephrolithiatic urine. Therefore, inflammation and fibrosis are suggested to be involved in the formation of kidney calculi. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Diagnostic accuracy of spot urine protein-to-creatinine ratio for proteinuria and its association with adverse pregnancy outcomes in Chinese pregnant patients with pre-eclampsia.

    PubMed

    Cheung, H C; Leung, K Y; Choi, C H

    2016-06-01

    International guidelines have endorsed spot urine protein-to-creatinine ratio of >30 mg protein/mmol creatinine as an alternative to a 24-hour urine sample to represent significant proteinuria. This study aimed to determine the accuracy of spot urine protein-to-creatinine ratio in predicting significant proteinuria and adverse pregnancy outcome. This case series was conducted in a regional obstetric unit in Hong Kong. A total of 120 Chinese pregnant patients with pre-eclampsia delivered at Queen Elizabeth Hospital from January 2011 to December 2013 were included. Relationship of spot urine protein-to-creatinine ratio and 24-hour proteinuria; accuracy of the ratio against 24-hour urine protein at different cut-offs; and relationship of such ratio and adverse pregnancy outcome were studied. Spot urine protein-to-creatinine ratio was correlated with 24-hour urine protein with Pearson correlation coefficient of 0.914 (P<0.0001) when the ratio was <200 mg/mmol. The optimal threshold of spot urine protein-to-creatinine ratio for diagnosing proteinuria in Chinese pregnant patients (33 mg/mmol) was similar to that stated in the international literature (30 mg/mmol). A cut-off of 20 mg/mmol provided a 100% sensitivity, and 52 mg/mmol provided a 100% specificity. There was no significant difference in spot urine protein-to-creatinine ratio between cases with and without adverse pregnancy outcome. Spot urine protein-to-creatinine ratio had a positive and significant correlation with 24-hour urine results in Chinese pre-eclamptic women when the ratio was <200 mg/mmol. Nonetheless, this ratio was not predictive of adverse pregnancy outcome.

  13. The Maillard protein cross-link pentosidine in urine from diabetic patients.

    PubMed

    Takahashi, M; Ohishi, T; Aoshima, H; Kawana, K; Kushida, K; Inoue, T; Horiuchi, K

    1993-07-01

    The Maillard protein cross-link pentosidine is a fluorescent condensation product of lysine, arginine and ribose. It accumulates in human tissues with age, and the accumulation process is accelerated in the tissues of diabetic patients. Using SP-Sephadex C-25 in the pretreatment for HPLC, we examined levels of pentosidine in urine without hydrolysis (free form) and levels of pentosidine in urine after hydrolysis (total forms), from 23 diabetic patients and 21 control subjects. The mean percentages of the values of free form per total forms (+/- SD) were 89 +/- 15% in diabetic patients, 88 +/- 16% in control subjects and 89 +/- 15% in total populations of diabetic patients and control subjects. There was a significant correlation between the values of free form and total forms in diabetic patients (r = 0.983, p = 0.0001), in control subjects (r = 0.820, p < 0.02) and in total populations of diabetic patients and control subjects (r = 0.951, p = 0.0001). The mean level of pentosidine per mol creatinine (+/- SD) was significantly elevated in urine from diabetic patients as compared to the level in control subjects (8.8 +/- 4.3 mumol/mol creatinine vs 4.2 +/- 1.4 mumol/mol creatinine, p = 0.0001 in free form; 10.1 +/- 5.3 mumol/mol creatinine vs 4.7 +/- 1.4 mumol/mol creatinine, p = 0.0001 in total forms). These results demonstrate that urinary pentosidine, especially in free form, could be a useful marker for the assessment of diabetes and diabetic complications.

  14. [Urine metabonomic study on long-term use of total ginsenosides in rats].

    PubMed

    Xie, Xie; Chen, Shao-Qiu; Lv, Ying-Fang; Wang, Xiao-Yan; Jia, Wei

    2014-12-01

    Due to its effect of systems regulation and promotion on body, Ginseng is always referred to be long-term used as a dietary supplement. But it was still unclear about its target of the tonic effects and also the side-effects long-term use may bring. Urine metabolomic method is suitable for long-term studies of pharmaco-dynamics, pharmacology and toxicology of traditional Chinese medicine because of its characteristics of non-invasive and monitoring the whole-body metabolism. This study was designed to detect the dynamic variation of rat urine metabolome along with a long-term administration of total ginsenosides using GC-TOF based metabolomic technology. Our result showed that either short-term or chronic administration of ginsenosides did not impact the rat urine metabolome significantly (as the PCA subgroup was not successful). By comparison, the short-term (1-3 w) dose of ginsenosides had the biggest metabolic influence including TCA cycle, catecholamines and neurotransmitter amino acids. Medium-term (6-10 w) dose had a gradually lower effect and long-term (27 w) dose almost had no effect. Our study indicates that both short and long-term administration of ginsenosides showed almost no obvious side-effect on the experimental animals.

  15. Exploratory study of proteins in urine of patients with histoplasma antigenuria.

    PubMed

    Kushnir, Mark M; Crockett, David K; Cloud, Joann L; Ashwood, Edward R; Rockwood, Alan L

    2012-02-01

    Disseminated histoplasmosis is an invasive fungal infection that can be fatal in patients with weak immune system. The goal of our exploratory study was to evaluate differences in urinary protein profiles among samples of healthy individuals, patients with proteinuria (PRU), and histoplasma antigenuria (HIS), and to identify physiological pathways associated with the excreted proteins. Urine samples were depleted of abundant proteins, deglycosylated, digested with trypsin, fractionated and analyzed by nano-LC-QTOF. The total number of human proteins identified in the samples was 117, of which 20 and 23 were unique to the samples from patients with PRU and HIS, respectively. Pathway analysis of proteins identified in samples of PRU and HIS patients suggested increased levels of proteins associated with acute response signaling, coagulation system, prothrombin activation, glucocorticoid regulation and the lipid antigen presentation signaling pathway networks. The obtained data provide information on protein expression associated with HIS, and suggest that further more rigorous studies aimed at the identification of proteins associated with proteinuria of different causes are feasible. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Total arsenic concentrations in Chinese children's urine by different geographic locations, ages, and genders.

    PubMed

    Zhang, Xuan; Wang, Beibei; Cui, Xiaoyong; Lin, Chunye; Liu, Xitao; Ma, Jin

    2018-06-01

    Little is known about the variation of Chinese children's exposure to arsenic by geography, age, gender, and other potential factors. The main objective of this study was to investigate the total arsenic concentration in Chinese children's urine by geographic locations, ages, and genders. In total, 259 24-h urine samples were collected from 210 2- to 12-year-old children in China and analyzed for total arsenic and creatinine concentrations. The results showed that the upper limit (upper limit of the 90% confidence interval for the 97.5 fractile) was 27.51 µg/L or 55.88 µg/g creatinine for Chinese children. The total urinary arsenic levels were significantly different for children in Guangdong, Hubei, and Gansu provinces (P < 0.05), where the upper limits were 24.29, 58.70, and 44.29 µg/g creatinine, respectively. In addition, the total urinary arsenic levels were higher for 2- to 7-year-old children than for 7- to 12-year-old children (P < 0.05; the upper limits were 59.06 and 44.29 µg/g creatinine, respectively) and higher for rural children than for urban children (P < 0.05; the upper limits were 59.06 and 50.44 µg/g creatinine, respectively). The total urinary arsenic levels for boys were not significantly different from those for girls (P > 0.05), although the level for boys (the upper limit was 59.30 µg/g) was slightly higher than that for girls (the upper limit was 58.64 µg/g creatinine). Because the total urinary arsenic concentrations are significantly different for general populations of children in different locations and age groups, the reference level of total urinary arsenic might be dependent on the geographic site and the child's age.

  17. The efficacy of semi-quantitative urine protein-to-creatinine (P/C) ratio for the detection of significant proteinuria in urine specimens in health screening settings.

    PubMed

    Chang, Chih-Chun; Su, Ming-Jang; Ho, Jung-Li; Tsai, Yu-Hui; Tsai, Wei-Ting; Lee, Shu-Jene; Yen, Tzung-Hai; Chu, Fang-Yeh

    2016-01-01

    Urine protein detection could be underestimated using the conventional dipstick method because of variations in urine aliquots. This study aimed to assess the efficacy of the semi-quantitative urine protein-to-creatinine (P/C) ratio compared with other laboratory methods. Random urine samples were requested from patients undergoing chronic kidney disease screening. Significant proteinuria was determined by the quantitative P/C ratio of at least 150 mg protein/g creatinine. The semi-quantitative P/C ratio, dipstick protein and quantitative protein concentrations were compared and analyzed. In the 2932 urine aliquots, 156 (5.3 %) urine samples were considered as diluted and 60 (39.2 %) were found as significant proteinuria. The semi-quantitative P/C ratio testing had the best sensitivity (70.0 %) and specificity (95.9 %) as well as the lowest underestimation rate (0.37 %) when compared to other laboratory methods in the study. In the semi-quantitative P/C ratio test, 19 (12.2 %) had positive, 52 (33.3 %) had diluted, and 85 (54.5 %) had negative results. Of those with positive results, 7 (36.8 %) were positive detected by traditional dipstick urine protein test, and 9 (47.4 %) were positive detected by quantitative urine protein test. Additionally, of those with diluted results, 25 (48.1 %) had significant proteinuria, and all were assigned as no significant proteinuria by both tests. The semi-quantitative urine P/C ratio is clinically applicable based on its better sensitivity and screening ability for significant proteinuria than other laboratory methods, particularly in diluted urine samples. To establish an effective strategy for CKD prevention, urine protein screening with semi-quantitative P/C ratio could be considered.

  18. Total Extracellular Small RNA Profiles from Plasma, Saliva, and Urine of Healthy Subjects

    PubMed Central

    Yeri, Ashish; Courtright, Amanda; Reiman, Rebecca; Carlson, Elizabeth; Beecroft, Taylor; Janss, Alex; Siniard, Ashley; Richholt, Ryan; Balak, Chris; Rozowsky, Joel; Kitchen, Robert; Hutchins, Elizabeth; Winarta, Joseph; McCoy, Roger; Anastasi, Matthew; Kim, Seungchan; Huentelman, Matthew; Van Keuren-Jensen, Kendall

    2017-01-01

    Interest in circulating RNAs for monitoring and diagnosing human health has grown significantly. There are few datasets describing baseline expression levels for total cell-free circulating RNA from healthy control subjects. In this study, total extracellular RNA (exRNA) was isolated and sequenced from 183 plasma samples, 204 urine samples and 46 saliva samples from 55 male college athletes ages 18–25 years. Many participants provided more than one sample, allowing us to investigate variability in an individual’s exRNA expression levels over time. Here we provide a systematic analysis of small exRNAs present in each biofluid, as well as an analysis of exogenous RNAs. The small RNA profile of each biofluid is distinct. We find that a large number of RNA fragments in plasma (63%) and urine (54%) have sequences that are assigned to YRNA and tRNA fragments respectively. Surprisingly, while many miRNAs can be detected, there are few miRNAs that are consistently detected in all samples from a single biofluid, and profiles of miRNA are different for each biofluid. Not unexpectedly, saliva samples have high levels of exogenous sequence that can be traced to bacteria. These data significantly contribute to the current number of sequenced exRNA samples from normal healthy individuals. PMID:28303895

  19. Dietary protein-induced increases in urine calcium are accompanied by similar increases in urine nitrogen and urine urea: a controlled clinical trial

    PubMed Central

    Bihuniak, Jessica D.; Simpson, Christine A.; Sullivan, Rebecca R.; Caseria, Donna M.; Kerstetter, Jane E.; Insogna, Karl L.

    2018-01-01

    To determine the usefulness of urine urea (UU) as an index of dietary protein intake 10 postmenopausal women were enrolled and completed a randomized, double-blind, cross-over feeding trial, from September 2008 to May 2010, comparing ten days of a 45g whey supplement to ten days of a 45 g maltodextrin control. Urine nitrogen (UN), calcium (UCa), UU and bone turnover markers were measured at days 0, 7, and 10. Paired sample t tests, Pearson’s correlation statistic, and simple linear regression were used to assess differences between treatments, and associations among urinary metabolites. UN/urinary creatinine (UCreat) rose from 12.3 ± 1.7 g/g (99.6 ± 13.8 mmol/mmol) to 16.8 ± 2.2 g/g (135.5 ± 17.8 mmol/mmol) with whey supplementation but did not change with maltodextrin. Whey supplementation caused UCa to rise by 4.76 ± 1.84 mg (1.19 ± 0.46 mmol) without a change in bone turnover markers. Since our goal was to estimate protein intake from UN/UCreat, we used our data to develop the following equation: protein intake (g/d) = 71.221 + 1.719×(UN, g)/Creat, g) (R = 0.46, R2 = 0.21). As a more rapid and less costly alternative to UN/UCreat, we next determined if urinary urea (UU) could predict protein intake and found that protein intake (g/d) = 63.844 + 1.11×(UU, g/Creat, g) (R = 0.58, R2 = 0.34). These data indicate that UU/UCreat is at least as good a marker of dietary protein intake as is urinary nitrogen and easier to quantitate in nutrition intervention trials. PMID:23438496

  20. Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine.

    PubMed

    Filip, Szymon; Vougas, Konstantinos; Zoidakis, Jerome; Latosinska, Agnieszka; Mullen, William; Spasovski, Goce; Mischak, Harald; Vlahou, Antonia; Jankowski, Joachim

    2015-01-01

    Proteome analysis of complex biological samples for biomarker identification remains challenging, among others due to the extended range of protein concentrations. High-abundance proteins like albumin or IgG of plasma and urine, may interfere with the detection of potential disease biomarkers. Currently, several options are available for the depletion of abundant proteins in plasma. However, the applicability of these methods in urine has not been thoroughly investigated. In this study, we compared different, commercially available immunodepletion and ion-exchange based approaches on urine samples from both healthy subjects and CKD patients, for their reproducibility and efficiency in protein depletion. A starting urine volume of 500 μL was used to simulate conditions of a multi-institutional biomarker discovery study. All depletion approaches showed satisfactory reproducibility (n=5) in protein identification as well as protein abundance. Comparison of the depletion efficiency between the unfractionated and fractionated samples and the different depletion strategies, showed efficient depletion in all cases, with the exception of the ion-exchange kit. The depletion efficiency was found slightly higher in normal than in CKD samples and normal samples yielded more protein identifications than CKD samples when using both initial as well as corresponding depleted fractions. Along these lines, decrease in the amount of albumin and other targets as applicable, following depletion, was observed. Nevertheless, these depletion strategies did not yield a higher number of identifications in neither the urine from normal nor CKD patients. Collectively, when analyzing urine in the context of CKD biomarker identification, no added value of depletion strategies can be observed and analysis of unfractionated starting urine appears to be preferable.

  1. Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine

    PubMed Central

    Filip, Szymon; Vougas, Konstantinos; Zoidakis, Jerome; Latosinska, Agnieszka; Mullen, William; Spasovski, Goce; Mischak, Harald; Vlahou, Antonia; Jankowski, Joachim

    2015-01-01

    Proteome analysis of complex biological samples for biomarker identification remains challenging, among others due to the extended range of protein concentrations. High-abundance proteins like albumin or IgG of plasma and urine, may interfere with the detection of potential disease biomarkers. Currently, several options are available for the depletion of abundant proteins in plasma. However, the applicability of these methods in urine has not been thoroughly investigated. In this study, we compared different, commercially available immunodepletion and ion-exchange based approaches on urine samples from both healthy subjects and CKD patients, for their reproducibility and efficiency in protein depletion. A starting urine volume of 500 μL was used to simulate conditions of a multi-institutional biomarker discovery study. All depletion approaches showed satisfactory reproducibility (n=5) in protein identification as well as protein abundance. Comparison of the depletion efficiency between the unfractionated and fractionated samples and the different depletion strategies, showed efficient depletion in all cases, with the exception of the ion-exchange kit. The depletion efficiency was found slightly higher in normal than in CKD samples and normal samples yielded more protein identifications than CKD samples when using both initial as well as corresponding depleted fractions. Along these lines, decrease in the amount of albumin and other targets as applicable, following depletion, was observed. Nevertheless, these depletion strategies did not yield a higher number of identifications in neither the urine from normal nor CKD patients. Collectively, when analyzing urine in the context of CKD biomarker identification, no added value of depletion strategies can be observed and analysis of unfractionated starting urine appears to be preferable. PMID:26208298

  2. A comparison of direct and indirect analytical approaches to measuring total nicotine equivalents in urine.

    PubMed

    Taghavi, Taraneh; Novalen, Maria; Lerman, Caryn; George, Tony P; Tyndale, Rachel F

    2018-05-31

    Total nicotine equivalents (TNE), the sum of nicotine and metabolites in urine, is a valuable tool for evaluating nicotine exposure. Most methods for measuring TNE involve two-step enzymatic hydrolysis for indirect quantification of glucuronide metabolites. Here, we describe a rapid, low-cost direct liquid chromatography - tandem mass spectrometry (LCMS) assay. In 139 smokers' urine samples, Bland-Altman, correlation, and regression analyses were used to investigate differences in quantification of nicotine and metabolites, TNE, and nicotine metabolite ratio (NMR) between direct and indirect LCMS methods. DNA from a subset (n=97 smokers) was genotyped for UGT2B10*2 and UGT2B17*2 and the known impact of these variants was evaluated using urinary ratios determined by the direct versus indirect method. The direct method showed high accuracy (0-9% bias) and precision (3-14% coefficient of variation) with similar distribution of nicotine metabolites to literary estimates and good agreement between the direct and indirect methods for nicotine, cotinine, and 3-hydroxycotinine (ratios 0.99-1.07), but less agreement for their respective glucuronides (ratios 1.16-4.17). The direct method identified urinary 3HC+3HC-GLUC/COT as having the highest concordance with plasma NMR and provided substantially better estimations of the established genetic impact of glucuronidation variants compared to the indirect method. Direct quantification of nicotine and metabolites is less time-consuming and less costly, and provides accurate estimates of nicotine intake, metabolism rate and the impact of genetic variation in smokers. Lower cost and maintenance combined with high accuracy and reproducibility make the direct method ideal for smoking biomarker, NMR and pharmacogenomics studies. Copyright ©2018, American Association for Cancer Research.

  3. Evaluation of the Urine Protein/Creatinine Ratio Measured with the Dipsticks Clinitek Atlas PRO 12.

    PubMed

    Hermida, Fernando J; Soto, Sonia; Benitez, Alfonso J

    2016-01-01

    Screening for urine proteins is recommended for the detection of albuminuria in high risk groups. The aim of this study was to compare the Clinitek Atlas PRO12 reagent urine strip with quantitative methods for the determination of protein/creatinine ratio and to evaluate the usefulness of the semi-quantitative Clinitek Atlas PRO12 reagent urine strip as a tool in the early detection of albuminuria among the general population. Six hundred first morning urine specimens were collected from outpatients with various clinical conditions. The results showed that the test data for the urine dipstick Clinitek Atlas PRO12 show good agreement with the quantitative measurement of protein, creatinine and protein/creatinine ratio. In addition, this study shows that 97.2% of the samples which gave "normal" protein/creatinine ratios by the semi-quantitative method, showed albumin/creatinine ratio < 30 mg/g by the quantitative methods. Our results show that Clinitek Atlas PRO12 reagent strips can be used for the purposes of albuminuria screening in the general population.

  4. Minimum urine flow rate during water deprivation: importance of the nonurea versus total osmolality in the inner medulla.

    PubMed

    Soroka, S D; Chayaraks, S; Cheema-Dhadli, S; Myers, J A; Rubin, S; Sonnenberg, H; Halperin, M L

    1997-06-01

    Antidiuretic hormone leads to an increase in the permeability for water and urea in the inner medullary collecting duct. Hence, urea may not be an "effective" osmole in the inner medulla during maximal renal water conservation. Accordingly, the purpose of this study was to evaluate whether differences in the rate of urea excretion would influence maximum renal water conservation in humans. In water-deprived rats, the concentration of urea and total osmolality were somewhat higher in the urine exiting the inner medullary collecting duct than in interstitial fluid obtained from the entire papillary tip. Nevertheless, the "nonurea" (total osmolality minus urea in millimolar terms) osmolality was virtually identical in both locations. Chronically fasted human subjects that were water-deprived for 16 h had a lower rate of urea excretion (71 +/- 7 versus 225 +/- 14 mumol/min) and a somewhat lower urine osmolality (745 +/- 53 versus 918 +/- 20 mosmol/kg H2O). Nevertheless, they had identical urine flow rates (0.5 +/- 0.01 and 0.5 +/- 0.02 ml/min, respectively), and their nonurea osmolality also was similar (587 +/- 25 and 475 +/- 14 mosmol/kg H2O, respectively) to the water-deprived normal subjects. The composition of their urine differed in that the principal nonurea osmoles became NH4+ and beta-hydroxybutyrate rather than Na and C1. During water deprivation in normal subjects, the ingestion of urea caused a twofold rise in urine flow rate, a fall in the nonurea osmolality, and a rise in the rate of excretion of nonurea osmoles. The nonurea osmolality of the urine, and presumably the medullary interstitial fluid as well, was inversely related to the urea excretion rate. In chronic fasting, the nature, but not the quantity, of nonurea osmoles changed. The similar minimum urine volume was predictable from an analysis based on nonurea osmole considerations.

  5. Interlaboratory trial for the measurement of total cobalt in equine urine and plasma by ICP-MS.

    PubMed

    Popot, Marie-Agnes; Ho, Emmie N M; Stojiljkovic, Natali; Bagilet, Florian; Remy, Pierre; Maciejewski, Pascal; Loup, Benoit; Chan, George H M; Hargrave, Sabine; Arthur, Rick M; Russo, Charlie; White, James; Hincks, Pamela; Pearce, Clive; Ganio, George; Zahra, Paul; Batty, David; Jarrett, Mark; Brooks, Lydia; Prescott, Lise-Anne; Bailly-Chouriberry, Ludovic; Bonnaire, Yves; Wan, Terence S M

    2017-09-01

    Cobalt is an essential mineral micronutrient and is regularly present in equine nutritional and feed supplements. Therefore, cobalt is naturally present at low concentrations in biological samples. The administration of cobalt chloride is considered to be blood doping and is thus prohibited. To control the misuse of cobalt, it was mandatory to establish an international threshold for cobalt in plasma and/or in urine. To achieve this goal, an international collaboration, consisting of an interlaboratory comparison between 5 laboratories for the urine study and 8 laboratories for the plasma study, has been undertaken. Quantification of cobalt in the biological samples was performed by inductively coupled plasma-mass spectrometry (ICP-MS). Ring tests were based on the analysis of 5 urine samples supplemented at concentrations ranging from 5 up to 500 ng/mL and 5 plasma samples spiked at concentrations ranging from 0.5 up to 25 ng/mL. The results obtained from the different laboratories were collected, compiled, and compared to assess the reproducibility and robustness of cobalt quantification measurements. The statistical approach for the ring test for total cobalt in urine was based on the determination of percentage deviations from the calculated means, while robust statistics based on the calculated median were applied to the ring test for total cobalt in plasma. The inter-laboratory comparisons in urine and in plasma were successful so that 97.6% of the urine samples and 97.5% of the plasma samples gave satisfactory results. Threshold values for cobalt in plasma and urine were established from data only obtained by laboratories involved in the ring test. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  6. Elevated urine levels of heparin-binding protein in children with urinary tract infection.

    PubMed

    Kjölvmark, Charlott; Akesson, Per; Linder, Adam

    2012-08-01

    Urinary tract infection (UTI) is a common infection diagnosis in children, and efficient diagnosis and treatment are important to avoid serious complications. In this study we investigated whether urinary levels of neutrophil-derived heparin-binding protein (HBP) can be used as a marker of UTI in children. These results were compared to those of dipstick analysis, interleukin-6 (IL-6) analysis in urine, and bacterial culturing. Seventy-eight children aged 0-18 years with fever and/or symptoms indicating UTI were enrolled in a prospective consecutive study. Urine samples were cultured and analyzed with dipstick, and concentrations of HBP and IL-6 were measured. Fifteen patients were classified as having UTI, 30 patients had fever but were diagnosed with a non-urinary tract infection, and 33 patients had neither UTI nor fever. Using a urine HBP (U-HBP) cut-off level of 32 ng/mL, the sensitivity and specificity for detecting UTI were 93.3 and 90.3 %, respectively. Receiver operating characteristic curves demonstrated that U-HBP levels were a higher specificity indicator of UTI than urine white blood cell counts or urine IL-6 levels; they also showed a higher sensitivity than the results of the urine nitrite test. All patients with significant growth of clinically relevant bacteria had elevated U-HBP levels. The results indicate that rapid analysis of U-HBP can provide helpful guidance in the management of children with suspected UTI.

  7. Effects of storage conditions on results for quantitative and qualitative evaluation of proteins in canine urine.

    PubMed

    Théron, Marie-Laure; Piane, Laetitia; Lucarelli, Laetitia; Henrion, Rémi; Layssol-Lamour, Catherine; Palanché, Florence; Concordet, Didier; Braun, Jean-Pierre D; Trumel, Catherine; Lavoué, Rachel

    2017-08-01

    OBJECTIVE To investigate effects of storage conditions on the canine urine protein-to-creatinine ratio (UPC) and on SDS-agarose gel electrophoresis (AGE) of urinary proteins. SAMPLE Urine specimens from 20 proteinuric (UPC > 0.5) and 20 nonproteinuric (UPC ≤ 0.2) dogs. PROCEDURES UPC and SDS-AGE were performed on urine specimens stored at room temperature (20°C) and 4°C for up to 5 days and at -20° and -80°C for up to 360 days; some specimens were subjected to 3 freeze-thaw cycles. Results were compared with those obtained for fresh urine specimens. RESULTS UPC was not affected by storage at room temperature or by freezing. A decrease in UPC was observed for specimens from nonproteinuric dogs after 5 days at 4°C (10%) and from both groups after 90 days at -20° and -80°C (≤ 20% and ≤ 15%, respectively). The SDS-AGE profiles revealed no visual changes regardless of duration of storage for specimens stored at room temperature, 4°C, and -80°C, except for 1 profile after 360 days at -80°C. Repeated freeze-thaw cycles did not affect SDS-AGE profiles. Appearance or strengthening of high-molecular-weight bands that could alter interpretation was evident in SDS-AGE profiles after storage at -20°C for ≥ 15 days (31/40 dogs). CONCLUSIONS AND CLINICAL RELEVANCE Storage of urine at -20° or -80°C for up to 1 year influenced the UPC without affecting clinical interpretation. Storage of urine specimens at -20°C impaired visual analysis of SDS-AGE. When SDS-AGE cannot be performed on fresh or recently refrigerated urine specimens, storage at -80°C is recommended.

  8. Total protein, albumin and low-molecular-weight protein excretion in HIV-positive patients.

    PubMed

    Campbell, Lucy J; Dew, Tracy; Salota, Rashim; Cheserem, Emily; Hamzah, Lisa; Ibrahim, Fowzia; Sarafidis, Pantelis A; Moniz, Caje F; Hendry, Bruce M; Poulton, Mary; Sherwood, Roy A; Post, Frank A

    2012-08-10

    Chronic kidney disease is common in HIV positive patients and renal tubular dysfunction has been reported in those receiving combination antiretroviral therapy (cART). Tenofovir (TFV) in particular has been linked to severe renal tubular disease as well as proximal tubular dysfunction. Markedly elevated urinary concentrations of retinal-binding protein (RBP) have been reported in patients with severe renal tubular disease, and low-molecular-weight proteins (LMWP) such as RBP may be useful in clinical practice to assess renal tubular function in patients receiving TFV. We analysed 3 LMWP as well as protein and albumin in the urine of a sample of HIV positive patients. In a cross-sectional fashion, total protein, albumin, RBP, cystatin C, and neutrophil gelatinase-associated lipocalin (NGAL) were quantified in random urine samples of 317 HIV positive outpatients and expressed as the ratio-to-creatinine (RBPCR, CCR and NGALCR). Exposure to cART was categorised as none, cART without TFV, and cART containing TFV and a non-nucleoside reverse-transcriptase-inhibitor (TFV/NNRTI) or TFV and a protease-inhibitor (TFV/PI). Proteinuria was present in 10.4 % and microalbuminuria in 16.7 % of patients. Albumin accounted for approximately 10 % of total urinary protein. RBPCR was within the reference range in 95 % of patients while NGALCR was elevated in 67 % of patients. No overall differences in urine protein, albumin, and LMWP levels were observed among patients stratified by cART exposure, although a greater proportion of patients exposed to TFV/PI had RBPCR >38.8 μg/mmol (343 μg/g) (p = 0.003). In multivariate analyses, black ethnicity (OR 0.43, 95 % CI 0.24, 0.77) and eGFR <75 mL/min/1.73 m2 (OR 3.54, 95 % CI 1.61, 7.80) were independently associated with upper quartile (UQ) RBPCR. RBPCR correlated well to CCR (r2 = 0.71), but not to NGALCR, PCR or ACR. In HIV positive patients, proteinuria was predominantly of tubular origin and microalbuminuria

  9. Effect of vitamin C and E supplementation on total antioxidant content of human breastmilk and infant urine.

    PubMed

    Zarban, Asghar; Toroghi, Mahsa Mostafavi; Asli, Marziye; Jafari, Masumeh; Vejdan, Morteza; Sharifzadeh, Gholamreza

    2015-05-01

    After delivery and birth, mothers and neonates are exposed to oxidative stress. The present study examined the effect of supplementation of the diet of breastfeeding mothers with vitamin C and E to improve the antioxidant content of breastmilk and evidence of antioxidant activity in infant urine. The subjects were 60 healthy lactating breastfeeding mothers and their infants 1-6 months of age. They were randomly allocated to a control group (n=30) consuming a free diet or an experimental group (n=30) consuming a free diet supplemented each day with effervescent tablets of vitamin C (500 mg) and chewable tablets of vitamin E (100 IU). After 30 days, the total antioxidant content of the mothers' breastmilk and evidence of antioxidant activity in the infants' urine were measured by the ferric reducing/antioxidant power assay. The free radical scavenging activity of the urine samples was measured by the α,α-diphenyl-β-picrylhydrazyl method. Differences pre- and postintervention were compared within and between the groups. Significantly higher levels of antioxidants in the breastmilk (610±295.5 to 716±237.5 μmol/L) and infant urine (43.2±21.8 to 75.0±49.2 μmol/mg creatinine) were observed in the experimental group over the control group (p<0.05). A significant increase in evidence of free radical scavenging in infant urine was observed in the experimental group after 30 days of supplementation by mothers (p<0.05). Consumption of vitamin C and E supplements appears to have a positive effect on total antioxidant content of breastmilk and evidence of antioxidant activity in infant urine.

  10. Identification of urine protein biomarkers with the potential for early detection of lung cancer.

    PubMed

    Zhang, Hongjuan; Cao, Jing; Li, Lin; Liu, Yanbin; Zhao, Hong; Li, Nan; Li, Bo; Zhang, Aiqun; Huang, Huanwei; Chen, She; Dong, Mengqiu; Yu, Lei; Zhang, Jian; Chen, Liang

    2015-07-02

    Lung cancer is the leading cause of cancer-related deaths and has an overall 5-year survival rate lower than 15%. Large-scale clinical trials have demonstrated a significant relative reduction in mortality in high-risk individuals with low-dose computed tomography screening. However, biomarkers capable of identifying the most at-risk population and detecting lung cancer before it becomes clinically apparent are urgently needed in the clinic. Here, we report the identification of urine biomarkers capable of detecting lung cancer. Using the well-characterized inducible Kras (G12D) mouse model of lung cancer, we identified alterations in the urine proteome in tumor-bearing mice compared with sibling controls. Marked differences at the proteomic level were also detected between the urine of patients and that of healthy population controls. Importantly, we identified 7 proteins commonly found to be significantly up-regulated in both tumor-bearing mice and patients. In an independent cohort, we showed that 2 of the 7 proteins were up-regulated in urine samples from lung cancer patients but not in those from controls. The kinetics of these proteins correlated with the disease state in the mouse model. These tumor biomarkers could potentially aid in the early detection of lung cancer.

  11. Random spot urine protein to creatinine ratio is a reliable measure of proteinuria in lupus nephritis in Koreans.

    PubMed

    Choi, In Ah; Park, Jin Kyun; Lee, Eun Young; Song, Yeong Wook; Lee, Eun Bong

    2013-01-01

    The accurate assessment of proteinuria is critical for the management of lupus nephritis. Measuring the protein to creatinine (P/C) ratio in random spot urine (RSU) samples has been introduced as an alternative to the 24-hour (24h) urine collection method. However, it remains unclear as to whether the RSU P/C ratio is reliable for assessing lupus nephritis (LN) in routine clinical practice. In total, 275 pairs of 24h urine and RSU samples from 102 patients with biopsy-proven LN were analysed. The correlation and concordance between the P/C ratios in the two sample types were assessed by Pearson or Spearman correlation and intra-class correlation coefficient (ICC) using mixed models for repeated measurements, respectively. The mean 24h urine P/C ratio was 3.2 ± 4.9. Overall, RSU P/C ratio correlated strongly with the 24h urine P/C ratio (r=0.944, p<0.001) with an excellent agreement (ICC=0.949, 95% confidence interval [CI]: 0.69-1.00). Subgroup analyses revealed that the correlation remained high in class II, III, IV, and V LN (rho=0.868, p<0.001; rho=0.649, p=0.007; r=0.945, p<0.001; and rho=0.900, p=0.001, respectively). The correlation between the 24h urine and RSU P/C ratio in the range of 0.5 to 3 was good (r=0.720, p<0.001) with ICC of 0.659 (95%CI 0.554-0.812). RSU P/C ratio ≥0.5 could predict 24h PCR ≥0.5 with 91.7% sensitivity and 70.2% specificity, whereas RSU P/C ratio ≥1.0 increased specificity up to 94.7%. The RSU P/C ratio is an excellent alternative to the 24 hour P/C ratio for assessing the presence of clinically significant proteinuria in LN. RSU P/C ratio >1.0 may prompt directly to a renal biopsy, whereas RSU P/C ratio between 0.5-1.0 should be followed by a confirmatory 24h urine collection.

  12. Simultaneous Analysis of Seven Biomarkers of Oxidative Damage to Lipids, Proteins, and DNA in Urine.

    PubMed

    Martinez, Maria P; Kannan, Kurunthachalam

    2018-06-05

    The determination of oxidative stress biomarkers (OSBs) is useful for the assessment of health status and progress of diseases in humans. Whereas previous methods for the determination of OSBs in urine were focused on a single marker, in this study, we present a method for simultaneous determination of biomarkers of oxidative damage to lipids, proteins, and DNA. 2,4-Dinitrophenylhydrazine (DNPH) derivatization followed by solid phase extraction (SPE) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) allowed the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG), o- o'-dityrosine (diY), malondialdehyde (MDA), and four F 2 -isoprostane isomers: 8-iso-prostaglandinF 2α (8-PGF 2α ), 11β-prostaglandinF 2α (11-PGF 2α ), 15( R)-prostaglandinF 2α (15-PGF 2α ), and 8-iso,15( R)-prostaglandinF 2α (8,15-PGF 2α ) in urine. Derivatization with DNPH and SPE was optimized to yield greater sensitivity and selectivity for the analysis of target chemicals. The limits of detection of target analytes in urine were below 30 pg mL -1 . The assay intra- and interday variability was below 16% of the relative standard deviation, and the recoveries of target chemicals spiked into synthetic urine were near 100%. The method was applied to the analysis of 21 real urine samples, and the analytes were found at a detection frequency of 85% for 8-PGF 2α and 15-PGF 2α , 71% for 11-PGF 2α , 81% for 8,15-PGF 2α , and 100% for diY, 8-OHdG, and MDA. This method offers simultaneous determination of multiple OSBs of different molecular origin in urine samples selectively with high accuracy and precision.

  13. [Validity of the assessment of urinary protein excretion by spot urine in patients with chronic kidney disease].

    PubMed

    Hayashi, Ami; Okada, Tomonari; Matsumoto, Hiroshi; Nagaoka, Yume; Wada, Toshikazu; Gondo, Asako; Nango, Tomoka; Miyaoka, Yoshitaka; Watanabe, Kanna; Iwata, Azusa; Nakao, Toshiyuki

    2013-01-01

    We investigate the validity of the assessment of urinary protein excretion by spot urine samples collected by different methods in outpatients with chronic kidney disease (CKD). SUBJECTS AND METHODS We obtained 24-hour urine and two spot urine samples, including the first morning urine and daytime urine in 159 CKD patients. Urinary protein excretion was assessed by the protein/creatinine ratio from spot urine samples (morning: m-UP (g/gCr), daytime: d-UP (g/gCr) ]. We examined the correlations and the differences among m-UP, d-UP and the actual urinary protein excretion obtained by 24-hour urine (a-UP(g/day) . Significant correlations were found between m-UP and a-UP, and between d-UP and a-UP (r = 0.88, 0.85; p < 0.001). Correlations between m-UP and a-UP were greater relative to those between d-UP and a-UP in patients with less than 3.5 g/day of a-UP and in patients with CKD stages 1 to approximately 3. The percent difference between m-UP and a-UP was--16.0 +/- 40.5%, and that between d-UP and a-UP was 27.1 +/- 72.9%. The absolute value of the percent difference between d-UP and a-UP tended to be greater than that between m-UP and a-UP (34.9 +/- 25.9% vs. 49.9 +/- 59.9%, p = 0.06). Urinary protein/creatinie ratio of the first morning urine is better approximate the urinary protein excretion obtained by 24-hour urine compared with that of spot urine in the daytime.

  14. Fluid consumption, total water intake and first morning urine osmolality in Spanish adolescents from Zaragoza: data from the HELENA study.

    PubMed

    Iglesia, I; Santaliestra-Pasías, A M; Bel-Serrat, S; Sadalla-Collese, T; Miguel-Berges, M L; Moreno, L A

    2016-05-01

    The objective of the study is to describe the hydration status and to assess the main food- and/or fluid intake-associated factors in healthy adolescents. A total of 194 adolescents from the city of Zaragoza aged 12.5-17.5 years (99 males) participating in the 'Healthy Lifestyle in Europe by Nutrition in Adolescence' (HELENA) cross-sectional study were included. First morning urine was collected, and osmolality was determined by freezing point depression osmometer. A self-reported computer-based 24-h dietary recall was applied the same day of the urine collection. Analysis of variance, Kruskal-Wallis procedure or Pearson's χ(2) analyses was used to examine the group associations. Seventy-one percent of adolescents did not meet the European Food Safety Agency (EFSA) recommendations for average total water intake (TWI), and 68% had high first morning urine osmolality values. TWI and the proportion of those meeting EFSA reference values significantly (P<0.05) decreased with increased osmolality. Males who met the EFSA recommendations consumed significantly (P<0.05) more plain water (1035.13 vs 582.68 ml) and dairy drinks (368.13 vs 226.68 ml) than those who did not. In females, the consumption of water (1359.41 vs 620.44 ml) and sugar-sweetened beverages (214.61 vs 127.42 ml) was significantly higher in those meeting the EFSA recommendations than in those who did not. First morning urine osmolality was associated with vegetables (unstandardized β: -0.6; 95% confidence interval (CI): -1.02 and -0.18) and fruits intake (β: -0.41; 95% CI: -0.63 and -0.19) in males and with dairy drinks (β: -0.39; 95% CI: -0.76 and -0.02) and fruits (β: -0.41; 95% CI: -0.73 and -0.10) in females. There was a high prevalence of inadequate TWI and high urine osmolality among these Spanish adolescents. Lower urine osmolality was associated with higher consumption of vegetables in males, dairy drinks in females and fruits in both males and females.

  15. Catecholamines - urine

    MedlinePlus

    Dopamine - urine test; Epinephrine - urine test; Adrenalin - urine test; Urine metanephrine; Normetanephrine; Norepinephrine - urine test; Urine catecholamines; VMA; HVA; Metanephrine; Homovanillic ...

  16. 21 CFR 862.1635 - Total protein test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... § 862.9. [52 FR 16122, May 1, 1987, as amended at 63 FR 59225, Nov. 3, 1998] ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Total protein test system. 862.1635 Section 862....1635 Total protein test system. (a) Identification. A total protein test system is a device intended to...

  17. 21 CFR 862.1635 - Total protein test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... § 862.9. [52 FR 16122, May 1, 1987, as amended at 63 FR 59225, Nov. 3, 1998] ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Total protein test system. 862.1635 Section 862....1635 Total protein test system. (a) Identification. A total protein test system is a device intended to...

  18. 21 CFR 862.1635 - Total protein test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... § 862.9. [52 FR 16122, May 1, 1987, as amended at 63 FR 59225, Nov. 3, 1998] ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Total protein test system. 862.1635 Section 862....1635 Total protein test system. (a) Identification. A total protein test system is a device intended to...

  19. 21 CFR 862.1635 - Total protein test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... § 862.9. [52 FR 16122, May 1, 1987, as amended at 63 FR 59225, Nov. 3, 1998] ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Total protein test system. 862.1635 Section 862....1635 Total protein test system. (a) Identification. A total protein test system is a device intended to...

  20. 21 CFR 862.1635 - Total protein test system.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... § 862.9. [52 FR 16122, May 1, 1987, as amended at 63 FR 59225, Nov. 3, 1998] ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Total protein test system. 862.1635 Section 862....1635 Total protein test system. (a) Identification. A total protein test system is a device intended to...

  1. Association between excess body weight and urine protein concentration in healthy dogs.

    PubMed

    Tefft, Karen M; Shaw, Darcy H; Ihle, Sherri L; Burton, Shelley A; Pack, LeeAnn

    2014-06-01

    Markedly overweight people can develop progressive proteinuria and kidney failure secondary to obesity-related glomerulopathy (ORG). Glomerular lesions in dogs with experimentally induced obesity are similar to those in people with ORG. The aim of this study was to evaluate if urine protein and albumin excretion is greater in overweight and obese dogs than in dogs of ideal body condition. Client-owned dogs were screened for underlying health conditions. These dogs were assigned a body condition score (BCS) using a 9-point scoring system. Dogs with a BCS of ≥ 6 were classified as being overweight/obese, and dogs with a BCS of 4 or 5 were classified as being of ideal body weight. The urine protein:creatinine ratio (UPC) and urine albumin:creatinine ratio (UAC) were then determined, and compared between 20 overweight/obese dogs and 22 ideal body weight control dogs. Median UPC (0.04 [range, 0.01-0.14; interquartile range, 0.07]) and UAC (0.41 [0-10.39; 3.21]) of overweight/obese dogs were not significantly different from median UPC (0.04 [0.01-0.32; 0.07]) and UAC (0.18 [0-7.04; 1.75]) in ideal body weight dogs. Clinicopathologic abnormalities consistent with ORG were absent from overweight/obese dogs in this study. © 2014 American Society for Veterinary Clinical Pathology and European Society for Veterinary Clinical Pathology.

  2. Urine heat shock protein 70 levels as a marker of urinary tract infection in children.

    PubMed

    Yilmaz, Alev; Yildirim, Zeynep Yuruk; Emre, Sevinc; Gedikbasi, Asuman; Yildirim, Tarik; Dirican, Ahmet; Ucar, Evren Onay

    2016-09-01

    Heat shock proteins (HSPs) are a multi-family group of proteins which are upregulated by the cell in response to exposure to hazardous (stress) factors, including infectious agents, to prevent changes in protein structure. The aim of our study was to assess whether urine levels of the 70-kDa family of HSPs (HSP70s) increase in children with urinary tract infection (UTI) and to determine the optimal urine (u) HSP70 cut-off level to predict UTI in children. Forty patients with symptomatic UTI (UTI group), 30 healthy children (control group), 21 asymptomatic patients with proven bacterial contamination in their urine culture (contamination group) and 30 patients with fever caused by other infections (non-UTI infection group) were enrolled in the study. Random urine samples were obtained for measurement of HSP70 and creatinine (Cr) from all groups. Urine was collected prior to the treatment of UTI at the time of presentation and after treatment. Urine HSP70 levels were measured by enzyme-linked immunosorbent analysis. A dimercaptosuccinic acid (DMSA) scan was performed at 5-7 days after presentation in UTI group to distinguish patients with acute pyelonephritis from those with cystitis; based on this scan, no patients had acute pyelonephritis. Patients were classified with pyelonephritis in the presence of all of the following signs: axillary fever of ≥39 °C, leukocytosis and positivity for C-reactive protein. The mean urine HSP70:Cr ratio (uHSP70/Cr) prior to treatment was significantly higher in the UTI group (449.86 ± 194.33 pg/mg) than in the control, contamination and non-UTI infection groups (39.93 ± 47.61, 32.43 ± 9.09 and 45.14 ± 19.76, respectively; p = 0.0001). Using a cut-off of 158 pg/mg uHSP70/Cr for the prediction of UTI, the sensitivity and specificity of the assay were 100 and 100 %, respectively (area under the time-concentration curve = 1). The uHSP70/Cr was highest in the patients with clinical pyelonephritis (p

  3. Quantitative label-free proteomic analysis of human urine to identify novel candidate protein biomarkers for schistosomiasis.

    PubMed

    Onile, Olugbenga Samson; Calder, Bridget; Soares, Nelson C; Anumudu, Chiaka I; Blackburn, Jonathan M

    2017-11-01

    Schistosomiasis is a chronic neglected tropical disease that is characterized by continued inflammatory challenges to the exposed population and it has been established as a possible risk factor in the aetiology of bladder cancer. Improved diagnosis of schistosomiasis and its associated pathology is possible through mass spectrometry to identify biomarkers among the infected population, which will influence early detection of the disease and its subtle morbidity. A high-throughput proteomic approach was used to analyse human urine samples for 49 volunteers from Eggua, a schistosomiasis endemic community in South-West, Nigeria. The individuals were previously screened for Schistosoma haematobium and structural bladder pathologies via microscopy and ultrasonography respectively. Samples were categorised into schistosomiasis, schistosomiasis with bladder pathology, bladder pathology, and a normal healthy control group. These samples were analysed to identify potential protein biomarkers. A total of 1306 proteins and 9701 unique peptides were observed in this study (FDR = 0.01). Fifty-four human proteins were found to be potential biomarkers for schistosomiasis and bladder pathologies due to schistosomiasis by label-free quantitative comparison between groups. Thirty-six (36) parasite-derived potential biomarkers were also identified, which include some existing putative schistosomiasis biomarkers that have been previously reported. Some of these proteins include Elongation factor 1 alpha, phosphopyruvate hydratase, histone H4 and heat shock proteins (HSP 60, HSP 70). These findings provide an in-depth analysis of potential schistosoma and human host protein biomarkers for diagnosis of chronic schistosomiasis caused by Schistosoma haematobium and its pathogenesis.

  4. Diurnal rhythms in the human urine metabolome during sleep and total sleep deprivation.

    PubMed

    Giskeødegård, Guro F; Davies, Sarah K; Revell, Victoria L; Keun, Hector; Skene, Debra J

    2015-10-09

    Understanding how metabolite levels change over the 24 hour day is of crucial importance for clinical and epidemiological studies. Additionally, the association between sleep deprivation and metabolic disorders such as diabetes and obesity requires investigation into the links between sleep and metabolism. Here, we characterise time-of-day variation and the effects of sleep deprivation on urinary metabolite profiles. Healthy male participants (n = 15) completed an in-laboratory study comprising one 24 h sleep/wake cycle prior to 24 h of continual wakefulness under highly controlled environmental conditions. Urine samples were collected over set 2-8 h intervals and analysed by (1)H NMR spectroscopy. Significant changes were observed with respect to both time of day and sleep deprivation. Of 32 identified metabolites, 7 (22%) exhibited cosine rhythmicity over at least one 24 h period; 5 exhibiting a cosine rhythm on both days. Eight metabolites significantly increased during sleep deprivation compared with sleep (taurine, formate, citrate, 3-indoxyl sulfate, carnitine, 3-hydroxyisobutyrate, TMAO and acetate) and 8 significantly decreased (dimethylamine, 4-DTA, creatinine, ascorbate, 2-hydroxyisobutyrate, allantoin, 4-DEA, 4-hydroxyphenylacetate). These data indicate that sampling time, the presence or absence of sleep and the response to sleep deprivation are highly relevant when identifying biomarkers in urinary metabolic profiling studies.

  5. Effect of urine urea nitrogen and protein intake adjusted by using the estimated urine creatinine excretion rate on the antiproteinuric effect of angiotensin II type I receptor blockers.

    PubMed

    Chin, Ho Jun; Kim, Dong Ki; Park, Jung Hwan; Shin, Sung Joon; Lee, Sang Ho; Choi, Bum Soon; Kim, Suhnggwon; Lim, Chun Soo

    2015-01-01

    The aim of this study was to determine the role of protein intake on proteinuria in chronic kidney disease (CKD), as it is presently not conclusive. This is a subanalysis of data from an open-label, case-controlled, randomized clinical trial on education about low-salt diets (NCT01552954). We estimated the urine excretion rate of parameters in a day, adjusted by using the equation for estimating urine creatinine excretion, and analyzed the effect of urine urea nitrogen (UUN), as well as estimating protein intake on the level of albuminuria in hypertensive patients with chronic kidney disease. Among 174 participants from whom complete 24-h urine specimens were collected, the estimates from the Tanaka equation resulted in the highest accuracy for the urinary excretion rate of creatinine, sodium, albumin, and UUN. Among 227 participants, the baseline value of estimated urine albumin excretion (eUalb) was positively correlated with the estimated UUN (eUUN) or protein intake according to eUUN (P = 0.012 and P = 0.038, respectively). We were able to calculate the ratios of eUalb and eUUN in 221 participants and grouped them according to the ratio of eUUN during 16-wk trial period. The proportion of patients that achieved a decrement of eUalb ≥25% during 16 wk with an angiotensin II type I receptor blocker (ARB) medication was 80% (24 of 30) in group 1, with eUUN ratio ≤-25%; 82.2% (111 of 135) in group 2, with eUUN ratio between -25% and 25%; and 66.1% (37 and 56) in group 3, with eUUN ratio ≥25% (P = 0.048). The probability of a decrease in albuminuria with ARB treatment was lower in patients with an increase of eUUN or protein intake during the 16 wk of ARB treatment, as observed in multiple logistic regression analysis as well. The estimated urine urea excretion rate showed a positive association with the level of albuminuria in hypertensive patients with chronic kidney disease. The increase of eUUN excretion ameliorated the antiproteinuric effect of ARB

  6. Day-to-day variability in spot urine protein-creatinine ratio measurements.

    PubMed

    Naresh, Chetana N; Hayen, Andrew; Craig, Jonathan C; Chadban, Steven J

    2012-10-01

    Accurate measurement of proteinuria is important in the diagnosis and management of chronic kidney disease (CKD). The reference standard test, 24-hour urinary protein excretion, is inconvenient and vulnerable to collection errors. Spot urine protein-creatinine ratio (PCR) is a convenient alternative and is in widespread use. However, day-to-day variability in PCR measurements has not been evaluated. Prospective cohort study of day-to-day variability in spot urine PCR measurement. Clinically stable outpatients with CKD (n = 145) attending a university hospital CKD clinic in Australia between July 2007 and April 2010. Spot urine PCR. Spot PCR variability was assessed and repeatability limits were determined using fractional polynomials. Spot PCRs were measured from urine samples collected at 9:00 am on consecutive days and 24-hour urinary protein excretion was collected concurrently. Paired results were analyzed from 145 patients: median age, 56 years; 59% men; and median 24-hour urinary protein excretion, 0.7 (range, 0.06-35.7) g/d. Day-to-day variability was substantial and increased in absolute terms, but decreased in relative terms with increasing baseline PCR. For patients with a low baseline PCR (20 mg/mmol [177 mg/g]), a change greater than ±160% (repeatability limits, 0-52 mg/mmol [0-460 mg/g]) is required to indicate a real change in proteinuria status with 95% certainty, whereas for those with a high baseline PCR (200 mg/mmol [1,768 mg/g]), a change of ±50% (decrease to <100 mg/mmol [<884 mg/g] or increase to >300 mg/mmol [>2,652 mg/g]) represents significant change. These study results need to be replicated in other ethnic groups. Changes in PCR observed in patients with CKD, ranging from complete resolution to doubling of PCR values, could be due to inherent biological variation and may not indicate a change in disease status. This should be borne in mind when using PCR in the diagnosis and management of CKD. Copyright © 2012 National Kidney Foundation

  7. Estimation of Daily Proteinuria in Patients with Amyloidosis by Using the Protein-To-Creatinine ratio in Random Urine Samples.

    PubMed

    Talamo, Giampaolo; Mir Muhammad, A; Pandey, Manoj K; Zhu, Junjia; Creer, Michael H; Malysz, Jozef

    2015-02-11

    Measurement of daily proteinuria in patients with amyloidosis is recommended at the time of diagnosis for assessing renal involvement, and for monitoring disease activity. Renal involvement is usually defined by proteinuria >500 mg/day. We evaluated the accuracy of the random urine protein-to-creatinine ratio (Pr/Cr) in predicting 24 hour proteinuria in patient with amyloidosis. We compared results of random urine Pr/Cr ratio and concomitant 24-hour urine collections in 44 patients with amyloidosis. We found a strong correlation (Spearman's ρ=0.874) between the Pr/Cr ratio and the 24 hour urine protein excretion. For predicting renal involvement, the optimal cut-off point of the Pr/Cr ratio was 715 mg/g. The sensitivity and specificity for this point were 91.8% and 95.5%, respectively, and the area under the curve value was 97.4%. We conclude that the random urine Pr/Cr ratio could be useful in the screening of renal involvement in patients with amyloidosis. If validated in a prospective study, the random urine Pr/Cr ratio could replace the 24 hour urine collection for the assessment of daily proteinuria and presence of nephrotic syndrome in patients with amyloidosis.

  8. Estimation of Daily Proteinuria in Patients with Amyloidosis by Using the Protein-To-Creatinine ratio in Random Urine Samples

    PubMed Central

    Talamo, Giampaolo; Mir Muhammad, A.; Pandey, Manoj K.; Zhu, Junjia; Creer, Michael H.; Malysz, Jozef

    2015-01-01

    Measurement of daily proteinuria in patients with amyloidosis is recommended at the time of diagnosis for assessing renal involvement, and for monitoring disease activity. Renal involvement is usually defined by proteinuria >500 mg/day. We evaluated the accuracy of the random urine protein-to-creatinine ratio (Pr/Cr) in predicting 24 hour proteinuria in patient with amyloidosis. We compared results of random urine Pr/Cr ratio and concomitant 24-hour urine collections in 44 patients with amyloidosis. We found a strong correlation (Spearman’s ρ=0.874) between the Pr/Cr ratio and the 24 hour urine protein excretion. For predicting renal involvement, the optimal cut-off point of the Pr/Cr ratio was 715 mg/g. The sensitivity and specificity for this point were 91.8% and 95.5%, respectively, and the area under the curve value was 97.4%. We conclude that the random urine Pr/Cr ratio could be useful in the screening of renal involvement in patients with amyloidosis. If validated in a prospective study, the random urine Pr/Cr ratio could replace the 24 hour urine collection for the assessment of daily proteinuria and presence of nephrotic syndrome in patients with amyloidosis. PMID:25918613

  9. Refractometric total protein concentrations in icteric serum from dogs.

    PubMed

    Gupta, Aradhana; Stockham, Steven L

    2014-01-01

    To determine whether high serum bilirubin concentrations interfere with the measurement of serum total protein concentration by refractometry and to assess potential biases among refractometer measurements. Evaluation study. Sera from 2 healthy Greyhounds. Bilirubin was dissolved in 0.1M NaOH, and the resulting solution was mixed with sera from 2 dogs from which food had been withheld to achieve various bilirubin concentrations up to 40 mg/dL. Refractometric total protein concentrations were estimated with 3 clinical refractometers. A biochemical analyzer was used to measure biuret assay-based total protein and bilirubin concentrations with spectrophotometric assays. No interference with refractometric measurement of total protein concentrations was detected with bilirubin concentrations up to 41.5 mg/dL. Biases in refractometric total protein concentrations were detected and were related to the conversion of refractive index values to total protein concentrations. Hyperbilirubinemia did not interfere with the refractometric estimation of serum total protein concentration. The agreement among total protein concentrations estimated by 3 refractometers was dependent on the method of conversion of refractive index to total protein concentration and was independent of hyperbilirubinemia.

  10. Optimization of analytical and pre-analytical conditions for MALDI-TOF-MS human urine protein profiles.

    PubMed

    Calvano, C D; Aresta, A; Iacovone, M; De Benedetto, G E; Zambonin, C G; Battaglia, M; Ditonno, P; Rutigliano, M; Bettocchi, C

    2010-03-11

    Protein analysis in biological fluids, such as urine, by means of mass spectrometry (MS) still suffers for insufficient standardization in protocols for sample collection, storage and preparation. In this work, the influence of these variables on healthy donors human urine protein profiling performed by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was studied. A screening of various urine sample pre-treatment procedures and different sample deposition approaches on the MALDI target was performed. The influence of urine samples storage time and temperature on spectral profiles was evaluated by means of principal component analysis (PCA). The whole optimized procedure was eventually applied to the MALDI-TOF-MS analysis of human urine samples taken from prostate cancer patients. The best results in terms of detected ions number and abundance in the MS spectra were obtained by using home-made microcolumns packed with hydrophilic-lipophilic balance (HLB) resin as sample pre-treatment method; this procedure was also less expensive and suitable for high throughput analyses. Afterwards, the spin coating approach for sample deposition on the MALDI target plate was optimized, obtaining homogenous and reproducible spots. Then, PCA indicated that low storage temperatures of acidified and centrifuged samples, together with short handling time, allowed to obtain reproducible profiles without artifacts contribution due to experimental conditions. Finally, interesting differences were found by comparing the MALDI-TOF-MS protein profiles of pooled urine samples of healthy donors and prostate cancer patients. The results showed that analytical and pre-analytical variables are crucial for the success of urine analysis, to obtain meaningful and reproducible data, even if the intra-patient variability is very difficult to avoid. It has been proven how pooled urine samples can be an interesting way to make easier the comparison between

  11. Effect of 21-day head down bed rest on urine proteins related to endothelium: Correlations with changes in carbohydrate metabolism

    NASA Astrophysics Data System (ADS)

    Kashirina, D.; Pastushkova, L.; Custaud, M. A.; Dobrokhotov, I.; Brzhozovsky, A.; Navasiolava, N.; Nosovsky, A.; Kononikhin, A.; Nikolaev, E.; Larina, I.

    2017-08-01

    We performed liquid chromatography-mass spectrometric study of the urine proteome in 8 healthy volunteers aged between 20 and 44 y.o. who have completed 21-day head-down bed rest. ANDSystem software which builds associative networks was used to identify the urinary proteins functionally related to the endothelium. We identified 7 endothelium-related biological processes, directly linked to 13 urine proteins. We performed manual annotation of the proteins which were the most important in terms of endothelial functions. Analysis of the correlations with biochemical variables revealed a positive correlation between fasting blood glucose and the following urine proteins: albumin, CD44 antigen, endothelial protein C receptor, mucin-1, osteopontin, receptor tyrosine kinase. As well, we found a positive correlation between HOMA-insulin resistance index and the following urine proteins: endothelial protein C receptor and syndecan-4. These results might suggest the involvement of above-mentioned proteins in glucose metabolism and their participation in the response to changes in blood glucose level.

  12. The role of obesity in the relation between total water intake and urine osmolality in US adults, 2009-2012.

    PubMed

    Rosinger, Asher Y; Lawman, Hannah G; Akinbami, Lara J; Ogden, Cynthia L

    2016-12-01

    Adequate water intake is critical to physiologic and cognitive functioning. Although water requirements increase with body size, it remains unclear whether weight status modifies the relation between water intake and hydration status. We examined how the association between water intake and urine osmolality, which is a hydration biomarker, varied by weight status. NHANES cross-sectional data (2009-2012) were analyzed in 9601 nonpregnant adults aged ≥20 y who did not have kidney failure. Weight status was categorized with the use of body mass index on the basis of measured height and weight (underweight or normal weight, overweight, and obesity). Urine osmolality was determined with the use of freezing-point depression osmometry. Hypohydration was classified according to the following age-dependent formula: ≥831 mOsm/kg - [3.4 × (age - 20 y)]. Total water intake was determined with the use of a 24-h dietary recall and was dichotomized as adequate or low on the basis of the Institute of Medicine's adequate intake recommendations for men and women (men: ≥3.7 or <3.7 L; nonlactating women: ≥2.7 or <2.7 L; lactating women: ≥3.8 or <3.8 L for adequate or low intakes, respectively). We tested interactions and conducted linear and log-binomial regressions. Total water intake (P = 0.002), urine osmolality (P < 0.001), and hypohydration prevalence (P < 0.001) all increased with higher weight status. Interactions between weight status and water intake status were significant in linear (P = 0.005) and log-binomial (P = 0.015) models, which were then stratified. The prevalence ratio of hypohydration between subjects with adequate water intake and those with low water intake was 0.56 (95% CI: 0.43, 0.73) in adults who were underweight or normal weight, 0.67 (95% CI: 0.57, 0.79) in adults who were overweight, and 0.78 (95% CI: 0.70, 0.88) in adults who were obese. On a population level, obesity modifies the association between water intake and hydration status. © 2016

  13. Spot urine protein measurements in normotensive pregnancies, pregnancies with isolated proteinuria and preeclampsia.

    PubMed

    Kattah, Andrea; Milic, Natasa; White, Wendy; Garovic, Vesna

    2017-10-01

    We performed a prospective, longitudinal study of pregnant women presenting to their first obstetrics visits to characterize the changes in spot urine protein-to-creatinine (UPCR) and albumin-to-creatinine ratios (UACR) in normotensive pregnancies, as well as identify clinical characteristics associated with isolated proteinuria and preeclampsia. We measured spot urinary albumin, protein, and creatinine at the first prenatal visit, end of the second trimester, and at delivery. In the normotensive pregnancies ( n = 142), we found that from the beginning of pregnancy to delivery, UACR increased by a median [interquartile range (IQR)] of 14.7 mg/g Cr (3.74-51.8) and UPCR by 60 mg/g Cr (30-130) ( P < 0.001 for both changes). Isolated proteinuria (defined as UPCR > 300 mg/g Cr in the absence of hypertension) was identified in 19/142 (13.4%) normotensive pregnancies. Increases in systolic and diastolic blood pressure from early pregnancy to delivery and increases in UACR from early to midpregnancy were associated with isolated proteinuria at delivery. Twelve women developed preeclampsia. Nulliparity, early, and midpregnancy diastolic blood pressures were strongly associated with the development of preeclampsia, but early changes in UACR were not. In conclusion, women who develop isolated proteinuria at delivery have a larger increase in blood pressure than women without proteinuria and have a "microalbuminuric" phase earlier in gestation, unlike women who develop preeclampsia. These findings suggest a different mechanism of urine protein excretion in women with isolated proteinuria as compared with women with preeclampsia, where proteinuria has a more abrupt onset. Copyright © 2017 the American Physiological Society.

  14. Can Tissue Cilia Lengths and Urine Cilia Proteins Be Markers of Kidney Diseases?

    PubMed

    Park, Kwon Moo

    2018-05-01

    The primary cilium is an organelle which consists of a microtubule in the core and a surrounding cilia membrane, and has long been recognized as a "vestigial organelle". However, new evidence demonstrates that the primary cilium has a notable effect on signal transduction in the cell and is associated with some genetic and non-genetic diseases. In the kidney, the primary cilium protrudes into the Bowman's space and the tubular lumen from the apical side of epithelial cells. The length of primary cilia is dynamically altered during the normal cell cycle, being shortened by retraction into the cell body at the entry of cell division and elongated at differentiation. Furthermore, the length of primary cilia is also dynamically changed in the cells, as a result and/or cause, during the progression of various kidney diseases including acute kidney injury and chronic kidney disease. Notably, recent data has demonstrated that the shortening of the primary cilium in the cell is associated with fragmentation, apart from retraction into the cell body, in the progression of diseases and that the fragmented primary cilia are released into the urine. This data reveals that the alteration of primary cilia length could be related to the progression of diseases. This review will consider if primary cilia length alteration is associated with the progression of kidney diseases and if the length of tissue primary cilia and the presence or increase of cilia proteins in the urine is indicative of kidney diseases.

  15. A validated inductively coupled plasma mass spectrometry (ICP-MS) method for the quantification of total platinum content in plasma, plasma ultrafiltrate, urine and peritoneal fluid.

    PubMed

    Lemoine, Lieselotte; Thijssen, Elsy; Noben, Jean-Paul; Adriaensens, Peter; Carleer, Robert; Speeten, Kurt Van der

    2018-04-15

    Oxaliplatin is a platinum (Pt) 1 containing antineoplastic agent that is applied in current clinical practice for the treatment of colon and appendiceal neoplasms. A fully validated, highly sensitive, high throughput inductively coupled plasma mass spectrometry (ICP-MS) method is provided to quantify the total Pt content in plasma, plasma ultrafiltrate, urine and peritoneal fluid. In this ICP-MS approach, the only step of sample preparation is a 1000-fold dilution in 0.5% nitric acid, allowing the analysis of 17 samples per hour. Detection of Pt was achieved over a linear range of 0.01-100 ng/mL. The limit of quantification was 18.0 ng/mL Pt in plasma, 8.0 ng/mL in ultrafiltrate and 6.1 ng/mL in urine and peritoneal fluid. The ICP-MS method was further validated for inter-and intraday precision and accuracy (≤15%), recovery, robustness and stability. Short-term storage of the biofluids, for 14 days, can be performed at -4 °C, -24 °C and -80 °C. As to long-term stability, up to 5 months, storage at -80 °C is encouraged. Furthermore, a timeline assessing the total and unbound Pt fraction in plasma and ultrafiltrate over a period of 45 h is provided. Following an incubation period of 5 h at 37 °C, 19-21% of Pt was recovered in the ultrafiltrate, emphasizing the extensive and rapid binding of oxaliplatin-derived Pt to plasma proteins. The described method can easily be implemented in a routine setting for pharmacokinetic studies in patients treated with oxaliplatin-based hyperthermic intraperitoneal perioperative chemotherapy. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Brain–blood amino acid correlates following protein restriction in murine maple syrup urine disease

    PubMed Central

    2014-01-01

    Background Conventional therapy for patients with maple syrup urine disease (MSUD) entails restriction of protein intake to maintain acceptable levels of the branched chain amino acid, leucine (LEU), monitored in blood. However, no data exists on the correlation between brain and blood LEU with protein restriction, and whether correction in blood is reflected in brain. Methods To address this question, we fed intermediate MSUD mice diets of 19% (standard) and 6% protein, with collection of sera (SE), striata (STR), cerebellum (CE) and cortex (CTX) for quantitative amino acid analyses. Results LEU and valine (VAL) levels in all brain regions improved on average 28% when shifting from 19% to 6% protein, whereas the same improvements in SE were on average 60%. Isoleucine (ILE) in brain regions did not improve, while the SE level improved 24% with low-protein consumption. Blood-branched chain amino acids (LEU, ILE, and VAL in sera (SE)) were 362-434 μM, consistent with human values considered within control. Nonetheless, numerous amino acids in brain regions remained abnormal despite protein restriction, including glutamine (GLN), aspartate (ASP), glutamate (GLU), gamma-aminobutyric acid (GABA), asparagine (ASN), citrulline (CIT) and serine (SER). To assess the specificity of these anomalies, we piloted preliminary studies in hyperphenylalaninemic mice, modeling another large neutral aminoacidopathy. Employing an identical dietary regimen, we found remarkably consistent abnormalities in GLN, ASP, and GLU. Conclusions Our results suggest that blood amino acid analysis may be a poor surrogate for assessing the outcomes of protein restriction in the large neutral amino acidopathies, and further indicate that chronic neurotransmitter disruptions (GLU, GABA, ASP) may contribute to long-term neurocognitive dysfunction in these disorders. PMID:24886632

  17. Brain-blood amino acid correlates following protein restriction in murine maple syrup urine disease.

    PubMed

    Vogel, Kara R; Arning, Erland; Wasek, Brandi L; McPherson, Sterling; Bottiglieri, Teodoro; Gibson, K Michael

    2014-05-08

    Conventional therapy for patients with maple syrup urine disease (MSUD) entails restriction of protein intake to maintain acceptable levels of the branched chain amino acid, leucine (LEU), monitored in blood. However, no data exists on the correlation between brain and blood LEU with protein restriction, and whether correction in blood is reflected in brain. To address this question, we fed intermediate MSUD mice diets of 19% (standard) and 6% protein, with collection of sera (SE), striata (STR), cerebellum (CE) and cortex (CTX) for quantitative amino acid analyses. LEU and valine (VAL) levels in all brain regions improved on average 28% when shifting from 19% to 6% protein, whereas the same improvements in SE were on average 60%. Isoleucine (ILE) in brain regions did not improve, while the SE level improved 24% with low-protein consumption. Blood-branched chain amino acids (LEU, ILE, and VAL in sera (SE)) were 362-434 μM, consistent with human values considered within control. Nonetheless, numerous amino acids in brain regions remained abnormal despite protein restriction, including glutamine (GLN), aspartate (ASP), glutamate (GLU), gamma-aminobutyric acid (GABA), asparagine (ASN), citrulline (CIT) and serine (SER). To assess the specificity of these anomalies, we piloted preliminary studies in hyperphenylalaninemic mice, modeling another large neutral aminoacidopathy. Employing an identical dietary regimen, we found remarkably consistent abnormalities in GLN, ASP, and GLU. Our results suggest that blood amino acid analysis may be a poor surrogate for assessing the outcomes of protein restriction in the large neutral amino acidopathies, and further indicate that chronic neurotransmitter disruptions (GLU, GABA, ASP) may contribute to long-term neurocognitive dysfunction in these disorders.

  18. Evaluation of the analytical variability of dipstick protein pads in canine urine.

    PubMed

    Giraldi, Marco; Paltrinieri, Saverio; Zatelli, Andrea

    2018-06-01

    The dipstick is a first-line and inexpensive test that can exclude the presence of proteinuria in dogs. However, no information is available about the analytical variability of canine urine dipstick analysis. The aim of this study was to assess the analytical variability in 2 dipsticks and the inter-operator variability in dipstick interpretation. Canine urine supernatants (n = 174) were analyzed with 2 commercially available dipsticks. Two observers evaluated each result blinded to the other observer and to the results of the other dipstick. Intra- and inter-assay variability was assessed in 5 samples (corresponding to the 5 different semi-quantitative results) tested 10 consecutive times over 5 consecutive days. The agreement between observers and between dipsticks was evaluated with Cohen's k test. Intra-assay repeatability was good (≤3/10 errors), whereas inter-assay variability was higher (from 1/5 to 4/5 discordant results). The concordance between the operators (k = 0.68 and 0.79 for the 2 dipsticks) and that of the dipsticks (k = 0.66 and 0.74 for the 2 operators) was good. However, 1 observer and 1 dipstick overestimated the results compared with the second observer or dipstick. In any case, discordant results accounted for a single unit of the semi-quantitative scale. As for any other method, analytic variability may affect the semi-quantitation of urinary proteins when using the dipstick method. Subjective interpretation of the pad and, to a lesser extent, intrinsic staining properties of the pads could affect the results. Further studies are warranted to evaluate the effect of this variability on clinical decisions. © 2018 American Society for Veterinary Clinical Pathology.

  19. Urine and Urination

    MedlinePlus

    Your kidneys make urine by filtering wastes and extra water from your blood. The waste is called urea. Your blood carries it to the kidneys. From the kidneys, urine travels down two thin tubes called ureters to ...

  20. Urine osmolality in the US population: Implications for environmental biomonitoring

    PubMed Central

    Yeh, Hung-Chieh; Lin, Yu-Sheng; Kuo, Chin-Chi; Weidemann, Darcy; Weaver, Virginia; Fadrowski, Jeffrey; Neu, Alicia; Navas-Acien, Ana

    2018-01-01

    Background For many environmental chemicals, concentrations in spot urine samples are considered valid surrogates of exposure and internal dose. To correct for urine dilution, spot urine concentrations are commonly adjusted for urinary creatinine. There are, however, several concerns about the use of urine creatinine. While urine osmolality is an attractive alternative; its characteristics and determinants in the general population remain unknown. Our objective was to describe the determinants of urine osmolality and to contrast the difference between osmolality and creatinine in urine. Methods From the National Health and Nutrition Examination Survey (NHANES) 2009–2012, 10,769 participants aged 16 years or older with measured urine osmolality and creatinine were used in the analysis. Very dilute and very concentrated urine was defined as urine creatinine lower than 0.3 g/l and higher than 3 g/l, respectively. Linear and logistic regression analyses were performed to investigate the associations of interest. Results Urine osmolality and creatinine were highly correlated (Pearson correlation coefficient = 0.75) and their respective median values were 648 mOsm/kg and 1.07 g/l. The prevalence of very dilute and very concentrated urine samples was 8.1% and 3.1%, respectively. Factors associated in the same direction with both urine osmolality and urine creatinine included age, sex, race, body mass index (BMI), hypertension, water intake, and blood osmolality. The magnitude of associations expressed as percent change was significantly stronger with creatinine than osmolality. Compared to urine creatinine, urine osmolality did not vary by diabetes status but was affected by daily total protein intake. Participants with chronic kidney disease (CKD) had significantly higher urine creatinine concentrations but lower urine osmolality. Both very dilute and concentrated urine were associated with a diverse array of sociodemographic, medical conditions, and dietary factors

  1. Immunoelectrophoresis - urine

    MedlinePlus

    Immunoglobulin electrophoresis - urine; Gamma globulin electrophoresis - urine; Urine immunoglobulin electrophoresis; IEP - urine ... A clean-catch urine sample is needed. The clean-catch method is used to prevent germs from the penis or vagina from getting ...

  2. Cy5 total protein normalization in Western blot analysis.

    PubMed

    Hagner-McWhirter, Åsa; Laurin, Ylva; Larsson, Anita; Bjerneld, Erik J; Rönn, Ola

    2015-10-01

    Western blotting is a widely used method for analyzing specific target proteins in complex protein samples. Housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. We present a new, more reliable method for normalization using Cy5-prelabeled total protein as a loading control. We used a prelabeling protocol based on Cy5 N-hydroxysuccinimide ester labeling that produces a linear signal response. We obtained a low coefficient of variation (CV) of 7% between the ratio of extracellular signal-regulated kinase (ERK1/2) target to Cy5 total protein control signals over the whole loading range from 2.5 to 20.0μg of Chinese hamster ovary cell lysate protein. Corresponding experiments using actin or tubulin as controls for normalization resulted in CVs of 13 and 18%, respectively. Glyceraldehyde-3-phosphate dehydrogenase did not produce a proportional signal and was not suitable for normalization in these cells. A comparison of ERK1/2 signals from labeled and unlabeled samples showed that Cy5 prelabeling did not affect antibody binding. By using total protein normalization we analyzed PP2A and Smad2/3 levels with high confidence. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea

    SciTech Connect

    Brown, Joseph N.; Brewer, Heather M.; Nicora, Carrie D.

    Background: We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Methods: Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbonmore » monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Results: Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Conclusions: Candidate proteins

  4. Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea

    PubMed Central

    2014-01-01

    Background We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Methods Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbon monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Results Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Conclusions Candidate proteins and mi

  5. Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea.

    PubMed

    Brown, Joseph N; Brewer, Heather M; Nicora, Carrie D; Weitz, Karl K; Morris, Michael J; Skabelund, Andrew J; Adkins, Joshua N; Smith, Richard D; Cho, Ji-Hoon; Gelinas, Richard

    2014-10-05

    We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbon monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Candidate proteins and miRNAs associated with the general diagnosis of

  6. Protein and microRNA biomarkers from lavage, urine, and serum in military personnel evaluated for dyspnea

    DOE PAGES

    Brown, Joseph N.; Brewer, Heather M.; Nicora, Carrie D.; ...

    2014-10-05

    Background: We have identified candidate protein and microRNA (miRNA) biomarkers for dyspnea by studying serum, lavage fluid, and urine from military personnel who reported serious respiratory symptoms after they were deployed to Iraq or Afghanistan. Methods: Forty-seven soldiers with the complaint of dyspnea who enrolled in the STudy of Active Duty Military Personnel for Environmental Dust Exposure (STAMPEDE) underwent comprehensive pulmonary evaluations at the San Antonio Military Medical Center. The evaluation included fiber-optic bronchoscopy with bronchoalveolar lavage. The clinical findings from the STAMPEDE subjects pointed to seven general underlying diagnoses or findings including airway hyperreactivity, asthma, low diffusivity of carbonmore » monoxide, and abnormal cell counts. The largest category was undiagnosed. As an exploratory study, not a classification study, we profiled proteins or miRNAs in lavage fluid, serum, or urine in this group to look for any underlying molecular patterns that might lead to biomarkers. Proteins in lavage fluid and urine were identified by accurate mass tag (database-driven) proteomics methods while miRNAs were profiled by a hybridization assay applied to serum, urine, and lavage fluid. Results: Over seventy differentially expressed proteins were reliably identified both from lavage and from urine in forty-eight dyspnea subjects compared to fifteen controls with no known lung disorder. Six of these proteins were detected both in urine and lavage. One group of subjects was distinguished from controls by expressing a characteristic group of proteins. A related group of dyspnea subjects expressed a unique group of miRNAs that included one miRNA that was differentially overexpressed in all three fluids studied. The levels of several miRNAs also showed modest but direct associations with several standard clinical measures of lung health such as forced vital capacity or gas exchange efficiency. Conclusions: Candidate proteins

  7. Protein and lipid damage in maple syrup urine disease patients: l-carnitine effect.

    PubMed

    Mescka, Caroline Paula; Wayhs, Carlos Alberto Yasin; Vanzin, Camila Simioni; Biancini, Giovana Brondani; Guerreiro, Gilian; Manfredini, Vanusa; Souza, Carolina; Wajner, Moacir; Dutra-Filho, Carlos Severo; Vargas, Carmen Regla

    2013-02-01

    Maple syrup urine disease (MSUD) is an inborn error of metabolism biochemically characterized by elevated levels of the branched chain amino acids (BCAA) leucine, isoleucine, valine and the corresponding branched-chain α-keto acids. This disorder is clinically characterized by ketoacidosis, seizures, coma, psychomotor delay and mental retardation whose pathophysiology is not completely understood. Recent studies have shown that oxidative stress may be involved in neuropathology of MSUD. l-Carnitine (l-Car) plays a central role in the cellular energy metabolism because it transports long-chain fatty acids for oxidation and ATP generation. In recent years many studies have demonstrated the antioxidant role of this compound. In this work, we investigated the effect of BCAA-restricted diet supplemented or not with l-Car on lipid peroxidation and in protein oxidation in MSUD patients. We found a significant increase of malondialdehyde and of carbonyl content in plasma of MSUD patients under BCAA-restricted diet compared to controls. Furthermore, patients under BCAA-restricted diet plus l-Car supplementation presented a marked reduction of malondialdehyde content in relation to controls, reducing the lipid peroxidation. In addition, free l-Car concentrations were negatively correlated with malondialdehyde levels. Our data show that l-Car may have an antioxidant effect, protecting against the lipid peroxidation and this could represent an additional therapeutic approach to the patients affected by MSUD. Copyright © 2012 ISDN. Published by Elsevier Ltd. All rights reserved.

  8. Effect of Major Royal Jelly Proteins on Spatial Memory in Aged Rats: Metabolomics Analysis in Urine.

    PubMed

    Chen, Di; Liu, Fang; Wan, Jian-Bo; Lai, Chao-Qiang; Shen, Li-Rong

    2017-04-19

    Royal jelly (RJ) produced by worker honeybees is the sole food for the queen bee throughout her life as well as the larvae of worker bees for the first 3 days after hatching. Supplementation of RJ in the diet has been shown to increase spatial memory in rodents. However, the key constituents in RJ responsible for improvement of cognitive function are unknown. Our objective was to determine if the major royal jelly proteins (MRJPs) extracted from RJ can improve the spatial memory of aged rats. The spatial memory assay using the Morris water maze test was administered once to rats after a 14-week feeding. Metabolomics analysis based on quadrupole time-of-flight mass spectrometry was conducted to examine the differences in compounds from urine. Aged male rats fed MRJPs showed improved spatial memory up to 48.5% when compared to the control male aged rats fed distilled water. The metabolite pattern of the MRJPs-fed aged rats was regressed to that of the young rats. Compounds altered by MRJPs were mapped to nicotinate and nicotinamide metabolism, cysteine taurine metabolism, and energy metabolism pathways. In summary, MRJPs may improve spatial memory and possess the potential for prevention of cognitive impairment via the cysteine and taurine metabolism and energy metabolism pathways in aged rats.

  9. Quantification Assays for Total and Polyglutamine-Expanded Huntingtin Proteins

    PubMed Central

    Boogaard, Ivette; Smith, Melanie; Pulli, Kristiina; Szynol, Agnieszka; Albertus, Faywell; Lamers, Marieke B. A. C.; Dijkstra, Sipke; Kordt, Daniel; Reindl, Wolfgang; Herrmann, Frank; McAllister, George; Fischer, David F.; Munoz-Sanjuan, Ignacio

    2014-01-01

    The expansion of a CAG trinucleotide repeat in the huntingtin gene, which produces huntingtin protein with an expanded polyglutamine tract, is the cause of Huntington's disease (HD). Recent studies have reported that RNAi suppression of polyglutamine-expanded huntingtin (mutant HTT) in HD animal models can ameliorate disease phenotypes. A key requirement for such preclinical studies, as well as eventual clinical trials, aimed to reduce mutant HTT exposure is a robust method to measure HTT protein levels in select tissues. We have developed several sensitive and selective assays that measure either total human HTT or polyglutamine-expanded human HTT proteins on the electrochemiluminescence Meso Scale Discovery detection platform with an increased dynamic range over other methods. In addition, we have developed an assay to detect endogenous mouse and rat HTT proteins in pre-clinical models of HD to monitor effects on the wild type protein of both allele selective and non-selective interventions. We demonstrate the application of these assays to measure HTT protein in several HD in vitro cellular and in vivo animal model systems as well as in HD patient biosamples. Furthermore, we used purified recombinant HTT proteins as standards to quantitate the absolute amount of HTT protein in such biosamples. PMID:24816435

  10. Total mercury levels in hair, toenail, and urine among women free from occupational exposure and their relations to renal tubular function

    SciTech Connect

    Ohno, Tomoko; Sakamoto, Mineshi; Kurosawa, Tomoko

    2007-02-15

    To investigate the relations among total mercury levels in hair, toenail, and urine, together with potential effects of methylmercury intake on renal tubular function, we determined their levels, and urinary N-acetyl-{beta}-d-glucosaminidase activity (NAG) and {alpha}{sub 1}-microglobulin (AMG) in 59 women free from occupational exposures, and estimated daily mercury intakes from fish and other seafood using a food frequency questionnaire. Mercury levels (mean+/-SD) in the women were 1.51+/-0.91{mu}g/g in hair, 0.59+/-0.32{mu}g/g in toenail, and 0.86+/-0.66{mu}g/g creatinine in urine; and, there were positive correlations among them (P<0.001). The daily mercury intake of 9.15+/-7.84{mu}g/day was significantly correlated with total mercury levels in hair,more » toenail, and urine (r=0.551, 0.537, and 0.604, P<0.001). Among the women, the NAG and AMG were positively correlated with both the daily mercury intake and mercury levels in hair, toenail, and urine (P<0.01); and, these relations were almost similar when using multiple regression analysis to adjust for possible confounders such as urinary cadmium (0.47+/-0.28{mu}g/g creatinine) and smoking status. In conclusion, mercury resulting from fish consumption can explain total mercury levels in hair, toenail, and urine to some degree (about 30%), partly through the degradation into the inorganic form, and it may confound the renal tubular effect of other nephrotoxic agents. Also, the following equation may be applicable to the population neither with dental amalgam fillings nor with occupational exposures: [hair mercury ({mu}g/g)]=2.44x[toenail mercury ({mu}g/g)].« less

  11. Evaluation of factors that affect analytic variability of urine protein-to-creatinine ratio determination in dogs.

    PubMed

    Rossi, Gabriele; Giori, Luca; Campagnola, Simona; Zatelli, Andrea; Zini, Eric; Paltrinieri, Saverio

    2012-06-01

    To determine whether preanalytic and analytic factors affect evaluation of the urinary protein-to-creatinine (UPC) ratio in dogs. 50 canine urine samples. The UPC ratio was measured to assess the intra-assay imprecision (20 measurements within a single session), the influence of predilution (1:10, 1:20, and 1:100) for urine creatinine concentration measurement, and the effect of storage at room temperature (approx 20°C), 4°C, and -20°C. The coefficient of variation at room temperature determined with the 1:20 predilution was < 10.0%, with the highest coefficients of variation found in samples with a low protein concentration or low urine specific gravity. This variability could result in misclassification of samples with UPC ratios close to the thresholds defined by the International Renal Interest Society to classify dogs as nonproteinuric (0.2), borderline proteinuric (0.21 to 0.50), or proteinuric (> 0.51). A proportional bias was found in samples prediluted 1:10, compared with samples prediluted 1:20 or 1:100. At room temperature, the UPC ratio did not significantly increase after 2 and 4 hours. After 12 hours at room temperature and at 4°C, the UPC ratio significantly increased. The UPC ratio did not significantly change during 3 months of storage at -20°C. The intra-assay precision of the UPC ratio was sufficiently low to avoid misclassification of samples, except for values close to 0.2 or 0.5. The optimal predilution ratio for urine creatinine concentration measurement was 1:20. A 1:100 predilution is recommended in samples with a urine specific gravity > 1.030. The UPC ratio must be measured as soon as samples are collected. Alternatively, samples should be immediately frozen to increase their stability and minimize the risk of misclassification of proteinuria.

  12. Urine epidermal growth factor, monocyte chemoattractant protein-1 or their ratio as predictors of complete remission in primary glomerulonephritis.

    PubMed

    Chanrat, Eakkapat; Worawichawong, Supanat; Radinahamed, Piyanuch; Sathirapongsasuti, Nuankanya; Nongnuch, Arkom; Assanatham, Montira; Udomsubpayakul, Umaporn; Kitiyakara, Chagriya

    2018-04-01

    The balance of several cytokines likely influences the resolution of glomerulonephritis. Monocyte chemoattractant protein-1(MCP-1) is a chemokine that promotes renal inflammation whereas epidermal growth factor (EGF) stimulates protective responses. Previously, high urine MCP-1(MCP-1) and low urine EGF (EGF) levels were found to be associated with tubulointerstitial fibrosis, but there is limited information on the value of these mediators as predictors of therapeutic responses or long term outcome in primary glomerulonephritis. To determine the performance of urine EGF, MCP-1 or their ratio at baseline as biomarkers to predict complete remission, and the relationship of these mediators with subsequent renal function 24 months later in primary glomerulonephritis. This is a prospective study of patients with biopsy-proven primary glomerulonephritis. Baseline urine samples were collected at biopsy before therapy. MCP-1 and EGF were analyzed by enzyme-linked immunosorbent assays and expressed as a ratio to urine creatinine (ng/mgCr) or as EGF/MCP-1 ratio (ng/ng). Proteinuria and estimated glomerular filtration rate (eGRF) were monitored after therapy. Complete remission (CR) was defined as proteinuria ≤ 0.3 g/gCr. Median follow-up was 20 months. Of all patients (n = 74), 38 patients (51.4%) subsequently achieved CR. Baseline urine EGF and EGF/MCP-1 levels were significantly higher in CR compared to Not CR. By contrast, MCP-1 was not different. High EGF (EGF > 75 ng/mgCr) was a significant predictor (OR 2.28) for CR by multivariate analysis after adjusting for proteinuria, blood pressure, baseline eGFR. In patients who completed 24 months follow-up (n = 43), baseline EGF correlated inversely with proteinuria and positively with eGFR at 24 months. High urine EGF level is a promising biomarker of CR. Baseline EGF levels correlated with kidney function at 2 years. EGF/MCP-1 was not superior to EGF alone. Further studies are necessary to

  13. New Method To Estimate Total Polyphenol Excretion: Comparison of Fast Blue BB versus Folin-Ciocalteu Performance in Urine.

    PubMed

    Hinojosa-Nogueira, Daniel; Muros, Joaquín; Rufián-Henares, José A; Pastoriza, Silvia

    2017-05-24

    Polyphenols are bioactive substances of vegetal origin with a significant impact on human health. The assessment of polyphenol intake and excretion is therefore important. The Folin-Ciocalteu (F-C) method is the reference assay to measure polyphenols in foods as well as their excretion in urine. However, many substances can influence the method, making it necessary to conduct a prior cleanup using solid-phase extraction (SPE) cartridges. In this paper, we demonstrate the use of the Fast Blue BB reagent (FBBB) as a new tool to measure the excretion of polyphenols in urine. Contrary to F-C, FBBB showed no interference in urine, negating the time-consuming and costly SPE cleanup. In addition, it showed excellent linearity (r 2 = 0.9997), with a recovery of 96.4% and a precision of 1.86-2.11%. The FBBB method was validated to measure the excretion of polyphenols in spot urine samples from Spanish children, showing a good correlation between polyphenol intake and excretion.

  14. The Human Urine Metabolome

    PubMed Central

    Bouatra, Souhaila; Aziat, Farid; Mandal, Rupasri; Guo, An Chi; Wilson, Michael R.; Knox, Craig; Bjorndahl, Trent C.; Krishnamurthy, Ramanarayan; Saleem, Fozia; Liu, Philip; Dame, Zerihun T.; Poelzer, Jenna; Huynh, Jessica; Yallou, Faizath S.; Psychogios, Nick; Dong, Edison; Bogumil, Ralf; Roehring, Cornelia; Wishart, David S.

    2013-01-01

    Urine has long been a “favored” biofluid among metabolomics researchers. It is sterile, easy-to-obtain in large volumes, largely free from interfering proteins or lipids and chemically complex. However, this chemical complexity has also made urine a particularly difficult substrate to fully understand. As a biological waste material, urine typically contains metabolic breakdown products from a wide range of foods, drinks, drugs, environmental contaminants, endogenous waste metabolites and bacterial by-products. Many of these compounds are poorly characterized and poorly understood. In an effort to improve our understanding of this biofluid we have undertaken a comprehensive, quantitative, metabolome-wide characterization of human urine. This involved both computer-aided literature mining and comprehensive, quantitative experimental assessment/validation. The experimental portion employed NMR spectroscopy, gas chromatography mass spectrometry (GC-MS), direct flow injection mass spectrometry (DFI/LC-MS/MS), inductively coupled plasma mass spectrometry (ICP-MS) and high performance liquid chromatography (HPLC) experiments performed on multiple human urine samples. This multi-platform metabolomic analysis allowed us to identify 445 and quantify 378 unique urine metabolites or metabolite species. The different analytical platforms were able to identify (quantify) a total of: 209 (209) by NMR, 179 (85) by GC-MS, 127 (127) by DFI/LC-MS/MS, 40 (40) by ICP-MS and 10 (10) by HPLC. Our use of multiple metabolomics platforms and technologies allowed us to identify several previously unknown urine metabolites and to substantially enhance the level of metabolome coverage. It also allowed us to critically assess the relative strengths and weaknesses of different platforms or technologies. The literature review led to the identification and annotation of another 2206 urinary compounds and was used to help guide the subsequent experimental studies. An online database containing

  15. Identification of potential bladder cancer markers in urine by abundant-protein depletion coupled with quantitative proteomics.

    PubMed

    Chen, Chien-Lun; Lin, Tsung-Shih; Tsai, Cheng-Han; Wu, Chih-Ching; Chung, Ting; Chien, Kun-Yi; Wu, Maureen; Chang, Yu-Sun; Yu, Jau-Song; Chen, Yi-Ting

    2013-06-24

    In this study, we evaluated the reproducibility of abundant urine protein depletion by hexapeptide-based library beads and an antibody-based affinity column using the iTRAQ technique. The antibody-based affinity-depletion approach, which proved superior, was then applied in conjunction with iTRAQ to discover proteins that were differentially expressed between pooled urine samples from hernia and bladder cancer patients. Several proteins, including seven apolipoproteins, TIM, SAA4, and proEGF were further verified in 111 to 203 individual urine samples from patients with hernia, bladder cancer, or kidney cancer. Six apolipoproteins (APOA1, APOA2, APOB, APOC2, APOC3, and APOE) were able to differentiate bladder cancer from hernia. SAA4 was significantly increased in bladder cancer subgroups, whereas ProEGF was significantly decreased in bladder cancer subgroups. Additionally, the combination of SAA4 and ProEGF exhibited higher diagnostic capacity (AUC=0.80 and p<0.001) in discriminating bladder cancer from hernia than either marker alone. Using MetaCore software to interpret global changes of the urine proteome caused by bladder cancer, we found that the most notable alterations were in immune-response/alternative complement and blood-coagulation pathways. This study confirmed the clinical significance of the urine proteome in the development of non-invasive biomarkers for the detection of bladder cancer. In this study, we evaluated the reproducibility of abundant urine protein depletion by hexapeptide-based library beads and an antibody-based affinity column using the iTRAQ technique. The antibody-based affinity-depletion approach, which proved superior, was then applied in conjunction with iTRAQ to discover proteins that were differentially expressed between pooled urine samples from hernia and bladder cancer patients. Several proteins, including seven apolipoproteins, TIM, SAA4, and proEGF were further verified in 111 to 203 individual urine samples from patients

  16. Urine protein/creatinine ratio as a mortality risk predictor in non-diabetics with normal renal function.

    PubMed

    Fulks, Michael; Stout, Robert L; Dolan, Vera F

    2012-01-01

    Determine the relative mortality in apparently healthy adults with various levels of urinary protein measured by urine protein/creatinine (p/c) ratio. By use of the Social Security Death Master File, mortality in 2010 was determined for 7.5 million life insurance applicants age 20 to 89 providing urine samples between 1992 and 2006. Relative mortality by Cox regression for bands of p/c ratios was determined using age and sex as covariates and with an age split at 60 after excluding those with hematuria (> 3 red cells/hpf), diabetes, evidence of blood sugar elevation, or eGFR < 60 mL/min. After the exclusions noted above, relative mortality increased to 160% beginning at a p/c ratio of 0.11 mg/mg and rose steadily above that value regardless of sex and age. Most of this risk was not explained by a history of hypertension or elevated systolic blood pressure. Albumin testing identified roughly a third of urine samples with elevated p/c ratios as not containing albumin; those cases appeared to be associated with much lower risk as long as the p/c ratio was < or = 1.0 mg/mg. Low levels of proteinuria identified as urine protein/creatinine ratios of 0.11 mg/mg or higher (much lower than the usual lower cut-off value of 0.21) are associated with substantial excess mortality risk, even after excluding diabetics and those with reduced kidney function or hematuria.

  17. Clinical utility of spot urine protein-to-creatinine ratio modified by estimated daily creatinine excretion in children.

    PubMed

    Yang, Eun Mi; Yoon, Bo Ae; Kim, Soo Wan; Kim, Chan Jong

    2017-06-01

    The spot urine protein-to-creatinine ratio (UPCR) is widely used to predict 24-h urine protein (24-h UP) excretion. In patients with low daily urine creatinine excretion (UCr), however, the UPCR may overestimate 24-h UP. The aim of this study was to predict 24-h UP using UPCR adjusted by estimated 24-h UCr in children. This study included 442 children whose 24-h UP and spot UPCR were measured concomitantly. Estimated 24-h UCr was calculated using three previously existing equations. We estimated the 24-h UP excretion from UPCR by multiplying the estimated UCr. The results were compared with the measured 24-h UP. There was a strong correlation between UPCR and 24-h UP (r = 0.801, P < 0.001), and the correlation improved after multiplying the UPCR by the measured UCr (r = 0.847, P < 0.001). Using the estimated UCr rather than the measured UCr, there was high accuracy and strong correlation between the estimated UPCR weighted by the Cockcroft-Gault equation and 24-h UP. Improvement was also observed in the subgroup (proteinuria vs. non-proteinuria) analysis, particularly in the proteinuria group. The spot UPCR multiplied by the estimated UCr improved the accuracy of prediction of the 24-h UP in children.

  18. Creatine ( methyl-d3) dilution in urine for estimation of total body skeletal muscle mass: accuracy and variability vs. MRI and DXA.

    PubMed

    Clark, Richard V; Walker, Ann C; Miller, Ram R; O'Connor-Semmes, Robin L; Ravussin, Eric; Cefalu, William T

    2018-01-01

    A noninvasive method to estimate muscle mass based on creatine ( methyl-d 3 ) (D 3 -creatine) dilution using fasting morning urine was evaluated for accuracy and variability over a 3- to 4-mo period. Healthy older (67- to 80-yr-old) subjects ( n = 14) with muscle wasting secondary to aging and four patients with chronic disease (58-76 yr old) fasted overnight and then received an oral 30-mg dose of D 3 -creatine at 8 AM ( day 1). Urine was collected during 4 h of continued fasting and then at consecutive 4- to 8-h intervals through day 5. Assessment was repeated 3-4 mo later in 13 healthy subjects and 1 patient with congestive heart failure. Deuterated and unlabeled creatine and creatinine were measured using liquid chromatography-tandem mass spectrometry. Total body creatine pool size and muscle mass were calculated from D 3 -creatinine enrichment in urine. Muscle mass was also measured by whole body MRI and 24-h urine creatinine, and lean body mass (LBM) was measured by dual-energy X-ray absorptiometry (DXA). D 3 -creatinine urinary enrichment from day 5 provided muscle mass estimates that correlated with MRI for all subjects ( r = 0.88, P < 0.0001), with less bias [difference from MRI = -3.00 ± 2.75 (SD) kg] than total LBM assessment by DXA, which overestimated muscle mass vs. MRI (+22.5 ± 3.7 kg). However, intraindividual variability was high with the D 3 -creatine dilution method, with intrasubject SD for estimated muscle mass of 2.5 kg vs. MRI (0.5 kg) and DXA (0.8 kg). This study supports further clinical validation of the D 3 -creatine method for estimating muscle mass. NEW & NOTEWORTHY Measurement of creatine ( methyl-d 3 ) (D 3 -creatine) and D 3 -creatinine excretion in fasted morning urine samples may be a simple, less costly alternative to MRI or dual-energy X-ray absorptiometry (DXA) to calculate total body muscle mass. The D 3 -creatine enrichment method provides estimates of muscle mass that correlate well with MRI, and with less

  19. Quantitative analysis of unconjugated and total bisphenol A in human urine using solid-phase extraction and UPLC-MS/MS: method implementation, method qualification and troubleshooting.

    PubMed

    Buscher, Brigitte; van de Lagemaat, Dick; Gries, Wolfgang; Beyer, Dieter; Markham, Dan A; Budinsky, Robert A; Dimond, Stephen S; Nath, Rajesh V; Snyder, Stephanie A; Hentges, Steven G

    2015-11-15

    The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Effects of Polyphenol, Measured by a Biomarker of Total Polyphenols in Urine, on Cardiovascular Risk Factors After a Long-Term Follow-Up in the PREDIMED Study

    PubMed Central

    Guo, Xiaohui; Tresserra-Rimbau, Anna; Estruch, Ramón; Martínez-González, Miguel A.; Medina-Remón, Alexander; Castañer, Olga; Corella, Dolores; Lamuela-Raventós, Rosa M.

    2016-01-01

    Several epidemiological studies have shown an inverse association between the consumption of polyphenol-rich foods and risk of cardiovascular diseases. However, accuracy and reliability of these studies may be increased using urinary total polyphenol excretion (TPE) as a biomarker for total polyphenol intake. Our aim was to assess if antioxidant activity, measured by a Folin-Ciocalteu assay in urine, is correlated with an improvement in cardiovascular risk factors (blood pressure and serum glucose, cholesterol, HDL-cholesterol, LDL-cholesterol, and triglyceride concentrations) in an elderly population at high risk. A longitudinal study was performed with 573 participants (aged 67.3 ± 5.9) from the PREDIMED study (ISRCTN35739639). We used Folin-Ciocalteu method to determine TPE in urine samples, assisting with solid phase extraction. Participants were categorized into three groups according to changes in TPE. Multiple linear regression models were used to assess relationships between TPE and clinical cardiovascular risk factors, adjusting for potential confounders. After a 5-year follow-up, significant inverse correlations were observed between changes in TPE and plasma triglyceride concentration (β = −8.563; P = 0.007), glucose concentration (β = −4.164; P = 0.036), and diastolic blood pressure (β = −1.316; P = 0.013). Our results suggest that the consumption of more polyphenols, measured as TPE in urine, could exert a protective effect against some cardiovascular risk factors. PMID:26881019

  1. Studies on the excretion of ascorbic acid 2-sulfate and total vitamin C into human urine after oral administration of ascorbic acid 2-sulfate.

    PubMed

    Tsujimura, M; Fukuda, T; Kasai, T

    1982-10-01

    The excretion of AsS and total vitamin C into urine after oral administration of AsS to humans was investigated. When 10 mmol of AsS was administered to the subjects, the excretion of AsS into urine continued for 60 hr in males and 48 hr in females. The average amount excreted per hour was less than 5 mg. These results differed from those for AsA and DAsA orally administered to humans. The determination of vitamin C after oral administration of AsS to the subjects consisting of ten males and six females showed no vitamin C effect in humans, similarly to the case with the guinea pig and the rhesus monkey.

  2. The effects of intracrystalline and surface-bound proteins on the attachment of calcium oxalate monohydrate crystals to renal cells in undiluted human urine

    PubMed Central

    Grover, Phulwinder K.; Thurgood, Lauren A.; Wang, Tingting; Ryall, Rosemary L.

    2010-01-01

    Objective To compare the binding to Madin-Darby canine kidney (MDCK)-II cells of: (i) inorganic calcium oxalate monohydrate (iCOM) crystals and COM crystals precipitated from urine containing different concentrations of protein; and (ii) urinary COM crystals containing intracrystalline and intracrystalline + surface-bound protein. Materials and methods Urinary COM crystals were generated in sieved (sCOM), centrifuged and filtered (cfCOM), and ultrafiltered (ufCOM) portions of a pooled human urine and their adhesion to MDCK-II cells was compared using six different ultrafiltered urine samples as the binding medium. Crystal matrix extract (CME) was prepared by demineralizing calcium oxalate crystals precipitated from human urine and used to prepare COM crystals with intracrystalline, and intracrystalline + surface-bound CME at protein concentrations of 0, 0.05, 0.1, 0.5 and 5.0 mg/L. The amount of protein associated with the crystals was qualitatively assessed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting, using prothrombin fragment 1 (PTF1) as a marker. Protein concentration was determined in sieved, centrifuged and filtered, and ultrafiltered fractions of 10 additional urine samples. Results The median crystal attachment in the six urine types decreased in the order iCOM > ufCOM > cfCOM = sCOM, in inverse proportion to the concentration of protein in the solution or urine from which they were precipitated. sCOM and cfCOM crystals bound ≈□ 23% less than iCOM crystals. The attachment of COM crystals generated in the presence of increasing concentrations of CME proteins was unaffected up to a concentration of 5 mg/L, but binding of crystals containing the same concentrations of intracrystalline + surface-bound proteins decreased proportionally at protein concentrations from 0 to 5.0 mg/L. Conclusion Inorganic COM crystals bind significantly more strongly to MDCK-II cells than urinary crystals precipitated from sieved

  3. Selected reaction monitoring as an effective method for reliable quantification of disease-associated proteins in maple syrup urine disease.

    PubMed

    Fernández-Guerra, Paula; Birkler, Rune I D; Merinero, Begoña; Ugarte, Magdalena; Gregersen, Niels; Rodríguez-Pombo, Pilar; Bross, Peter; Palmfeldt, Johan

    2014-09-01

    Selected reaction monitoring (SRM) mass spectrometry can quantitatively measure proteins by specific targeting of peptide sequences, and allows the determination of multiple proteins in one single analysis. Here, we show the feasibility of simultaneous measurements of multiple proteins in mitochondria-enriched samples from cultured fibroblasts from healthy individuals and patients with mutations in branched-chain α-ketoacid dehydrogenase (BCKDH) complex. BCKDH is a mitochondrial multienzyme complex and its defective activity causes maple syrup urine disease (MSUD), a rare but severe inherited metabolic disorder. Four different genes encode the catalytic subunits of BCKDH: E1α (BCKDHA), E1β (BCKDHB), E2 (DBT), and E3 (DLD). All four proteins were successfully quantified in healthy individuals. However, the E1α and E1β proteins were not detected in patients carrying mutations in one of those genes, whereas mRNA levels were almost unaltered, indicating instability of E1α and E1β monomers. Using SRM we elucidated the protein effects of mutations generating premature termination codons or misfolded proteins. SRM is a complement to transcript level measurements and a valuable tool to shed light on molecular mechanisms and on effects of pharmacological therapies at protein level. SRM is particularly effective for inherited disorders caused by multiple proteins such as defects in multienzyme complexes.

  4. Selected reaction monitoring as an effective method for reliable quantification of disease-associated proteins in maple syrup urine disease

    PubMed Central

    Fernández-Guerra, Paula; Birkler, Rune I D; Merinero, Begoña; Ugarte, Magdalena; Gregersen, Niels; Rodríguez-Pombo, Pilar; Bross, Peter; Palmfeldt, Johan

    2014-01-01

    Selected reaction monitoring (SRM) mass spectrometry can quantitatively measure proteins by specific targeting of peptide sequences, and allows the determination of multiple proteins in one single analysis. Here, we show the feasibility of simultaneous measurements of multiple proteins in mitochondria-enriched samples from cultured fibroblasts from healthy individuals and patients with mutations in branched-chain α-ketoacid dehydrogenase (BCKDH) complex. BCKDH is a mitochondrial multienzyme complex and its defective activity causes maple syrup urine disease (MSUD), a rare but severe inherited metabolic disorder. Four different genes encode the catalytic subunits of BCKDH: E1α (BCKDHA), E1β (BCKDHB), E2 (DBT), and E3 (DLD). All four proteins were successfully quantified in healthy individuals. However, the E1α and E1β proteins were not detected in patients carrying mutations in one of those genes, whereas mRNA levels were almost unaltered, indicating instability of E1α and E1β monomers. Using SRM we elucidated the protein effects of mutations generating premature termination codons or misfolded proteins. SRM is a complement to transcript level measurements and a valuable tool to shed light on molecular mechanisms and on effects of pharmacological therapies at protein level. SRM is particularly effective for inherited disorders caused by multiple proteins such as defects in multienzyme complexes. PMID:25333063

  5. Significant increase in cultivation of Gardnerella vaginalis, Alloscardovia omnicolens, Actinotignum schaalii, and Actinomyces spp. in urine samples with total laboratory automation.

    PubMed

    Klein, Sabrina; Nurjadi, Dennis; Horner, Susanne; Heeg, Klaus; Zimmermann, Stefan; Burckhardt, Irene

    2018-04-13

    While total laboratory automation (TLA) is well established in laboratory medicine, only a few microbiological laboratories are using TLA systems. Especially in terms of speed and accuracy, working with TLA is expected to be superior to conventional microbiology. We compared in total 35,564 microbiological urine cultures with and without incubation and processing with BD Kiestra TLA for a 6-month period each retrospectively. Sixteen thousand three hundred thirty-eight urine samples were analyzed in the pre-TLA period and 19,226 with TLA. Sixty-two percent (n = 10,101/16338) of the cultures processed without TLA and 68% (n = 13,102/19226) of the cultures processed with TLA showed growth. There were significantly more samples with two or more species per sample and with low numbers of colony forming units (CFU) after incubation with TLA. Regarding the type of bacteria, there were comparable amounts of Enterobacteriaceae in the samples, slightly less non-fermenting Gram-negative bacteria, but significantly more Gram-positive cocci, and Gram-positive rods. Especially Alloscardivia omnicolens, Gardnerella vaginalis, Actinomyces spp., and Actinotignum schaalii were significantly more abundant in the samples incubated and processed with TLA. The time to report was significantly lower in the TLA processed samples by 1.5 h. We provide the first report in Europe of a large number of urine samples processed with TLA. TLA showed enhanced growth of non-classical and rarely cultured bacteria from urine samples. Our findings suggest that previously underestimated bacteria may be relevant pathogens for urinary tract infections. Further studies are needed to confirm our findings.

  6. Calcium - urine

    MedlinePlus

    Urinary Ca+2; Kidney stones - calcium in urine; Renal calculi - calcium in your urine; Parathyroid - calcium in urine ... A 24-hour urine sample is most often needed: On day 1, urinate into the toilet when you wake up in the morning. ...

  7. Urine Color

    MedlinePlus

    ... during urinary tract infections caused by pseudomonas bacteria. Dark brown or cola-colored urine Brown urine can ... of fava beans, rhubarb or aloe can cause dark brown urine. Medications. A number of drugs can ...

  8. Total chemical synthesis of human matrix Gla protein

    PubMed Central

    Hackeng, Tilman M.; Rosing, Jan; Spronk, Henri M.H.; Vermeer, Cees

    2001-01-01

    Human matrix Gla protein (MGP) is a vitamin K–dependent extracellular matrix protein that binds Ca2+ ions and that is involved in the prevention of vascular calcification. MGP is a 10.6-kD protein (84 amino acids) containing five γ-carboxyglutamic acid (Gla) residues and one disulfide bond. Studies of the mechanism by which MGP prevents calcification of the arterial media are hampered by the low solubility of the protein (<10 μg/mL). Because of solubility problems, processing of a recombinantly expressed MGP-fusion protein chimera to obtain MGP was unsuccessful. Here we describe the total chemical synthesis of MGP by tBoc solid-phase peptide synthesis (SPPS) and native chemical ligation. Peptide Tyr1-Ala53 was synthesized on a derivatized resin yielding a C-terminal thioester group. Peptide Cys54-Lys84 was synthesized on Lys-PAM resin yielding a C-terminal carboxylic acid. Subsequent native chemical ligation of the two peptides resulted in the formation of a native peptide bond between Ala53 and Cys54. Folding of the 1–84-polypeptide chain in 3 M guanidine (pH 8) resulted in a decrease of molecular mass from 10,605 to 10,603 (ESI-MS), representing the loss of two protons because of the formation of the Cys54-Cys60 internal disulfide bond. Like native MGP, synthetic MGP had the same low solubility when brought into aqueous buffer solutions with physiological salt concentrations, confirming its native like structure. However, the solubility of MGP markedly increased in borate buffer at pH 7.4 in the absence of sodium chloride. Ca2+-binding to MGP was confirmed by analytical HPLC, on which the retention time of MGP was reduced in the presence of CaCl2. Circular dichroism studies revealed a sharp increase in α-helicity at 0.2 mM CaCl2 that may explain the Ca2+-dependent shift in high-pressure liquid chromatography (HPLC)-retention time of MGP. In conclusion, facile and efficient chemical synthesis in combination with native chemical ligation yielded MGP

  9. Effect of a grain challenge on ruminal, urine, and fecal pH, apparent total-tract starch digestibility, and milk composition of Holstein and Jersey cows.

    PubMed

    Luan, S; Cowles, K; Murphy, M R; Cardoso, F C

    2016-03-01

    The effects of a grain challenge on ruminal, urine, and fecal pH, apparent total-tract starch digestibility, and milk composition were determined. Six Holstein cows, 6 rumen-cannulated Holstein cows, and 6 Jersey cows were used in a replicated 3 × 3 Latin square design balanced to measure carryover effects. Periods (10 d) were divided into 4 stages (S): S1, d 1 to 3, served as baseline with regular total mixed ration ad libitum; S2, d 4, served as restricted feeding, with cows offered 50% of the total mixed ration fed on S1 (dry matter basis); S3, d 5, a grain challenge was performed, in which cows were fed total mixed ration ad libitum and not fed (CON) or fed an addition of 10% (MG) or 20% (HG) pellet wheat-barley (1:1) top-dressed onto the total mixed ration, based on dry matter intake obtained in S1; S4, d 6 to 10, served as recovery stage with regular total mixed ration fed ad libitum. Overall, cows had a quadratic treatment effect for milk yield where CON (22.6 kg/d) and HG (23.5 kg/d) had lower milk yield than cows in MG (23.7 kg/d). Jersey cows had a quadratic treatment effect for dry matter intake where cows in CON (13.2 kg/d) and HG (12.4 kg/d) had lower dry matter intake than cows in MG (14 kg/d). Holstein cows had a linear treatment effect for dry matter intake (17.7, 18.4, and 18.6 kg/d for CON, MG, and HG, respectively). Rumen pH for the rumen-cannulated cows had a linear treatment effect (6.45, 6.35, and 6.24 for CON, MG, and HG, respectively). Cows in HG spent more time with rumen pH below 5.8 (4.33 h) than MG (2 h) or CON (2.17 h) as shown by the quadratic treatment effect. Holstein cows in HG (8.46) had lower urine pH than MG (8.51) or CON (8.54) as showed by the linear treatment effect for urine pH. Apparent total-tract starch digestibility had a tendency for a linear treatment effect on S3 (97.62 ± 1.5, 97.47 ± 1.5, and 91.84 ± 1.6%, for CON, MG, and HG, respectively). Fecal pH was associated with rumen pH depression as early as 15 h after

  10. Lime powder treatment reduces urinary excretion of total protein and transferrin but increases uromodulin excretion in patients with urolithiasis.

    PubMed

    Tosukhowong, Piyaratana; Kulpradit, Pimsuda; Chaiyarit, Sakdithep; Ungjareonwattana, Wattanachai; Kalpongnukul, Nuttiya; Ratchanon, Supoj; Thongboonkerd, Visith

    2018-06-01

    Our previous study has shown that lime powder (LP) had an inhibitory effect against calcium oxalate stone formation. However, the precise mechanisms underlying such beneficial effect remained unclear. Our present study thus aimed to address the effect of LP on excretory level and compositions of urinary proteins using a proteomics approach. From a total of 80 calcium oxalate stone formers recruited into our 2-year randomized clinical trial of LP effect, 10 patients with comparable age and clinical parameters were selected for this proteomic study. 24-h urine specimens were collected from all subjects, at baseline (before) and after LP treatment for 6 months, and then subjected to quantitative proteomics analysis and subsequent validation by ELISA. Total urinary protein excretion was significantly decreased by LP treatment, but unaffected by placebo. Nanoflow liquid chromatography coupled to tandem mass spectrometry (nanoLC-MS/MS) followed by quantitative analysis revealed 17 proteins whose levels were significantly altered (16 decreased and 1 increased) exclusively by LP treatment. Among these, the decrease of transferrin and increase of uromodulin were validated by ELISA. Moreover, there was a significant correlation between microalbuminuria and urinary transferrin level by Pearson's correlation test. In summary, LP treatment caused significant reduction in total urinary protein excretion and changes in urinary protein compositions that could be linked to stone inhibitory effects and might be relevant mechanisms responsible for the beneficial effects of LP to prevent kidney stone formation and recurrence.

  11. Elevation of urinary adipsin in preeclampsia: correlation with urine protein concentration and the potential use for a rapid diagnostic test.

    PubMed

    Wang, Tao; Zhou, Rong; Gao, Linbo; Wang, Yanyun; Song, Changping; Gong, Yunhui; Jia, Jin; Xiong, Wei; Dai, Li; Zhang, Lin; Hu, Huaizhong

    2014-10-01

    Early diagnosis and treatment of preeclampsia are essential for prevention of seizure development and fetus maturation. Although various methods have been developed for predicting or monitoring the onset of preeclampsia, a simple assay that can be used as a home or point of care test remains unavailable. We attempted to find a urinary protein that could be used as a biomarker for developing such a test. Urinary samples were collected from 124 preeclampsia and 135 healthy pregnant women for screening using a protein array technology and quantification by ELISA. A urinary protein, adipsin, was found significantly increased, and the adipsin creatinine ratio was closely correlated with the urinary 24-hour protein in patients with preeclampsia. When combined with the increased diastolic blood pressure (≥90 mm Hg), the sensitivity was 90.3% and the specificity reached 100.0% for preeclampsia diagnosis. We then developed a laminar flow immunoassay for rapid diagnosis, and the sensitivity and specificity were 89.04% and 100%, respectively, when combined with increased diastolic blood pressure. Because of the easiness of sample collection, assay conduction, and result interpretation, this urine test can be potentially used as a home test for monitoring preeclampsia onset for high-risk pregnant women and as a rapid test for a preliminary diagnosis for emergency patients at hospitals. © 2014 American Heart Association, Inc.

  12. Comparison of synovial fluid, urine, and serum ion levels in metal-on-metal total hip arthroplasty at a minimum follow-up of 18 years.

    PubMed

    Lass, Richard; Grübl, Alexander; Kolb, Alexander; Stelzeneder, David; Pilger, Alexander; Kubista, Bernd; Giurea, Alexander; Windhager, Reinhard

    2014-09-01

    Diagnosis of adverse reactions to metal debris in metal-on-metal hip arthroplasty is a multifactorial process. Systemic ion levels are just one factor in the evaluation and should not be relied upon solely to determine the need for revision surgery. Furthermore, the correlation between cobalt or chromium serum, urine, or synovial fluid levels and adverse local tissue reactions is still incompletely understood. The hypothesis was that elevated serum and urine metal-ion concentrations are associated with elevated local metal-ion concentrations in primary total hip arthroplasties (THA) and with failure of metal-on-metal articulations in the long-term. In our present study, we evaluated these concentrations in 105 cementless THA with metal-on-metal articulating surfaces with small head diameter at a minimum of 18 years postoperatively. Spearman correlation showed a high correlation between the joint fluid aspirate concentration of cobalt and chromium with the serum cobalt (r = 0.81) and chromium level (r = 0.77) in patients with the THA as the only source of metal-ions. In these patients serum metal-ion analysis is a valuable method for screening. In patients with more than one source of metal or renal insufficiency additional investigations, like joint aspirations are an important tool for evaluation of wear and adverse tissue reactions in metal-on-metal THA. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  13. Leucine aminopeptidase - urine

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003617.htm Leucine aminopeptidase - urine To use the sharing features on this page, please enable JavaScript. Leucine aminopeptidase is a type of protein called an ...

  14. Identification of L. infantum chagasi proteins in VL patients' urine: a promising antigen discovery approach of vaccine candidates

    PubMed Central

    Kashino, Suely S.; Abeijon, Claudia; Qin, Lizeng; Kanunfre, Kelly A.; Kubrusly, Flávia S.; Silva, Fernando O.; Costa, Dorcas L.; Campos, Dioclécio; Costa, Carlos H.N.; Raw, Isaias; Campos-Neto, Antonio

    2012-01-01

    Visceral leishmaniasis (VL) is a serious lethal parasitic disease caused by Leishmania donovani in Asia and by Leishmania infantum chagasi in Southern Europe and South America. VL is endemic in 47 countries with an annual incidence estimated to be 500,000 cases. This high incidence is due in part to the lack of an efficacious vaccine. Here, we introduce an innovative approach to directly identify parasite vaccine candidate antigens that are abundantly produced in vivo in humans with VL. We combined RP-HPLC and mass spectrometry and categorized three L. infantum chagasi proteins, presumably produced in spleen, liver, and bone marrow lesions and excreted in the patients’ urine. Specifically, these proteins were the following: Li-isd1 (XP_001467866.1), Li-txn1 (XP_001466642.1), and Li-ntf2 (XP_001463738.1). Initial vaccine validation studies were performed with the rLi-ntf2 protein produced in E. coli mixed with the adjuvant BpMPLA-SE. This formulation stimulated potent Th1 response in BALB/c mice. Compared to control animals, mice immunized with Li-ntf2 + BpMPLASE had a marked parasite burden reduction in spleens at 40 days post-challenge with virulent L. infantum chagasi. These results strongly support the proposed antigen discovery strategy of vaccine candidates to kala-azar and opens novel possibilities for vaccine development to other serious infectious diseases. PMID:22443237

  15. Amylase - urine

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003607.htm Amylase - urine To use the sharing features on this page, ... test that measures the amount of amylase in urine. Amylase is an enzyme that helps digest carbohydrates. ...

  16. Cortisol - urine

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003703.htm Cortisol urine test To use the sharing features on this page, please enable JavaScript. The cortisol urine test measures the level of cortisol in the ...

  17. Urine Cytology

    MedlinePlus

    ... types of cells were found in your urine sample. You may need to repeat the test. Negative. This means no cancer cells were identified in your urine sample. Atypical. This indicates that some abnormalities were found ...

  18. Urination - painful

    MedlinePlus

    ... the urine ? Are there any changes in the volume or frequency of urination? Do you feel the ... Walls RM, et al, eds. Rosen's Emergency Medicine: Concepts and Clinical Practice . 8th ed. Philadelphia, PA: Elsevier ...

  19. Myoglobin urine test

    MedlinePlus

    Urine myoglobin; Heart attack - myoglobin urine test; Myositis - myoglobin urine test; Rhabdomyolysis - myoglobin urine test ... The test involves only normal urination, which should cause no discomfort.

  20. Measurements of C-reactive protein (CRP) and nerve-growth-factor (NGF) concentrations in serum and urine samples of dogs with neurologic disorders.

    PubMed

    Kordass, Ulrike; Carlson, Regina; Stein, Veronika Maria; Tipold, Andrea

    2016-01-08

    The purpose of this study was to prove the hypothesis that C-reactive protein (CRP) and nerve growth factor (NGF) may be potential biomarkers for lower urinary tract disorders and may be able to distinguish between micturition dysfunctions of different origin in dogs with spinal cord diseases. NGF- and CRP- concentrations were measured in serum and urine samples using specific ELISA-Kits. Results in urine were standardized by urine-creatinine levels. CRP in serum was detectable in 32/76 and in urine samples in 40/76 patients. NGF could be measured in all serum and in 70/76 urine samples. Urinary CRP concentrations were significantly higher in dogs with micturition dysfunction (p = 0.0009) and in dogs with different neurological diseases (p = 0.0020) compared to the control group. However, comparing dogs with spinal cord disorders with and without associated micturition dysfunction no significant difference could be detected for NGF and CRP values in urine or serum samples. Additionally, levels did not decrease significantly, when measured at the time when the dogs regained the ability to urinate properly (urinary NGF p = 0.7962; urinary CRP p = 0.078). Urine samples with bacteria and/or leukocytes had no significant increase in urinary NGF (p = 0.1112) or CRP (p = 0.0534) concentrations, but higher CRP-levels in urine from dogs with cystitis were found compared to dogs without signs of cystitis. From these data we conclude that neither CRP nor NGF in urine or serum can be considered as reliable biomarkers for micturition disorders in dogs with spinal cord disorders in a clinical setting, but their production might be part of the pathogenesis of such disorders. Significantly higher levels of CRP could be found in the urine of dogs with micturition dysfunctions compared to control dogs. This phenomenon could potentially be explained by unspecific extrahepatic CRP production by smooth muscle cells in the dilated bladder.

  1. Urine protein profiling identified alpha-1-microglobulin and haptoglobin as biomarkers for early diagnosis of acute allograft rejection following kidney transplantation.

    PubMed

    Stubendorff, Beatrice; Finke, Stephanie; Walter, Martina; Kniemeyer, Olaf; von Eggeling, Ferdinand; Gruschwitz, Torsten; Steiner, Thomas; Ott, Undine; Wolf, Gunter; Wunderlich, Heiko; Junker, Kerstin

    2014-12-01

    Early diagnosis of acute rejection and effective immunosuppressive therapy lead to improvement in graft survival following kidney transplantation. In this study, we aimed to establish a urinary protein profile suitable to distinguish between patients with rejection and stable graft function and to predict acute rejection based on postoperatively collected urine samples. A further objective was to identify candidate proteins for the use as biomarkers in clinical practice. Urine samples of 116 kidney recipients were included. Rejection was proven by biopsy (n = 58), and stable transplant function was monitored for at least 2 years (n = 58). Postoperative urine samples were collected between 3rd and 10th day following transplantation. Urinary protein profiles were obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Protein identification and validation were performed using multiplex fluorescence 2DE, peptide mass fingerprinting and enzyme-linked immunosorbent assay. A protein profile including four mass peaks differentiated acute rejection from stable transplants at the time point of rejection and at the postoperative state with 73 % sensitivity and 88 % specificity. Alpha-1-microglobulin (A1MG) and Haptoglobin (Hp) were identified as putative rejection biomarkers. Protein levels were significantly higher in postoperative urine from patients with rejection (A1MG 29.13 vs. 22.06 μg/ml, p = 0.001; Hp 628.34 vs. 248.57 ng/ml, p = 0.003). The combination of both proteins enabled the diagnosis of early rejection with 85 % sensitivity and 80 % specificity. Protein profiling using mass spectrometry is suitable for noninvasive detection of rejection-specific changes following kidney transplantation. A specific protein profile enables the prediction of early acute allograft rejection in the immediate postoperative period. A1MG and Hp appear to be reliable rejection biomarkers.

  2. Quantification of proteins in urine samples using targeted mass spectrometry methods.

    PubMed

    Khristenko, Nina; Domon, Bruno

    2015-01-01

    Numerous clinical proteomics studies are focused on the development of biomarkers to improve either diagnostics for early disease detection or the monitoring of the response to the treatment. Although, a wealth of biomarker candidates are available, their evaluation and validation in a true clinical setup remains challenging. In biomarkers evaluation studies, a panel of proteins of interest are systematically analyzed in a large cohort of samples. However, in spite of the latest progresses in mass spectrometry, the consistent detection of pertinent proteins in high complex biological samples is still a challenging task. Thus, targeted LC-MS/MS methods are better suited for the systematic analysis of biomarkers rather than shotgun approaches. This chapter describes the workflow used to perform targeted quantitative analyses of proteins in urinary samples. The peptides, as surrogates of the protein of interest, are commonly measured using a triple quadrupole mass spectrometers operated in selected reaction monitoring (SRM) mode. More recently, the advances in targeted LC-MS/MS analysis based on parallel reaction monitoring (PRM) performed on a quadrupole-orbitrap instrument have allowed to increase the specificity and selectivity of the measurements.

  3. Holding Thermal Receipt Paper and Eating Food after Using Hand Sanitizer Results in High Serum Bioactive and Urine Total Levels of Bisphenol A (BPA)

    PubMed Central

    Hormann, Annette M.; vom Saal, Frederick S.; Nagel, Susan C.; Stahlhut, Richard W.; Moyer, Carol L.; Ellersieck, Mark R.; Welshons, Wade V.; Toutain, Pierre-Louis; Taylor, Julia A.

    2014-01-01

    Bisphenol A (BPA) is an endocrine disrupting environmental contaminant used in a wide variety of products, and BPA metabolites are found in almost everyone’s urine, suggesting widespread exposure from multiple sources. Regulatory agencies estimate that virtually all BPA exposure is from food and beverage packaging. However, free BPA is applied to the outer layer of thermal receipt paper present in very high (∼20 mg BPA/g paper) quantities as a print developer. Not taken into account when considering thermal paper as a source of BPA exposure is that some commonly used hand sanitizers, as well as other skin care products, contain mixtures of dermal penetration enhancing chemicals that can increase by up to 100 fold the dermal absorption of lipophilic compounds such as BPA. We found that when men and women held thermal receipt paper immediately after using a hand sanitizer with penetration enhancing chemicals, significant free BPA was transferred to their hands and then to French fries that were eaten, and the combination of dermal and oral BPA absorption led to a rapid and dramatic average maximum increase (Cmax) in unconjugated (bioactive) BPA of ∼7 ng/mL in serum and ∼20 µg total BPA/g creatinine in urine within 90 min. The default method used by regulatory agencies to test for hazards posed by chemicals is intra-gastric gavage. For BPA this approach results in less than 1% of the administered dose being bioavailable in blood. It also ignores dermal absorption as well as sublingual absorption in the mouth that both bypass first-pass liver metabolism. The elevated levels of BPA that we observed due to holding thermal paper after using a product containing dermal penetration enhancing chemicals have been related to an increased risk for a wide range of developmental abnormalities as well as diseases in adults. PMID:25337790

  4. Holding thermal receipt paper and eating food after using hand sanitizer results in high serum bioactive and urine total levels of bisphenol A (BPA).

    PubMed

    Hormann, Annette M; Vom Saal, Frederick S; Nagel, Susan C; Stahlhut, Richard W; Moyer, Carol L; Ellersieck, Mark R; Welshons, Wade V; Toutain, Pierre-Louis; Taylor, Julia A

    2014-01-01

    Bisphenol A (BPA) is an endocrine disrupting environmental contaminant used in a wide variety of products, and BPA metabolites are found in almost everyone's urine, suggesting widespread exposure from multiple sources. Regulatory agencies estimate that virtually all BPA exposure is from food and beverage packaging. However, free BPA is applied to the outer layer of thermal receipt paper present in very high (∼20 mg BPA/g paper) quantities as a print developer. Not taken into account when considering thermal paper as a source of BPA exposure is that some commonly used hand sanitizers, as well as other skin care products, contain mixtures of dermal penetration enhancing chemicals that can increase by up to 100 fold the dermal absorption of lipophilic compounds such as BPA. We found that when men and women held thermal receipt paper immediately after using a hand sanitizer with penetration enhancing chemicals, significant free BPA was transferred to their hands and then to French fries that were eaten, and the combination of dermal and oral BPA absorption led to a rapid and dramatic average maximum increase (Cmax) in unconjugated (bioactive) BPA of ∼7 ng/mL in serum and ∼20 µg total BPA/g creatinine in urine within 90 min. The default method used by regulatory agencies to test for hazards posed by chemicals is intra-gastric gavage. For BPA this approach results in less than 1% of the administered dose being bioavailable in blood. It also ignores dermal absorption as well as sublingual absorption in the mouth that both bypass first-pass liver metabolism. The elevated levels of BPA that we observed due to holding thermal paper after using a product containing dermal penetration enhancing chemicals have been related to an increased risk for a wide range of developmental abnormalities as well as diseases in adults.

  5. PAROTID FLUID TOTAL PROTEIN IN PATIENTS WITH UREMIA AND PROTEINURIA.

    DTIC Science & Technology

    Stimulated parotid fluid samples (238) were collected from 32 patients to determine if altered renal function was associated with deviations in...tubular necrosis, and 15 had normal renal function. There were no significant differences in parotid fluid protein concentration or minute secretion associated with the state of renal function. (Author)

  6. Triiodothyronine and thyroxine in urine. II. Renal handling, and effect of urinary protein.

    PubMed

    Burke, C W; Shakespear, R A

    1976-03-01

    Mean urinary clearances of T3 were 164 ml/min in normal subjects, 177 in pregnancy, 221 in thyrotoxicosis, 174 in hypothyroidism, and 194 in 3 persons with undetectable T4 but normal T3 levels. T4 clearances were 38 ml/min in normal subjects, 48 in thyrotoxicosis, and 138 in hypothyroidism. Low creatinine clearance was associated with low clearances of T4 and T3. The data suggest urinary excretion of T3 by glomerular filtration of serum unbound T3 with added tubular excretion; and T4 excretion by glomerular filtration of unbound T4 and tubular reabsorption. However, 3-9% of urinary T3 and 5-12% of urinary T4 were bound to urinary proteins, and increased protein excretion caused markedly increased T4 excretion. In addition, 52% of urinary T3 and 68% of urinary T4 were bound to other substances of approximate mol wt 500-2,000, which may influence tubular handling of T3 or T4.

  7. [Protein content in urine of male and female water vole (Arvicola amphibious) at the period of spring growth and sexual maturation].

    PubMed

    Nazarova, G G; Proskurniak, L P

    2012-01-01

    The study was carried out on the captive bread water voles Arvicola amphibious kept in vivarium. At the first decade of January, March, and June, the body length and anogenital distance were measured, the body mass was determined, and urine was collected for determination of its protein content. The obtained results have shown that the protein content depends on sex of the animals and is connected with the reproductive status of males and their dimension-weight characteristics. The urinary protein excretion level in females remained stable at different months, whereas in males its sharp rise was noted at the period of spring growth and sexual maturation. The significant sexual differences were established in March and enhanced in June. In March the urine protein content in males was noted to correlate positively with the body mass and length and with the anogenital distance. The males reached sexual maturity at the earlier calendar terms than the females did; in sexually mature males the urine protein content was significantly higher than in the sexually immature ones.

  8. Proximal tubule proteins are significantly elevated in bladder urine of patients with ureteropelvic junction obstruction and may represent novel biomarkers: A pilot study.

    PubMed

    Gerber, Claire; Harel, Miriam; Lynch, Miranda L; Herbst, Katherine W; Ferrer, Fernando A; Shapiro, Linda H

    2016-04-01

    Ureteropelvic junction obstruction (UPJO) is the major cause of hydronephrosis in children and may lead to renal injury and early renal dysfunction. However, diagnosis of the degree of obstruction and severity of renal injury relies on invasive and often inconclusive renal scans. Biomarkers from voided urine that detect early renal injury are highly desirable because of their noninvasive collection and their potential to assist in earlier and more reliable diagnosis of the severity of obstruction. Early in response to UPJO, increased intrarenal pressure directly impacts the proximal tubule brush border. We hypothesize that single-pass, apically expressed proximal tubule brush border proteins will be shed into the urine early and rapidly and will be reliable noninvasive urinary biomarkers, providing the tools for a more reliable stratification of UPJO patients. We performed a prospective cohort study at Connecticut Children's Medical Center. Bladder urine samples from 12 UPJO patients were obtained prior to surgical intervention. Control urine samples were collected from healthy pediatric patients presenting with primary nocturnal enuresis. We determined levels of NGAL, KIM-1 (previously identified biomarkers), CD10, CD13, and CD26 (potentially novel biomarkers) by ELISA in control and experimental urine samples. Urinary creatinine levels were used to normalize the urinary protein levels measured by ELISA. Each of the proximal tubule proteins outperformed the previously published biomarkers. No differences in urinary NGAL and KIM-1 levels were observed between control and obstructed patients (p = 0.932 and p = 0.799, respectively). However, levels of CD10, CD13, and CD26 were significantly higher in the voided urine of obstructed individuals when compared with controls (p = 0.002, p = 0.024, and p = 0.007, respectively) (Figure). Targeted identification of reliable, noninvasive biomarkers of renal injury is critical to aid in diagnosing patients at risk, guiding

  9. Characterization of protein-bound gold in rat urine following aurothiomalate administration and of rat and human albumin-gold-thiomalate.

    PubMed

    Shaw, C F; Schaeffer-Memmel, N; Krawczak, D

    1986-03-01

    The metabolites of gold in the urine of rats given the antiarthritic drug aurothiomalate were investigated by gel permeation chromatography, electrophoresis, and chemical studies. Following a single dose of aurtothiomalate, the excreted gold was protein-bound in the high-molecular-weight (greater than or equal to 150,000 dalton) and serum albumin fractions. Electrophoresis confirmed the presence of albumin, but showed that the other proteins present differ from those in normal or in vitro aurothiomalate-incubated rat sera. The pattern of the proteins establishes that the proteinuria was of the glomerular type. The alterations in the gold distribution produced by incubation of the urine with the low-molecular-weight thiol penicillamine and with exogenously added aurothiomalate indicated the existence of a labile equilibrium of gold among protein binding sites in the urine. Incubation of rat and human sera and commercially prepared serum albumins with aurothiomalate increased the electrophoretic mobility of the albumin. The significance of this change in electrophoretic mobility with respect to two models of gold binding by serum albumin is discussed.

  10. Urine Odor

    MedlinePlus

    ... doctor. Brunzel NA. Physical examination of urine. In: Fundamentals of Urine and Body Fluid Analysis. 3rd ed. St. Louis, Mo.: Saunders Elsevier; 2013:97. McPherson RA, et al., eds. Henry's Clinical Diagnosis and Management by Laboratory Methods. 23rd ed. St. Louis, Mo.: ...

  11. Detecting early kidney damage in horses with colic by measuring matrix metalloproteinase -9 and -2, other enzymes, urinary glucose and total proteins

    PubMed Central

    Arosalo, Bela M; Raekallio, Marja; Rajamäki, Minna; Holopainen, Elina; Kastevaara, Tuulia; Salonen, Hanna; Sankari, Satu

    2007-01-01

    Background The aim of the study was to investigate urine matrix metalloproteinase (MMP-2 and -9) activity, alkaline phosphatase/creatinine (U-AP/Cr) and gamma-glutamyl-transpeptidase/creatinine (U-GGT/Cr) ratios, glucose concentration, and urine protein/creatinine (U-Prot/Cr) ratio and to compare data with plasma MMP-2 and -9 activity, cystatin-C and creatinine concentrations in colic horses and healthy controls. Horses with surgical colic (n = 5) were compared to healthy stallions (n = 7) that came for castration. Blood and urine samples were collected. MMP gelatinolytic activity was measured by zymography. Results We found out that horses with colic had significantly higher urinary MMP-9 complex and proMMP-9 activities than horses in the control group. Colic horses also had higher plasma MMP-2 activity than the control horses. Serum creatinine, although within reference range, was significantly higher in the colic horses than in the control group. There was no significant increase in urinary alkaline phosphatase, gamma-glutamyltranspeptidase or total proteins in the colic horses compared to the control group. A human cystatin-C test (Dako Cytomation latex immunoassay® based on turbidimetry) did not cross react with equine cystatin-C. Conclusion The results indicate that plasma MMP-2 may play a role in the pathogenesis of equine colic and urinary MMP-9 in equine kidney damage. PMID:17244354

  12. HPLC-MS/MS analysis of anthocyanins in human plasma and urine using protein precipitation and dilute-and-shoot sample preparation methods, respectively.

    PubMed

    Liu, Junguo; Song, Jiuxue; Huang, Karen; Michel, Deborah; Fang, Jim

    2018-05-01

    A high-performance liquid chromatography tandem-mass spectrometry (HPLC-MS/MS) method has been developed to analyze anthocyanins in urine and plasma to further understand their absorption, distribution, metabolism and excretion. The method employed a Synergi RP-Max column (250 × 4.6 mm, 4 μm) and an API 4000 mass spectrometer. A gradient elution system consisted of mobile phase A (water-1% formic acid) and mobile phase B (acetonitrile) with a flow rate of 0.60 mL/min. The gradient was initiated at 5% B, increased to 21% B at 20 min, and then increased to 40% B at 35 min. The analysis of anthocyanins presents a challenge because of the poor stability of anthocyanins during sample preparation, especially during solvent evaporation. In this method, the degradation of anthocyanins was minimized using protein precipitation and dilute-and-shoot and sample preparation methods for plasma and urine, respectively. No interferences were observed from endogenous compounds. The method has been used to analyze anthocyanin concentrations in urine and plasma samples from volunteers administered saskatoon berries. Cyanidin-3-galactoside, cyanidin-3-glucoside, cyanidin-3-arabinoside, cyanidin-3-xyloside and quercetin-3-galactoside, the five major flavonoid components in saskatoon berries, were identified in plasma and urine samples. Copyright © 2017 John Wiley & Sons, Ltd.

  13. Monocyte chemoattractant protein-1 and interleukin-8 levels in urine and serum of patents with hemolytic uremic syndrome.

    PubMed

    van Setten, P A; van Hinsbergh, V W; van den Heuvel, L P; Preyers, F; Dijkman, H B; Assmann, K J; van der Velden, T J; Monnens, L A

    1998-06-01

    The epidemic form of the hemolytic uremic syndrome (HUS) in children is hallmarked by endothelial cell damage, most predominantly displayed by the glomerular capillaries. The influx of mononuclear (MO) and polymorphonuclear cells (PMNs) into the glomeruli may be an important event in the initiation, prolongation, and progression of glomerular endothelial cell damage in HUS patients. The molecular mechanisms for the recruitment of these leukocytes into the kidney are unclear, but monocyte chemoattractant protein-1 (MCP-1) and IL-8 are suggested to be prime candidates. In this study, we analyzed the presence of both chemokines in 24-h urinary (n = 15) and serum (n = 14) samples of HUS children by specific ELISAs. Furthermore, kidney biopsies of three different HUS children were examined for MO and PMN cell infiltration by histochemical techniques and electron microscopy. Whereas the chemokines MCP-1 and IL-8 were present in only very limited amounts in urine of 17 normal control subjects, serial samples of HUS patients demonstrated significantly elevated levels of both chemokines. HUS children with anuria showed higher initial and maximum chemokine levels than their counterparts without anuria. A strong positive correlation was observed between urinary MCP-1 and IL-8 levels. Whereas initial serum IL-8 levels were significantly increased in HUS children, serum MCP-1 levels were only slightly elevated compared with serum MCP-1 in control children. No correlation was found between urinary and serum chemokine concentrations. Histologic and EM studies of HUS biopsy specimens clearly showed the presence of MOs and to a lesser extent of PMNs in the glomeruli. The present data suggest an important local role for MOs and PMNs in the process of glomerular endothelial-cell damage. The chemokines MCP-1 and IL-8 may possibly be implicated in the pathogenesis of HUS through the recruitment and activation of MOs and PMNs, respectively.

  14. A dye binding method for measurement of total protein in microalgae.

    PubMed

    Servaites, Jerome C; Faeth, Julia L; Sidhu, Sukh S

    2012-02-01

    Protein is a large component of the standing biomass of algae. The total protein content of algae is difficult to measure because of the problems encountered in extracting all of the protein from the cells. Here we modified an existing protein assay to measure total protein in microalgae cells that involves little or no extraction of protein from the cells. Aliquots of fresh or pretreated cells were spotted onto filter paper strips. After drying, the strips were stained in a 0.1% (w/v) solution of the protein stain Coomassie Brilliant Blue R-250 for 16 to 24 h and then destained. The stained protein spots were cut out from the paper, and dye was eluted in 1% (w/v) sodium dodecyl sulfate (SDS). Absorbance at 600 nm was directly proportional to protein concentration. Cells that were recalcitrant to taking up the dye could be either heated at 80°C for 10 min in 1% SDS or briefly sonicated for 3 min to facilitate penetration of the dye into the cells. Total protein measured in Chlorella vulgaris using this method compared closely with that measured using the total N method. Total protein concentrations were measured successfully in 12 algal species using this dye binding method. Copyright © 2011 Elsevier Inc. All rights reserved.

  15. Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm-Horsfall protein in human urine.

    PubMed

    Akimoto, Masaru; Hokazono, Eisaku; Ota, Eri; Tateishi, Takiko; Kayamori, Yuzo

    2016-01-01

    Tamm-Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm-Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm-Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results. We developed a novel, quantitative assay for Tamm-Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm-Horsfall protein from other major urine components. Our method exhibited high precision (based on the relative standard deviations of intraday [≤2.77%] and interday [≤5.35%] repetitions). The Tamm-Horsfall protein recovery rate was 100.0-104.2%. The mean Tamm-Horsfall protein concentration in 25 healthy individuals was 31.6 ± 18.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r = 0.906), but enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm-Horsfall protein concentrations. The high sensitivity and reproducibility of our Tamm-Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high-performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm-Horsfall protein. © The Author(s) 2015.

  16. Urine Preservative

    NASA Technical Reports Server (NTRS)

    Smith, Scott M. (Inventor); Nillen, Jeannie (Inventor)

    2001-01-01

    Disclosed is CPG, a combination of a chlorhexidine salt (such as chlorhexidine digluconate, chlorhexidine diacetate, or chlorhexidine dichloride) and n-propyl gallate that can be used at ambient temperatures as a urine preservative.

  17. Urine - bloody

    MedlinePlus

    ... movement The urine can also turn a red color from certain drugs, beets, or other foods. ... surgery or an injury? Have you recently eaten foods that may cause a change in color, like beets, berries, or rhubarb? Tests that may ...

  18. Bilirubin - urine

    MedlinePlus

    ... Direct bilirubin - urine Images Male urinary system References Berk PD, Korenblat KM. Approach to the patient with ... Review Date 5/21/2017 Updated by: Laura J. Martin, MD, MPH, ABIM Board Certified in Internal ...

  19. Comparison between urine albumin-to-creatinine ratio and urine protein dipstick testing for prevalence and ability to predict the risk for chronic kidney disease in the general population (Iwate-KENCO study): a prospective community-based cohort study.

    PubMed

    Koeda, Yorihiko; Tanaka, Fumitaka; Segawa, Toshie; Ohta, Mutsuko; Ohsawa, Masaki; Tanno, Kozo; Makita, Shinji; Ishibashi, Yasuhiro; Itai, Kazuyoshi; Omama, Shin-Ichi; Onoda, Toshiyuki; Sakata, Kiyomi; Ogasawara, Kuniaki; Okayama, Akira; Nakamura, Motoyuki

    2016-05-12

    This study compared the combination of estimated glomerular filtration rate (eGFR) and urine albumin-to-creatinine ratio (UACR) vs. eGFR and urine protein reagent strip testing to determine chronic kidney disease (CKD) prevalence, and each method's ability to predict the risk for cardiovascular events in the general Japanese population. Baseline data including eGFR, UACR, and urine dipstick tests were obtained from the general population (n = 22 975). Dipstick test results (negative, trace, positive) were allocated to three levels of UACR (<30, 30-300, >300), respectively. In accordance with Kidney Disease Improving Global Outcomes CKD prognosis heat mapping, the cohort was classified into four risk grades (green: grade 1; yellow: grade 2; orange: grade 3, red: grade 4) based on baseline eGFR and UACR levels or dipstick tests. During the mean follow-up period of 5.6 years, 708 new onset cardiovascular events were recorded. For CKD identified by eGFR and dipstick testing (dipstick test ≥ trace and eGFR <60 mL/min/1.73 m(2)), the incidence of CKD was found to be 9 % in the general population. In comparison to non-CKD (grade 1), although cardiovascular risk was significantly higher in risk grades ≥3 (relative risk (RR) = 1.70; 95 % CI: 1.28-2.26), risk predictive ability was not significant in risk grade 2 (RR = 1.20; 95 % CI: 0.95-1.52). When CKD was defined by eGFR and UACR (UACR ≥30 mg/g Cr and eGFR <60 mL/min/1.73 m(2)), prevalence was found to be 29 %. Predictive ability in risk grade 2 (RR = 1.41; 95 % CI: 1.19-1.66) and risk grade ≥3 (RR = 1.76; 95 % CI: 1.37-2.28) were both significantly greater than for non-CKD. Reclassification analysis showed a significant improvement in risk predictive abilities when CKD risk grading was based on UACR rather than on dipstick testing in this population (p < 0.001). Although prevalence of CKD was higher when detected by UACR rather than urine dipstick testing, the predictive

  20. Rapid changes in the serum total protein and globulin levels in complications caused by facultatively pathogenic Gram-negative bacteria.

    PubMed

    Petrás, G; Kiss, S; Juraszek, J; Merétey, K

    1978-01-01

    The changes in the levels of total protein and four globulin fractions were followed up throughout the entire course of complications caused by Gram-negative facultative pathogens in 37 acute cases of respiratory insufficiency accompanying different underlying illnesses and in 9 chronic, bedridden patients given artificial ventilation. At the onset of the infectious complications, in the first place in septic shock, the levels of various globulin fractions showed a decrease corresponding to a half-life of 2 to 4 days. Neither the increased catabolism, nor the protein losses by the urine and tracheal secretions offer a sufficient explanation for the escape of globulins of this extent from the plasma. It seems that this is a consequence of the increase in capillary permeability due to the effect of antigen-antibody reactions and that of endotoxin. As a result, in the critical phase of the infectious complications, at the point of culmination, e.g. in septic shock, diminished amount of different globulins is transported to the site of utilization, that is, to the inflammatory area.

  1. Colorimetric protein determination in microalgae (Chlorophyta): association of milling and SDS treatment for total protein extraction.

    PubMed

    Mota, Maria Fernanda S; Souza, Marcella F; Bon, Elba P S; Rodrigues, Marcoaurelio A; Freitas, Suely Pereira

    2018-05-24

    The use of colorimetric methods for protein quantification in microalgae is hindered by their elevated amounts of membrane-embedded intracellular proteins. In this work, the protein content of three species of microalgae was determined by the Lowry method after the cells were dried, ball-milled, and treated with the detergent sodium dodecyl sulfate (SDS). Results demonstrated that the association of milling and SDS treatment resulted in a 3- to 7-fold increase in protein quantification. Milling promoted microalgal disaggregation and cell wall disruption enabling access of the SDS detergent to the microalgal intracellular membrane proteins and their efficient solubilization and quantification. © 2018 Phycological Society of America.

  2. Mining the human urine proteome for monitoring renal transplant injury

    SciTech Connect

    Sigdel, Tara K.; Gao, Yuqian; He, Jintang

    The human urinary proteome reflects systemic and inherent renal injury perturbations and can be analyzed to harness specific biomarkers for different kidney transplant injury states. 396 unique urine samples were collected contemporaneously with an allograft biopsy from 396 unique kidney transplant recipients. Centralized, blinded histology on the graft was used to classify matched urine samples into categories of acute rejection (AR), chronic allograft nephropathy (CAN), BK virus nephritis (BKVN), and stable graft (STA). Liquid chromatography–mass spectrometry (LC-MS) based proteomics using iTRAQ based discovery (n=108) and global label-free LC-MS analyses of individual samples (n=137) for quantitative proteome assessment were used inmore » the discovery step. Selected reaction monitoring (SRM) was applied to identify and validate minimal urine protein/peptide biomarkers to accurately segregate organ injury causation and pathology on unique urine samples (n=151). A total of 958 proteins were initially quantified by iTRAQ, 87% of which were also identified among 1574 urine proteins detected in LC-MS validation. 103 urine proteins were significantly (p<0.05) perturbed in injury and enriched for humoral immunity, complement activation, and lymphocyte trafficking. A set of 131 peptides corresponding to 78 proteins were assessed by SRM for their significance in an independent sample cohort. A minimal set of 35 peptides mapping to 33 proteins, were modeled to segregate different injury groups (AUC =93% for AR, 99% for CAN, 83% for BKVN). Urinary proteome discovery and targeted validation identified urine protein fingerprints for non-invasive differentiation of kidney transplant injuries, thus opening the door for personalized immune risk assessment and therapy.« less

  3. HCG in urine

    MedlinePlus

    Beta-HCG - urine; Human chorionic gonadotropin - urine; Pregnancy test - hCG in urine ... To collect a urine sample, you urinate into a special (sterile) cup. Home pregnancy tests require the test strip to be dipped into ...

  4. Ketones urine test

    MedlinePlus

    Ketone bodies - urine; Urine ketones; Ketoacidosis - urine ketones test; Diabetic ketoacidosis - urine ketones test ... Urine ketones are usually measured as a "spot test." This is available in a test kit that ...

  5. Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression.

    PubMed

    Draisci, Rosa; Montesissa, Clara; Santamaria, Barbara; D'Ambrosio, Chiara; Ferretti, Giovanni; Merlanti, Roberta; Ferranti, Carolina; De Liguoro, Marco; Cartoni, Claudia; Pistarino, Erika; Ferrara, Lino; Tiso, Micaela; Scaloni, Andrea; Cosulich, M Elisabetta

    2007-09-01

    Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-Q-MS/MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone alpha- and beta-epimers were detected in plasma and urine only within 2 and 24 h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE, MALDI-TOF-MS and muLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasma samples collected before treatment, was found upregulated even 36 h after hormone treatment. Extensive mass mapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed by Western blot analysis confirmed a significant and time dependent increase of this ApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine.

  6. Latent class evaluation of a milk test, a urine test, and the fat-to-protein percentage ratio in milk to diagnose ketosis in dairy cows.

    PubMed

    Krogh, M A; Toft, N; Enevoldsen, C

    2011-05-01

    In this study, 3 commonly used tests to diagnose ketosis were evaluated with a latent class model to avoid the assumption of an available perfect test. The 3 tests were the KetoLac BHB (Sanwa Kagaku Kenkyusho Co. Ltd., Nagoya, Japan) test strip that tests milk for β-hydroxybutyrate, the KetoStix (Bayer Diagnostics Europe Ltd., Dublin, Ireland) test strip that tests urine for acetoacetate, and the fat-to-protein percentage ratio (FPR) in milk. A total of 8,902 cows were included in the analysis. The cows were considered to be a random sample from the population of Danish dairy cattle under intensive management, thus representing a natural spectrum of ketosis as a disease. All cows had a recorded FPR between 7 and 21 d postpartum. The KetoLac BHB recordings were available from 2,257 cows and 6,645 cows had a KetoStix recording. The recordings were analyzed with a modified Hui-Walter model, in a Bayesian framework. The specificity of the KetoLac BHB test and the KetoStix test were both high [0.99 (0.97-0.99)], whereas the specificity of FPR was somewhat lower [0.79 (0.77-0.81)]. The best sensitivity was for the KetoStix test [0.78 (0.55-0.98)], followed by the FPR [0.63 (0.58-0.71)] and KetoLac BHB test [0.58 (0.35-0.93)]. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  7. Age and gender related changes of salivary total protein levels for forensic application.

    PubMed

    Bhuptani, D; Kumar, S; Vats, M; Sagav, R

    2018-05-30

    Saliva is one of the most commonly encountered biological fluids found at the crime scene. Forensic science including forensic odontology is focused on the positive identification of individuals. The salivary protein profiling can help in personalization by the changes associated with age throughout life and gender. These changes also seem to vary with the dietary habits, environmental factors and geographical areas. Thus, the aim of present study is to estimate these changes in salivary total protein concentration and profiling in individuals of Gujarat, India. The association of total protein concentration and protein content with the age, gender, tooth eruption, functions of the protein and its physiological significance are also intended for study in this population. One hundred unstimulated whole saliva samples from study subjects of Gujarat population were collected and grouped based on age and gender. Total protein concentration was determined by Bradford assay; also protein was separated and analyzed using Sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS PAGE). T Test and ANOVA were used for statistical analysis. The concentration of Total Protein was found to be between 2-4 mg/ml. It showed a positive correlation with age and gender. It can be concluded more protein bands were prominently present in the adolescents group followed by children and lastly in the adults groups.More high (more than 80 kDa) and low (less than 30 kDa) molecular weight proteins are seen in children and adolescents than adults. SDS PAGE allowed identification and comparison of group variabilities in protein profiles. The total salivary protein showed an association between the parameters under this study which will aid in the individual identification in the field of forensics.

  8. Urine alkalization facilitates uric acid excretion

    PubMed Central

    2010-01-01

    Background Increase in the incidence of hyperuricemia associated with gout as well as hypertension, renal diseases and cardiovascular diseases has been a public health concern. We examined the possibility of facilitated excretion of uric acid by change in urine pH by managing food materials. Methods Within the framework of the Japanese government's health promotion program, we made recipes which consist of protein-rich and less vegetable-fruit food materials for H+-load (acid diet) and others composed of less protein but vegetable-fruit rich food materials (alkali diet). Healthy female students were enrolled in this consecutive 5-day study for each test. From whole-day collected urine, total volume, pH, organic acid, creatinine, uric acid and all cations (Na+,K+,Ca2+,Mg2+,NH4+) and anions (Cl-,SO42-,PO4-) necessary for the estimation of acid-base balance were measured. Results Urine pH reached a steady state 3 days after switching from ordinary daily diets to specified regimens. The amount of acid generated ([SO42-] +organic acid-gut alkai) were linearly related with those of the excretion of acid (titratable acidity+ [NH4+] - [HCO3-]), indicating that H+ in urine is generated by the metabolic degradation of food materials. Uric acid and excreted urine pH retained a linear relationship, where uric acid excretion increased from 302 mg/day at pH 5.9 to 413 mg/day at pH 6.5, despite the fact that the alkali diet contained a smaller purine load than the acid diet. Conclusion We conclude that alkalization of urine by eating nutritionally well-designed food is effective for removing uric acid from the body. PMID:20955624

  9. Total Protein Content Determination of Microalgal Biomass by Elemental Nitrogen Analysis and a Dedicated Nitrogen-to-Protein Conversion Factor

    SciTech Connect

    Laurens, Lieve M; Olstad-Thompson, Jessica L; Templeton, David W

    Accurately determining protein content is important in the valorization of algal biomass in food, feed, and fuel markets, where these values are used for component balance calculations. Conversion of elemental nitrogen to protein is a well-accepted and widely practiced method, but depends on developing an applicable nitrogen-to-protein conversion factor. The methodology reported here covers the quantitative assessment of the total nitrogen content of algal biomass and a description of the methodology that underpins the accurate de novo calculation of a dedicated nitrogen-to-protein conversion factor.

  10. The effect of urea on refractometric total protein measurement in dogs and cats with azotemia.

    PubMed

    Legendre, Kelsey P; Leissinger, Mary; Le Donne, Viviana; Grasperge, Britton J; Gaunt, Stephen D

    2017-03-01

    While protein is the predominant solute measured in plasma or serum by a refractometer, nonprotein substances also contribute to the angle of refraction. There is debate in the current literature regarding which nonprotein substances cause factitiously high refractometric total protein measurements, as compared to the biuret assay. The purpose of the study was to determine if the blood of azotemic animals, specifically with increased blood urea concentration, will have significantly higher refractometric total protein concentrations compared to the total protein concentrations measured by biuret assay. A prospective case series was conducted by collecting data from azotemic (n = 26) and nonazotemic (n = 34) dogs and cats. In addition, an in vitro study was performed where urea was added to an enhanced electrolyte solution at increasing concentrations, and total protein was assessed by both the refractometer and spectrophotometer. Statistical analysis was performed to determine the effect of urea. The refractometric total protein measurement showed a positive bias when compared to the biuret protein measurement in both groups, but the bias was higher in the azotemic group vs the nonazotemic group. The mean difference in total protein measurements of the nonazotemic group (0.59 g/dL) was significantly less (P < .01) than the mean difference of the azotemic group (0.95 g/dL). The in vitro experiment revealed a positive bias with a proportional error. This study demonstrated that increasing concentrations of urea significantly increased the total protein concentration measured by the refractometer as compared to the biuret assay, both in vivo and in vitro. © 2017 American Society for Veterinary Clinical Pathology.

  11. Utility of Urinary Protein-Creatinine Ratio and Protein Content in a 24-Hour Urine Collection in Systemic Lupus Erythematosus: A Systematic Review and Meta-Analysis.

    PubMed

    Medina-Rosas, Jorge; Yap, Kristy S; Anderson, Melanie; Su, Jiandong; Touma, Zahi

    2016-09-01

    To systematically review literature on the utility of spot urinary protein-creatinine ratio (PCR) as a screening test for proteinuria and its ability to accurately measure proteinuria compared with 24-hour urine collection (24H-P) in patients with systemic lupus erythematosus (SLE). We conducted a literature search (1900-2015) for articles comparing PCR and 24H-P in SLE patients in the databases Medline, Web of Science, and Embase. Included studies and their results were critically appraised and analyzed. Thirteen studies (1,001 patients; 84.01% women) were included. Ten studies reported on Pearson's correlation (range 0.67-0.94), and 3 studies reported on Spearman's correlation (range 0.78-1.00). The meta-analysis of studies with Pearson's correlation showed a high overall correlation of 0.80 between 24H-P and PCR, yet with high heterogeneity (I(2)  = 97.2%). Correlation analysis is not sufficient to evaluate the utility of a new test against the gold standard test, and analysis on agreement is required. Seven studies reported on agreement: 3 studies analyzed concordance correlation coefficient (0.48-0.94), 3 analyzed intraclass correlation coefficient (0.66-0.95), and 1 analyzed kappa coefficient (0.58). These results confirmed that the agreement between 24H-P and PCR was inappropriate. Three studies included Bland-Altman plots, and the results also demonstrated poor agreement between both tests. The PCR has a utility as a screening test for proteinuria in SLE patients. The studies' results of 24H-P and PCR showed poor agreement between both tests, signifying that PCR should not be a substitute for the gold standard test (24H-P) to accurately measure proteinuria. © 2016, American College of Rheumatology.

  12. [Analysis of total proteins in the seed of almond (Prunus dulcis) by two-dimensional electrophoresis].

    PubMed

    Li, Dong-dong; He, Shao-heng

    2004-07-01

    To analyse the total proteins in the seeds of almond (Prunus dulcis), one of the popular ingestent allergens in China, by two-dimensional electrophoresis. The total proteins of the seeds were extracted by trichloracetic acid (TCA) method, and then separated by isoelectric focusing as first dimension and SDS-PAGE as the second dimension. The spots of proteins were visualized by staining with Coomassie Brilliant Blue R-250. After analysis with software (ImageMaster 2D), 188 different proteins were detected. The isoelectric points (pI) for approximately 28% of total proteins were between 4.5-5.5, and the relative molecular mass (M(r)) of approximately 62% total proteins were between (20-25)x10(3). This was the first high-resolution, two-dimensional protein map of the seed of almond (Prunus dulcis) in China. Our finding has laid a solid foundation for further identification, characterization, gene cloning and standardization of allergenic proteins in the seed of almond (Prunus dulcis).

  13. A double stain for total and oxidized proteins from two-dimensional fingerprints.

    PubMed

    Talent, J M; Kong, Y; Gracy, R W

    1998-10-01

    Oxidative modification of proteins plays a major role in the etiology of aging and age-related diseases. For example, in Alzheimer's disease, although evidence points to oxidation of proteins as a causative factor in loss of cognitive abilities, it is not known which specific proteins of the brain are most susceptible to these modifications. Thus, it is of interest to identify the specific proteins which are susceptible to oxidation in vivo. Two-dimensional protein fingerprint methods offer the analytical potential for resolution of thousands of individual proteins from tissues, and the oxidized proteins can be visualized with immunological probes. Sensitive methods permit recovery and sufficient amino acid sequencing to identify these proteins. However, for such analyses it is essential to simultaneously analyze both protein content and level of oxidation. We have evaluated several approaches, identified the sources of artifacts and interferences, and developed a double-staining procedure that allows visualization and quantitation of total protein patterns as well as the specific oxidized proteins from two-dimensional protein fingerprints. The method has been applied to cells grown in culture and to tissue extracts from young and old animals. Copyright 1998 Academic Press.

  14. Comparison of biuret and refractometry methods for the serum total proteins measurement in ruminants.

    PubMed

    Katsoulos, Panagiotis D; Athanasiou, Labrini V; Karatzia, Maria A; Giadinis, Nektarios; Karatzias, Harilaos; Boscos, Constantin; Polizopoulou, Zoe S

    2017-12-01

    Determination of serum total protein concentration is commonly performed by the biuret method. Refractometric measurement is a faster and less expensive alternative but its accuracy has not been determined in ruminants. The purpose of the study was to compare the serum total protein concentrations in cattle, sheep, and goats measured by the biuret method with those obtained by refractometry. Serum total protein concentration was determined in 120 cattle, 67 sheep, and 58 goat blood samples refractometrically and with the biuret method. The data were analyzed with a paired samples t-test, and Passing and Bablok regression equations and Bland and Altman plots were generated. There was a strong linear relationship between the total protein values determined with the refractometer and the biuret method in cattle, sheep, and goats. The statistical accuracy, which represents a bias correction factor that measures the deviation of the best-fit line from the 45° line through the origin, was 90.63% for cattle, 93.05% for sheep, and 91.76% for goats. The mean protein values determined with the refractometer were significantly lower than those measured with the biuret method in cattle and goats (P < .05) but not in sheep (P > .05). The evaluated refractometer was sufficiently accurate for the determination of serum total proteins in cattle, sheep, and goats, although it cannot be used interchangeably with the biuret method. The RIs should be corrected for negative bias based on the created equations. © 2017 American Society for Veterinary Clinical Pathology.

  15. Comparison of total protein concentration in skeletal muscle as measured by the Bradford and Lowry assays.

    PubMed

    Seevaratnam, Rajini; Patel, Barkha P; Hamadeh, Mazen J

    2009-06-01

    The Lowry and Bradford assays are the most commonly used methods of total protein quantification, yet vary in several aspects. To date, no comparisons have been made in skeletal muscle. We compared total protein concentrations of mouse red and white gastrocnemius, reagent stability, protein stability and range of linearity using both assays. The Lowry averaged protein concentrations 15% higher than the Bradford with a moderate correlation (r = 0.36, P = 0.01). However, Bland-Altman analysis revealed considerable bias (15.8 +/- 29.7%). Both Lowry reagents and its protein-reagent interactions were less stable over time than the Bradford. The linear range of concentration was smaller for the Lowry (0.05-0.50 mg/ml) than the Bradford (0-2.0 mg/ml). We conclude that the Bradford and Lowry measures of total protein concentration in skeletal muscle are not interchangeable. The Bradford and Lowry assays have various strengths and weaknesses in terms of substance interference and protein size. However, the Bradford provides greater reagent stability, protein-reagent stability and range of linearity, and requires less time to analyse compared to the Lowry assay.

  16. Validated age-specific reference values for CSF total protein levels in children.

    PubMed

    Kahlmann, V; Roodbol, J; van Leeuwen, N; Ramakers, C R B; van Pelt, D; Neuteboom, R F; Catsman-Berrevoets, C E; de Wit, M C Y; Jacobs, B C

    2017-07-01

    To define age-specific reference values for cerebrospinal fluid (CSF) total protein levels for children and validate these values in children with Guillain-Barré syndrome (GBS), acute disseminated encephalomyelitis (ADEM) and multiple sclerosis (MS). Reference values for CSF total protein levels were determined in an extensive cohort of diagnostic samples from children (<18 year) evaluated at Erasmus Medical Center/Sophia Children's Hospital. These reference values were confirmed in children diagnosed with disorders unrelated to raised CSF total protein level and validated in children with GBS, ADEM and MS. The test results of 6145 diagnostic CSF samples from 3623 children were used to define reference values. The reference values based on the upper limit of the 95% CI (i.e. upper limit of normal) were for 6 months-2 years 0.25 g/L, 2-6 years 0.25 g/L, 6-12 years 0.28 g/L, 12-18 years 0.34 g/L. These reference values were confirmed in a subgroup of 378 children diagnosed with disorders that are not typically associated with increased CSF total protein. In addition, the CSF total protein levels in these children in the first 6 months after birth were highly variable (median 0.47 g/L, IQR 0.26-0.65). According to these new reference values, CSF total protein level was elevated in 85% of children with GBS, 66% with ADEM and 23% with MS. More accurate age-specific reference values for CSF total protein levels in children were determined. These new reference values are more sensitive than currently used values for diagnosing GBS and ADEM in children. Copyright © 2017 European Paediatric Neurology Society. Published by Elsevier Ltd. All rights reserved.

  17. Prediction of Cortical Defect Using C-Reactive Protein and Urine Sodium to Potassium Ratio in Infants with Febrile Urinary Tract Infection

    PubMed Central

    Jung, Su Jin

    2016-01-01

    Purpose We investigated whether C-reactive protein (CRP) levels, urine protein-creatinine ratio (uProt/Cr), and urine electrolytes can be useful for discriminating acute pyelonephritis (APN) from other febrile illnesses or the presence of a cortical defect on 99mTc dimercaptosuccinic acid (DMSA) scanning (true APN) from its absence in infants with febrile urinary tract infection (UTI). Materials and Methods We examined 150 infants experiencing their first febrile UTI and 100 controls with other febrile illnesses consecutively admitted to our hospital from January 2010 to December 2012. Blood (CRP, electrolytes, Cr) and urine tests [uProt/Cr, electrolytes, and sodium-potassium ratio (uNa/K)] were performed upon admission. All infants with UTI underwent DMSA scans during admission. All data were compared between infants with UTI and controls and between infants with or without a cortical defect on DMSA scans. Using multiple logistic regression analysis, the ability of the parameters to predict true APN was analyzed. Results CRP levels and uProt/Cr were significantly higher in infants with true APN than in controls. uNa levels and uNa/K were significantly lower in infants with true APN than in controls. CRP levels and uNa/K were relevant factors for predicting true APN. The method using CRP levels, u-Prot/Cr, u-Na levels, and uNa/K had a sensitivity of 94%, specificity of 65%, positive predictive value of 60%, and negative predictive value of 95% for predicting true APN. Conclusion We conclude that these parameters are useful for discriminating APN from other febrile illnesses or discriminating true APN in infants with febrile UTI. PMID:26632389

  18. High dietary protein intake is associated with an increased body weight and total death risk.

    PubMed

    Hernández-Alonso, Pablo; Salas-Salvadó, Jordi; Ruiz-Canela, Miguel; Corella, Dolores; Estruch, Ramón; Fitó, Montserrat; Arós, Fernando; Gómez-Gracia, Enrique; Fiol, Miquel; Lapetra, José; Basora, Josep; Serra-Majem, Lluis; Muñoz, Miguel Ángel; Buil-Cosiales, Pilar; Saiz, Carmen; Bulló, Mònica

    2016-04-01

    High dietary protein diets are widely used to manage overweight and obesity. However, there is a lack of consensus about their long-term efficacy and safety. Therefore, the aim of this study was to assess the effect of long-term high-protein consumption on body weight changes and death outcomes in subjects at high cardiovascular risk. A secondary analysis of the PREDIMED trial was conducted. Dietary protein was assessed using a food-frequency questionnaire during the follow-up. Cox proportional hazard models were used to estimate the multivariate-adjusted hazard ratio (HR) and 95% confidence intervals (95%CI) for protein intake in relation to the risk of body weight and waist circumference changes, cardiovascular disease, cardiovascular death, cancer death and total death. Higher total protein intake, expressed as percentage of energy, was significantly associated with a greater risk of weight gain when protein replaced carbohydrates (HR: 1.90; 95%CI: 1.05, 3.46) but not when replaced fat (HR: 1.69; 95%CI: 0.94, 3.03). However, no association was found between protein intake and waist circumference. Contrary, higher total protein intake was associated with a greater risk of all-cause death in both carbohydrate and fat substitution models (HR: 1.59; 95%CI: 1.08, 2.35; and HR: 1.66; 95%CI: 1.13, 2.43, respectively). A higher consumption of animal protein was associated with an increased risk of fatal and non-fatal outcomes when protein substituted carbohydrates or fat. Higher dietary protein intake is associated with long-term increased risk of body weight gain and overall death in a Mediterranean population at high cardiovascular risk. Copyright © 2015 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  19. Porphyrins - urine test

    MedlinePlus

    Urine uroporphyrin; Urine coproporphyrin; Porphyria - uroporphyrin ... After you provide a urine sample, it is tested in the lab. This is called a random urine sample. If needed, your health care provider ...

  20. Urine metabolomics.

    PubMed

    Zhang, Aihua; Sun, Hui; Wu, Xiuhong; Wang, Xijun

    2012-12-24

    Metabolomics is a powerful technique for the discovery of novel biomarkers and elucidation of biochemical pathways to improve diagnosis, prognosis and therapy. An advantage of this approach is its ability to assess global metabolic profiles to enhance pathologic characterization. Urine is an ideal bio-medium for disease study because it is readily available, easily obtained and less complex than other body fluids. Ease of collection allows for serial sampling to monitor disease and therapeutic response. Because of this potential, this paper will review urine metabolomic analysis, discuss its significance in the post-genomic era and highlight the specific roles of endogenous small molecule metabolites in this emerging field. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Comparison of Spot Urine Protein to Creatinine Ratio to 24-Hour Proteinuria to Identify Important Change Over Time in Proteinuria in Lupus.

    PubMed

    Medina-Rosas, Jorge; Su, Jiandong; Cook, Richard J; Sabapathy, Arthy; Touma, Zahi

    2017-09-01

    The aim of this study was to determine whether spot urine protein-to-creatinine ratio (PCR) accurately measures the change in proteinuria compared with 24-hour proteinuria (24H-P). This was a retrospective analysis on patients' paired visits and paired urine samples for PCR and 24H-P. Patients with both abnormal 24H-P (>0.5 g/d) and PCR (>0.05 g/mmol) or both normal 24H-P (≤0.5 g/d) and PCR (≤0.05 g/mmol) at baseline visit were identified.The first follow-up visit with partial recovery (50% decrease in proteinuria) or complete recovery (≤0.5 g/d) was identified for those with abnormal baseline 24H-P, and new proteinuria (>0.5 g/d) was identified for those with normal 24H-P. Twenty-four-hour urine collection and PCR end-point frequencies were compared. Twenty-four-hour urine collection results were converted to 24H-PCR. Twenty-four-hour PCR and PCR were utilized to measure the magnitude of change (by standardized response mean [SRM]) in patients who achieved the end points. Of 230 patients, at baseline, 95 patients had abnormal and 109 had normal 24H-P and PCR. On follow-up, 57 achieved partial recovery, and 53 achieved complete recovery by 24H-P. Standardized response mean was -1.03 and -1.10 for 24H-PCR and PCR, respectively. By PCR, 53 patients had partial recovery, and 27 had complete recovery. Standardized response mean was -1.25 and -0.86 by 24H-PCR and PCR, respectively.For new proteinuria, 28 patients were identified by 24H-P and 21 by PCR. Twenty-four-hour PCR SRM was 0.80, and PCR SRM was 0.68. Protein-to-creatinine ratio does not have sufficient accuracy compared with 24H-P for improvement and worsening to be used in lieu of 24H-P.

  2. Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics

    PubMed Central

    2015-01-01

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ∼10 μg of total protein were processed by 96FASP and LC-MS resulting in 700–900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics. PMID:24797144

  3. Protein Losses and Urea Nitrogen Underestimate Total Nitrogen Losses in Peritoneal Dialysis and Hemodialysis Patients.

    PubMed

    Salame, Clara; Eaton, Simon; Grimble, George; Davenport, Andrew

    2018-04-28

    Muscle wasting is associated with increased mortality and is commonly reported in dialysis patients. Hemodialysis (HD) and peritoneal dialysis (PD) treatments lead to protein losses in effluent dialysate. We wished to determine whether changes in current dialysis practice had increased therapy-associated nitrogen losses. Cross-sectional cohort study. Measurement of total protein, urea and total nitrogen in effluent dialysate from 24-hour collections from PD patients, and during haemodiafiltration (HDF) and haemodialysis (HD) sessions. One hundred eight adult dialysis patients. Peritoneal dialysis, high-flux haemodialysis and haemodiafiltration. Total nitrogen and protein losses. Dialysate protein losses were measured in 68 PD and 40 HD patients. Sessional losses of urea (13.9 [9.2-21.1] vs. 4.8 [2.8-7.8] g); protein (8.6 [7.2-11.1] vs. 6.7 [3.9-11.1] g); and nitrogen (11.5 [8.7-17.7] vs. 4.9 [2.6-9.5] g) were all greater for HD than PD, P < .001. Protein-derived nitrogen was 71.9 (54.4-110.4) g for HD and 30.8 (16.1-59.6) g for PD. Weekly protein losses were lower with HD 25.9 (21.5-33.4) versus 46.6 (27-77.6) g/week, but nitrogen losses were similar. We found no difference between high-flux HD and HDF: urea (13.5 [8.8-20.6] vs. 15.3 [10.5-25.5] g); protein (8.8 [7.3-12.2] vs. 7.6 [5.8-9.0] g); and total nitrogen (11.6 [8.3-17.3] vs. 10.8 [8.9-22.5] g). Urea nitrogen (UN) only accounted for 45.1 (38.3-51.0)% PD and 63.0 (55.3-62.4)% HD of total nitrogen losses. Although sessional losses of protein and UN were greater with HD, weekly losses were similar between modalities. We found no differences between HD and HDF. However, total nitrogen losses were much greater than the combination of protein and UN, suggesting greater nutritional losses with dialysis than previously reported. Copyright © 2018 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

  4. Protein extraction from human anagen head hairs 1-millimeter or less in total length.

    PubMed

    Carlson, Traci L; Moini, Mehdi; Eckenrode, Brian A; Allred, Brent M; Donfack, Joseph

    2018-04-01

    A simple method for extracting protein from human anagen (i.e., actively growing hair stage) head hairs was developed in this study for cases of limited sample availability and/or studies of specific micro-features within a hair. The distinct feature segments of the hair from one donor were divided lengthwise (i.e., each of ∼200-400 μm) and then pooled for three individual hairs to form a total of eight composite hair samples (i.e., each of ∼1 mm or less in total length). The proteins were extracted, digested using trypsin, and characterized via nano-flow liquid chromatography tandem-mass spectrometry (nLC-MS/MS). A total of 63 proteins were identified from all eight protein samples analyzed of which 60% were keratin and keratin-associated proteins. The major hair keratins identified are consistent with previous studies using fluorescence in situ hybridization and nLC-MS/MS while requiring over 400-8000-fold less sample. The protein extraction method from micro-sized human head hairs described in this study will enable proteomic analysis of biological evidence for cases of limited sample availability and will complement hair research. For example, research seeking to develop alternative non-DNA based techniques for comparing questioned to known hairs, and understanding the biochemistry of hair decomposition.

  5. Comparison of five methods for determination of total plasma protein concentration.

    PubMed

    Okutucu, Burcu; Dinçer, Ayşşe; Habib, Omer; Zihnioglu, Figen

    2007-08-01

    Quantitation of exact total protein content is often a key step and is common to many applications in general biochemistry research and routine clinical laboratory practice. Before embarking on any type of protein analysis, particularly comparative techniques, it is important to accurately quantitate the amount of protein in the sample. In order to assess the quality of total protein estimation results, five methods were tested and were applied to the same pooled plasma sample. For this aim, Bradford (Coomassie Brilliant Blue), Lowry (Folin-Ciocalteau), Biüret, Pesce and Strande (Ponceau-S/TCA), and modified method of Schaffner-Weismann (Amido Black 10B) were used. The last two methods employ simultaneous precipitation of proteins with the acid containing dye solutions followed by dissolution of precipitate in a NaOH solution. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in literature, accuracy and reproducibility/coefficient of variation. All of the methods tested show a CV %<6. Besides pooled plasma, a known concentration of human serum albumin was also analyzed and discussed by means of standardization of plasma total protein content.

  6. Evidence that the ZNT3 protein controls the total amount of elemental zinc in synaptic vesicles

    USGS Publications Warehouse

    Linkous, D.H.; Flinn, J.M.; Koh, J.Y.; Lanzirotti, A.; Bertsch, P.M.; Jones, B.F.; Giblin, L.J.; Frederickson, C.J.

    2008-01-01

    The ZNT3 protein decorates the presynaptic vesicles of central neurons harboring vesicular zinc, and deletion of this protein removes staining for zinc. However, it has been unclear whether only histochemically reactive zinc is lacking or if, indeed, total elemental zinc is missing from neurons lacking the Slc30a3 gene, which encodes the ZNT3 protein. The limitations of conventional histochemical procedures have contributed to this enigma. However, a novel technique, microprobe synchrotron X-ray fluorescence, reveals that the normal 2- to 3-fold elevation of zinc concentration normally present in the hippocampal mossy fibers is absent in Slc30a3 knockout (ZNT3) mice. Thus, the ZNT3 protein evidently controls not only the "stainability" but also the actual mass of zinc in mossy-fiber synaptic vesicles. This work thus confirms the metal-transporting role of the ZNT3 protein in the brain. ?? The Histochemical Society, Inc.

  7. [A novel protein equalizer based on single chain variable fragment display M13 phage library for nephropathy patient urine study].

    PubMed

    Zhao, Peng; Tao, Dingyin; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

    2009-05-01

    A novel protein equalizer was developed with single chain variable fragment (scFv) library displaying M13 phage covalently bonded on monolithic cryogel. Due to the great number and various kinds of displayed scFv fragments, as well as strong and specific binding capacity between scFv fragments and proteins, a new protein equalizer technology is preferable in the pretreatment of complex protein samples. After the sample dissolved in phosphate buffer solution (PBS), it was repeatedly loaded onto the equalizer for five times, the bound proteins were in sequence eluted by 2 mol/L NaCl and 50 mmol/L Gly-HC1 (pH 2.5) solution, followed by digestion with thrombin. All proteins or peptides collected from each fraction were further analyzed by high performance liquid chromatography-electrospray tandem mass spectrometry (RPLC-ESI-MS/MS) with a serially coupled long microcolumn. Compared with the untreated samples, the identified protein number was increased from 142 to 396. Furthermore, from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis results, it was found that the protein concentration difference was reduced obviously in the eluant of direct sample loading, and most high abundance proteins were identified in the eluant of NaCl. All these results demonstrate that the novel protein equalizer with scFv display M13 phage library immobilized on cyrogel could effectively reduce the dynamic range of proteins in complex samples, enabling the identification of more low abundance proteins.

  8. Urine Test: Microalbumin-to-Creatinine Ratio (For Parents)

    MedlinePlus

    ... involves measuring the amount of a protein called albumin in the urine (pee). The amount of urine albumin is compared with the quantity of a waste ... steady rate, so comparing the ratio of urine albumin with creatinine in the same urine specimen helps ...

  9. Urine Collected From Diapers Can Be Used for 2-Dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) in Infants and Young Children

    PubMed Central

    Kennedy, Mary Jayne; Griffin, Angela; Su, Ruifeng; Merchant, Michael; Klein, Jon

    2011-01-01

    Urinary proteomic profiling has potential to identify candidate biomarkers of renal injury in infants provided an adequate urine sample can be obtained. Although diapers are used to obtain urine for clinical evaluation, their use for proteomic analysis has not been investigated. We therefore performed feasibility studies on the use of diaper-extracted urine for 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Pediatric waste urine (2–20 mL) was applied to gel-containing, non-gel and cotton-gauze diapers and then mechanically expressed. Urine volume and total protein were measured pre- and post-extraction. Proteins were separated via 2D-PAGE following application of urine (20–40 mL) to each matrix. 2D-PAGE was also performed on clinical specimens collected using each diaper type. Differences in the adsorption and retention of urine volume and protein were noted between matrices. Non-gel and cotton-gauze diapers provided the best protein/volume recovery and the lowest interference with the Bradford assay. 2D-PAGE was also successfully completed using urine samples from both cotton fiber matrices. Conversely, samples from low-gel diapers demonstrated poor protein separation and reproducibility. Diapers containing cotton-fiber matrices appear adequate for 2D-PAGE. Qualitative and quantitative analyses of resolved proteins using replicate, high resolution gels will be required, however, before diaper-extracted urine can be applied in proteomic profiling. PMID:21137001

  10. Total Protein of Whole Saliva as a Biomarker of Anaerobic Threshold

    ERIC Educational Resources Information Center

    Bortolini, Miguel Junior Sordi; De Agostini, Guilherme Gularte; Reis, Ismair Teodoro; Lamounier, Romeu Paulo Martins Silva; Blumberg, Jeffrey B.; Espindola, Foued Salmen

    2009-01-01

    Saliva provides a convenient and noninvasive matrix for assessing specific physiological parameters, including some biomarkers of exercise. We investigated whether the total protein concentration of whole saliva (TPWS) would reflect the anaerobic threshold during an incremental exercise test. After a warm-up period, 13 nonsmoking men performed a…

  11. Total chemical synthesis and X-ray structure of kaliotoxin by racemic protein crystallography.

    PubMed

    Pentelute, Brad L; Mandal, Kalyaneswar; Gates, Zachary P; Sawaya, Michael R; Yeates, Todd O; Kent, Stephen B H

    2010-11-21

    Here we report the total synthesis of kaliotoxin by 'one pot' native chemical ligation of three synthetic peptides. A racemic mixture of D- and L-kaliotoxin synthetic protein molecules gave crystals in the centrosymmetric space group P1 that diffracted to atomic-resolution (0.95 Å), enabling the X-ray structure of kaliotoxin to be determined by direct methods.

  12. Total Protein Extraction for Metaproteomics Analysis of Methane Producing Biofilm: The Effects of Detergents

    PubMed Central

    Huang, Hung-Jen; Chen, Wei-Yu; Wu, Jer-Horng

    2014-01-01

    Protein recovery is crucial for shotgun metaproteomics to study the in situ functionality of microbial populations from complex biofilms but still poorly addressed by far. To fill this knowledge gap, we systematically evaluated the sample preparation with extraction buffers comprising four detergents for the metaproteomics analysis of a terephthalate-degrading methanogenic biofilm using an on-line two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) system. Totally, 1018 non-repeated proteins were identified with the four treatments. On the whole, each treatment could recover the biofilm proteins with specific distributions of molecular weight, hydrophobicity, and isoelectric point. The extraction buffers containing zwitterionic and anionic detergents were found to harvest the proteins with better efficiency and quality, allowing identification up to 76.2% of total identified proteins with the LC-MS/MS analysis. According to the annotation with a relevant metagenomic database, we further observed different taxonomic profiles of bacterial and archaeal members and discriminable patterns of the functional expression among the extraction buffers used. Overall, the finding of the present study provides first insight to the effect of the detergents on the characteristics of extractable proteins from biofilm and the developed protocol combined with nano 2D-LC/MS/MS analysis can improve the metaproteomics studies on microbial functionality of biofilms in the wastewater treatment systems. PMID:24914765

  13. Hydrodynamic size-based separation and characterization of protein aggregates from total cell lysates

    PubMed Central

    Tanase, Maya; Zolla, Valerio; Clement, Cristina C; Borghi, Francesco; Urbanska, Aleksandra M; Rodriguez-Navarro, Jose Antonio; Roda, Barbara; Zattoni, Andrea; Reschiglian, Pierluigi; Cuervo, Ana Maria; Santambrogio, Laura

    2016-01-01

    Herein we describe a protocol that uses hollow-fiber flow field-flow fractionation (FFF) coupled with multiangle light scattering (MALS) for hydrodynamic size-based separation and characterization of complex protein aggregates. The fractionation method, which requires 1.5 h to run, was successfully modified from the analysis of protein aggregates, as found in simple protein mixtures, to complex aggregates, as found in total cell lysates. In contrast to other related methods (filter assay, analytical ultracentrifugation, gel electrophoresis and size-exclusion chromatography), hollow-fiber flow FFF coupled with MALS allows a flow-based fractionation of highly purified protein aggregates and simultaneous measurement of their molecular weight, r.m.s. radius and molecular conformation (e.g., round, rod-shaped, compact or relaxed). The polyethersulfone hollow fibers used, which have a 0.8-mm inner diameter, allow separation of as little as 20 μg of total cell lysates. In addition, the ability to run the samples in different denaturing and nondenaturing buffer allows defining true aggregates from artifacts, which can form during sample preparation. The protocol was set up using Paraquat-induced carbonylation, a model that induces protein aggregation in cultured cells. This technique will advance the biochemical, proteomic and biophysical characterization of molecular-weight aggregates associated with protein mutations, as found in many CNS degenerative diseases, or chronic oxidative stress, as found in aging, and chronic metabolic and inflammatory conditions. PMID:25521790

  14. Quantitation of total protein deposits on contact lenses by means of amino acid analysis.

    PubMed

    Yan, G; Nyquist, G; Caldwell, K D; Payor, R; McCraw, E C

    1993-04-01

    This study was done to characterize and quantify the protein deposits on worn contact lenses and to measure the residual deposits after extraction in 2% sodium dodecyl sulfate and the total protein deposits on worn vifilcon, atlafilcon, and tefilcon lenses (Food and Drug Administration Types IV, II, and I, respectively). Contact lens extracts were separated with gel electrophoresis, and the amount of protein was estimated after silver staining and densitometry. To determine the residual deposits, the contact lenses were hydrolyzed, and amino acid analysis was carried out by reverse-phase high-performance liquid chromatography after precolumn derivatization with phenylisothiocyanate. Refinement of the hydrolysis conditions was undertaken to minimize interference by the lens polymers. The extraction removed only approximately 25% of the protein deposits. Mild hydrolytic conditions, 20 hr in 6 N HCl at 105 degrees C, were found to cause minimal polymer interference. Of the 350, 10, and 20 micrograms of protein typically determined on whole vifilcon, atlafilcon, and tefilcon lenses, the polymers were estimated to account for 4, 0.5, and less than 0.4 micrograms, respectively. Hydrolysis of worn contact lenses with subsequent amino acid separation can be applied to determine the total protein deposits without the uncertainty inherent in extraction of the deposits.

  15. Evaluation of on-line desalter-inductively coupled plasma-mass spectrometry system for determination of Cr(III), Cr(VI), and total chromium concentrations in natural water and urine samples

    NASA Astrophysics Data System (ADS)

    Sun, Y. C.; Lin, C. Y.; Wu, S. F.; Chung, Y. T.

    2006-02-01

    We have developed a simple and convenient method for the determination of Cr(III), Cr(VI), and the total chromium concentrations in natural water and urine samples that use a flow injection on-line desalter-inductively coupled plasma-mass spectrometry system. When using aqueous ammonium chloride (pH 8) as the stripping solution, the severe interference from sodium in the matrix can be eliminated prior to inductively coupled plasma-mass spectrometry measurement, and the Cr(VI) level can be determined directly. To determine the total concentration of Cr in natural water and urine samples, we used H 2O 2 or HNO 3 to decompose the organic matter and convert all chromium species into the Cr(VI) oxidation state. To overcome the spectral interference caused by the matrix chloride ion in the resulting solutions, we employed cool plasma to successfully suppress chloride-based molecular ion interference during the inductively coupled plasma-mass spectrometry measurement. By significantly eliminating interference from the cationic and anionic components in the matrices prior to the inductively coupled plasma-mass spectrometry measurement, we found that the detection limit reached 0.18 μg L - 1 (based on 3 sigma). We validated this method through the analysis of the total chromium content in two reference materials (NIST 1643c and 2670E) and through measuring the recovery in spiked samples.

  16. A High-throughput Screening Assay for Determining Cellular Levels of Total Tau Protein

    PubMed Central

    Dehdashti, Seameen J.; Zheng, Wei; Gever, Joel R.; Wilhelm, Robert; Nguyen, Dac-Trung; Sittampalam, Gurusingham; McKew, John C.; Austin, Christopher P.; Prusiner, Stanley B.

    2014-01-01

    The microtubule-associated protein (MAP) tau has been implicated in the pathology of numerous neurodegenerative diseases. In the past decade, the hyperphosphorylated and aggregated states of tau protein have been important targets in the drug discovery field for the potential treatment of Alzheimer’s disease. Although several compounds have been reported to reduce the hyperphosphorylated state of tau or impact the stabilization of tau, their therapeutic activities are still to be validated. Recently, reduction of total cellular tau protein has emerged as an alternate intervention point for drug development and a potential treatment of tauopathies. We have developed and optimized a homogenous assay, using the AlphaLISA and HTRF assay technologies, for the quantification of total cellular tau protein levels in the SH-SY5Y neuroblastoma cell line. The signal-to-basal ratios were 375 and 5.3, and the Z’ factors were 0.67 and 0.60 for the AlphaLISA and HTRF tau assays, respectively. The clear advantages of this homogeneous tau assay over conventional total tau assays, such as ELISA and Western blot, are the elimination of plate wash steps and miniaturization of the assay into 1536-well plate format for the ultra–high-throughput screening of large compound libraries. PMID:23905996

  17. The Presence Of Strange Males' Odor Induces Behavioral Responses And Elevated Levels Of Low Molecular Weight Proteins Excreted In The Urine Of Mature Water Vole Males (Arvicola amphibius L).

    PubMed

    Nazarova, Galina G; Proskurniak, Lyudmila P; Yuzhik, Ekaterina I

    2016-03-01

    We hypothesized that low molecular weight urinary proteins play a role in male-male chemical communication in the water vole, Arvicola ampibius L. We studied the effect of placing soiled litter from strange males into the cage of another sexually mature male on the intensity of its digging and scattering, urination on the litter, and alteration in the levels of low molecular weight proteins (15-25 kDa) excreted in the urine before and after 4 days of exposure as determined by chip electrophoresis. The intensity of digging and scattering was positively correlated with levels of testosterone in serum of males exposed to strange male odors (r = 0.56; P < 0.01), as well as with the concentration of low molecular weight proteins in the donor's urine (r = 0.52, P < 0.05). At the end of the experiment, the level of low molecular weight protein in excreted urine was elevated in the males exposed to the strange male's litter. These results highlight the importance of quantitative inter-individual variation of low molecular weight urinary proteins in the modulation of the physiology and behavior of conspecifics.

  18. Self-Renewal and Differentiation Capacity of Urine-Derived Stem Cells after Urine Preservation for 24 Hours

    PubMed Central

    Shi, Yingai; Bharadwaj, Shantaram; Leng, Xiaoyan; Zhou, Xiaobo; Liu, Hong; Atala, Anthony; Zhang, Yuanyuan

    2013-01-01

    Despite successful approaches to preserve organs, tissues, and isolated cells, the maintenance of stem cell viability and function in body fluids during storage for cell distribution and transportation remains unexplored. The aim of this study was to characterize urine-derived stem cells (USCs) after optimal preservation of urine specimens for up to 24 hours. A total of 415 urine specimens were collected from 12 healthy men (age range 20–54 years old). About 6×104 cells shed off from the urinary tract system in 24 hours. At least 100 USC clones were obtained from the stored urine specimens after 24 hours and maintained similar biological features to fresh USCs. The stored USCs had a “rice grain” shape in primary culture, and expressed mesenchymal stem cell surface markers, high telomerase activity, and normal karyotypes. Importantly, the preserved cells retained bipotent differentiation capacity. Differentiated USCs expressed myogenic specific proteins and contractile function when exposed to myogenic differentiation medium, and they expressed urothelial cell-specific markers and barrier function when exposed to urothelial differentiation medium. These data demonstrated that up to 75% of fresh USCs can be safely persevered in urine for 24 hours and that these cells stored in urine retain their original stem cell properties, indicating that preserved USCs could be available for potential use in cell-based therapy or clinical diagnosis. PMID:23349776

  19. Identification of Leishmania infantum chagasi proteins in urine of patients with visceral leishmaniasis: a promising antigen discovery approach of vaccine candidates.

    PubMed

    Kashino, S S; Abeijon, C; Qin, L; Kanunfre, K A; Kubrusly, F S; Silva, F O; Costa, D L; Campos, D; Costa, C H N; Raw, I; Campos-Neto, A

    2012-07-01

    Visceral leishmaniasis (VL) is a serious lethal parasitic disease caused by Leishmania donovani in Asia and by Leishmania infantum chagasi in southern Europe and South America. VL is endemic in 47 countries with an annual incidence estimated to be 500,000 cases. This high incidence is due in part to the lack of an efficacious vaccine. Here, we introduce an innovative approach to directly identify parasite vaccine candidate antigens that are abundantly produced in vivo in humans with VL. We combined RP-HPLC and mass spectrometry and categorized three L. infantum chagasi proteins, presumably produced in spleen, liver and bone marrow lesions and excreted in the patients' urine. Specifically, these proteins were the following: Li-isd1 (XP_001467866.1), Li-txn1 (XP_001466642.1) and Li-ntf2 (XP_001463738.1). Initial vaccine validation studies were performed with the rLi-ntf2 protein produced in Escherichia coli mixed with the adjuvant BpMPLA-SE. This formulation stimulated potent Th1 response in BALB/c mice. Compared to control animals, mice immunized with Li-ntf2+ BpMPLA-SE had a marked parasite burden reduction in spleens at 40 days post-challenge with virulent L. infantum chagasi. These results strongly support the proposed antigen discovery strategy of vaccine candidates to VL and opens novel possibilities for vaccine development to other serious infectious diseases. © 2012 Blackwell Publishing Ltd.

  20. Quantification of free and total bisphenol A and bisphenol B in human urine by dispersive liquid-liquid microextraction (DLLME) and heart-cutting multidimensional gas chromatography-mass spectrometry (MD-GC/MS).

    PubMed

    Cunha, S C; Fernandes, J O

    2010-11-15

    A novel method combining dispersive liquid-liquid microextraction (DLLME) and heart-cutting multidimensional gas chromatography coupled to mass spectrometry was developed for the determination of free and total bisphenol A (BPA) and bisphenol B (BPB) in human urine samples. The DLLME procedure combines extraction, derivatization and concentration of the analytes into one step. Several important variables influencing the extraction efficiency and selectivity such as nature and volume of extractive and dispersive solvents as well as the amount of acetylating reagent were investigated. The temperature and time to hydrolyze BPA and BPB conjugates with a β-glucuronidase and sulfatase enzyme preparation were also studied. Under the optimized conditions good efficiency extraction (71-93%) and acceptable total DLLME yields (56-77%) were obtained for both analytes. Matrix-matched calibration curves were linear with correlation coefficients higher than 0.996 in the range level 0.1-5 μg/l, and the relative standard deviations (%RSD) were lower than 20% (n=6). The limits of detection were 0.03 and 0.05 μg/l for BPA and BPB, respectively. The applicability of the proposed method for determining urinary free and total BPA and BPB was assessed by analyzing the human urine of a group of 20 volunteers. Free BPA was detected in 45% of the sample whereas total BPA was detected in 85% of the samples at concentrations ranging between 0.39 and 4.99 μg/l. BPB was detected in conjugated form in two samples. Copyright © 2010 Elsevier B.V. All rights reserved.

  1. Glucose urine test

    MedlinePlus

    Urine sugar test; Urine glucose test; Glucosuria test; Glycosuria test ... After you provide a urine sample, it is tested right away. The health care provider uses a dipstick made with a color-sensitive pad. The ...

  2. Urine output - decreased

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003147.htm Urine output - decreased To use the sharing features on this page, please enable JavaScript. Decreased urine output means that you produce less urine than ...

  3. Urine drainage bags

    MedlinePlus

    ... this page: //medlineplus.gov/ency/patientinstructions/000142.htm Urine drainage bags To use the sharing features on this page, please enable JavaScript. Urine drainage bags collect urine. Your bag will attach ...

  4. Osmolality urine test

    MedlinePlus

    ... balance and urine concentration. Osmolality is a more exact measurement of urine concentration than the urine specific ... must be authorized in writing by ADAM Health Solutions. About MedlinePlus Site Map FAQs Customer Support Get ...

  5. Urine - abnormal color

    MedlinePlus

    ... medlineplus.gov/ency/article/003139.htm Urine - abnormal color To use the sharing features on this page, please enable JavaScript. The usual color of urine is straw-yellow. Abnormally colored urine ...

  6. Electrolytic pretreatment of urine

    NASA Technical Reports Server (NTRS)

    1977-01-01

    Electrolysis has been under evaluation for several years as a process to pretreat urine for ultimate recovery of potable water in manned spacecraft applications. The conclusions that were drawn from this investigation are the following: (1) A platinum alloy containing 10 percent rhodium has been shown to be an effective, corrosion-resistant anode material for the electrolytic pretreatment of urine. Black platinum has been found to be suitable as a cathode material. (2) The mechanism of the reactions occurring during the electrolysis of urine is two-stage: (a) a total Kjeldahl nitrogen and total organic carbon (TOC) removal in the first stage is the result of electrochemical oxidation of urea to CO2, H2O, and ammonia followed by chloride interaction to produce N2 from ammonia, (b) after the urea has been essentially removed and the chloride ions have no more ammonia to interact with, the chloride ions start to oxidize to higher valence states, thus producing perchlorates. (3) Formation of perchlorates can be suppressed by high/low current operation, elevated temperature, and pH adjustment. (4) UV-radiation showed promise in assisting electrolytic TOC removal in beaker tests, but was not substantiated in limited single cell testing. This may have been due to non-optimum configurations of the single cell test rig and the light source.

  7. Modified Cut-Off Value of the Urine Protein-To-Creatinine Ratio Is Helpful for Identifying Patients at High Risk for Chronic Kidney Disease: Validation of the Revised Japanese Guideline.

    PubMed

    Yamamoto, Hiroyuki; Yamamoto, Kyoko; Yoshida, Katsumi; Shindoh, Chiyohiko; Takeda, Kyoko; Monden, Masami; Izumo, Hiroko; Niinuma, Hiroyuki; Nishi, Yutaro; Niwa, Koichiro; Komatsu, Yasuhiro

    2015-11-01

    Chronic kidney disease (CKD) is a global public health issue, and strategies for its early detection and intervention are imperative. The latest Japanese CKD guideline recommends that patients without diabetes should be classified using the urine protein-to-creatinine ratio (PCR) instead of the urine albumin-to-creatinine ratio (ACR); however, no validation studies are available. This study aimed to validate the PCR-based CKD risk classification compared with the ACR-based classification and to explore more accurate classification methods. We analyzed two previously reported datasets that included diabetic and/or cardiovascular patients who were classified into early CKD stages. In total, 860 patients (131 diabetic patients and 729 cardiovascular patients, including 193 diabetic patients) were enrolled. We assessed the CKD risk classification of each patient according to the estimated glomerular filtration rate and the ACR-based or PCR-based classification. The use of the cut-off value recommended in the current guideline (PCR 0.15 g/g creatinine) resulted in risk misclassification rates of 26.0% and 16.6% for the two datasets. The misclassification was primarily caused by underestimation. Moderate to substantial agreement between each classification was achieved: Cohen's kappa, 0.56 (95% confidence interval, 0.45-0.69) and 0.72 (0.67-0.76) in each dataset, respectively. To improve the accuracy, we tested various candidate PCR cut-off values, showing that a PCR cut-off value of 0.08-0.10 g/g creatinine resulted in improvement in the misclassification rates and kappa values. Modification of the PCR cut-off value would improve its efficacy to identify high-risk populations who will benefit from early intervention.

  8. Effect of injected rotenone on the production and composition of urine from the rainbow trout (Salmo gairdneri)

    USGS Publications Warehouse

    Erickson, D.A.; Gingerich, W.H.

    1986-01-01

    Renal function was evaluated in adult rainbow trout (Salmo gairdneri) dosed i.a. with rotenone at 225 and 275 μg/kg. The chemical composition of urine samples and urine flow rates collected over a 5-h pretreatment period were compared with hourly urine samples collected over a 5-h posttreatment period. Significant increases in osmolality and in concentrations of sodium, potassium, chloride, glucose, and total protein were observed in the urine of treated fish. Urine solute concentrations reached maximum values within 1 to 3 h after treatment and decreased thereafter, indicating that the effects were reversible. Concentrations of sodium and chloride were highly correlated in 2-h posttreatment urine samples at the low (r = 0.922) and high (r = 0.981) rotenone treatments. Urine flow rates were reduced in trout at each dose of rotenone but the decrease in volume of urine voided was not dose-dependent. In a separate study, [14C]polyethylene glycol was used as a filtration marker to determine the effect of rotenone treatment (225 &mu:g/kg) on urine flow rate, glomerular filtration rate, and renal water reabsorption. We showed that posttreatment urine flow rates were reduced partly by reduced glomerular filtration and partly by increased water reabsorption. Transient increases in plasma osmolality and hematocrit also were observed 0.5 h after rotenone treatment.

  9. A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence

    PubMed Central

    Asanov, Alexander; Zepeda, Angélica; Vaca, Luis

    2012-01-01

    We have developed a novel microarray technology based on total internal reflection fluorescence (TIRF) in combination with DNA and protein bioassays immobilized at the TIRF surface. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing and stringency control, and measure only end-point results, our TIRF microarray technology provides several orders of magnitude better signal-to-background ratio, performs analysis rapidly in one step, and measures the entire course of association and dissociation kinetics between target DNA and protein molecules and the bioassays. In many practical cases detection of only DNA or protein markers alone does not provide the necessary accuracy for diagnosing a disease or detecting a pathogen. Here we describe TIRF microarrays that detect DNA and protein markers simultaneously, which reduces the probabilities of false responses. Supersensitive and multiplexed TIRF DNA and protein microarray technology may provide a platform for accurate diagnosis or enhanced research studies. Our TIRF microarray system can be mounted on upright or inverted microscopes or interfaced directly with CCD cameras equipped with a single objective, facilitating the development of portable devices. As proof-of-concept we applied TIRF microarrays for detecting molecular markers from Bacillus anthracis, the pathogen responsible for anthrax. PMID:22438738

  10. Multiexcitation Fluorogenic Labeling of Surface, Intracellular, and Total Protein Pools in Living Cells

    PubMed Central

    2016-01-01

    Malachite green (MG) is a fluorogenic dye that shows fluorescence enhancement upon binding to its engineered cognate protein, a fluorogen activating protein (FAP). Energy transfer donors such as cyanine and rhodamine dyes have been conjugated with MG to modify the spectral properties of the fluorescent complexes, where the donor dyes transfer energy through Förster resonance energy transfer to the MG complex resulting in binding-conditional fluorescence emission in the far-red region. In this article, we use a violet-excitable dye as a donor to sensitize the far-red emission of the MG-FAP complex. Two blue emitting fluorescent coumarin dyes were coupled to MG and evaluated for energy transfer to the MG-FAP complex via its secondary excitation band. 6,8-Difluoro-7-hydroxycoumarin-3-carboxylic acid (Pacific blue, PB) showed the most efficient energy transfer and maximum brightness in the far-red region upon violet (405 nm) excitation. These blue-red (BluR) tandem dyes are spectrally varied from other tandem dyes and are able to produce fluorescence images of the MG-FAP complex with a large Stokes shift (>250 nm). These dyes are cell-permeable and are used to label intracellular proteins. Used together with a cell-impermeable hexa-Cy3-MG (HCM) dye that labels extracellular proteins, we are able to visualize extracellular, intracellular, and total pools of cellular protein using one fluorogenic tag that combines with distinct dyes to effect different spectral characteristics. PMID:27159569

  11. [Determination of total protein content in soya-bean milk via visual moving reaction boundary titration].

    PubMed

    Guo, Chengye; Wang, Houyu; Zhang, Lei; Fan, Liuyin; Cao, Chengxi

    2013-11-01

    A visual, rapid and accurate moving reaction boundary titration (MRBT) method was used for the determination of the total protein in soya-bean milk. During the process, moving reaction boundary (MRB) was formed by hydroxyl ions in the catholyte and soya-bean milk proteins immobilized in polyacrylamide gel (PAG), and an acid-base indicator was used to denote the boundary motion. The velocity of MRB has a relationship with protein concentration, which was used to obtain a standard curve. By paired t-test, there was no significant difference of the protein content between MRBT and Kjeldahl method at 95% confidence interval. The procedure of MRBT method required about 10 min, and it had linearity in the range of 2.0-14.0 g/L, low limit of detection (0.05 g/L), good precision (RSD of intra-day < 1.90% and inter-day < 4.39%), and high recoveries (97.41%-99.91%). In addition, non-protein nitrogen (NPN) such as melamine added into the soya-bean milk had weak influence on MRBT results.

  12. Conversion to Sirolimus Ameliorates Cyclosporine-Induced Nephropathy in the Rat: Focus on Serum, Urine, Gene, and Protein Renal Expression Biomarkers

    PubMed Central

    Sereno, José; Nunes, Sara; Rodrigues-Santos, Paulo; Rocha-Pereira, Petronila; Fernandes, João; Teixeira, Frederico; Reis, Flávio

    2014-01-01

    Protocols of conversion from cyclosporin A (CsA) to sirolimus (SRL) have been widely used in immunotherapy after transplantation to prevent CsA-induced nephropathy, but the molecular mechanisms underlying these protocols remain nuclear. This study aimed to identify the molecular pathways and putative biomarkers of CsA-to-SRL conversion in a rat model. Four animal groups (n = 6) were tested during 9 weeks: control, CsA, SRL, and conversion (CsA for 3 weeks followed by SRL for 6 weeks). Classical and emergent serum, urinary, and kidney tissue (gene and protein expression) markers were assessed. Renal lesions were analyzed in hematoxylin and eosin, periodic acid-Schiff, and Masson's trichrome stains. SRL-treated rats presented proteinuria and NGAL (serum and urinary) as the best markers of renal impairment. Short CsA treatment presented slight or even absent kidney lesions and TGF-β, NF-κ β, mTOR, PCNA, TP53, KIM-1, and CTGF as relevant gene and protein changes. Prolonged CsA exposure aggravated renal damage, without clear changes on the traditional markers, but with changes in serums TGF-β and IL-7, TBARs clearance, and kidney TGF-β and mTOR. Conversion to SRL prevented CsA-induced renal damage evolution (absent/mild grade lesions), while NGAL (serum versus urine) seems to be a feasible biomarker of CsA replacement to SRL. PMID:24971338

  13. Estimation of salivary flow rate, pH, buffer capacity, calcium, total protein content and total antioxidant capacity in relation to dental caries severity, age and gender.

    PubMed

    Pandey, Pallavi; Reddy, N Venugopal; Rao, V Arun Prasad; Saxena, Aditya; Chaudhary, C P

    2015-03-01

    The aim of the study was to evaluate salivary flow rate, pH, buffering capacity, calcium, total protein content and total antioxidant capacity in relation to dental caries, age and gender. The study population consisted of 120 healthy children aged 7-15 years that was further divided into two groups: 7-10 years and 11-15 years. In this 60 children with DMFS/dfs = 0 and 60 children with DMFS/dfs ≥5 were included. The subjects were divided into two groups; Group A: Children with DMFS/dfs = 0 (caries-free) Group B: Children with DMFS/dfs ≥5 (caries active). Unstimulated saliva samples were collected from all groups. Flow rates were determined, and samples analyzed for pH, buffer capacity, calcium, total protein and total antioxidant status. Salivary antioxidant activity is measured with spectrophotometer by an adaptation of 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulphonate) assays. The mean difference of the two groups; caries-free and caries active were proved to be statistically significant (P < 0.05) for salivary calcium, total protein and total antioxidant level for both the sexes in the age group 7-10 years and for the age 11-15 years the mean difference of the two groups were proved to be statistically significant (P < 0.05) for salivary calcium level for both the sexes. Salivary total protein and total antioxidant level were proved to be statistically significant for male children only. In general, total protein and total antioxidants in saliva were increased with caries activity. Calcium content of saliva was found to be more in caries-free group and increased with age.

  14. Seasonal changes in amino acids, protein and total nitrogen in needles of fertilized Scots pine trees.

    PubMed

    Näsholm, T; Ericsson, A

    1990-09-01

    Seasonal changes in amino acids, protein and total nitrogen in needles of 30-year-old, fertilized Scots pine (Pinus sylvestris L.) trees growing in Northern Sweden were investigated over two years in field experiments. The studied plots had been fertilized annually for 17 years with (i) a high level of N, (ii) a medium level of N, or (iii) a medium level of N, P and K. Trees growing on unfertilized plots served as controls. In control trees, glutamine, glutamic acid, gamma-aminobutyric acid, aspartic acid and proline represented 50-70% of the total free amino acids determined. Arginine was present only in low concentrations in control trees throughout the year, but it was usually the most abundant amino acid in fertilized trees. Glutamine concentrations were high during the spring and summer in both years of study, whereas proline concentrations were high in the spring but otherwise low throughout the year. In the first year of study, glutamic acid concentrations were high during the spring and summer, whereas gamma-aminobutyric acid was present in high concentrations during the winter months. This pattern was less pronounced in the second year of investigation. The concentrations of most amino acids, except glutamic acid, increased in response to fertilization. Nitrogen fertilization increased the foliar concentration of arginine from < 1 micromol g(dw) (-1) in control trees to a maximum of 110 micromol g(dw) (-1). Trees fertilized with nitrogen, phosphorus and potassium had significantly lower arginine concentrations than trees fertilized with the same amount of nitrogen only. Protein concentrations were similar in all fertilized trees but higher than those in control trees. For all treatments, protein concentrations were high in winter and at a minimum in early spring. In summer, the protein concentration remained almost constant except for a temporary decrease which coincided with the expansion of new shoots. Apart from arginine, the amino acid composition of

  15. Quantification of free and total desmosine and isodesmosine in human urine by liquid chromatography tandem mass spectrometry: a comparison of the surrogate-analyte and the surrogate-matrix approach for quantitation.

    PubMed

    Ongay, Sara; Hendriks, Gert; Hermans, Jos; van den Berge, Maarten; ten Hacken, Nick H T; van de Merbel, Nico C; Bischoff, Rainer

    2014-01-24

    In spite of the data suggesting the potential of urinary desmosine (DES) and isodesmosine (IDS) as biomarkers for elevated lung elastic fiber turnover, further validation in large-scale studies of COPD populations, as well as the analysis of longitudinal samples is required. Validated analytical methods that allow the accurate and precise quantification of DES and IDS in human urine are mandatory in order to properly evaluate the outcome of such clinical studies. In this work, we present the development and full validation of two methods that allow DES and IDS measurement in human urine, one for the free and one for the total (free+peptide-bound) forms. To this end we compared the two principle approaches that are used for the absolute quantification of endogenous compounds in biological samples, analysis against calibrators containing authentic analyte in surrogate matrix or containing surrogate analyte in authentic matrix. The validated methods were employed for the analysis of a small set of samples including healthy never-smokers, healthy current-smokers and COPD patients. This is the first time that the analysis of urinary free DES, free IDS, total DES, and total IDS has been fully validated and that the surrogate analyte approach has been evaluated for their quantification in biological samples. Results indicate that the presented methods have the necessary quality and level of validation to assess the potential of urinary DES and IDS levels as biomarkers for the progression of COPD and the effect of therapeutic interventions. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Total protein analysis as a reliable loading control for quantitative fluorescent Western blotting.

    PubMed

    Eaton, Samantha L; Roche, Sarah L; Llavero Hurtado, Maica; Oldknow, Karla J; Farquharson, Colin; Gillingwater, Thomas H; Wishart, Thomas M

    2013-01-01

    using total protein analysis on samples run in parallel with stains such as Coomassie blue provides a more robust approach.

  17. Determination of fat and total protein content in milk using conventional digital imaging.

    PubMed

    Kucheryavskiy, Sergey; Melenteva, Anastasiia; Bogomolov, Andrey

    2014-04-01

    The applicability of conventional digital imaging to quantitative determination of fat and total protein in cow's milk, based on the phenomenon of light scatter, has been proved. A new algorithm for extracting features from digital images of milk samples has been developed. The algorithm takes into account spatial distribution of light, diffusely transmitted through a sample. The proposed method has been tested on two sample sets prepared from industrial raw milk standards, with variable fat and protein content. Partial Least-Squares (PLS) regression on the features calculated from images of monochromatically illuminated milk samples resulted in models with high prediction performance when analysed the sets separately (best models with cross-validated R(2)=0.974 for protein and R(2)=0.973 for fat content). However when analysed the sets jointly with the obtained results were significantly worse (best models with cross-validated R(2)=0.890 for fat content and R(2)=0.720 for protein content). The results have been compared with previously published Vis/SW-NIR spectroscopic study of similar samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. INFLUENCE OF TOTAL BODY X-IRRADIATION ON THE LEVELS OF CREATINE PHOSPHATE, INORGANIC PHOSPHORUS AND ATP IN MUSCLE AND ON THE LEVELS OF CREATINE, CREATININE, N'-METHYL-NICOTINAMIDE AND NITROGEN IN URINE

    SciTech Connect

    Kumta, U.S.; Gurnani, S.U.; Sahasrabudhe, M.B.

    1957-09-01

    The influence of total-body irradiation on the levels of creatine phosphate (CP), adenosine triphosphate (ATP) and inorganic phosphorus (IP) in muscle has been investigated in rats. CP and ATP levels decrease by about 33% while those of 1P increase 4 times in irradiated rats. Studies on the influence of irradiation on the excretion of creatine, creatinine, and N'-methyl- nicotinamide in urine show that the excretion of creatine and N'-methyl- nlcotinamide is increased two-fold while that of creatinine is increased by 160%. It is suggested that the low levels of creatine phosphate are probably due to an impairment in the phosphorylationmore » of creatine or due to an adaptive breakdown of creatine phosphate leading to increased excretion of creatine and creatinine. (auth)« less

  19. Urine melanin test

    MedlinePlus

    Thormahlen's test; Melanin - urine ... A clean-catch urine sample is needed. ... this substance that it shows up in the urine. ... Normally, melanin is not present in urine. Normal value ranges may ... measurements or test different samples. Talk to your health ...

  20. Utility of untimed single urine protein/creatinine ratio as a substitute for 24-h proteinuria for assessment of proteinuria in systemic lupus erythematosus.

    PubMed

    Medina-Rosas, Jorge; Gladman, Dafna D; Su, Jiandong; Sabapathy, Arthy; Urowitz, Murray B; Touma, Zahi

    2015-10-24

    In this study, we determined: (1) the utility of an untimed sample of urine protein/creatinine ratio (PCR) as a screening test for proteinuria, (2) its ability to accurately measure proteinuria, and (3) cutoff values for PCR predicting protein content in a 24-h urine collection sample (24hP) of 0.5, 1.0, and 2.0 g/day. Analysis was performed on data from a single lupus cohort (2008-2014). Proteinuria was measured in a 24hP and with PCR. On the basis of 24hP, samples were divided into 4 groups: group 1, <0.5 g/day; group 2, 0.5-0.99 g/day; group 3, 1-1.99 g/day; and group 4, ≥2 g/day. To determine the validity of PCR in screening for proteinuria, the Pearson correlation coefficient was determined for the urine samples with normal PCR (<0.05 g/mmol) and normal 24hP (<0.5 g/day). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of PCR were calculated. To determine the ability of PCR to accurately measure the level of proteinuria, in addition to the correlation between 24hP and PCR, agreement was determined by intraclass correlation coefficient, concordance correlation coefficient, and Bland-Altman plot between 24hP/24hC and PCR. The best cutoffs for PCR predicting a 24hP of 0.5, 1.0, and 2.0 g/day were determined with the receiver operating characteristic curve. The correlation of the samples with normal PCR as well as 24hP (n = 552) was 0.29 (p < 0.0001). PCR sensitivity and specificity against 24hP were 91 % and 83 %, respectively. The PPV was 82.5 %, and the NPV was 91.4 %. The correlation for all samples (n = 1233) was high, but low to moderate for groups 1, 2, 3, and 4. The agreement for all samples was appropriate but poor for groups 1, 2, 3, and 4. PCR cutoffs for 24hP of 0.5, 1.0, and 2.0 g/day were 0.08, 0.16, and 0.35 g/mmol, respectively. PCR can be used as a screening test for proteinuria, and the best cutoff value to predict a 24hP of 0.5 g/day is 0.08 g/mmol (800

  1. Quantitative Analysis of Protein Translocations by Microfluidic Total Internal Reflection Fluorescence Flow Cytometry

    PubMed Central

    Wang, Jun; Fei, Bei; Geahlen, Robert L.

    2010-01-01

    Protein translocation, or the change in a protein’s location between different subcellular compartments, is a critical process by which intracellular proteins carry out their cellular functions. Aberrant translocation events contribute to various diseases ranging from metabolic disorders to cancer. In this study, we demonstrate the use of a newly developed single-cell tool, microfluidic total internal reflection fluorescence flow cytometry (TIRF-FC), for detecting both cytosol to plasma membrane and cytosol to nucleus translocations using the tyrosine kinase Syk and the transcription factor NF-κB as models. This technique detects fluorescent molecules at the plasma membrane and in the membrane-proximal cytosol in single cells. We were able to record quantitatively changes in the fluorescence density in the evanescent field associated with these translocation processes for large cell populations with single cell resolution. We envision that TIRF-FC will provide a new approach to explore the molecular biology and clinical relevance of protein translocations. PMID:20820633

  2. Interferences of homogentisic acid (HGA) on routine clinical chemistry assays in serum and urine and the implications for biochemical monitoring of patients with alkaptonuria.

    PubMed

    Curtis, S L; Roberts, N B; Ranganath, L R

    2014-05-01

    We have assessed the effect of elevated concentrations of homogentisic acid (HGA) as in alkaptonuria (AKU), on a range of routine chemistry tests in serum and urine. HGA was added to pooled serum and a range of assays was analysed with Roche Modular chemistries. Effects on urine were assessed by diluting normal urine with urine from a patient with AKU, adding HGA to urine and after lowering output of urinary HGA with nitisinone treatment. Serum enzymatic creatinine showed 30% negative interference with 100μmol/L HGA and >50% at 400μmol/L. Serum urate 100 to 480μmol/L was reduced up to 20% at 100 and to 50% with 400μmol/L HGA. Serum cholesterol between 3 and 11mmol/L was reduced by 0.5mmol/L with 400μmol/L HGA. Urine enzymatic creatinine and urate with >2mmol/L HGA showed concentration dependent negative interference up to 80%. A positive interference in urine total protein by benzethonium turbidometric assay was observed, with 10mmol/L HGA equivalent to 1g/L protein. Jaffe creatinine, Na, K, Cl, Mg, Ca, phosphate, ALT, GGT, ALP activities and urea in serum and or urine were not affected by increases in HGA. To avoid interferences by HGA in alkaptonuria concentration of HGA should be established before samples are assayed with peroxidase assays and benzethonium urine protein. Copyright © 2013 The Canadian Society of Clinical Chemists. All rights reserved.

  3. Urine circulating-tumor DNA (ctDNA) detection of acquired EGFR T790M mutation in non-small-cell lung cancer: An outcomes and total cost-of-care analysis.

    PubMed

    Sands, Jacob; Li, Qianyi; Hornberger, John

    2017-08-01

    Third-generation tyrosine kinase inhibitors (TKIs) have proven effective in patients with the acquired EGFR T790M resistance mutation who progress on prior EGFR TKI therapy. Median progression-free survival (PFS) on a 3rd-gen TKI was 9-10 months for T790M+ patients compared to 2.8 months for T790M- patients. PFS is similar regardless of the specimen used to assess T790M, such as tissue, plasma, or urine ctDNA. This study aimed to assess the total cost of care of a urine-testing strategy (UTS) versus a tissue-testing strategy (TTS) for T790M detection, in patients with EGFR-mutation positive lung adenocarcinoma and progression on prior TKI therapy. Long-term outcomes and economic implications were assessed from a US payer perspective. Endpoints were PFS, overall survival (OS), medical resource use and related costs. We included published randomized drug trials and Medicare fee schedules. A state-transition analysis and Markov model tracked patients from stable disease to progression and death. Univariate and multivariate sensitivity analyses were performed to assess the robustness of findings and identify factors that most influenced outcomes and costs. UTS increased the rate of detection of patients with T790M mutation eligible for treatment with 3rd generation TKI by 7% compared with TTS; urine ctDNA testing detected T790M mutation in some patients for whom biopsy could not be performed or when tissue testing yielded indeterminate results. Due to enhanced targeting of TKI therapy, UTS increased PFS and OS by 0.44 and 0.35 months, respectively. UTS yields a savings of $1243-$1680 per patient due to avoidance of biopsy, potential biopsy-associated complications, and tissue-based molecular testing in approximately 55.6% of patients. Probability of T790M detection by tissue and cost of biopsy procedure were the most influential factors. UTS prolonged PFS/OS due to increased detection of T790M mutation and decreased biopsies and complication-related costs. Copyright

  4. Achieving second order advantage with multi-way partial least squares and residual bi-linearization with total synchronous fluorescence data of monohydroxy-polycyclic aromatic hydrocarbons in urine samples.

    PubMed

    Calimag-Williams, Korina; Knobel, Gaston; Goicoechea, H C; Campiglia, A D

    2014-02-06

    An attractive approach to handle matrix interference in samples of unknown composition is to generate second- or higher-order data formats and process them with appropriate chemometric algorithms. Several strategies exist to generate high-order data in fluorescence spectroscopy, including wavelength time matrices, excitation-emission matrices and time-resolved excitation-emission matrices. This article tackles a different aspect of generating high-order fluorescence data as it focuses on total synchronous fluorescence spectroscopy. This approach refers to recording synchronous fluorescence spectra at various wavelength offsets. Analogous to the concept of an excitation-emission data format, total synchronous data arrays fit into the category of second-order data. The main difference between them is the non-bilinear behavior of synchronous fluorescence data. Synchronous spectral profiles change with the wavelength offset used for sample excitation. The work presented here reports the first application of total synchronous fluorescence spectroscopy to the analysis of monohydroxy-polycyclic aromatic hydrocarbons in urine samples of unknown composition. Matrix interference is appropriately handled by processing the data either with unfolded-partial least squares and multi-way partial least squares, both followed by residual bi-linearization. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Biomarkers of Oxidative Stress Study V: Ozone exposure of rats and its effect on lipids, proteins and DNA in plasma and urine

    PubMed Central

    Kadiiska, Maria B.; Basu, Samar; Brot, Nathan; Cooper, Christopher; Csallany, A. Saari; Davies, Michael J.; George, Magdalene M.; Murray, Dennis M.; Roberts, L. Jackson; Shigenaga, Mark K.; Sohal, Rajindar S.; Stocker, Roland; Van Thiel, David H.; Wiswedel, Ingrid; Hatch, Gary E.; Mason, Ronald P.

    2014-01-01

    Ozone exposure effect on free radical-catalyzed oxidation products of lipids, proteins and DNA in the plasma and urine of rats was studied as a continuation of the international Biomarker of Oxidative Stress Study (BOSS) sponsored by NIEHS/NIH. The goal was to identify a biomarker for ozone-induced oxidative stress and to assess whether inconsistent results often reported in the literature might be due to the limitations of the available methods for measuring the various types of oxidative products. The time and dose-dependent effects of ozone exposure on rat plasma lipid hydroperoxides, malondialdehyde, F2-isoprostanes, protein carbonyls, methionine oxidation, tyrosine- and phenylalanine oxidation products, as well as urinary malondialdehyde and F2-isoprostanes were investigated with various techniques. The criterion used to recognize a marker in the model of ozone exposure was that a significant effect could be identified and measured in a biological fluid seen at both doses at more than one time point. No statistically significant differences between the experimental and control groups at either ozone dose and time point studied could be identified in this study. Tissue samples were not included. Despite all the work accomplished in the BOSS study of ozone, no available product of oxidation in biological fluid has yet met the required criteria of being a biomarker. The current negative findings as a consequence of ozone exposure are of great importance, because they document that in complex systems, as the present in vivo experiment, the assays used may not provide meaningful data of ozone oxidation, especially in human studies. PMID:23608465

  6. Biomarkers of oxidative stress study V: ozone exposure of rats and its effect on lipids, proteins, and DNA in plasma and urine.

    PubMed

    Kadiiska, Maria B; Basu, Samar; Brot, Nathan; Cooper, Christopher; Saari Csallany, A; Davies, Michael J; George, Magdalene M; Murray, Dennis M; Jackson Roberts, L; Shigenaga, Mark K; Sohal, Rajindar S; Stocker, Roland; Van Thiel, David H; Wiswedel, Ingrid; Hatch, Gary E; Mason, Ronald P

    2013-08-01

    Ozone exposure effect on free radical-catalyzed oxidation products of lipids, proteins, and DNA in the plasma and urine of rats was studied as a continuation of the international Biomarker of Oxidative Stress Study (BOSS) sponsored by NIEHS/NIH. The goal was to identify a biomarker for ozone-induced oxidative stress and to assess whether inconsistent results often reported in the literature might be due to the limitations of the available methods for measuring the various types of oxidative products. The time- and dose-dependent effects of ozone exposure on rat plasma lipid hydroperoxides, malondialdehyde, F2-isoprostanes, protein carbonyls, methionine oxidation, and tyrosine- and phenylalanine oxidation products, as well as urinary malondialdehyde and F2-isoprostanes were investigated with various techniques. The criterion used to recognize a marker in the model of ozone exposure was that a significant effect could be identified and measured in a biological fluid seen at both doses at more than one time point. No statistically significant differences between the experimental and the control groups at either ozone dose and time point studied could be identified in this study. Tissue samples were not included. Despite all the work accomplished in the BOSS study of ozone, no available product of oxidation in biological fluid has yet met the required criteria of being a biomarker. The current negative findings as a consequence of ozone exposure are of great importance, because they document that in complex systems, as the present in vivo experiment, the assays used may not provide meaningful data of ozone oxidation, especially in human studies. Published by Elsevier Inc.

  7. Comparison of Associations of Urine Protein-Creatinine Ratio Versus Albumin-Creatinine Ratio With Complications of CKD: A Cross-sectional Analysis

    PubMed Central

    Fisher, Herrick; Hsu, Chi-yuan; Vittinghoff, Eric; Lin, Feng; Bansal, Nisha

    2013-01-01

    Background Urine albumin-creatinine ratio (ACR) and protein-creatinine ratio (PCR) are important markers of kidney damage and are utilized for prognosis in persons with chronic kidney disease (CKD). Despite how commonly these measurements are done in clinical practice, relatively few studies have directly compared the performance of these two measures with regard to associations with clinical outcomes, which may inform clinicians about which measure of urinary protein excretion is best. We studied the association of ACR and PCR with common complications of CKD. Study Design Cross-sectional study. Setting & Participants 3,481 participants with CKD in the Chronic Renal Insufficiency Cohort (CRIC) study. Predictors ACR and PCR. Outcomes We examined the association between ACR and PCR with measures of common CKD complications: serum hemoglobin, bicarbonate, parathyroid hormone, phosphorus, potassium and albumin. Measurements Restricted cubic spline analyses adjusted for estimated glomerular filtration rate (eGFR; calculated by the MDRD [Modification of Diet in Renal Disease] Study Equation) were performed to study the continuous association with our predictors with each outcome. Results Mean eGFR was 43 ± 13 (SD) ml/min/1.73 m2 and median levels of PCR and ACR were 140 and 46 mg/g, respectively. In continuous analyses adjusted for eGFR, higher ACR and PCR were comparable and both were associated with lower levels of serum hemoglobin, bicarbonate, and albumin and higher levels of parathyroid hormone, phosphorus, and potassium. Across all outcomes, the associations of ACR and PCR were comparable with only small, absolute differences in the outcome measure. Similar associations were seen in patients with diabetes mellitus. Limitations Participants largely had moderate CKD with low levels of ACR and PCR, so results may not be generalizable to all CKD populations. Conclusions In persons with CKD, ACR and PCR are relatively comparable in their associations with common

  8. Genetics Home Reference: maple syrup urine disease

    MedlinePlus

    ... disease is an inherited disorder in which the body is unable to process certain protein building blocks (amino acids) properly. The condition gets its name from the distinctive sweet odor of affected infants' urine. It is also characterized ...

  9. Clinical performance evaluation of total protein measurement by digital refractometry and characterization of non-protein solute interferences.

    PubMed

    Hunsaker, Joshua J H; Wyness, Sara P; Snow, Taylor M; Genzen, Jonathan R

    2016-12-01

    Refractometric methods to measure total protein (TP) in serum and plasma specimens have been replaced by automated biuret methods in virtually all routine clinical testing. A subset of laboratories, however, still report using refractometry to measure TP in conjunction with serum protein electrophoresis. The objective of this study was therefore to conduct a modern performance evaluation of a digital refractometer for TP measurement. Performance evaluation of a MISCO Palm Abbe™ digital refractometer was conducted through device familiarization, carryover, precision, accuracy, linearity, analytical sensitivity, analytical specificity, and reference interval verification. Comparison assays included a manual refractometer and an automated biuret assay. Carryover risk was eliminated using a demineralized distilled water (ddH 2 O) wash step. Precision studies demonstrated overall imprecision of 2.2% CV (low TP pool) and 0.5% CV (high TP pool). Accuracy studies demonstrated correlation to both manual refractometry and the biuret method. An overall positive bias (+5.0%) was observed versus the biuret method. On average, outlier specimens had an increased triglyceride concentration. Linearity was verified using mixed dilutions of: a) low and high concentration patient pools, or b) albumin-spiked ddH 2 O and high concentration patient pool. Decreased recovery was observed using ddH 2 O dilutions at low TP concentrations. Significant interference was detected at high concentrations of glucose (>267 mg/dL) and triglycerides (>580 mg/dL). Current laboratory reference intervals for TP were verified. Performance characteristics of this digital refractometer were validated in a clinical laboratory setting. Biuret method remains the preferred assay for TP measurement in routine clinical analyses.

  10. Total protein of whole saliva as a biomarker of anaerobic threshold.

    PubMed

    Bortolini, Miguel Junior Sordi; De Agostini, Guilherme Gularte; Reis, Ismair Teodoro; Lamounier, Romeu Paulo Martins Silva; Blumberg, Jeffrey B; Espindola, Foued Salmen

    2009-09-01

    Saliva provides a convenient and noninvasive matrix for assessing specific physiological parameters, including some biomarkers of exercise. We investigated whether the total protein concentration of whole saliva (TPWS) would reflect the anaerobic threshold during an incremental exercise test. After a warm-up period, 13 nonsmoking men performed a maximum incremental exercise on a cycle ergometer. Blood and stimulated saliva were collected during the test. The TPWS anaerobic threshold (PAT) was determined using the Dmax method. The PAT was correlated with the blood lactate anaerobic threshold (AT; r = .93, p < .05). No significant difference (p = .16) was observed between PAT and AT. Thus, TPWS provides a convenient and noninvasive matrix for determining the anaerobic threshold during incremental exercise tests.

  11. Total solids content and degree of hydrolysis influence proteolytic inactivation kinetics following whey protein hydrolysate manufacture.

    PubMed

    Conesa, Celia; FitzGerald, Richard J

    2013-10-23

    The kinetics and thermodynamics of the thermal inactivation of Corolase PP in two different whey protein concentrate (WPC) hydrolysates with degree of hydrolysis (DH) values of ~10 and 21%, and at different total solids (TS) levels (from 5 to 30% w/v), were studied. Inactivation studies were performed in the temperature range from 60 to 75 °C, and residual enzyme activity was quantified using the azocasein assay. The inactivation kinetics followed a first-order model. Analysis of the activation energy, thermodynamic parameters, and D and z values, demonstrated that the inactivation of Corolase PP was dependent on solution TS. The intestinal enzyme preparation was more heat sensitive at low TS. Moreover, it was also found that the enzyme was more heat sensitive in solutions at higher DH.

  12. Blood count and C-reactive protein evolution in gastric cancer patients with total gastrectomy surgery

    PubMed Central

    CSENDES J., Attila; MUÑOZ Ch., Andrea; BURGOS L., Ana María

    2014-01-01

    Background The complete blood count (CBC) and C-reactive protein (CRP) are useful inflammatory parameters for ruling out acute postoperative inflammatory complications. Aim To determine their changes in gastric cancer patients submitted to total gastrectomy. Methods This is a prospective study, with 36 patients with gastric cancer who were submitted to elective total gastrectomy. On the first, third and fifth postoperative day (POD), blood count and CRP changes were assessed. Patients with postoperative complications were excluded. Results Twenty-one (58%) were men and 15 (42%) women. The mean age was 65 years. The leukocytes peaked on the 1st POD with a mean of 13,826 u/mm³, and decreased to 8,266 u/mm³ by the 5th POD. The bacilliforms peaked on the 1st POD with a maximum value of 1.48%. CRP reached its maximum level on the 3rd POD with a mean of 144.64 mg/l±44.84. Preoperative hematocrit (HCT) was 35% and 33.67% by the 5th POD. Hemoglobin, showed similar values. Conclusions Leukocytes increased during the 1st POD but reached normal values by the 5th POD. CRP peaked on the 3rd POD but did not reach normal values by the 5th POD. PMID:25626929

  13. Urine concentration test

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003608.htm Urine concentration test To use the sharing features on this page, please enable JavaScript. A urine concentration test measures the ability of the kidneys ...

  14. Nitrites in Urine

    MedlinePlus

    ... Why do I need a nitrites in urine test? Your health care provider may have ordered a urinalysis as part ... Fever What happens during a nitrites in urine test? Your health care provider will need to collect a sample of ...

  15. Urobilinogen in Urine

    MedlinePlus

    ... Why do I need a urobilinogen in urine test? Your health care provider may have ordered this test as part ... skin What happens during a urobilinogen in urine test? Your health care provider will need to collect a sample of ...

  16. Urine drug screen

    MedlinePlus

    Drug screen - urine ... detect the presence of illegal and some prescription drugs in your urine. Their presence may indicate that you recently used these drugs. Some drugs may remain in your system for ...

  17. Leukocyte esterase urine test

    MedlinePlus

    ... the urine. This may mean you have a urinary tract infection . If this test is positive, the urine should ... Results Mean An abnormal result indicates a possible urinary tract infection. The following may turn the test abnormal even ...

  18. Sodium urine test

    MedlinePlus

    ... or monitor many types of kidney diseases. Normal Results For adults, normal urine sodium values are generally ... meaning of your specific test result. What Abnormal Results Mean A higher than normal urine sodium level ...

  19. Cytology exam of urine

    MedlinePlus

    Urine cytology; Bladder cancer - cytology; Urethral cancer - cytology; Renal cancer - cytology ... The test is done to detect cancer of the urinary tract. It is often done when blood is seen in the urine. It is also useful for monitoring people ...

  20. Urine specific gravity test

    MedlinePlus

    ... medlineplus.gov/ency/article/003587.htm Urine specific gravity test To use the sharing features on this page, please enable JavaScript. Urine specific gravity is a laboratory test that shows the concentration ...

  1. Uric acid - urine

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003616.htm Uric acid urine test To use the sharing features on this page, please enable JavaScript. The uric acid urine test measures the level of uric acid ...

  2. Fluid balance, glomerular filtration rate, and urine output in dogs anesthetized for an orthopedic surgical procedure.

    PubMed

    Boscan, Pedro; Pypendop, Bruno H; Siao, Kristine T; Francey, Thierry; Dowers, Kristy; Cowgill, Larry; Ilkiw, Jan E

    2010-05-01

    To determine fluid retention, glomerular filtration rate, and urine output in dogs anesthetized for a surgical orthopedic procedure. 23 dogs treated with a tibial plateau leveling osteotomy. 12 dogs were used as a control group. Cardiac output was measured in 5 dogs, and 6 dogs received carprofen for at least 14 days. Dogs received oxymorphone, atropine, propofol, and isoflurane for anesthesia (duration, 4 hours). Urine and blood samples were obtained for analysis every 30 minutes. Lactated Ringer's solution was administered at 10 mL/kg/h. Urine output was measured and glomerular filtration rate was estimated. Fluid retention was measured by use of body weight, fluid balance, and bioimpedance spectroscopy. No difference was found among control, cardiac output, or carprofen groups, so data were combined. Median urine output and glomerular filtration rate were 0.46 mL/kg/h and 1.84 mL/kg/min. Dogs retained a large amount of fluids during anesthesia, as indicated by increased body weight, positive fluid balance, increased total body water volume, and increased extracellular fluid volume. The PCV, total protein concentration, and esophageal temperature decreased in a linear manner. Dogs anesthetized for a tibial plateau leveling osteotomy retained a large amount of fluids, had low urinary output, and had decreased PCV, total protein concentration, and esophageal temperature. Evaluation of urine output alone in anesthetized dogs may not be an adequate indicator of fluid balance.

  3. Urine sample (image)

    MedlinePlus

    A "clean-catch" urine sample is performed by collecting the sample of urine in midstream. Men or boys should wipe clean the head ... water and rinse well. A small amount of urine should initially fall into the toilet bowl before ...

  4. Determination of total mercury in seafood and other protein-rich products

    SciTech Connect

    Landi, S.; Fagioli, F.; Locatelli, C.

    1992-11-01

    A previously developed wet-digestion method for the determination of total mercury in plants by cold vapor atomic absorption spectroscopy (CVAAS) was extended to the analysis of seafood and other products rich in proteins. Oxidation of matrixes is accomplished by K{sub 2}Cr{sub 2}O{sub 7} in the presence of diluted H{sub 2}SO{sub 4}; a simple air condenser is used to reflux vapors released from the boiling mixture. The original procedure (A) and 2 modifications (B and C), which differ with respect to the mode of acidification and/or digestion time and the types of condensers used, were compared for precision and accuracy bymore » means of National Institute of Standards and Technology Research Material 50 Albacore Tuna and proved to be reliable (Hg present, 0.95{plus_minus}0.1 {mu}g/g; Hg found, 0.97 {plus_minus} 0.029 {mu}g/g [A], 0.98 {plus_minus} 0.018 {mu}g/g [B], and 0.94 {plus_minus} 0.025 {mu}g/g [C]). The modified procedures were tested further in Hg recovery experiments on a variety of biological matrixes with different spiking substances and again showed good analytical characteristics (overall average recoveries = 98 {plus_minus} 5.1% for seafood and 100 {plus_minus} 3.6 for protein-rich baby foods). 22 refs., 1 fig., 5 tabs.« less

  5. Total replacement of corn by mesquite pod meal considering nutritional value, performance, feeding behavior, nitrogen balance, and microbial protein synthesis of Holstein-Zebu crossbred dairy steers.

    PubMed

    de Oliveira Moraes, Gláucia Sabrine; de Souza, Evaristo Jorge Oliveira; Véras, Antonia Sherlânea Chaves; de Paula Almeida, Marina; da Cunha, Márcio Vieira; Torres, Thaysa Rodrigues; da Silva, Camila Sousa; Pereira, Gerfesson Felipe Cavalcanti

    2016-10-01

    The objective of the present study to assess the effects of mesquite pod addition replacing corn (0, 250, 500, 750, and 1000 g/kg in the dry matter basis) on nutrient intake, animal performance, feeding behavior, nutrient digestibility, nitrogen balance, and microbial protein synthesis. Twenty-five Holstein-Zebu crossbred dairy steers at 219 ± 22 kg initial body weight and 18 months of age were used. The experiment lasted 84 days, divided into three periods of 28 days. A completely randomized design was used, and data were submitted to analysis using PROC GLM for analysis of variance and PROC REG for regression analysis using the software Statistical Analysis Systems version 9.1. Experimental diets were composed of Tifton 85 hay, soybean meal, ground corn, mesquite pod meal, and mineral salt. Samples of food offered were collected during the last 3 days of each period, and the leftovers were collected daily, with samples bulked per week. At the end of each 28-day period, the remaining animals were weighed to determine total weight gain and average daily gain. The assessment of behavioral patterns was performed through instantaneous scans in 5-min intervals for three consecutive 12-h days. A single urine sample from each animal was collected on the last day of each collection period at about 4 h after the first feeding. The replacement of corn by mesquite pod meal did not significantly influence treatments regarding nutrients intake, animal performance, and feeding behavior. Retained and consumed nitrogen ratio did not statistically differ between replacement levels. Likewise, there were no statistical differences regarding microbial protein synthesis and efficiency between replacement levels. Mesquite pod meal can be used in Holstein-Zebu crossbred dairy steers' diet with total corn replacement.

  6. Serial-omics characterization of equine urine

    PubMed Central

    Yuan, Min; Breitkopf, Susanne B.

    2017-01-01

    Horse urine is easily collected and contains molecules readily measurable using mass spectrometry that can be used as biomarkers representative of health, disease or drug tampering. This study aimed at analyzing microliter levels of horse urine to purify, identify and quantify proteins, polar metabolites and non-polar lipids. Urine from a healthy 12 year old quarter horse mare on a diet of grass hay and vitamin/mineral supplements with limited pasture access was collected for serial-omics characterization. The urine was treated with methyl tert-butyl ether (MTBE) and methanol to partition into three distinct layers for protein, non-polar lipid and polar metabolite content from a single liquid-liquid extraction and was repeated two times. Each layer was analyzed by high performance liquid chromatography—high resolution tandem mass spectrometry (LC-MS/MS) to obtain protein sequence and relative protein levels as well as identify and quantify small polar metabolites and lipids. The results show 46 urine proteins, many related to normal kidney function, structural and circulatory proteins as well as 474 small polar metabolites but only 10 lipid molecules. Metabolites were mostly related to urea cycle and ammonia recycling as well as amino acid related pathways, plant diet specific molecules, etc. The few lipids represented triglycerides and phospholipids. These data show a complete mass spectrometry based—omics characterization of equine urine from a single 333 μL mid-stream urine aliquot. These omics data help serve as a baseline for healthy mare urine composition and the analyses can be used to monitor disease progression, health status, monitor drug use, etc. PMID:29028822

  7. Plasma protein hydroperoxides during aging in humans: correlation with paraoxonase 1 (PON1) arylesterase activity and plasma total thiols.

    PubMed

    Mehdi, Mohammad Murtaza; Rizvi, Syed Ibrahim

    2013-02-01

    Oxidative stress is thought to play a major role in the development of several age-dependent diseases. Proteins are major targets for oxidative attack. Protein hydroperoxides are formed by hydroxyl and singlet oxygen attack on protein, forming relatively stable hydroperoxides on histidine, tyrosine and tryptophan residues. This study investigated the levels of plasma protein hydroperoxides and antioxidant potential of plasma during aging in humans. We correlated the protein hydroperoxide formation with plasma antioxidant potential, paraoxonase 1 (PON1) arylesterase activity and plasma total thiols. The protein hydroperoxides and antioxidant potential were measured in plasma of human subjects aged between 20 and 81 years of both genders. Increase in plasma protein hydroperoxides and decrease in plasma antioxidant potential were observed as function of human age. This study provides strong correlation between plasma protein hydroperoxides formation and decrease in plasma antioxidant potential during aging. PON1 arylesterase activity and plasma total thiols levels were also found to show significant correlation with increasing levels of plasma protein hydroperoxides during aging. The plasma protein hydroperoxides provide a reliable marker of long-term redox balance and degree of oxidative stress during aging process. Copyright © 2013 IMSS. Published by Elsevier Inc. All rights reserved.

  8. Effects of synchronization of carbohydrate and protein supply in total mixed ration with korean rice wine residue on ruminal fermentation, nitrogen metabolism and microbial protein synthesis in holstein steers.

    PubMed

    Piao, Min Yu; Kim, Hyun J; Seo, J K; Park, T S; Yoon, J S; Kim, K H; Ha, Jong K

    2012-11-01

    Three Holstein steers in the growing phase, each with a ruminal cannula, were used to test the hypothesis that the synchronization of the hourly rate of carbohydrate and nitrogen (N) released in the rumen would increase the amount of retained nitrogen for growth and thus improve the efficiency of microbial protein synthesis (EMPS). In Experiment 1, in situ degradability coefficients of carbohydrate and N in feeds including Korean rice wine residue (RWR) were determined. In Experiment 2, three total mixed ration (TMR) diets having different rates of carbohydrate and N release in the rumen were formulated using the in situ degradability of the feeds. All diets were made to contain similar contents of crude protein (CP) and neutral detergent fiber (NDF) but varied in their hourly pattern of nutrient release. The synchrony index of the three TMRs was 0.51 (LS), 0.77 (MS) and 0.95 (HS), respectively. The diets were fed at a restricted level (2% of the animal's body weight) in a 3×3 Latin-square design. Synchronizing the hourly supply of energy and N in the rumen did not significantly alter the digestibility of dry matter, organic matter, crude protein, NDF or acid detergent fiber (ADF) (p>0.05). The ruminal NH3-N content of the LS group at three hours after feeding was significantly higher (p<0.05) than that of the other groups; however, the mean values of ruminal NH3-N, pH and VFA concentration among the three groups were not significantly different (p>0.05). In addition, the purine derivative (PD) excretion in urine and microbial-N production (MN) among the three groups were not significantly different (p>0.05). In conclusion, synchronizing dietary energy and N supply to the rumen did not have a major effect on nutrient digestion or microbial protein synthesis (MPS) in Holstein steers.

  9. Reference intervals for 24 laboratory parameters determined in 24-hour urine collections.

    PubMed

    Curcio, Raffaele; Stettler, Helen; Suter, Paolo M; Aksözen, Jasmin Barman; Saleh, Lanja; Spanaus, Katharina; Bochud, Murielle; Minder, Elisabeth; von Eckardstein, Arnold

    2016-01-01

    Reference intervals for many laboratory parameters determined in 24-h urine collections are either not publicly available or based on small numbers, not sex specific or not from a representative sample. Osmolality and concentrations or enzymatic activities of sodium, potassium, chloride, glucose, creatinine, citrate, cortisol, pancreatic α-amylase, total protein, albumin, transferrin, immunoglobulin G, α1-microglobulin, α2-macroglobulin, as well as porphyrins and their precursors (δ-aminolevulinic acid and porphobilinogen) were determined in 241 24-h urine samples of a population-based cohort of asymptomatic adults (121 men and 120 women). For 16 of these 24 parameters creatinine-normalized ratios were calculated based on 24-h urine creatinine. The reference intervals for these parameters were calculated according to the CLSI C28-A3 statistical guidelines. By contrast to most published reference intervals, which do not stratify for sex, reference intervals of 12 of 24 laboratory parameters in 24-h urine collections and of eight of 16 parameters as creatinine-normalized ratios differed significantly between men and women. For six parameters calculated as 24-h urine excretion and four parameters calculated as creatinine-normalized ratios no reference intervals had been published before. For some parameters we found significant and relevant deviations from previously reported reference intervals, most notably for 24-h urine cortisol in women. Ten 24-h urine parameters showed weak or moderate sex-specific correlations with age. By applying up-to-date analytical methods and clinical chemistry analyzers to 24-h urine collections from a large population-based cohort we provide as yet the most comprehensive set of sex-specific reference intervals calculated according to CLSI guidelines for parameters determined in 24-h urine collections.

  10. Estimation of lactose interference in vaccines and a proposal of methodological adjustment of total protein determination by the lowry method.

    PubMed

    Kusunoki, Hideki; Okuma, Kazu; Hamaguchi, Isao

    2012-01-01

    For national regulatory testing in Japan, the Lowry method is used for the determination of total protein content in vaccines. However, many substances are known to interfere with the Lowry method, rendering accurate estimation of protein content difficult. To accurately determine the total protein content in vaccines, it is necessary to identify the major interfering substances and improve the methodology for removing such substances. This study examined the effects of high levels of lactose with low levels of protein in freeze-dried, cell culture-derived Japanese encephalitis vaccine (inactivated). Lactose was selected because it is a reducing sugar that is expected to interfere with the Lowry method. Our results revealed that concentrations of ≥ 0.1 mg/mL lactose interfered with the Lowry assays and resulted in overestimation of the protein content in a lactose concentration-dependent manner. On the other hand, our results demonstrated that it is important for the residual volume to be ≤ 0.05 mL after trichloroacetic acid precipitation in order to avoid the effects of lactose. Thus, the method presented here is useful for accurate protein determination by the Lowry method, even when it is used for determining low levels of protein in vaccines containing interfering substances. In this study, we have reported a methodological adjustment that allows accurate estimation of protein content for national regulatory testing, when the vaccine contains interfering substances.

  11. Accessibility of selenomethionine proteins by total chemical synthesis: structural studies of human herpesvirus-8 MIP-II.

    PubMed

    Shao, W; Fernandez, E; Wilken, J; Thompson, D A; Siani, M A; West, J; Lolis, E; Schweitzer, B I

    1998-12-11

    The determination of high resolution three-dimensional structures by X-ray crystallography or nuclear magnetic resonance (NMR) is a time-consuming process. Here we describe an approach to circumvent the cloning and expression of a recombinant protein as well as screening for heavy atom derivatives. The selenomethionine-modified chemokine macrophage inflammatory protein-II (MIP-II) from human herpesvirus-8 has been produced by total chemical synthesis, crystallized, and characterized by NMR. The protein has a secondary structure typical of other chemokines and forms a monomer in solution. These results indicate that total chemical synthesis can be used to accelerate the determination of three-dimensional structures of new proteins identified in genome programs.

  12. Fast and selective determination of total protein in milk powder via titration of moving reaction boundary electrophoresis.

    PubMed

    Guo, Cheng-ye; Wang, Hou-yu; Liu, Xiao-ping; Fan, Liu-yin; Zhang, Lei; Cao, Cheng-xi

    2013-05-01

    In this paper, moving reaction boundary titration (MRBT) was developed for rapid and accurate quantification of total protein in infant milk powder, from the concept of moving reaction boundary (MRB) electrophoresis. In the method, the MRB was formed by the hydroxide ions and the acidic residues of milk proteins immobilized via cross-linked polyacrylamide gel (PAG), an acid-base indicator was used to denote the boundary motion. As a proof of concept, we chose five brands of infant milk powders to study the feasibility of MRBT method. The calibration curve of MRB velocity versus logarithmic total protein content of infant milk powder sample was established based on the visual signal of MRB motion as a function of logarithmic milk protein content. Weak influence of nonprotein nitrogen (NPN) reagents (e.g., melamine and urea) on MRBT method was observed, due to the fact that MRB was formed with hydroxide ions and the acidic residues of captured milk proteins, rather than the alkaline residues or the NPN reagents added. The total protein contents in infant milk powder samples detected via the MRBT method were in good agreement with those achieved by the classic Kjeldahl method. In addition, the developed method had much faster measuring speed compared with the Kjeldahl method. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. PBG urine test

    MedlinePlus

    ... test. Alternative Names Porphobilinogen test; Porphyria - urine; PBG Images Male urinary system References Fuller SJ, Wiley JS. Heme biosynthesis and its disorders: porphyrias and sideroblastic ...

  14. Urine Pretreat Injection System

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A new method of introducing the OXONE (Registered Trademark) Monopersulfate Compound for urine pretreat into a two-phase urine/air flow stream has been successfully tested and evaluated. The feasibility of this innovative method has been established for purposes of providing a simple, convenient, and safe method of handling a chemical pretreat required for urine processing in a microgravity space environment. Also, the Oxone portion of the urine pretreat has demonstrated the following advantages during real time collection of 750 pounds of urine in a Space Station design two-phase urine Fan/Separator: Eliminated urine precipitate buildup on internal hardware and plumbing; Minimized odor from collected urine; and Virtually eliminated airborne bacteria. The urine pretreat, as presently defined for the Space Station program for proper downstream processing of urine, is a two-part chemical treatment of 5.0 grams of Oxone and 2.3 ml of H2SO4 per liter of urine. This study program and test demonstrated only the addition of the proper ratio of Oxone into the urine collection system upstream of the Fan/Separator. This program was divided into the following three major tasks: (1) A trade study, to define and recommend the type of Oxone injection method to pursue further; (2) The design and fabrication of the selected method; and (3) A test program using high fidelity hardware and fresh urine to demonstrate the method feasibility. The trade study was conducted which included defining several methods for injecting Oxone in different forms into a urine system. Oxone was considered in a liquid, solid, paste and powered form. The trade study and the resulting recommendation were presented at a trade study review held at Hamilton Standard on 24-25 October 94. An agreement was reached at the meeting to continue the solid tablet in a bag concept which included a series of tablets suspended in the urine/air flow stream. These Oxone tablets would slowly dissolve at a controlled rate

  15. Determination of protein concentration in raw milk by mid-infrared fourier transform infrared/attenuated total reflectance spectroscopy.

    PubMed

    Etzion, Y; Linker, R; Cogan, U; Shmulevich, I

    2004-09-01

    This study investigates the potential use of attenuated total reflectance spectroscopy in the mid-infrared range for determining protein concentration in raw cow milk. The determination of protein concentration is based on the characteristic absorbance of milk proteins, which includes 2 absorbance bands in the 1500 to 1700 cm(-1) range, known as the amide I and amide II bands, and absorbance in the 1060 to 1100 cm(-1) range, which is associated with phosphate groups covalently bound to casein proteins. To minimize the influence of the strong water band (centered around 1640 cm(-1)) that overlaps with the amide I and amide II bands, an optimized automatic procedure for accurate water subtraction was applied. Following water subtraction, the spectra were analyzed by 3 methods, namely simple band integration, partial least squares (PLS) and neural networks. For the neural network models, the spectra were first decomposed by principal component analysis (PCA), and the neural network inputs were the spectra principal components scores. In addition, the concentrations of 2 constituents expected to interact with the protein (i.e., fat and lactose) were also used as inputs. These approaches were tested with 235 spectra of standardized raw milk samples, corresponding to 26 protein concentrations in the 2.47 to 3.90% (weight per volume) range. The simple integration method led to very poor results, whereas PLS resulted in prediction errors of about 0.22% protein. The neural network approach led to prediction errors of 0.20% protein when based on PCA scores only, and 0.08% protein when lactose and fat concentrations were also included in the model. These results indicate the potential usefulness of Fourier transform infrared/attenuated total reflectance spectroscopy for rapid, possibly online, determination of protein concentration in raw milk.

  16. Determination of the X-ray structure of the snake venom protein omwaprin by total chemical synthesis and racemic protein crystallography.

    PubMed

    Banigan, James R; Mandal, Kalyaneswar; Sawaya, Michael R; Thammavongsa, Vilasak; Hendrickx, Antoni P A; Schneewind, Olaf; Yeates, Todd O; Kent, Stephen B H

    2010-10-01

    The 50-residue snake venom protein L-omwaprin and its enantiomer D-omwaprin were prepared by total chemical synthesis. Radial diffusion assays were performed against Bacillus megaterium and Bacillus anthracis; both L- and D-omwaprin showed antibacterial activity against B. megaterium. The native protein enantiomer, made of L-amino acids, failed to crystallize readily. However, when a racemic mixture containing equal amounts of L- and D-omwaprin was used, diffraction quality crystals were obtained. The racemic protein sample crystallized in the centrosymmetric space group P2(1)/c and its structure was determined at atomic resolution (1.33 A) by a combination of Patterson and direct methods based on the strong scattering from the sulfur atoms in the eight cysteine residues per protein. Racemic crystallography once again proved to be a valuable method for obtaining crystals of recalcitrant proteins and for determining high-resolution X-ray structures by direct methods.

  17. The characterisation of novel secreted Ly-6 proteins from rat urine by the combined use of two-dimensional gel electrophoresis, microbore high performance liquid chromatography and expressed sequence tag data.

    PubMed

    Southan, Christopher; Cutler, Paul; Birrell, Helen; Connell, John; Fantom, Kenneth G M; Sims, Matthew; Shaikh, Narjis; Schneider, Klaus

    2002-02-01

    A proteomic study of rat urine was undertaken using two-dimensional gel electrophoresis, microbore high performance liquid chromatography, mass spectrometry and N-terminal sequencing. Five known urinary proteins were identified but two novel peptide fragments matched a large number of rat expressed sequence tags (ESTs) from a liver library. By combining protein chemical and nucleotide data, two 101-residue open reading frames with 90% amino acid identity were determined, rat urinary protein 1 (RUP-1) and RUP-2. The data established signal peptide removal and provided evidence for N-glycosylation. A third related sequence, rat spleen protein (RSP-1) was confirmed from EST searches. These three proteins have been submitted to SWISS-PROT as P81827, P81828 and Q9QXN2, respectively. A fourth novel homologue was found in porcine and bovine ESTs from embryo libraries. Alignment with known homologues showed conserved cysteine positions characteristic of a secreted subfamily of Ly-6 proteins. In two cases, antineoplastic urinary protein and caltrin, these homologues have unverified functional annotations. The RUP sequences showed high scoring matches to three unrelated rat mRNAs subsequently established to be chimeric. Two of these share extended sectional identity to RUP-1 but the third may represent another novel Ly-6 homologue. These chimeras have caused serious annotation errors in secondary databases.

  18. Urine: Waste product or biologically active tissue?

    PubMed

    2018-03-01

    Historically, urine has been viewed primarily as a waste product with little biological role in the overall health of an individual. Increasingly, data suggest that urine plays a role in human health beyond waste excretion. For example, urine might act as an irritant and contribute to symptoms through interaction with-and potential compromise of-the urothelium. To explore the concept that urine may be a vehicle for agents with potential or occult bioactivity and to discuss existing evidence and novel research questions that may yield insight into such a role, the National Institute of Diabetes and Digestive and Kidney Disease invited experts in the fields of comparative evolutionary physiology, basic science, nephrology, urology, pediatrics, metabolomics, and proteomics (among others) to a Urinology Think Tank meeting on February 9, 2015. This report reflects ideas that evolved from this meeting and current literature, including the concept of urine quality, the biological, chemical, and physical characteristics of urine, including the microbiota, cells, exosomes, pH, metabolites, proteins, and specific gravity (among others). Additionally, the manuscript presents speculative, and hopefully testable, ideas about the functional roles of urine constituents in health and disease. Moving forward, there are several questions that need further understanding and pursuit. There were suggestions to consider actively using various animal models and their biological specimens to elaborate on basic mechanistic information regarding human bladder dysfunction. Published 2018. This article is a U.S. Government work and is in the public domain in the USA.

  19. Total leaf crude protein, amino acid composition and elemental content in the USDA-ARS bamboo germplasm collection

    USDA-ARS?s Scientific Manuscript database

    Bamboo shoots and leaves are valuable food sources for both humans and livestock. The USDA-ARS National Plant Germplasm System (NPGS) collections hold 93 bamboo species in 20 genera. Total leaf protein, amino acid composition and elemental content for these important genetic resources had never bee...

  20. Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips.

    PubMed

    Jara, Rocío F; Sepúlveda, Carolina; Ip, Hon S; Samuel, Michael D

    2015-04-01

    Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

  1. Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips

    USGS Publications Warehouse

    Jara, Rocio F.; Sepúlveda, Carolina; Ip, Hon S.; Samuel, Michael D.

    2015-01-01

    Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

  2. Urine Monitoring System

    NASA Technical Reports Server (NTRS)

    Feedback, Daniel L.; Cibuzar, Branelle R.

    2009-01-01

    The Urine Monitoring System (UMS) is a system designed to collect an individual crewmember's void, gently separate urine from air, accurately measure void volume, allow for void sample acquisition, and discharge remaining urine into the Waste Collector Subsystem (WCS) onboard the International Space Station. The Urine Monitoring System (UMS) is a successor design to the existing Space Shuttle system and will resolve anomalies such as: liquid carry-over, inaccurate void volume measurements, and cross contamination in void samples. The crew will perform an evaluation of airflow at the ISS UMS urinal hose interface, a calibration evaluation, and a full user interface evaluation. o The UMS can be used to facilitate non-invasive methods for monitoring crew health, evaluation of countermeasures, and implementation of a variety of biomedical research protocols on future exploration missions.

  3. Urine collection device

    NASA Technical Reports Server (NTRS)

    Michaud, R. B. (Inventor)

    1981-01-01

    A urine collection device for females is described. It is comprised of a collection element defining a urine collection chamber and an inlet opening into the chamber and is adapted to be disposed in surrounding relation to the urethral opening of the user. A drainage conduit is connected to the collection element in communication with the chamber whereby the chamber and conduit together comprise a urine flow pathway for carrying urine generally away from the inlet. A first body of wicking material is mounted adjacent the collection element and extends at least partially into the flow pathway. The device preferably also comprise a vaginal insert element including a seal portion for preventing the entry of urine into the vagina.

  4. [Preliminary proteomics analysis of the total proteins of HL Type cytoplasmic male sterility rice anther].

    PubMed

    Wen, Li; Liu, Gai; Zhang, Zai-Jun; Tao, Jun; Wan, Cui-Xiang; Zhu, Ying-Guo

    2006-03-01

    The proteins of HL type cytoplasmic male sterility rice anther of YTA (CMS) and YTB (maintenance line) were separated by two-dimensional electrophoresis with immobilized ph (3-10 non-linear) gradients as the first dimension and SDS-PAGE as the second. The silver-stained proteins spots were analyzed using Image Master 2D software, there were about 1800 detectable spots on each 2D-gel, and about 85 spots were differential expressed. With direct MALDI-TOF mass spectrometry analysis and protein database searching, 9 protein spots out of 16 were identified. Among those proteins, there were Putative nucleic acid binding protein, glucose-1-phosphate adenylyltransferase (ADP-glucose pyrophosphorylase, AGPase) (EC: 2.7.7.27) large chain, UDP-glucuronic acid decarboxylase, putative calcium-binding protein annexin, putative acetyl-CoA synthetase and putative lipoamide dehydrogenase etc. They were closely associated with metabolism, protein biosynthesis, transcription, signal transduction and so on, all of which are cell activities that are essential to pollen development. Some of the identified proteins, i.e. AGPase, putative lipoamide dehydrogenase and putative acetyl-CoA synthetase were deeply discussed on the relationship to CMS. AGPase catalyzes a very important step in the biosynthesis of alpha 1,4-glucans (glycogen or starch) in bacteria and plants: synthesis of the activated glucosyl donor, ADP-glucose, from glucose-1-phosphate and ATP. The lack of the AGPase in male sterile line might directly result in the reduction of starch, and the synthesis of starch was the most important processes during the development of pollen. In present research, the descent or reduction of putative lipoamide dehydrogenase and putative acetyl-CoA synthetase seemed involved in pollen sterility in rice. The degeneration and formation of various tissues during pollen development may impose high demands for energy and key biosynthetic intermediates. Under such conditions, the TCA cycle needs

  5. Immobilization methods for the rapid total chemical synthesis of proteins on microtiter plates.

    PubMed

    Zitterbart, Robert; Krumrey, Michael; Seitz, Oliver

    2017-07-01

    The chemical synthesis of proteins typically involves the solid-phase peptide synthesis of unprotected peptide fragments that are stitched together in solution by native chemical ligation (NCL). The process is slow, and throughput is limited because of the need for repeated high performance liquid chromatography purification steps after both solid-phase peptide synthesis and NCL. With an aim to provide faster access to functional proteins and to accelerate the functional analysis of synthetic proteins by parallelization, we developed a method for the high performance liquid chromatography-free synthesis of proteins on the surface of microtiter plates. The method relies on solid-phase synthesis of unprotected peptide fragments, immobilization of the C-terminal fragment and on-surface NCL with an unprotected peptide thioester in crude form. Herein, we describe the development of a suitable immobilization chemistry. We compared (i) formation of nickel(II)-oligohistidine complexes, (ii) Cu-based [2 + 3] alkine-azide cycloaddition and (iii) hydrazone ligation. The comparative study identified the hydrazone ligation as most suitable. The sequence of immobilization via hydrazone ligation, on-surface NCL and radical desulfurization furnished the targeted SH3 domains in near quantitative yield. The synthetic proteins were functional as demonstrated by an on-surface fluorescence-based saturation binding analysis. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

  6. Adulteration of urine by "Urine Luck".

    PubMed

    Wu, A H; Bristol, B; Sexton, K; Cassella-McLane, G; Holtman, V; Hill, D W

    1999-07-01

    In vitro adulterants are used to invalidate assays for urine drugs of abuse. The present study examined the effect of pyridinium chlorochromate (PCC) found in the product "Urine Luck". PCC was prepared and added to positive urine controls at concentrations of 0, 10, 50, and 100 g/L. The controls were assayed for methamphetamine, benzoylecgonine (BE), codeine and morphine, tetrahydrocannabinol (THC), and phencyclidine (PCP) with the Emit II (Syva) and Abuscreen Online (Roche) immunoassays, and by gas chromatography/mass spectrometry (GC/MS). Two tests were also developed to detect PCC in urine: a spot test to detect chromate ions using 10 g/L 1,5-diphenylcarbazide as the indicator, and a GC/MS assay for pyridine. We tested 150 samples submitted for routine urinalysis, compliance, and workplace drug testing for PCC, using these assays. Response rates decreased at 100 g/L PCC for all Emit II drug assays and for the Abuscreen morphine and THC assays. In contrast, the Abuscreen amphetamine assay produced apparently higher results, and no effect was seen on the results for BE or PCP. The PCC did not affect the GC/MS recovery of methamphetamine, BE, PCP, or their deuterated internal standards, but decreased GC/MS recovery of the opiates at both intermediate (50 g/L) and high (100 g/L) PCC concentrations and apparent concentrations of THC and THC-d3 at all PCC concentrations. Two of 50 samples submitted for workplace drug testing under chain-of-custody conditions were positive for PCC, whereas none of the remaining 100 specimens submitted for routine urinalysis or compliance drug testing were positive. PCC is an effective adulterant for urine drug testing of THC and opiates. Identification of PCC use can be accomplished with use of a spot test for the oxidant.

  7. Total amino acid stabilization during cell-free protein synthesis reactions.

    PubMed

    Calhoun, Kara A; Swartz, James R

    2006-05-17

    Limitations in amino acid supply have been recognized as a substantial problem in cell-free protein synthesis reactions. Although enzymatic inhibitors and fed-batch techniques have been beneficial, the most robust way to stabilize amino acids is to remove the responsible enzymatic activities by genetically modifying the source strain used for cell extract preparation. Previous work showed this was possible for arginine, serine, and tryptophan, but cysteine degradation remained a major limitation in obtaining high protein synthesis yields. Through radiolabel techniques, we confirmed that cysteine degradation was caused by the activity of glutamate-cysteine ligase (gene gshA) in the cell extract. Next, we created Escherichia coli strain KC6 that combines a gshA deletion with previously described deletions for arginine, serine, and tryptophan stabilization. Strain KC6 grows well, and active cell extract can be produced from it for cell-free protein synthesis reactions. The extract from strain KC6 maintains stable amino acid concentrations of all 20 amino acids in a 3-h batch reaction. Yields for three different proteins improved 75-250% relative to cell-free expression using the control extract.

  8. Development of a strategy for the total chemical synthesis of an allergenic protein: the peach LTP Pru p 3.

    PubMed

    Buhler, Sofie; Akkerdaas, Jaap H; A Pertinhez, Thelma; Van Ree, Ronald; Dossena, Arnaldo; Sforza, Stefano; Tedeschi, Tullia

    2017-04-01

    The possibility to obtain allergenic proteins by means of total chemical synthesis would be a big step forward in the development of cures to food allergy and in the study of the mechanism of allergic reactions, because this would allow to achieve control at the molecular level over the structure of the product and to study its relationship with the allergenic activity in fine details. This is instead not possible by using allergens produced by extraction from natural sources or by recombinant DNA techniques. In this work, we aimed to test for the first time the feasibility of the total chemical synthesis of an allergenic protein. Pru p 3, the most studied member of the family of lipid transfer proteins, relevant plant food pan-allergens, was used as model target. Strategies for the convergent assembly of the target protein, starting from five peptide fragments to be bound by means of either native chemical ligation or peptide hydrazide ligation, followed by desulfurization, to achieve ligations at alanine, were developed and tested. All the reaction conditions were set up and optimized. Two large peptides covering the two halves of the protein sequence were synthesized and structurally characterized by means of circular dichroism, and their immunogenicity was proved by means of immunoblot, using antibodies against Pru p 3, and immunoCAP inhibition tests. Finally, the five peptides were bound together to produce the whole protein stretch. The obtained results demonstrate the feasibility of total chemical synthesis as a new way to obtain pure allergens. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

  9. Bringing the science of proteins into the realm of organic chemistry: total chemical synthesis of SEP (synthetic erythropoiesis protein).

    PubMed

    Kent, Stephen B H

    2013-11-11

    Erythropoietin, commonly known as EPO, is a glycoprotein hormone that stimulates the production of red blood cells. Recombinant EPO has been described as "arguably the most successful drug spawned by the revolution in recombinant DNA technology". Recently, the EPO glycoprotein molecule has re-emerged as a major target of synthetic organic chemistry. In this article I will give an account of an important body of earlier work on the chemical synthesis of a designed EPO analogue that had full biological activity and improved pharmacokinetic properties. The design and synthesis of this "synthetic erythropoiesis protein" was ahead of its time, but has gained new relevance in recent months. Here I will document the story of one of the major accomplishments of synthetic chemistry in a more complete way than is possible in the primary literature, and put the work in its contemporaneous context. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Validation of protein and potassium intakes assessed from 24 h recalls against levels estimated from 24 h urine samples in children and adolescents of Turkish descent living in Germany: results from the EVET! Study.

    PubMed

    Bokhof, Beate; Buyken, Anette E; Doğan, Canan; Karaboğa, Arzu; Kaiser, Josa; Sonntag, Antje; Kroke, Anja

    2012-04-01

    Nutrition-related health problems such as obesity are frequent among children and adolescents of Turkish descent living in Germany, yet data on their dietary habits are scarce. One reason might be the lack of validated assessment tools for this target group. We therefore aimed to validate protein and K intakes from one 24 h recall against levels estimated from one 24 h urine sample in children and adolescents of Turkish descent living in Germany. Cross-sectional analyses comprised estimation of mean differences, Pearson correlation coefficients, cross-classifications and Bland-Altman plots to assess the agreement between the nutritional intake estimated from a single 24 h recall and a single 24 h urine sample collected on the previous day. Dortmund, Germany. Data from forty-three study participants (aged 5-18 years; 26% overweight) with a traditional Turkish background were included. The 24 h recall significantly overestimated mean protein and K intake by 10·7 g/d (95% CI of mean difference: 0·6, 20·7 g/d) and 344 mg/d (95% CI 8, 680 mg/d), respectively. Correlations between intake estimates were r = 0·25 (P = 0·1) and 0·31 (P = 0·05). Both methods classified 70% and 69% of the participants into the same/adjacent quartile of protein and K intake and misclassified 7% and 7%, respectively, into the opposite quartile. Bland-Altman plots indicated a wide scattering of differences in both protein and K intake. Among children and adolescents of traditional Turkish descent living in Germany, one 24 h recall may only be valid for categorizing subjects into high, medium or low consumers.

  11. A Rapid and Reliable Method for Total Protein Extraction from Succulent Plants for Proteomic Analysis.

    PubMed

    Lledías, Fernando; Hernández, Felipe; Rivas, Viridiana; García-Mendoza, Abisaí; Cassab, Gladys I; Nieto-Sotelo, Jorge

    2017-08-01

    Crassulacean acid metabolism plants have some morphological features, such as succulent and reduced leaves, thick cuticles, and sunken stomata that help them prevent excessive water loss and irradiation. As molecular constituents of these morphological adaptations to xeric environments, succulent plants produce a set of specific compounds such as complex polysaccharides, pigments, waxes, and terpenoids, to name a few, in addition to uncharacterized proteases. Since all these compounds interfere with the analysis of proteins by electrophoretic techniques, preparation of high quality samples from these sources represents a real challenge. The absence of adequate protocols for protein extraction has restrained the study of this class of plants at the molecular level. Here, we present a rapid and reliable protocol that could be accomplished in 1 h and applied to a broad range of plants with reproducible results. We were able to obtain well-resolved SDS/PAGE protein patterns in extracts from different members of the subfamilies Agavoideae (Agave, Yucca, Manfreda, and Furcraea), Nolinoideae (Dasylirion and Beucarnea), and the Cactaceae family. This method is based on the differential solubility of contaminants and proteins in the presence of acetone and pH-altered solutions. We speculate about the role of saponins and high molecular weight carbohydrates to produce electrophoretic-compatible samples. A modification of the basic protocol allowed the analysis of samples by bidimensional electrophoresis (2DE) for proteomic analysis. Furostanol glycoside 26-O-β-glucosidase (an enzyme involved in steroid saponin synthesis) was successfully identified by mass spectrometry analysis and de novo sequencing of a 2DE spot from an Agave attenuata sample.

  12. Nonhazardous Urine Pretreatment Method

    NASA Technical Reports Server (NTRS)

    Akse, James R.; Holtsnider, John T.

    2012-01-01

    A method combines solid phase acidification with two non-toxic biocides to prevent ammonia volatilization and microbial proliferation. The safe, non-oxidizing biocide combination consists of a quaternary amine and a food preservative. This combination has exhibited excellent stabilization of both acidified and unacidified urine. During pretreatment tests, composite urine collected from donors was challenged with a microorganism known to proliferate in urine, and then was processed using the nonhazardous urine pre-treatment method. The challenge microorganisms included Escherichia coli, a common gram-negative bacteria; Enterococcus faecalis, a ureolytic gram-positive bacteria; Candida albicans, a yeast commonly found in urine; and Aspergillus niger, a problematic mold that resists urine pre-treatment. Urine processed in this manner remained microbially stable for over 57 days. Such effective urine stabilization was achieved using non-toxic, non-oxidizing biocides at higher pH (3.6 to 5.8) than previous methods in use or projected for use aboard the International Space Station (ISS). ISS urine pretreatment methods employ strong oxidants including ozone and hexavalent chromium (Cr(VI)), a carcinogenic material, under very acidic conditions (pH = 1.8 to 2.4). The method described here offers a much more benign chemical environment than previous pretreatment methods, and will lower equivalent system mass (ESM) by reducing containment volume and mass, system complexity, and crew time needed to handle pre-treatment chemicals. The biocides, being non-oxidizing, minimize the potential for chemical reactions with urine constituents to produce volatile, airborne contaminants such as cyanogen chloride. Additionally, the biocides are active under significantly less acidic conditions than those used in the current system, thereby reducing the degree of required acidification. A simple flow-through solid phase acidification (SPA) bed is employed to overcome the natural buffering

  13. Protein-surfactant interactions at hydrophobic interfaces studied with total internal reflection fluorescence correlation spectroscopy (TIR-FCS).

    PubMed

    Sonesson, Andreas W; Blom, Hans; Hassler, Kai; Elofsson, Ulla M; Callisen, Thomas H; Widengren, Jerker; Brismar, Hjalmar

    2008-01-15

    The aim of this work was to study the dynamics of proteins near solid surfaces in the presence or absence of competing surfactants by means of total internal reflection fluorescence correlation spectroscopy (TIR-FCS). Two different proteins were studied, bovine serum albumin (BSA) and Thermomyces lanuginosus lipase (TLL). A nonionic/anionic (C12E6/LAS) surfactant composition was used to mimic a detergent formulation and the surfaces used were C18 terminated glass. It was found that with increasing surfactant concentrations the term in the autocorrelation function (ACF) representing surface binding decreased. This suggested that the proteins were competed off the hydrophobic surface by the surfactant. When fitting the measured ACF to a model for surface kinetics, it was seen that with raised C12E6/LAS concentration, the surface interaction rate increased for both proteins. Under these experimental conditions this meant that the time the protein was bound to the surface decreased. At 10 microM C12E6/LAS the surface interaction was not visible for BSA, whereas it was still distinguishable in the ACF for TLL. This indicated that TLL had a higher affinity than BSA for the C18 surface. The study showed that TIR-FCS provides a useful tool to quantify the surfactant effect on proteins adsorption.

  14. Nonenzymatic chemiluminescent detection and quantitation of total protein on Western and slot blots allowing subsequent immunodetection and sequencing.

    PubMed

    Alba, F J; Daban, J R

    1997-10-01

    We have studied the light emission efficiency of proteins labeled with different fluorescent dyes chemically excited by the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H2O2 reaction. Using this peroxyoxalate chemiluminescence system, the best results were obtained with proteins covalently labeled with 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF). Blotted proteins on polyvinylidene difluoride (PVDF) membranes can be labeled rapidly with MDPF. Our results demonstrate that energy from the excited intermediate produced in the TCPO-H2O2 reaction can be efficiently transferred to MDPF-labeled proteins in solution and on PVDF membranes. Although this nonenzymatic chemiluminescent system produces a background emission that reduces the sensitivity, the method developed in this work allows detection of 5 ng of protein in blots after 5 min exposure to X-ray film. Chemiluminescence of MDPF-labeled proteins on Western and slot blots may also be detected and quantified using a charge-coupled device (CCD) camera or a storage phosphor imaging system. This chemiluminescent method allows the staining of the total electrophoretic pattern but does not preclude further N-terminal sequencing and immunodetection of specific bands.

  15. Low molecular weight proteins in urines from healthy subjects as well as diabetic, nephropathic and diabetic-nephropathic patients: a MALDI study.

    PubMed

    Lapolla, Annunziata; Seraglia, Roberta; Molin, Laura; Williams, Katherine; Cosma, Chiara; Reitano, Rachele; Sechi, Annalisa; Ragazzi, Eugenio; Traldi, Pietro

    2009-03-01

    Urine samples from healthy subjects as well as diabetic, nephropathic and diabetic-nephropathic patients were analyzed by matrix assisted laser desorption/ionization (MALDI) mass spectrometry in order to establish evidence of some possible differences in the peptide profile related to the pathological states. Multivariate analysis suggested the possibility of a distinction among the considered groups of patients. Some differences have been found, in particular, in the relative abundances of three ions at m/z 1912, 1219 and 2049. For these reasons, further investigation was carried out by MALDI/TOF/TOF to determine the sequence of these peptides and, consequently, to individuate their possible origin. By this approach, the peptide at m/z 1912 was found to originate from uromodulin, and its lower expression in the case of nephropathy can be well related to the pathological condition. Ions at m/z 2049 and 1219 originate from the collagen alpha-1(I) chain precursor and from the collagen alpha-5 (IV) chain precursor, respectively, and, also in this case, their different expressions can be related to the pathologies under investigation. The obtained data seem to indicate that urine is an interesting biological fluid to investigate on the peptide profile and to obtain, consequently, information on the dismetabolism activated by specific pathologies. Copyright (c) 2009 John Wiley & Sons, Ltd.

  16. Turbid white urine

    PubMed Central

    Vera, Manel; Molano, Alejandra; Rodríguez, Patricia

    2010-01-01

    Turbid white urine ‘albinuria’ is defined as a urine discoloration described as milky or cloudy. One of the most frequent causes of turbid white urine is chyluria complicating filariasis (Table 1). The extant causes of albinuria are non parasitic and rare. Amongst their aetiologies stand excessive mineral sediment excretion such as calciuria and phosphaturia, massive pyuria and fungal infections, and rarely congenital malformations of the lymphatic vessels. Malingering is also possible, in patients adding milk to their urine. We observed a case of albinuria in which the diagnostic work up led to diagnosing an exceptional cause of chyluria in a patient living in a region of Colombia where filariasis is not endemic. PMID:25949403

  17. Urinating more at night

    MedlinePlus

    ... blood calcium level Certain medicines, including water pills (diuretics) Diabetes insipidus Waking often during the night to ... cause. If excessive nighttime urination is due to diuretic medicines, you may be told to take your ...

  18. Potassium urine test

    MedlinePlus

    ... in the clinical evaluation of electrolyte, water, and acid-base disorders. In: Alpern RJ, Orson WM, Caplan M, ... Lin S-H, Halperin ML. Interpretation of electrolyte and acid-base parameters in blood and urine. In: Skorecki K, ...

  19. Urination - difficulty with flow

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003143.htm Urination - difficulty with flow To use the sharing features on ... Duplication for commercial use must be authorized in writing by ADAM Health Solutions. About MedlinePlus Site Map ...

  20. Frequent or urgent urination

    MedlinePlus

    Urgent urination; Urinary frequency or urgency; Urgency-frequency syndrome; Overactive bladder (OAB) syndrome; Urge syndrome ... Medicines such as water pills (diuretics) Overactive bladder syndrome Radiation therapy to the pelvis, which is used ...

  1. Urine Tests (For Parents)

    MedlinePlus

    ... a doctor suspects that a child has a urinary tract infection (UTI) or a health problem that can cause an ... to-Creatinine Ratio Kidney Diseases in Childhood Recurrent Urinary Tract Infections and Related Conditions Urinary Tract Infections Urine Test: ...

  2. Tracer techniques for urine volume determination and urine collection and sampling back-up system

    NASA Technical Reports Server (NTRS)

    Ramirez, R. V.

    1971-01-01

    The feasibility, functionality, and overall accuracy of the use of lithium were investigated as a chemical tracer in urine for providing a means of indirect determination of total urine volume by the atomic absorption spectrophotometry method. Experiments were conducted to investigate the parameters of instrumentation, tracer concentration, mixing times, and methods for incorporating the tracer material in the urine collection bag, and to refine and optimize the urine tracer technique to comply with the Skylab scheme and operational parameters of + or - 2% of volume error and + or - 1% accuracy of amount of tracer added to each container. In addition, a back-up method for urine collection and sampling system was developed and evaluated. This back-up method incorporates the tracer technique for volume determination in event of failure of the primary urine collection and preservation system. One chemical preservative was selected and evaluated as a contingency chemical preservative for the storage of urine in event of failure of the urine cooling system.

  3. Aligator: A computational tool for optimizing total chemical synthesis of large proteins.

    PubMed

    Jacobsen, Michael T; Erickson, Patrick W; Kay, Michael S

    2017-09-15

    The scope of chemical protein synthesis (CPS) continues to expand, driven primarily by advances in chemical ligation tools (e.g., reversible solubilizing groups and novel ligation chemistries). However, the design of an optimal synthesis route can be an arduous and fickle task due to the large number of theoretically possible, and in many cases problematic, synthetic strategies. In this perspective, we highlight recent CPS tool advances and then introduce a new and easy-to-use program, Aligator (Automated Ligator), for analyzing and designing the most efficient strategies for constructing large targets using CPS. As a model set, we selected the E. coli ribosomal proteins and associated factors for computational analysis. Aligator systematically scores and ranks all feasible synthetic strategies for a particular CPS target. The Aligator script methodically evaluates potential peptide segments for a target using a scoring function that includes solubility, ligation site quality, segment lengths, and number of ligations to provide a ranked list of potential synthetic strategies. We demonstrate the utility of Aligator by analyzing three recent CPS projects from our lab: TNFα (157 aa), GroES (97 aa), and DapA (312 aa). As the limits of CPS are extended, we expect that computational tools will play an increasingly important role in the efficient execution of ambitious CPS projects such as production of a mirror-image ribosome. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Characterization of ultrafiltration of undiluted and diluted stored urine.

    PubMed

    Ouma, J; Septien, S; Velkushanova, K; Pocock, J; Buckley, C

    2016-11-01

    Urine ultrafiltration (UF) was studied in terms of flux, permeability, resistance and fouling. Two types of samples were used: stored urine representing the feedstock obtained from urine diversion dry toilets; and diluted stored urine representing the feedstock obtained from urinals. Three different filtration experiment sets were adopted in this study. For the first case, pressure was set in an ascending order, i.e. from 10 to 60 kPa during filtration of stored urine. For the second case, pressure was set in a descending order, i.e. from 60 to 10 kPa for the same feed stream. The third case involved filtration of diluted urine with pressure in ascending order, i.e. from 10 to 60 kPa. The results indicated that diluted urine had higher flux than undiluted urine with maximum values of 43 and 26 L·m -2 ·h -1 respectively. Cake formation was the dominating fouling mechanism during urine filtration with a contribution of about 90% to the total hydraulic resistance. The contribution of chemically irreversible fouling was low (-2%), unless operating from high to low pressures. Indeed, irreversible fouling appeared to be greater during the experiments starting at higher pressure. Although undiluted urine had a higher fouling potential compared to diluted urine, the specific cake resistance was higher for diluted urine, probably due to a denser cake caused by lower particle sizes in that sample. The permeate obtained after urine filtration had much lower suspended solids content compared to the feedstock, with rejections up to 99%. The concentration of the ionic species remained unchanged, and 75% of the organic compounds and dissolved solids remained in the permeate. Urine UF could then be used as pre-treatment to remove suspended solids.

  5. BAG3 regulates total MAP1LC3B protein levels through a translational but not transcriptional mechanism.

    PubMed

    Rodríguez, Andrea E; López-Crisosto, Camila; Peña-Oyarzún, Daniel; Salas, Daniela; Parra, Valentina; Quiroga, Clara; Morawe, Tobias; Chiong, Mario; Behl, Christian; Lavandero, Sergio

    2016-01-01

    Autophagy is mainly regulated by post-translational and lipid modifications of ATG proteins. In some scenarios, the induction of autophagy is accompanied by increased levels of certain ATG mRNAs such as MAP1LC3B/LC3B, ATG5 or ATG12. However, little is known about the regulation of ATG protein synthesis at the translational level. The cochaperone of the HSP70 system BAG3 (BCL2-associated athanogene 3) has been associated to LC3B lipidation through an unknown mechanism. In the present work, we studied how BAG3 controls autophagy in HeLa and HEK293 cells. Our results showed that BAG3 regulates the basal amount of total cellular LC3B protein by controlling its mRNA translation. This effect was apparently specific to LC3B because other ATG protein levels were not affected. BAG3 knockdown did not affect LC3B lipidation induced by nutrient deprivation or proteasome inhibition. We concluded that BAG3 maintains the basal amount of LC3B protein by controlling the translation of its mRNA in HeLa and HEK293 cells.

  6. BAG3 regulates total MAP1LC3B protein levels through a translational but not transcriptional mechanism

    PubMed Central

    Rodríguez, Andrea E.; López-Crisosto, Camila; Peña-Oyarzún, Daniel; Salas, Daniela; Parra, Valentina; Quiroga, Clara; Morawe, Tobias; Chiong, Mario; Behl, Christian; Lavandero, Sergio

    2016-01-01

    ABSTRACT Autophagy is mainly regulated by post-translational and lipid modifications of ATG proteins. In some scenarios, the induction of autophagy is accompanied by increased levels of certain ATG mRNAs such as MAP1LC3B/LC3B, ATG5 or ATG12. However, little is known about the regulation of ATG protein synthesis at the translational level. The cochaperone of the HSP70 system BAG3 (BCL2-associated athanogene 3) has been associated to LC3B lipidation through an unknown mechanism. In the present work, we studied how BAG3 controls autophagy in HeLa and HEK293 cells. Our results showed that BAG3 regulates the basal amount of total cellular LC3B protein by controlling its mRNA translation. This effect was apparently specific to LC3B because other ATG protein levels were not affected. BAG3 knockdown did not affect LC3B lipidation induced by nutrient deprivation or proteasome inhibition. We concluded that BAG3 maintains the basal amount of LC3B protein by controlling the translation of its mRNA in HeLa and HEK293 cells. PMID:26654586

  7. LC-MS display of the total modified amino acids in cataract lens proteins and in lens proteins glycated by ascorbic acid in vitro.

    PubMed

    Cheng, Rongzhu; Feng, Qi; Ortwerth, Beryl J

    2006-05-01

    We previously reported chromatographic evidence supporting the similarity of yellow chromophores isolated from aged human lens proteins, early brunescent cataract lens proteins and calf lens proteins ascorbylated in vitro [Cheng, R. et al. Biochimica et Biophysica Acta 1537, 14-26, 2001]. In this paper, new evidence supporting the chemical identity of the modified amino acids in these protein populations were collected by using a newly developed two-dimensional LC-MS mapping technique supported by tandem mass analysis of the major species. The pooled water-insoluble proteins from aged normal human lenses, early stage brunescent cataract lenses and calf lens proteins reacted with or without 20 mM ascorbic acid in air for 4 weeks were digested with a battery of proteolytic enzymes under argon to release the modified amino acids. Aliquots equivalent to 2.0 g of digested protein were subjected to size-exclusion chromatography on a Bio-Gel P-2 column and four major A330nm-absorbing peaks were collected. Peaks 1, 2 and 3, which contained most of the modified amino acids were concentrated and subjected to RP-HPLC/ESI-MS, and the mass elution maps were determined. The samples were again analyzed and those peaks with a 10(4) - 10(6) response factor were subjected to MS/MS analysis to identify the daughter ions of each modification. Mass spectrometric maps of peaks 1, 2 and 3 from cataract lenses showed 58, 40 and 55 mass values, respectively, ranging from 150 to 600 Da. Similar analyses of the peaks from digests of the ascorbylated calf lens proteins gave 81, 70 and 67 mass values, respectively, of which 100 were identical to the peaks in the cataract lens proteins. A total of 40 of the major species from each digest were analyzed by LC-MS/MS and 36 were shown to be identical. Calf lens proteins incubated without ascorbic acid showed several similar mass values, but the response factors were 100 to 1000-fold less for every modification. Based upon these data, we conclude

  8. Determination of total antioxidant capacity of milk by CUPRAC and ABTS methods with separate characterisation of milk protein fractions.

    PubMed

    Çekiç, Sema Demirci; Demir, Aslı; Başkan, Kevser Sözgen; Tütem, Esma; Apak, Reşat

    2015-05-01

    Most milk-applied antioxidant assays in literature are based on the isolation and quantification of individual antioxidative compounds, whereas total antioxidant capacity (TAC) gives a more holistic picture due to cooperative action of antioxidants. Recently, the cupric reducing antioxidant capacity (CUPRAC) method has been modified to measure the antioxidant capacities of thiol-containing proteins, where the classical ammonium acetate buffer - that may otherwise precipitate proteins- was replaced with concentrated urea buffer (able to expose embedded thiol groups of proteins to oxidative attack) adjusted to pH 7.0. Thus, antioxidant capacity of milk was investigated with two competing TAC assays, namely CUPRAC and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid))/persulphate, because only these assays were capable of evaluating protein contribution to the observed TAC value. As milk fat caused turbidity, experiments were carried out with skim milk or defatted milk samples. To determine TAC, modified CUPRAC method was applied to whole milk, separated and redissolved protein fractions, and the remaining liquid phase after necessary operations. Both TAC methods were investigated for their dilution sensitivity and antioxidant power assessment of separate milk fractions such as casein and whey. Proteins like β-lactoglobulin and casein (but not simple thiols) exhibited enhanced CUPRAC reactivity with surfactant (SDS) addition. Addition of milk protein fractions to whole skim milk produced significant 'negative-biased' deviations (up to -26% relative standard error) from TAC absorbance additivity in the application of the ABTS method, as opposed to that of the CUPRAC method less affected by chemical deviations from Beer's law thereby producing much smaller deviations from additivity (i.e. the property of additivity is valid when the measured TAC of a mixture is equal to the sum of individual antioxidant capacities of its constituents).

  9. Immunological changes following protein losing enteropathy after surgery total cavopulmonary connection (TCPC) by cytomics

    NASA Astrophysics Data System (ADS)

    Bocsi, József; Lenz, Dominik; Mittag, Anja; Sauer, Ursula; Wild, Lena; Hess, John; Schranz, Dietmar; Hambsch, Jörg; Schneider, Peter; Tárnok, Attila

    2008-02-01

    Complex immunophenotyping single-cell analysis are essential for systems biology and cytomics. The application of cytomics in immunology and cardiac research and diagnostics is very broad, ranging from the better understanding of the cardiovascular cell biology to the identification of heart function and immune consequences after surgery. TCPC or Fontan-type circulation is an accepted palliative surgery for patients with a functionally univentricular heart. Protein-losing enteropathy (PLE), the enteric loss of proteins, is a potential late complication after TCPC surgery. PLE etiology is poorly understood, but immunological factors seem to play a role. This study was aimed to gain insight into immune phenotype alterations following post-TCPC PLE. Patients were studied during routine follow-up up to 5yrs after surgery, blood samples of TCPC patients without (n=21, age 6.8+/-2.6 years at surgery; mean+/-SD) and with manifest PLE (n=12, age 12.8+/- 4.5 years at sampling) and age matched healthy children (control, n=22, age 8.6+/-2.5 years) were collected. Routine laboratory, immune phenotype and serological parameters were determined. Following PLE the immune phenotype dramatically changed with signs of acute inflammation (increased neutrophil and monocyte count, CRP, IL-8). In contrast, lymphocyte count (NK-cells, αβTCR +CD4 +, αβTCR +CD8 + cells) decreased (p<0.001). The residual T-cells had elevated CD25 and CD69 expression. In PLE-patients unique cell populations with CD3 +αβ/γδTCR - and αβTCR +CD4 -8 - phenotype were present in increased frequencies. Our studies show dramatically altered leukocyte phenotype after PLE in TCPC patients. These alterations resemble to changes in autoimmune diseases. We conclude that autoimmune processes may play a role in etiology and pathophysiology of PLE.

  10. A Whole-Cell Surface Plasmon Resonance Sensor Based on a Leucine Auxotroph of Escherichia coli Displaying a Gold-Binding Protein: Usefulness for Diagnosis of Maple Syrup Urine Disease.

    PubMed

    Woo, Min-Ah; Park, Jung Hun; Cho, Daeyeon; Sim, Sang Jun; Kim, Moon Il; Park, Hyun Gyu

    2016-03-01

    We developed a whole-cell surface plasmon resonance (SPR) sensor based on a leucine auxotroph of Escherichia coli displaying a gold-binding protein (GBP) in response to cell growth and applied this sensor to the diagnosis of maple syrup urine disease, which is represented by the elevated leucine level in blood. The leucine auxotroph was genetically engineered to grow displaying GBP in a proportion to the concentration of target amino acid leucine. The GBP expressed on the surface of the auxotrophs directly bound to the golden surface of an SPR chip without the need for any additional treatment or reagents, which consequently produced SPR signals used to determine leucine levels in a test sample. Gold nanoparticles (GNPs) were further applied to the SPR system, which significantly enhanced the signal intensity up to 10-fold by specifically binding to GBP expressed on the cell surface. Finally, the diagnostic utility of our system was demonstrated by its employment in reliably determining different statuses of maple syrup urine disease based on a known cutoff level of leucine. This new approach based on an amino acid-auxotrophic E. coli strain expressing a GBP that binds to an SPR sensor holds great promise for detection of other metabolic diseases of newborn babies including homocystinuria and phenylketonuria, which are also associated with abnormal levels of amino acids.

  11. Combined nitrogen limitation and cadmium stress stimulate total carbohydrates, lipids, protein and amino acid accumulation in Chlorella vulgaris (Trebouxiophyceae).

    PubMed

    Chia, Mathias Ahii; Lombardi, Ana Teresa; da Graça Gama Melão, Maria; Parrish, Christopher C

    2015-03-01

    Metals have interactive effects on the uptake and metabolism of nutrients in microalgae. However, the effect of trace metal toxicity on amino acid composition of Chlorella vulgaris as a function of varying nitrogen concentrations is not known. In this research, C. vulgaris was used to investigate the influence of cadmium (10(-7) and 2.0×10(-8)molL(-1) Cd) under varying nitrogen (2.9×10(-6), 1.1×10(-5) and 1.1×10(-3)molL(-1)N) concentrations on its growth rate, biomass and biochemical composition. Total carbohydrates, total proteins, total lipids, as well as individual amino acid proportions were determined. The combination of Cd stress and N limitation significantly inhibited growth rate and cell density of C. vulgaris. However, increasing N limitation and Cd stress stimulated higher dry weight and chlorophyll a production per cell. Furthermore, biomolecules like total proteins, carbohydrates and lipids increased with increasing N limitation and Cd stress. Ketogenic and glucogenic amino acids were accumulated under the stress conditions investigated in the present study. Amino acids involved in metal chelation like proline, histidine and glutamine were significantly increased after exposure to combined Cd stress and N limitation. We conclude that N limitation and Cd stress affects the physiology of C. vulgaris by not only decreasing its growth but also stimulating biomolecule production. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Inhibition of glycogen synthase kinase-3beta downregulates total tau proteins in cultured neurons and its reversal by the blockade of protein phosphatase-2A.

    PubMed

    Martin, Ludovic; Magnaudeix, Amandine; Esclaire, Françoise; Yardin, Catherine; Terro, Faraj

    2009-02-03

    In tauopathies such as Alzheimer's disease (AD), the molecular mechanisms of tau protein aggregation into neurofibrillary tangles (NFTs) and their contribution to neurodegeneration remain not understood. It was recently demonstrated that tau, regardless of its aggregation, might represent a key mediator of neurodegeneration. Therefore, reduction of tau levels might represent a mechanism of neuroprotection. Glycogen synthase kinase-3beta (GSK3beta) and protein phosphatase-2A (PP2A) are key enzymes involved in the regulation of tau phosphorylation, and have been suggested to be involved in the abnormal tau phosphorylation and aggregation in AD. Connections between PP2A and GSK3beta signaling have been reported. We have previously demonstrated that exposure of cultured cortical neurons to lithium decreased tau protein expression and provided neuroprotection against Abeta. Since lithium is not a specific inhibitor of GSK3beta (ID50=2.0 mM), whether or not the lithium-induced tau decrease involves GSK3beta remained to be determined. For that purpose, cultured cortical neurons were exposed to 6-bromo-indirubin-3'-oxime (6-BIO), a more selective and potent GSK3beta inhibitor (ID50=1.5 microM) or to lithium. Analysis of tau levels and phosphorylation by western-blot assays showed that lithium and 6-BIO dose-dependently decreased both tau protein levels and tau phosphorylation. Conversely, inhibition of cyclin-dependent kinase-5 (CDK5) by roscovitine decreased phosphorylated tau but failed to alter tau protein levels. These data indicate that GSK3beta might be selectively involved in the regulation of tau protein levels. Moreover, inhibition of PP2A by okadaic acid, but not that of PP2B (protein phosphatase-2B)/calcineurin by FK506, dose-dependently reversed lithium-induced tau decrease. These data indicate that GSK3beta regulates both tau phosphorylation and total tau levels through PP2A.

  13. Exposure to Total and Protein-Unbound Rifampin Is Not Affected by Malnutrition in Indonesian Tuberculosis Patients

    PubMed Central

    Ruslami, R.; Later-Nijland, H.; Mooren, F.; Teulen, M.; Apriani, L.; Koenderink, J. B.; Russel, F. G.; Burger, D. M.; Alisjahbana, B.; Wieringa, F.; van Crevel, R.; Aarnoutse, R. E.

    2015-01-01

    Nutritional status may have a profound impact on the pharmacokinetics of drugs, yet only few data are available for tuberculosis (TB) drugs. As malnutrition occurs frequently among TB patients, we assessed the effect of malnutrition on the steady-state pharmacokinetics of total and protein-unbound rifampin during the intensive phase of TB treatment. In a descriptive pharmacokinetic study in Bandung, Indonesia, patients received a fixed standard rifampin dose of 450 mg once daily during the intensive phase of TB treatment. A full pharmacokinetic curve for rifampin was recorded, and total and unbound concentrations of rifampin were analyzed in all samples. Rifampin pharmacokinetic parameters were compared between severely malnourished (BMI of <16.0 kg/m2), malnourished (BMI of <18.5 kg/m2), and well-nourished (BMI of ≥18.5 kg/m2) individuals. No difference in total and protein-unbound pharmacokinetic parameters between severely malnourished (n = 7), malnourished (n = 11), and well-nourished (n = 25) patients could be demonstrated. In addition, no significant correlation between BMI and exposure (area under the concentration-time curve from 0 to 24 h [AUC0–24] and maximum concentration of drug in serum [Cmax]) was found. Females had significantly higher total AUC0–24 (geometric mean, 59.2 versus 48.2 h · mg/liter; P = 0.02) and higher unbound AUC0–24 (geometric mean, 6.2 versus 4.8 h · mg/liter; P = 0.02) than males. Overall, a marked 2-fold interindividual variation in the free fraction was observed (7.6 to 15.0%; n = 36). Nutritional status and BMI do not appear to have a major effect on total and protein-unbound pharmacokinetic parameters of rifampin in Indonesian subjects. The large interindividual variability in the free fraction of rifampin suggests that protein-unbound rather than total rifampin concentrations should preferably be used to study exposure-response relationships. PMID:25801554

  14. Influence of condensing equipment and temperature on exhaled breath condensate pH, total protein and leukotriene concentrations.

    PubMed

    Czebe, Krisztina; Barta, Imre; Antus, Balázs; Valyon, Márta; Horváth, Ildikó; Kullmann, Tamás

    2008-05-01

    Exhaled breath condensate analysis is an attractive but still not fully standardised method for investigating airway pathology. Adherence of biomarkers to various condensing surfaces and changes in condensing temperature has been considered to be responsible for the variability of the results. Our aims were to compare the efficacy of different types of condensers and to test the influence of condensing temperature on condensate composition. Breath condensates from 12 healthy persons were collected in two settings: (1) by using three condensers of different type (EcoScreen, R-Tube, Anacon) and (2) by using R-Tube condenser either cooled to -20 or -70 degrees C. Condensate pH at standardised CO(2) level was determined; protein content was measured by the Bradford method and leukotrienes by EIA. Breath condensates collected using EcoScreen were more alkaline (6.45+/-0.20 vs. 6.19+/-0.23, p<0.05 and 6.10+/-0.26, p<0.001) and contained more protein (3.89+/-2.03 vs. 2.65+/-1.98, n.s. and 1.88+/-1.99 microg/ml, p<0.004) as compared to the other devices. Only parameters obtained with R-Tube and Anacon correlated. Condensing temperature affected condensate pH (5.99+/-0.20 at -20 degrees C and 5.82+/-0.07 at -70 degrees C, p<0.05) but not protein content. Leukotriene B(4) was not found in any sample and cysteinyl-leukotriene was not found in condensates collected with R-Tube or Anacon. Condenser type influences sample pH, total protein content and cysteinyl-leukotriene concentration. Condensing temperature influences condensate pH but not total protein content. These results suggest that adherence of the biomarkers to condenser surface and condensing temperature may play a role but does not fully explain the variability of EBC biomarker levels.

  15. The Urine Proteome as a Biomarker of Radiation Injury

    PubMed Central

    Sharma, Mukut; Halligan, Brian D.; Wakim, Bassam T.; Savin, Virginia J.; Cohen, Eric P.; Moulder, John E.

    2009-01-01

    Terrorist attacks or nuclear accidents could expose large numbers of people to ionizing radiation, and early biomarkers of radiation injury would be critical for triage, treatment and follow-up of such individuals. However, no such biomarkers have yet been proven to exist. We tested the potential of high throughput proteomics to identify protein biomarkers of radiation injury after total body X-ray irradiation in a rat model. Subtle functional changes in the kidney are suggested by an increased glomerular permeability for macromolecules measured within 24 hours after TBI. Ultrastructural changes in glomerular podocytes include partial loss of the interdigitating organization of foot processes. Analysis of urine by LC-MS/MS and 2D-GE showed significant changes in the urine proteome within 24 hours after TBI. Tissue kallikrein 1-related peptidase, cysteine proteinase inhibitor cystatin C and oxidized histidine were found to be increased while a number of proteinase inhibitors including kallikrein-binding protein and albumin were found to be decreased post-irradiation. Thus, TBI causes immediately detectable changes in renal structure and function and in the urinary protein profile. This suggests that both systemic and renal changes are induced by radiation and it may be possible to identify a set of biomarkers unique to radiation injury. PMID:19746194

  16. FAMBE-pH: A Fast and Accurate Method to Compute the Total Solvation Free Energies of Proteins

    PubMed Central

    Vorobjev, Yury N.; Vila, Jorge A.

    2009-01-01

    A fast and accurate method to compute the total solvation free energies of proteins as a function of pH is presented. The method makes use of a combination of approaches, some of which have already appeared in the literature; (i) the Poisson equation is solved with an optimized fast adaptive multigrid boundary element (FAMBE) method; (ii) the electrostatic free energies of the ionizable sites are calculated for their neutral and charged states by using a detailed model of atomic charges; (iii) a set of optimal atomic radii is used to define a precise dielectric surface interface; (iv) a multilevel adaptive tessellation of this dielectric surface interface is achieved by using multisized boundary elements; and (v) 1:1 salt effects are included. The equilibrium proton binding/release is calculated with the Tanford–Schellman integral if the proteins contain more than ∼20–25 ionizable groups; for a smaller number of ionizable groups, the ionization partition function is calculated directly. The FAMBE method is tested as a function of pH (FAMBE-pH) with three proteins, namely, bovine pancreatic trypsin inhibitor (BPTI), hen egg white lysozyme (HEWL), and bovine pancreatic ribonuclease A (RNaseA). The results are (a) the FAMBE-pH method reproduces the observed pKa's of the ionizable groups of these proteins within an average absolute value of 0.4 pK units and a maximum error of 1.2 pK units and (b) comparison of the calculated total pH-dependent solvation free energy for BPTI, between the exact calculation of the ionization partition function and the Tanford–Schellman integral method, shows agreement within 1.2 kcal/mol. These results indicate that calculation of total solvation free energies with the FAMBE-pH method can provide an accurate prediction of protein conformational stability at a given fixed pH and, if coupled with molecular mechanics or molecular dynamics methods, can also be used for more realistic studies of protein folding, unfolding, and dynamics

  17. Chemical Method of Urine Volume Measurement

    NASA Technical Reports Server (NTRS)

    Petrack, P.

    1967-01-01

    A system has been developed and qualified as flight hardware for the measurement of micturition volumes voided by crewmen during Gemini missions. This Chemical Urine Volume Measurement System (CUVMS) is used for obtaining samples of each micturition for post-flight volume determination and laboratory analysis for chemical constituents of physiological interest. The system is versatile with respect to volumes measured, with a capacity beyond the largest micturition expected to be encountered, and with respect to mission duration of inherently indefinite length. The urine sample is used for the measurement of total micturition volume by a tracer dilution technique, in which a fixed, predetermined amount of tritiated water is introduced and mixed into the voided urine, and the resulting concentration of the tracer in the sample is determined with a liquid scintillation spectrometer. The tracer employed does not interfere with the analysis for the chemical constituents of the urine. The CUVMS hardware consists of a four-way selector valve in which an automatically operated tracer metering pump is incorporated, a collection/mixing bag, and tracer storage accumulators. The assembled system interfaces with a urine receiver at the selector valve inlet, sample bags which connect to the side of the selector valve, and a flexible hose which carries the excess urine to the overboard drain connection. Results of testing have demonstrated system volume measurement accuracy within the specification limits of +/-5%, and operating reliability suitable for system use aboard the GT-7 mission, in which it was first used.

  18. Effect of Cell Phone Use on Salivary Total Protein, Enzymes and Oxidative Stress Markers in Young Adults: A Pilot Study

    PubMed Central

    Joy, Jasmi; Sunitha, Venkatesh; Rai, Manoj P.; Rao, Suresh; Nambranathayil, Shafeeque; Baliga, Manjeshwar Shrinath

    2015-01-01

    Introduction: The present study aimed to assess the levels of salivary enzymes, protein and oxidant-antioxidant system in young college-going cell phone users. Materials and Methods: The cell users (students) were categorized in to two groups – less mobile users and high mobile users, based on the duration and frequency of cell use. Unstimulated whole saliva samples of the volunteers were analysed for amylase, lactate dehydrogenase (LDH), malondialdehdye (MDA) and glutathione (GSH). Results: High mobile users had significantly higher levels of amylase (p = 0.001), LDH (p = 0.002) and MDA (p = 0.002) in saliva, when compared to less mobile users. The marginal decrease in salivary total proteins, GSH and flow rate were statistically not significant (p >0.05). Conclusion: Significant changes in salivary enzymes and MDA suggest adverse effect of high use of cell phones on cell health. PMID:25859446

  19. In-vivo assessment of total body protein in rats by prompt-γ neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Stamatelatos, Ion E.; Boozer, Carol N.; Ma, Ruimei; Yasumura, Seiichi

    1997-02-01

    A prompt-(gamma) neutron activation analysis facility for in vivo determination of total body protein (TBP) in rats has been designed. TBP is determined in vivo by assessment of total body nitrogen. The facility is based on a 252Cf radionuclide neutron source within a heavy water moderator assembly and two NaI(Tl) scintillation detectors. The in vivo precision of the technique, as estimated by three repeated measurements of 15 rats is 6 percent, for a radiation dose equivalent of 60 mSv. The radiation dose per measurement is sufficiently low to enable serial measurements on the same animal. MCNP-4A Monte Carlo transport code was utilized to calculate thermal neutron flux correction factors to account for differences in size and shape of the rats and calibration phantoms. Good agrement was observed in comparing body nitrogen assessment by prompt-(gamma) neutron activation and chemical carcass analysis.

  20. A history of urine microscopy.

    PubMed

    Cameron, J Stewart

    2015-11-01

    The naked-eye appearance of the urine must have been studied by shamans and healers since the Stone Age, and an elaborate interpretation of so-called Uroscopy began around 600 AD as a form of divination. A 1000 years later, the first primitive monocular and compound microscopes appeared in the Netherlands, and along with many other objects and liquids, urine was studied from around 1680 onwards as the enlightenment evolved. However, the crude early instruments did not permit fine study because of chromatic and linear/spherical blurring. Only after complex multi-glass lenses which avoided these problems had been made and used in the 1820s in London by Lister, and in Paris by Chevalier and Amici, could urinary microscopy become a practical, clinically useful tool in the 1830s. Clinical urinary microscopy was pioneered by Rayer and his pupils in Paris (especially Vigla), in the late 1830s, and spread to UK and Germany in the 1840s, with detailed descriptions and interpretations of cells and formed elements of the urinary sediment by Nasse, Henle, Robinson and Golding Bird. Classes were held, most notably by Donné in Paris. After another 50 years, optical microscopy had reached its apogee, with magnifications of over 1000 times obtainable free of aberration, using immersion techniques. Atlases of the urinary sediment were published in all major European countries and in the US. Polarised light and phase contrast was used also after 1900 to study urine, and by the early 20th century, photomicroscopy (pioneered by Donné and Daguerre 50 years previously, but then ignored) became usual for teaching and recording. In the 1940s electron microscopy began, followed by detection of specific proteins and cells using immunofluorescent antibodies. All this had been using handheld methodology. Around 1980, machine-assisted observations began, and have dominated progress since.

  1. Rapid determination of total protein and wet gluten in commercial wheat flour using siSVR-NIR.

    PubMed

    Chen, Jia; Zhu, Shipin; Zhao, Guohua

    2017-04-15

    The determination of total protein and wet gluten is of critical importance when screening commercial flour for desired processing suitability. To this end, a near-infrared spectroscopy (NIR) method with support vector regression was developed in the present study. The effects of spectral preprocessing and the synergy interval on model performance were investigated. The results showed that the models from raw spectra were not acceptable, but they were substantially improved by properly applying spectral preprocessing methods. Meanwhile, the synergy interval was validated with a good ability to improve the performance of models based on the whole spectrum. The coefficient of determination (R 2 ), the root mean square error of prediction (RMSEP) and the standard deviation ratio (SDR) of the best models for total protein (wet gluten) were 0.906 (0.850), 0.425 (1.024) and 3.065 (2.482), respectively. These two best models have similar and lower relative errors (approximately 8.8%), which indicates their feasibility. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Leucine, Not Total Protein, Content of a Supplement Is the Primary Determinant of Muscle Protein Anabolic Responses in Healthy Older Women.

    PubMed

    Devries, Michaela C; McGlory, Chris; Bolster, Douglas R; Kamil, Alison; Rahn, Maike; Harkness, Laura; Baker, Steven K; Phillips, Stuart M

    2018-06-13

    Older adults show a blunted muscle protein synthesis (MPS) response to postprandial hyperaminoacidemia relative to younger adults. Evidence suggests that this anabolic resistance can be overcome by consuming greater quantities of leucine. The purpose of this trial was to determine whether the addition of leucine to a smaller dose (10 g) of milk proteins would, when compared with a larger dose (25 g) of whey protein isolate (WPI), result in similar increases in acute (hourly) and integrated (daily) myofibrillar protein synthesis (myoPS). Healthy older (mean ± SD age: 69 ± 1 y) women (n = 11/group) were randomly assigned with the use of a single-blind, parallel-group design to twice-daily consumption of either WPI [25 g WPI (3 g l-leucine)] or leucine (LEU; 10 g milk protein with 3 g total l-leucine) for 6 d. Participants performed unilateral resistance exercise to allow assessment of the impact of the supplement alone and with resistance exercise. We determined acute (13C6-phenylanine) and integrated [using deuterated water (D2O)] rates of myoPS in the fasting (acute), basal (integrated), nonexercised, and exercised states. Acute myoPS increased in both legs in response to LEU (fed: 45%; fed+exercise: 71%; P < 0.001) and WPI (fed: 29%; fed+exercise: 47%; P < 0.001) compared with fasting; the increase was greater with LEU than with WPI in the exercised leg (46%; P = 0.04) but not in the rested leg (P = 0.07). The acute myoPS response was greater in the exercised leg than in the rested leg for both WPI (63%) and LEU (58%) (P < 0.001). Integrated myoPS increased with WPI and LEU in the exercised leg (both 9%; P < 0.001) during supplementation, and with WPI (3%; P = 0.02) but not LEU (2%, P = 0.1) in the rested leg compared with the basal state. A lower-protein (10 compared with 25 g/dose), leucine-matched beverage induced similar increases in acute and integrated myoPS in healthy older women. Lower-protein supplements with added leucine may

  3. Citric acid urine test

    MedlinePlus

    Urine - citric acid test; Renal tubular acidosis - citric acid test; Kidney stones - citric acid test; Urolithiasis - citric acid test ... No special preparation is necessary for this test. But the results ... test is usually done while you are on a normal diet. Ask your ...

  4. Automated color classification of urine dipstick image in urine examination

    NASA Astrophysics Data System (ADS)

    Rahmat, R. F.; Royananda; Muchtar, M. A.; Taqiuddin, R.; Adnan, S.; Anugrahwaty, R.; Budiarto, R.

    2018-03-01

    Urine examination using urine dipstick has long been used to determine the health status of a person. The economical and convenient use of urine dipstick is one of the reasons urine dipstick is still used to check people health status. The real-life implementation of urine dipstick is done manually, in general, that is by comparing it with the reference color visually. This resulted perception differences in the color reading of the examination results. In this research, authors used a scanner to obtain the urine dipstick color image. The use of scanner can be one of the solutions in reading the result of urine dipstick because the light produced is consistent. A method is required to overcome the problems of urine dipstick color matching and the test reference color that have been conducted manually. The method proposed by authors is Euclidean Distance, Otsu along with RGB color feature extraction method to match the colors on the urine dipstick with the standard reference color of urine examination. The result shows that the proposed approach was able to classify the colors on a urine dipstick with an accuracy of 95.45%. The accuracy of color classification on urine dipstick against the standard reference color is influenced by the level of scanner resolution used, the higher the scanner resolution level, the higher the accuracy.

  5. Study on furundu, a traditional Sudanese fermented roselle (Hibiscus sabdariffa L.) seed: effect on in vitro protein digestibility, chemical composition, and functional properties of the total proteins.

    PubMed

    Yagoub, Abu El-Gasim A; Mohamed, Babiker E; Ahmed, Abdel Halim R; El Tinay, Abdullahi H

    2004-10-06

    Furundu, a meat substitute, is traditionally prepared by cooking the karkade (Hibiscus sabdariffa L.) seed and then fermenting it for 9 days. Physicochemical and functional properties of raw and cooked seed and of furundu ferments were analyzed. Furundu preparation resulted in significant changes in karkade seed major nutrients. Total polyphenols and phytic acid were also reduced. The increase in total acidity and fat acidity coupled with a decrease in pH indicates microbial hydrolysis of the major nutrients; proteins, carbohydrates, and fats. In vitro digestibility of the seed proteins reached the maximum value (82.7%) at the sixth day of fermentation, but thereafter it significantly decreased. The effect of furundu preparation on N solubility profiles and functional properties, such as emulsification and foaming properties and other related parameters, is investigated in water and in 1 M NaCl extracts from defatted flour samples. The results show that cooking followed by fermentation affects proteins solubility in water and 1 M NaCl. The foaming capacity (FC) from the flour of raw seed decreased as a result of cooking. Fermentation for 9 days significantly increased the FC of the cooked seed, restoring the inherent value. Foam from fermented samples collapsed more rapidly during a period of 120 min as compared to the foam from raw and cooked karkade seeds; stability in 1 M NaCl was lower as compared to those in water. In water, the emulsion stability (ES) from the fermented samples was significantly higher than that of the raw seed flour. Addition of 1 M NaCl significantly decreased the ES of the fermented samples.

  6. 24-hour urine copper test

    MedlinePlus

    ... medlineplus.gov/ency/article/003604.htm 24-hour urine copper test To use the sharing features on this page, please enable JavaScript. The 24-hour urine copper test measures the amount of copper in ...

  7. Circulating vitamin D binding protein, total, free and bioavailable 25-hydroxyvitamin D and risk of colorectal cancer

    PubMed Central

    Ying, Hou-Qun; Sun, Hui-Ling; He, Bang-Shun; Pan, Yu-Qin; Wang, Feng; Deng, Qi-Wen; Chen, Jie; Liu, Xian; Wang, Shu-Kui

    2015-01-01

    Epidemiological investigation have suggested that there is a significantly inverse association between circulating 25-hydroxyvitamin D (25(OH)D) and the risk for developing colorectal cancer (CRC) in humans. However, little is known about the role of vitamin D binding protein (VDBP) in colorectal carcinogenesis. Blood samples were collected from 212 CRC patients and 212 controls matched with age, gender and blood collection time. We used logistic regression to calculate the odds ratios and 95% confidence intervals for further estimation of the association of the quartiles of VDBP, total, free and bioavailable 25(OH)D with CRC risk. The results revealed that there was no significant association between circulating VDBP concentrations and CRC in the present study, and that a negative association existed between total 25(OH)D and the risk of CRC, which was unchanged after adjustment for VDBP. Higher levels of free and bioavailable 25(OH)D were significantly associated with decreased risk of CRC. After stratifying by VDBP, high levels of total, free and bioavailable 25(OH)D were associated significantly with decreased CRC risk among participants with circulating VDBP below the median. These findings indicate that VDBP is not directly associated with the risk of CRC, but it modulates circulating free and bioavailable 25(OH)D concentration. PMID:25609140

  8. Estimation of Daily Sodium and Potassium Excretion Using Spot Urine and 24-Hour Urine Samples in a Black Population (Benin).

    PubMed

    Mizéhoun-Adissoda, Carmelle; Houehanou, Corine; Chianéa, Thierry; Dalmay, François; Bigot, André; Preux, Pierre-Marie; Bovet, Pascal; Houinato, Dismand; Desport, Jean-Claude

    2016-07-01

    The 24-hour urine collection method is considered the gold standard for the estimation of ingested potassium and sodium. Because of the impracticalities of collecting all urine over a 24-hour period, spot urine is often used for epidemiological investigations. This study aims to assess the agreement between spot urine and 24-hour urine measurements to determine sodium and potassium intake. A total of 402 participants aged 25 to 64 years were randomly selected in South Benin. Spot urine was taken during the second urination of the day. Twenty-four-hour urine was also collected. Samples (2-mL) were taken and then stored at -20°C. The analysis was carried out using potentiometric dosage. The agreement between spot urine and 24-hour urine measurements was established using Bland-Altman plots. A total of 354 results were analyzed. Daily sodium chloride and potassium chloride urinary excretion means were 10.2±4.9 g/24 h and 2.9±1.4 g/24 h, respectively. Estimated daily sodium chloride and potassium chloride means from the spot urine were 10.7±7.0 g/24 h and 3.9±2.1 g/24 h, respectively. Concordance coefficients were 0.61 at d=-0.5 g, (d±2SD=-11 g and 10.1 g) for sodium chloride and 0.61 at d=-1 g, (d±2SD=-3.8 g and 1.8 g) for potassium chloride. Spot urine method is acceptable for estimating 24-hour urinary sodium and potassium excretion to assess sodium and potassium intake in a black population. However, the confidence interval for the mean difference, which is too large, makes the sodium chloride results inadmissible at a clinical level. © 2015 Wiley Periodicals, Inc.

  9. Quantitative analysis of total proteins and carbohydrates in the digestive gland-gonad complex (DGG) and hemolymph of the freshwater prosobranch snail Lanistes carinatus.

    PubMed

    Hassan, Amaal Hassan M; Mahmoud, Salim; El-Hamidy, A

    2010-08-01

    Laboratory investigations were carried out to quantify the amount of total proteins, carbohydrates and reducing sugar in the hemolymph and Digestive Gland Gonad- complex (DGG) of infected and uinfected lanistes arinatus. Snails were naturally infected with two different types of trematode la:val stages (rediae of gymnocepahalus cercaria and sporocyst of xiphidiocercaria), collected from the River Nile at Sohag governorate, Egypt. Analysis was carried out using an extraction of the DGGs tissue with buffer solution, while snails' hemolymph was applied directly. The results revealed that snail infection by rediae of gymnocephalus cercariae led to non significant increasing in both total carbohydrates and protein in hemolymph. However, infection by sporocysts of xiphidiocercariae caused a significant increasing only in hemolymph total protein. On the other hand, the amount of both reducing sugar and total proteins in DGG did not increase significantly whenever the infection caused by both types of trematode larvae. However, total carbohydrates in DGG increased significantly.

  10. Higher Total Protein Intake and Change in Total Protein Intake Affect Body Composition but Not Metabolic Syndrome Indexes in Middle-Aged Overweight and Obese Adults Who Perform Resistance and Aerobic Exercise for 36 Weeks.

    PubMed

    Campbell, Wayne W; Kim, Jung Eun; Amankwaah, Akua F; Gordon, Susannah L; Weinheimer-Haus, Eileen M

    2015-09-01

    Studies assessing the effects of protein supplementation on changes in body composition (BC) and health rarely consider the impact of total protein intake (TPro) or the change in TPro (CTPro) from participants' usual diets. This secondary data analysis assessed the impact of TPro and CTPro on changes in BC and metabolic syndrome (MetS) indexes in overweight and obese middle-aged adults who participated in an exercise training program. Men and women [n = 117; age: 50 ± 0.7 y, body mass index (BMI; in kg/m(2)): 30.1 ± 0.3; means ± SEs] performed resistance exercise 2 d/wk and aerobic exercise 1 d/wk and consumed an unrestricted diet along with 200-kcal supplements (0, 10, 20, or 30 g whey protein) twice daily for 36 wk. Protein intake was assessed via 4-d food records. Multiple linear regression model and stratified analysis were applied for data analyses. Among all subjects, TPro and CTPro were inversely associated (P < 0.05) with changes in body mass, fat mass (FM), and BMI. Changes in BC were different (P < 0.05) among groups that consumed <1.0 (n = 43) vs. ≥1.0 to <1.2 (n = 29) vs. ≥1.2 g · kg(-1) · d(-1) (n = 45). The TPro group with ≥1.0 to <1.2 g ·: kg(-1) ·: d(-1) reduced FM and %FM and increased percentage of LM (%LM) compared with the lowest TPro group, whereas the TPro group with ≥1.2 g ·: kg(-1) ·: d(-1) presented intermediate responses on changes in FM, %FM, and %LM. The gain in LM was not different among groups. In addition, MetS indexes were not influenced by TPro and CTPro. In conjunction with exercise training, higher TPro promoted positive changes in BC but not in MetS indexes in overweight and obese middle-aged adults. Changes in TPro from before to during the intervention also influenced BC responses and should be considered in future research when different TPro is achieved via diet or supplements. This trial was registered at clinicaltrials.gov as NCT00812409. © 2015 American Society for Nutrition.

  11. Urine Galactomannan-to-Creatinine Ratio for Detection of Invasive Aspergillosis in Patients with Hematological Malignancies.

    PubMed

    Reischies, Frederike M J; Raggam, Reinhard B; Prattes, Juergen; Krause, Robert; Eigl, Susanne; List, Agnes; Quehenberger, Franz; Strenger, Volker; Wölfler, Albert; Hoenigl, Martin

    2016-03-01

    Galactomannan (GM) testing of urine specimens may provide important advantages, compared to serum testing, such as easy noninvasive sample collection. We evaluated a total of 632 serial urine samples from 71 patients with underlying hematological malignancies and found that the urine GM/creatinine ratio, i.e., (urine GM level × 100)/urine creatinine level, which takes urine dilution into account, reliably detected invasive aspergillosis and may be a promising diagnostic tool for patients with hematological malignancies. (This study has been registered at ClinicalTrials.gov under registration no. NCT01576653.). Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  12. Use of the Vettest 8008 and refractometry for determination of total protein, albumin, and globulin concentrations in feline effusions.

    PubMed

    Papasouliotis, Kostas; Murphy, Kate; Dodkin, Steve; Torrance, Andy G

    2002-01-01

    Pleural and peritoneal effusion is a common clinical finding in feline practice. Determination of fluid albumin (ALB) and globulin (GLOB) concentrations in addition to total protein (TP) concentration can be helpful in diagnosing or ruling out certain diseases in cats, especially feline infectious peritonitis (FIP). The objective of this study was to compare effusion TP, ALB, and GLOB results obtained by a refractometer and a bench-top dry chemistry analyzer with those results obtained by a reference method. Twenty-six pleural and 14 peritoneal effusion samples were analyzed from 40 cats with various diseases. TP and ALB concentrations were determined by a reference automated wet chemistry analyzer (Kone Specific, Kone Instruments, Espoo, Finland), a bench-top dry chemistry analyzer (Vettest 8008, IDEXX Laboratories Ltd, Chalfont St Peter, UK), and a refractometer (Atago SPR-T2, Atago Co, Tokyo, Japan). GLOB, albumin to globulin (A/G) ratio, and globulins as a percentage of total proteins (GLOB%) were calculated. Results were analyzed by paired t tests, difference plots, and Deming s regression analysis. Correlation coefficients (r) for TP with Vettest versus Kone and refractometer versus Kone methods were.97 and.94, respectively. GLOB and GLOB% values were significantly higher and A/G ratios were significantly lower with Vettest versus Kone methods. Correlation coefficients for ALB, GLOB, GLOB% and A/G ratio with Vettest versus Kone methods were.86,.93,.82, and.73, respectively. Although correlation with other methods was good, the refractometer underestimated TP concentrations in 3 samples. The refractometer is an acceptable method for determination of TP concentration in feline effusions. The Vettest 8008 also is an acceptable method for the determination of TP and ALB concentrations, however, calculated A/G ratios obtained with the Vettest are unacceptable.

  13. [Influence of extremely low frequency magnetic field on total protein and -sh groups concentrations in liver homogenates].

    PubMed

    Ciejka, Elżbieta; Kowalczyk, Agata; Gorąca, Anna

    2014-01-01

    Free radicals are atoms, molecules or their fragments, whose excess leads to the development of oxidative stress, the cause of many neoplastic, neurodegenerative and inflammatory diseases, as well as aging of organisms. Industrial pollution, tobacco smoke, ionizing radiation, ultrasound and magnetic fields are the major exogenous sources of free radicals. The low frequency mag- netic field is commonly applied in physiotherapy. The aim of the present study was to evaluate the effect of extremely low frequency magnetic field (1L.F-MF) on the concentration ofsullhydryl groups (-SH) and proteins in liver tissues of experimental animals de- pending on the time of exposure to the field. Twenty one Sprague-D)awley male rats, aged 3-4 months were randomly divided into 3 experimental groups (each containing 7 animals): controls (group I), the rats exposed to IEI.F-MF of 40 Hz, 7 mT (this kind of the ELF-MF is mostly used in magnetotherapy), 30 min/day for 2 weeks (group II) and the rats exposed to 40 Hz, 7 mT for 60 min/day for 2 weeks (group III). The concentrations of proteins and sulfhydryl groups in the liver tissues were determined after exposure to magnetic fields. Exposure to low magnetic field: 40 Hz, 7 mT for 30 min/day and 60 min/day for 2 weeks caused a significant increase in the concentration of-SH groups and total protein levels in the liver tissues. The study results suggest that exposure to magnetic fields leads to the development of adaptive mechanisms to maintain the balance in the body oxidation-reduction and in the case of the studied parameters does not depend on the time of exposure.

  14. The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane.

    PubMed

    Russo, Alessandra; Bonci, Paola; Bonci, Paolo

    2012-06-01

    The aim of this work is to quantify the total protein and growth factors content in a tissue-suspension obtained from processed human amniotic membrane (hAM). hAM was collected, frozen, freeze dried, powdered and sterilized by γ-irradiation. At each step of the process, samples were characterized for the total protein amounts by a Bradford protein assay and for the growth factor concentrations by ELISA test of the tissue suspensions. Frozen-hAM samples show higher release of total proteins and specific growth factors in the tissue suspension in comparison with freeze-dried hAM. We observed that even if the protein extraction is hindered once the tissue is dried, the powdering process allows a greater release in the tissue suspension of total proteins and growth factors after tissue re-solubilization in comparison with only the freeze-drying process (+91 ± 13% for EGF, +16 ± 4% for HGF, +11 ± 5% for FGF, +16 ± 9% for TGF-β1), and a greater release of EGF (85 ± 10%) in comparison with only the freezing process, because proteins become much readily solubilized in the solution. According with these results, we describe a protocol to obtain a new sterile biological product from hAM tissue, with well-known effects of thermal, mechanical and physical processes on the total protein and grow factors contents.

  15. The effect of balneotherapy on C-reactive protein, serum cholesterol, triglyceride, total antioxidant status and HSP-60 levels.

    PubMed

    Oláh, Mihály; Koncz, Agnes; Fehér, Judit; Kálmánczhey, Judit; Oláh, Csaba; Balogh, Sándor; Nagy, György; Bender, Tamás

    2010-05-01

    An increasing body of evidence substantiating the effectiveness of balneotherapy has accumulated during recent decades. In the present study, 42 ambulatory patients (23 males and 19 females, mean age 59.5 years) with degenerative musculoskeletal disease were randomised into one of two groups-bathing in tap water or in mineral water at the same temperature-and subjected to 30-min balneotherapy sessions on 15 occasions. Study parameters comprised serum levels of sensitised C-reactive protein (CRP), plasma lipids, heat shock protein (HSP-60) and total antioxidant status (TAS). In both groups, CRP levels followed a decreasing tendency, which still persisted 3 months later. At 3 months after balneotherapy, serum cholesterol levels were still decreasing in patients who had used medicinal water, but exhibited a trend towards an increase in the control group. Triglyceride levels followed a decreasing trend in both patient groups. TAS showed a declining tendency in both groups. No changes of HSP-60 levels were observed in either group. Balneotherapy with the thermal water from Hajdúszoboszló spa had a more pronounced physiological effect compared to that seen in the control group treated with tap water in a 3 month period.

  16. Estimation of salivary glucose, salivary amylase, salivary total protein and salivary flow rate in diabetics in India.

    PubMed

    Panchbhai, Arati S; Degwekar, Shirish S; Bhowte, Rahul R

    2010-09-01

    Diabetes is known to influence salivary composition and function, eventually affecting the oral cavity. We thus evaluated saliva samples for levels of glucose, amylase and total protein, and assessed salivary flow rate in diabetics and healthy non-diabetics. We also analyzed these parameters with regard to duration and type of diabetes mellitus and gender, and aimed to assess the interrelationships among the variables included in the study. A total of 120 age- and sex-matched participants were divided into 3 groups of 40 each; the uncontrolled diabetic group, the controlled diabetic group and the healthy non-diabetic group. Salivary investigations were performed using unstimulated whole saliva. Mean salivary glucose levels were found to be significantly elevated in both uncontrolled and controlled diabetics, as compared to healthy non-diabetics. There were significant decreases in mean salivary amylase levels in controlled diabetics when compared to healthy non-diabetics. Other than salivary glucose, no other parameters were found to be markedly affected in diabetes mellitus. Further research is needed to explore the clinical implications of these study results.

  17. Ultrasonic-based membrane aided sample preparation of urine proteomes.

    PubMed

    Jesus, Jemmyson Romário; Santos, Hugo M; López-Fernández, H; Lodeiro, Carlos; Arruda, Marco Aurélio Zezzi; Capelo, J L

    2018-02-01

    A new ultrafast ultrasonic-based method for shotgun proteomics as well as label-free protein quantification in urine samples is developed. The method first separates the urine proteins using nitrocellulose-based membranes and then proteins are in-membrane digested using trypsin. The enzymatic digestion process is accelerated from overnight to four minutes using a sonoreactor ultrasonic device. Overall, the sample treatment pipeline comprising protein separation, digestion and identification is done in just 3h. The process is assessed using urine of healthy volunteers. The method shows that male can be differentiated from female using the protein content of urine in a fast, easy and straightforward way. 232 and 226 proteins are identified in urine of male and female, respectively. From this, 162 are common to both genders, whilst 70 are unique to male and 64 to female. From the 162 common proteins, 13 are present at levels statistically different (p < 0.05). The method matches the analytical minimalism concept as outlined by Halls, as each stage of this analysis is evaluated to minimize the time, cost, sample requirement, reagent consumption, energy requirements and production of waste products. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Effect of freeze/thaw cycles on several biomarkers in urine from patients with kidney disease.

    PubMed

    Zhang, Yinan; Luo, Yi; Lu, Huijuan; Wang, Niansong; Shen, Yixie; Chen, Ruihua; Fang, Pingyan; Yu, Hong; Wang, Congrong; Jia, Weiping

    2015-04-01

    Urine samples were collected from eleven randomly selected patients with kidney disease, including diabetic nephropathy, chronic nephritis, and nephritic syndrome. Urine samples were treated with one of four protocols for freezing and thawing: freeze directly and thaw directly; freeze directly and thaw by temperature gradient; freeze by temperature gradient and thaw directly; and freeze by temperature gradient and thaw by temperature gradient. After one to six freeze/thaw cycles at -20°C or -80°C, different biomarkers showed differential stabilities. The concentrations of total protein, calcium, and potassium did not change significantly after five freeze/thaw cycles at either -20°C or -80°C. Albumin could only sustain three freeze/thaw cycles at -20°C before it started to degrade. We recommend that urine be stored at -80°C as albumin and the organic ions could sustain five and six freeze/thaw cycles, respectively, using the simple "direct freeze and direct thaw" protocol. Furthermore, in most cases, gradient freeze/thaw cycles are not necessary for urine sample storage.

  19. The use of semi-quantitative tests at Cesarean section delivery for the differentiation of canine fetal fluids from maternal urine on the basis of biochemical characteristics.

    PubMed

    Balogh, Orsolya; Roch, Marie; Keller, Stefanie; Michel, Erika; Reichler, Iris M

    2017-01-15

    In dogs, there is no diagnostic test to identify and differentiate fetal fluids from maternal urine in the event that a clear-yellowish vulvar discharge is observed pre-whelping. The objective of this study was to find a test that could easily and accurately identify rupture of the fetal membranes preceding parturition. Maternal urine, and amniotic fluid (AMF) and allantoic fluid (ALF) from only one fetus per bitch, were collected intraoperatively during Cesarean section. Specific gravity (SG) was analyzed with a refractometer, whereas the presence of leukocytes, protein, glucose, ketones, bilirubin, urobilinogen, nitrite, erythrocyte/hemoglobin (Hb), and the pH were assessed using a urine dipstick (Combur-Test ® ). Combined calcium and magnesium (Ca/Mg) content were evaluated with the Total Hardness Test. The AmniSure test, which detects rupture of fetal membranes in women on the basis of the presence of human placental alpha microglobulin-1, was also performed on canine AMF, ALF, and urine. Data were analyzed using the Fisher's exact test, Wilcoxon signed-rank test, and Pearson's correlation. Sensitivity, specificity, and positive and negative likelihood ratios (LR) were calculated for parameters with significant difference between urine and both fetal fluids. Maternal urine had higher SG and lower leukocyte, protein, Hb, and Ca/Mg content than AMF and ALF. Glucose was more often present in AMF (n = 17) and ALF (n = 12) than in urine (n = 1), whereas ketone bodies were rarely detected in ALF compared with urine. Bilirubin content was higher in urine and ALF than in AMF. AMF pH was less variable and higher than the pH of ALF or urine. The AmniSure was negative in all samples tested. Sensitivity and specificity for SG and for the detection of leukocytes, protein, glucose, Hb, Ca/Mg, and glucose without ketones in urine and fetal fluids were between 42% to 100% and 65% to 100%, respectively. Best positive LR was achieved for the detection of glucose without ketones

  20. Urine albumin/creatinine ratio, high sensitivity C-reactive protein and N-terminal pro brain natriuretic peptide--three new cardiovascular risk markers--do they improve risk prediction and influence treatment?

    PubMed

    Olsen, Michael H; Sehestedt, Thomas; Lyngbaek, Stig; Hansen, Tine W; Rasmussen, Susanne; Wachtell, Kristian; Torp-Pedersen, Christian; Hildebrandt, Per R; Ibsen, Hans

    2010-01-01

    In order to prioritize limited health resources in a time of increasing demands optimal cardiovascular risk stratification is essential. We tested the additive prognostic value of 3 relatively new, but established cardiovascular risk markers: N-terminal pro brain natriuretic peptide (Nt-proBNP), related to hemodynamic cardiovascular risk factors, high sensitivity C-reactive protein (hsCRP), related to metabolic cardiovascular risk factors and urine albumin/creatinine ratio (UACR), related to hemodynamic as well as metabolic risk factors. In healthy subjects with a 10-year risk of cardiovascular death lower than 5% based on HeartScore and therefore not eligible for primary prevention, the actual 10-year risk of cardiovascular death exceeded 5% in a small subgroup of subjects with UACR higher than the 95-percentile of approximately 1.6 mg/mmol. Combined use of high UACR or high hsCRP identified a larger subgroup of 16% with high cardiovascular risk in which primary prevention may be advised despite low-moderate cardiovascular risk based on HeartScore. Furthermore, combined use of high UACR or high Nt-proBNP in subjects with known cardiovascular disease or diabetes identified a large subgroup of 48% with extremely high cardiovascular risk who should be referred for specialist care to optimize treatment.

  1. Adulterants in Urine Drug Testing.

    PubMed

    Fu, S

    Urine drug testing plays an important role in monitoring licit and illicit drug use for both medico-legal and clinical purposes. One of the major challenges of urine drug testing is adulteration, a practice involving manipulation of a urine specimen with chemical adulterants to produce a false negative test result. This problem is compounded by the number of easily obtained chemicals that can effectively adulterate a urine specimen. Common adulterants include some household chemicals such as hypochlorite bleach, laundry detergent, table salt, and toilet bowl cleaner and many commercial products such as UrinAid (glutaraldehyde), Stealth® (containing peroxidase and peroxide), Urine Luck (pyridinium chlorochromate, PCC), and Klear® (potassium nitrite) available through the Internet. These adulterants can invalidate a screening test result, a confirmatory test result, or both. To counteract urine adulteration, drug testing laboratories have developed a number of analytical methods to detect adulterants in a urine specimen. While these methods are useful in detecting urine adulteration when such activities are suspected, they do not reveal what types of drugs are being concealed. This is particularly the case when oxidizing urine adulterants are involved as these oxidants are capable of destroying drugs and their metabolites in urine, rendering the drug analytes undetectable by any testing technology. One promising approach to address this current limitation has been the use of unique oxidation products formed from reaction of drug analytes with oxidizing adulterants as markers for monitoring drug misuse and urine adulteration. This novel approach will ultimately improve the effectiveness of the current urine drug testing programs. © 2016 Elsevier Inc. All rights reserved.

  2. Dietary total antioxidant capacity from different assays in relation to serum C-reactive protein among young Japanese women.

    PubMed

    Kobayashi, Satomi; Murakami, Kentaro; Sasaki, Satoshi; Uenishi, Kazuhiro; Yamasaki, Mitsuyo; Hayabuchi, Hitomi; Goda, Toshinao; Oka, Jun; Baba, Keiko; Ohki, Kazuko; Watanabe, Reiko; Sugiyamama, Yoshiko

    2012-10-30

    The association between dietary total antioxidant capacity (TAC) from different assays and serum C-reactive protein (CRP) has not been assessed in non-Western populations. We examined the association between dietary TAC and serum CRP concentration in young Japanese women using different four TAC assays. The subjects were 443 young Japanese women aged 18-22 years. Dietary TAC was assessed with a self-administered diet history questionnaire and the TAC value of each food using the following four assays: ferric reducing ability of plasma (FRAP); oxygen radical absorbance capacity (ORAC); Trolox equivalent antioxidant capacity (TEAC); and total radical-trapping antioxidant parameter (TRAP). Serum CRP concentrations were measured by highly sensitive nephelometry. The major contributor to dietary TAC was green, barley, and oolong tea (FRAP: 53%, ORAC: 45%, TEAC: 36%, and TRAP: 44%). The prevalence of elevated CRP concentrations (≥ 1 mg/L) was 5.6%. TAC from FRAP was inversely associated with serum CRP concentrations (adjusted odds ratio [OR] for elevated CRP concentration in high [compared with low] dietary TAC group: 0.39 [95% confidence interval (CI): 0.16-0.98]; P = 0.04). TAC from ORAC was inversely associated with CRP, although the association was not significant (OR: 0.48 [95% CI: 0.20-1.14]; P = 0.10). TAC from TEAC was inversely associated with CRP (OR: 0.32 [95% CI: 0.12-0.82]; P = 0.02), as was TAC from TRAP (OR: 0.31 [95% CI: 0.12-0.81]; P = 0.02). Dietary TAC was inversely associated with serum CRP concentration in young Japanese women regardless of assay. Further studies are needed in other populations to confirm these results.

  3. Dietary total antioxidant capacity from different assays in relation to serum C-reactive protein among young Japanese women

    PubMed Central

    2012-01-01

    Background The association between dietary total antioxidant capacity (TAC) from different assays and serum C-reactive protein (CRP) has not been assessed in non-Western populations. We examined the association between dietary TAC and serum CRP concentration in young Japanese women using different four TAC assays. Methods The subjects were 443 young Japanese women aged 18–22 years. Dietary TAC was assessed with a self-administered diet history questionnaire and the TAC value of each food using the following four assays: ferric reducing ability of plasma (FRAP); oxygen radical absorbance capacity (ORAC); Trolox equivalent antioxidant capacity (TEAC); and total radical-trapping antioxidant parameter (TRAP). Serum CRP concentrations were measured by highly sensitive nephelometry. Results The major contributor to dietary TAC was green, barley, and oolong tea (FRAP: 53%, ORAC: 45%, TEAC: 36%, and TRAP: 44%). The prevalence of elevated CRP concentrations (≥ 1 mg/L) was 5.6%. TAC from FRAP was inversely associated with serum CRP concentrations (adjusted odds ratio [OR] for elevated CRP concentration in high [compared with low] dietary TAC group: 0.39 [95% confidence interval (CI): 0.16-0.98]; P = 0.04). TAC from ORAC was inversely associated with CRP, although the association was not significant (OR: 0.48 [95% CI: 0.20-1.14]; P = 0.10). TAC from TEAC was inversely associated with CRP (OR: 0.32 [95% CI: 0.12-0.82]; P = 0.02), as was TAC from TRAP (OR: 0.31 [95% CI: 0.12-0.81]; P = 0.02). Conclusions Dietary TAC was inversely associated with serum CRP concentration in young Japanese women regardless of assay. Further studies are needed in other populations to confirm these results. PMID:23110638

  4. [Animal experiment studies on the changes in lipid and protein metabolism in L-carnitine-supplemented total parenteral nutrition].

    PubMed

    Böhles, H; Segerer, H; Fekl, W; Stehr, K

    1983-02-01

    The influence of i.v. L-carnitine on parameters of lipid- and nitrogen metabolism was studied during total parenteral nutrition of mini pigs (x: 4077; n = 9). The infusion protocol was divided into isocaloric and isonitrogenous 48-hour-periods. Amino acids (3 g/kg/day) were administered throughout all three periods. 140 Cal/kg/day were given as non-protein calories, consisting only of glucose during period 1. During periods 2 and 3 an amount of glucose calorically equivalent to 4 g fat/kg/day was substituted with a lipid emulsion. In period 3, L-carnitine (1,5 mg/kg/day) was added. During the entire regime key parameters of fat and nitrogen metabolism were determined. During all three periods indirect calorimetry was performed and the respiratory quotient calculated. The results demonstrate a more effective lipolysis and oxydation of fatty acids during L-carnitine supplementation. This results in an increased energy gain from exogenously administered fat and a distinct improvement of nitrogen balance.

  5. Albumin adsorption onto surfaces of urine collection and analysis containers☆

    PubMed Central

    Robinson, Mary K.; Caudill, Samuel P.; Koch, David D.; Ritchie, James; Hortin, Glen; Eckfeldt, John H.; Sandberg, Sverre; Williams, Desmond; Myers, Gary; Miller, W. Greg

    2017-01-01

    Background Adsorption of albumin onto urine collection and analysis containers may cause falsely low concentrations. Methods We added 125I-labeled human serum albumin to urine and to phosphate buffered solutions, incubated them with 22 plastic container materials and measured adsorption by liquid scintillation counting. Results Adsorption of urine albumin (UA) at 5–6 mg/l was <0.9%; and at 90 mg/l was <0.4%. Adsorption was generally less at pH 8 than pH 5 but only 3 cases had p <0.05. Adsorption from 11 unaltered urine samples with albumin 5–333 mg/l was <0.8%. Albumin adsorption for the material with greatest binding was extrapolated to the surface areas of 100 ml and 2 l collection containers, and to instrument sample cups and showed <1% change in concentration at 5 mg/l and <0.5% change at 20 mg/l or higher concentrations. Adsorption of albumin from phosphate buffered solutions (2–28%) was larger than that from urine. Conclusions Albumin adsorption differed among urine samples and plastic materials, but the total influence of adsorption was <1% for all materials and urine samples tested. Adsorption of albumin from phosphate buffered solutions was larger than that from urine and could be a limitation for preparations used as calibrators. PMID:24513540

  6. Albumin adsorption onto surfaces of urine collection and analysis containers.

    PubMed

    Robinson, Mary K; Caudill, Samuel P; Koch, David D; Ritchie, James; Hortin, Glen; Eckfeldt, John H; Sandberg, Sverre; Williams, Desmond; Myers, Gary; Miller, W Greg

    2014-04-20

    Adsorption of albumin onto urine collection and analysis containers may cause falsely low concentrations. We added (125)I-labeled human serum albumin to urine and to phosphate buffered solutions, incubated them with 22 plastic container materials and measured adsorption by liquid scintillation counting. Adsorption of urine albumin (UA) at 5-6 mg/l was <0.9%; and at 90 mg/l was <0.4%. Adsorption was generally less at pH8 than pH5 but only 3 cases had p<0.05. Adsorption from 11 unaltered urine samples with albumin 5-333 mg/l was <0.8%. Albumin adsorption for the material with greatest binding was extrapolated to the surface areas of 100 ml and 2l collection containers, and to instrument sample cups and showed <1% change in concentration at 5 mg/l and <0.5% change at 20 mg/l or higher concentrations. Adsorption of albumin from phosphate buffered solutions (2-28%) was larger than that from urine. Albumin adsorption differed among urine samples and plastic materials, but the total influence of adsorption was <1% for all materials and urine samples tested. Adsorption of albumin from phosphate buffered solutions was larger than that from urine and could be a limitation for preparations used as calibrators. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Advances in Urine Microscopy.

    PubMed

    Becker, Gavin J; Garigali, Giuseppe; Fogazzi, Giovanni B

    2016-06-01

    Urine microscopy is an important tool for the diagnosis and management of several conditions affecting the kidneys and urinary tract. In this review, we describe the automated instruments, based either on flow cytometry or digitized microscopy, that are currently in use in large clinical laboratories. These tools allow the examination of large numbers of samples in short periods. We also discuss manual urinary microscopy commonly performed by nephrologists, which we encourage. After discussing the advantages of phase contrast microscopy over bright field microscopy, we describe the advancements of urine microscopy in various clinical conditions. These include persistent isolated microscopic hematuria (which can be classified as glomerular or nonglomerular on the basis of urinary erythrocyte morphology), drug- and toxin-related cystalluria (which can be a clue for the diagnosis of acute kidney injury associated with intrarenal crystal precipitation), and some inherited conditions (eg, adenine phosphoribosyltransferase deficiency, which is associated with 2,8-dihydroxyadenine crystalluria, and Fabry disease, which is characterized by unique urinary lamellated fatty particles). Finally, we describe the utility of identifying "decoy cells" and atypical malignant cells, which can be easily done with phase contrast microscopy in unfixed samples. Copyright © 2016 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

  8. Basic or extended urine sampling to analyse urine production?

    PubMed

    Denys, Marie-Astrid; Kapila, Vansh; Weiss, Jeffrey; Goessaert, An-Sofie; Everaert, Karel

    2017-09-01

    Frequency volume charts are valuable tools to objectify urine production in patients with nocturia, enuresis or nocturnal incontinence. Analyses of daytime and nighttime urine (=basic collection) or analyses of urine samples collected every 3 h (=extended collection) extend this evaluation by describing circadian patterns of water and solute diuresis (=renal function profiles). To assess intra-individual correlation and agreement between renal function profiles provided using basic and extended urine collections, and using two extended urine collections. To create a short-form of the extended collection. This prospective observational study was executed at Ghent University Hospital, Belgium. Study participation was open for anyone visiting the hospital. Participants collected one basic and two extended 24-h urine collections. Urinary levels of osmolality, sodium and creatinine were determined. There was a moderate to strong correlation between results of basic and extended urinalyses. Comparing both extended urinalyses showed a moderate correlation between the eight individual samples and a weak to strong correlation between the mean daytime and nighttime values of renal functions. Different samples could be considered as most representative for mean daytime values, while all samples collected between 03 and 05am showed the highest agreement with mean nighttime values of renal function. Since there is a good correlation and agreement between basic and extended urine collections to study the mechanisms underlying urine production, the choice of urine sampling method to evaluate urine production depends on the purpose. A nighttime-only urine sample collected between 03 and 05am may be the most practical approach. © 2017 Wiley Periodicals, Inc.

  9. Effect of urine creatinine level during pregnancy on dipstick test.

    PubMed

    Baba, Yosuke; Furuta, Itsuko; Zhai, Tianyue; Ohkuchi, Akihide; Yamada, Takahiro; Takahashi, Kayo; Matsubara, Shigeki; Minakami, Hisanori

    2017-06-01

    Dipstick results for proteinuria are affected by urine concentration, and thus urine creatinine concentration ([Cr]). This study was performed to determine whether spot urine [Cr] changes significantly during pregnancy, leading to a significantly different false-negative rate (FNR) on dipstick test between trimester. The [Cr] and protein concentrations ([P]) were analyzed in 631 spot urine samples with negative/equivocal dipstick from 425 pregnant women. False-negative dipstick was defined as [P] : [Cr] ratio (P/Cr) > 0.27 mg/mg. Median [Cr] was 117 mg/dL (range, 6.5-326 mg/dL), 72 mg/dL (range, 4.3-477 mg/dL), and 73 mg/dL (range, 8.4-396 mg/dL) in the first (n = 96), second (n = 344), and third (n = 191) trimester urine samples, respectively (P = 0.000, Kruskal-Wallis). Both [P] and P/Cr increased significantly with advancing gestation. FNR 9.4% (18/191) in the third trimester was significantly higher than that of 0.0% (0/96) in the second trimester and that of 0.5% (2/344) in the third trimester. In the 20 urine samples with false-negative dipstick, median [Cr] was 47.0 mg/dL (range, 11.0-358 mg/dL) and the proportion of samples with dilute urine, that is, [Cr] <47 mg/dL, was significantly higher than in the remaining 611 urine samples (50%, 10/20 vs 28%, 174/611, respectively, P = 0.046). Urine samples in the second and third trimesters were more likely to be diluted compared with the first trimester. This was associated with high FNR in third trimester urine samples. © 2017 Japan Society of Obstetrics and Gynecology.

  10. Association of translocator protein total distribution volume with duration of untreated major depressive disorder: a cross-sectional study.

    PubMed

    Setiawan, Elaine; Attwells, Sophia; Wilson, Alan A; Mizrahi, Romina; Rusjan, Pablo M; Miler, Laura; Xu, Cynthia; Sharma, Sarita; Kish, Stephen; Houle, Sylvain; Meyer, Jeffrey H

    2018-04-01

    People with major depressive disorder frequently exhibit increasing persistence of major depressive episodes. However, evidence for neuroprogression (ie, increasing brain pathology with longer duration of illness) is scarce. Microglial activation, which is an important component of neuroinflammation, is implicated in neuroprogression. We examined the relationship of translocator protein (TSPO) total distribution volume (V T ), a marker of microglial activation, with duration of untreated major depressive disorder, and with total illness duration and antidepressant exposure. In this cross-sectional study, we recruited participants aged 18-75 years from the Toronto area and the Centre for Addiction and Mental Health (Toronto, ON, Canada). Participants either had major depressive episodes secondary to major depressive disorder or were healthy, as confirmed with a structured clinical interview and consultation with a study psychiatrist. To be enrolled, participants with major depressive episodes had to score a minimum of 17 on the 17-item Hamilton Depression Rating Scale, and had to be medication free or taking a stable dose of medication for at least 4 weeks before PET scanning. Eligible participants were non-smokers; had no history of or concurrent alcohol or substance dependence, neurological illness, autoimmune disorder, or severe medical problems; and were free from acute medical illnesses for the previous 2 weeks before PET scanning. Participants were excluded if they had used brain stimulation treatments within the 6 months before scanning, had used anti-inflammatory drugs lasting at least 1 week within the past month, were taking hormone replacement therapy, had psychotic symptoms, had bipolar disorder (type I or II) or borderline antisocial personality disorder, or were pregnant or breastfeeding. We scanned three primary grey-matter regions of interest (prefrontal cortex, anterior cingulate cortex, and insula) and 12 additional regions and subregions using 18

  11. The association of 83 plasma proteins with CHD mortality, BMI, HDL-, and total-cholesterol in men: applying multivariate statistics to identify proteins with prognostic value and biological relevance.

    PubMed

    Heidema, A Geert; Thissen, Uwe; Boer, Jolanda M A; Bouwman, Freek G; Feskens, Edith J M; Mariman, Edwin C M

    2009-06-01

    In this study, we applied the multivariate statistical tool Partial Least Squares (PLS) to analyze the relative importance of 83 plasma proteins in relation to coronary heart disease (CHD) mortality and the intermediate end points body mass index, HDL-cholesterol and total cholesterol. From a Dutch monitoring project for cardiovascular disease risk factors, men who died of CHD between initial participation (1987-1991) and end of follow-up (January 1, 2000) (N = 44) and matched controls (N = 44) were selected. Baseline plasma concentrations of proteins were measured by a multiplex immunoassay. With the use of PLS, we identified 15 proteins with prognostic value for CHD mortality and sets of proteins associated with the intermediate end points. Subsequently, sets of proteins and intermediate end points were analyzed together by Principal Components Analysis, indicating that proteins involved in inflammation explained most of the variance, followed by proteins involved in metabolism and proteins associated with total-C. This study is one of the first in which the association of a large number of plasma proteins with CHD mortality and intermediate end points is investigated by applying multivariate statistics, providing insight in the relationships among proteins, intermediate end points and CHD mortality, and a set of proteins with prognostic value.

  12. Urine cup for collection of urine from cows.

    PubMed

    Fellner, V; Weiss, M F; Belo, A T; Belyea, R L; Martz, F A; Orma, A H

    1988-08-01

    A urine cup for continuous and complete collection of urine from cows was constructed from Plastisol, cotton webb strapping, Velcro Brand touch fasteners [corrected], snap-fasteners, denim patches, weather stripping, and vacuum hose. The urine cup was made from Plastisol using a heated lead mold. It was large enough to enclose a 9 cm x 6 cm area around the vulva of a cow and was attached by strapping and Velcro Brand touch fasteners [corrected] to patches glued to the rump. Urine cups were used repeatedly and provided for long-term collection of urine from cows, eliminating the need for indwelling catheters. Applications include long-term nutrient balance, radioisotope, and metabolism studies.

  13. Comparison of refractometry and biuret assay for measurement of total protein concentration in canine abdominal and pleural fluid specimens.

    PubMed

    Rose, Alexandra; Funk, Deborah; Neiger, Reto

    2016-04-01

    To compare total protein (TP) concentrations in canine pleural and abdominal fluid specimens as measured by refractometry and biuret assay. Diagnostic test evaluation. Data regarding 92 pleural and 148 abdominal fluid specimens from dogs with various diseases. TP concentrations in fluid specimens as measured by refractometry and biuret assay were recorded. Strength of association between sets of measurements was assessed by Spearman rank correlations and Bland-Altman plots. Optimal concentration cutoff for diagnostic discrimination between exudate and nonexudate was identified by construction of receiver operating characteristic curves. Median TP concentration in pleural fluid specimens was 2.7 g/dL (range, 0.3 to 4.8 g/dL) for refractometry and 2.9 g/dL (range, 0.7 to 5.8 g/dL) for biuret assay. Median TP concentration in abdominal fluid specimens was 3.5 g/dL (range, 0.1 to 6.0 g/dL) for refractometry and 3.5 g/dL (range, 0.6 to 5.7 g/dL) for biuret assay. Correlation was significant between refractometric and biuret results for pleural (ρ = 0.921) and abdominal (ρ = 0.908) fluid. Bland-Altman plots revealed bias of -0.18 g/dL for pleural fluid and -0.03 g/dL for abdominal fluid for refractometry versus biuret assay. With a TP concentration of ≥ 3 g/dL used to distinguish exudate from nonexudate, sensitivity of refractometry was 77% for pleural fluid and 80% for abdominal fluid. Specificity was 100% and 94%, respectively. Refractometry yielded acceptable results for measurement of TP concentration in canine pleural and abdominal fluid specimens, providing a more rapid and convenient method than biuret assay.

  14. Total external reflection X-ray fluorescence analysis of protein-metal ion interactions in biological systems

    NASA Astrophysics Data System (ADS)

    Novikova, N. N.; Kovalchuk, M. V.; Yur'eva, E. A.; Konovalov, O. V.; Rogachev, A. V.; Stepina, N. D.; Sukhorukov, V. S.; Tsaregorodtsev, A. D.; Chukhrai, E. S.; Yakunin, S. N.

    2012-09-01

    This paper presents the results of an investigation into hemoglobin-based protein films that were formed on a liquid surface. X-ray standing wave measurements were performed at the ID 10 beamline of the European Synchrotron Radiation Facility (ESRF) and at the Langmuir station of the Kurchatov Synchrotron Radiation Source. It was found that the ability of the protein to bind metal ions is substantially increased due to the conformational rearrangements of protein macromolecules caused by various damaging effects. The elemental composition of protein preparations, which were isolated from children and adults with chronic metabolic diseases accompanied by endogenous intoxication, was analyzed. The results of the investigations offer evidence that an increase in the ligand-binding properties of the protein molecules, which was observed in model experiments using protein films, is a common trait and corresponds to in vivo processes accompanying metabolic disturbances in the body.

  15. Intake of total, animal and plant protein and subsequent changes in weight or waist circumference in European men and women: the Diogenes project.

    PubMed

    Halkjær, J; Olsen, A; Overvad, K; Jakobsen, M U; Boeing, H; Buijsse, B; Palli, D; Tognon, G; Du, H; van der A, D L; Forouhi, N G; Wareham, N J; Feskens, E J M; Sørensen, T I A; Tjønneland, A

    2011-08-01

    As protein is considered to increase thermogenesis and satiety more than other macronutrients, it may have beneficial effects on prevention of weight gain and weight maintenance. The objective of this study is to assess the association between the amount and type of dietary protein, and subsequent changes in weight and waist circumference (WC). 89,432 men and women from five countries participating in European Prospective Investigation into Cancer and Nutrition (EPIC) were followed for a mean of 6.5 years. Associations between the intake of protein or subgroups of protein (from animal and plant sources) and changes in weight (g per year) or WC (cm per year) were investigated using gender and centre-specific multiple regression analyses. Adjustments were made for other baseline dietary factors, baseline anthropometrics, demographic and lifestyle factors and follow-up time. We used random effect meta-analyses to obtain pooled estimates across centres. Higher intake of total protein, and protein from animal sources was associated with subsequent weight gain for both genders, strongest among women, and the association was mainly attributable to protein from red and processed meat and poultry rather than from fish and dairy sources. There was no overall association between intake of plant protein and subsequent changes in weight. No clear overall associations between intakes of total protein or any of the subgroups and changes in WC were present. The associations showed some heterogeneity between centres, but pooling of estimates was still considered justified. A high intake of protein was not found associated with lower weight or waist gain in this observational study. In contrast, protein from food items of animal origin, especially meat and poultry, seemed to be positively associated with long-term weight gain. There were no clear associations for waist changes.

  16. Urine sampling and collection system

    NASA Technical Reports Server (NTRS)

    Fogal, G. L.; Mangialardi, J. K.; Reinhardt, C. G.

    1971-01-01

    This specification defines the performance and design requirements for the urine sampling and collection system engineering model and establishes requirements for its design, development, and test. The model shall provide conceptual verification of a system applicable to manned space flight which will automatically provide for collection, volume sensing, and sampling of urine.

  17. Urine pH test

    MedlinePlus

    ... 2015:chap 4. Remer T, Manz F. Potential renal acid load of foods and its influence on urine pH. J Am Diet Assoc . 1995;95(7):791-797. PMID: 7797810 www.ncbi.nlm.nih.gov/pubmed/7797810 . Riley RS, McPherson RA. Basic examination of urine. In: McPherson RA, Pincus MR, ...

  18. Blood in the Urine (Hematuria)

    MedlinePlus

    ... 2 weeks later to make sure the urine is free of red blood cells. If hematuria only happens once, there's usually ... follow-up tests to make sure your urine is free of red blood cells. If the hematuria goes away, your doctor ...

  19. Chemical Functionalization of Germanium with Dextran Brushes for Immobilization of Proteins Revealed by Attenuated Total Reflection Fourier Transform Infrared Difference Spectroscopy.

    PubMed

    Schartner, Jonas; Hoeck, Nina; Güldenhaupt, Jörn; Mavarani, Laven; Nabers, Andreas; Gerwert, Klaus; Kötting, Carsten

    2015-07-21

    Protein immobilization studied by attenuated total reflection Fourier transform infrared (ATR-FT-IR) difference spectroscopy is an emerging field enabling the study of proteins at atomic detail. Gold or glass surfaces are frequently used for protein immobilization. Here, we present an alternative method for protein immobilization on germanium. Because of its high refractive index and broad spectral window germanium is the best material for ATR-FT-IR spectroscopy of thin layers. So far, this technique was mainly used for protein monolayers, which lead to a limited signal-to-noise ratio. Further, undesired protein-protein interactions can occur in a dense layer. Here, the germanium surface was functionalized with thiols and stepwise a dextran brush was generated. Each step was monitored by ATR-FT-IR spectroscopy. We compared a 70 kDa dextran with a 500 kDa dextran regarding the binding properties. All surfaces were characterized by atomic force microscopy, revealing thicknesses between 40 and 110 nm. To analyze the capability of our system we utilized N-Ras on mono-NTA (nitrilotriacetic acid) functionalized dextran, and the amount of immobilized Ras corresponded to several monolayers. The protein stability and loading capacity was further improved by means of tris-NTA for immobilization. Small-molecule-induced changes were revealed with an over 3 times higher signal-to-noise ratio compared to monolayers. This improvement may allow the observation of very small and so far hidden changes in proteins upon stimulus. Furthermore, we immobilized green fluorescent protein (GFP) and mCherry simultaneously enabling an analysis of the surface by fluorescence microscopy. The absence of a Förster resonance energy transfer (FRET) signal demonstrated a large protein-protein distance, indicating an even distribution of the protein within the dextran.

  20. Stability of Synthetic Cathinones in Urine.

    PubMed

    Glicksberg, Lindsay; Kerrigan, Sarah

    2018-03-01

    In this report, we evaluate the concentration, pH, temperature and analyte-dependent effects on cathinone stability in preserved human urine. A total of 22 synthetic cathinones were evaluated at 100 ng/mL and 1,000 ng/mL in pH 4 and pH 8 urine over 6 months. Specimens were stored at -20°C, 4°C, 20°C and 32°C. The stability of synthetic cathinones was highly dependent on urine pH and storage temperature. Cathinones were considerably more stable in acidic urine (pH 4) at low temperature. In alkaline urine (pH 8) at 32°C, significant losses (>20%) were observed within hours for the majority of drugs. In contrast, all drugs were stable in frozen and refrigerated urine at pH 4 for the duration of the study. These results highlight the importance of sample storage and the potential for pre-analytical changes in concentration during routine shipping and handling of specimens. Significant structural influence was also observed. Cathinones bearing a tertiary amine (pyrrolidine group) were significantly more stable than their secondary amine counterparts. The methylenedioxy group also exerted a significant stabilizing effect on both the tertiary and secondary amines. In the absence of the methylenedioxy group, no significant differences in stability were observed between the unsubstituted and ring substituted secondary amines. Half-lives at ambient temperature in pH 8 urine ranged from 9 h (3-fluoromethcathinone) to 4.3 months (methylenedioxypyrovalerone and 3,4-methylenedioxy-α-pyrrolidinobutiophenone), demonstrating the importance of analyte dependence, and the dual stabilizing effect of both the pyrollidine and methylenedioxy groups. Biological evidence may be subjected to a variety of environmental conditions prior to, and during transport to the forensic laboratory. These findings demonstrate the inherent instability of certain cathinone species in biological evidence under some conditions. Moreover, this study highlights the need for quantitative drug findings in

  1. Intake of total, animal and plant proteins, and their food sources in 10 countries in the European Prospective Investigation into Cancer and Nutrition.

    PubMed

    Halkjaer, J; Olsen, A; Bjerregaard, L J; Deharveng, G; Tjønneland, A; Welch, A A; Crowe, F L; Wirfält, E; Hellstrom, V; Niravong, M; Touvier, M; Linseisen, J; Steffen, A; Ocké, M C; Peeters, P H M; Chirlaque, M D; Larrañaga, N; Ferrari, P; Contiero, P; Frasca, G; Engeset, D; Lund, E; Misirli, G; Kosti, M; Riboli, E; Slimani, N; Bingham, S

    2009-11-01

    To describe dietary protein intakes and their food sources among 27 redefined centres in 10 countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). Between 1995 and 2000, 36 034 persons, aged between 35 and 74 years, were administered a standardized 24-h dietary recall (24-HDR) using a computerized interview software programme (EPIC-SOFT). Intakes (g/day) of total, animal and plant proteins were estimated using the standardized EPIC Nutrient Database (ENDB). Mean intakes were adjusted for age, and weighted by season and day of recall. Mean total and animal protein intakes were highest in the Spanish centres among men, and in the Spanish and French centres among women; the lowest mean intakes were observed in the UK health-conscious group, in Greek men and women, and in women in Potsdam. Intake of plant protein was highest among the UK health-conscious group, followed by some of the Italian centres and Murcia, whereas Sweden and Potsdam had the lowest intake. Cereals contributed to the highest proportion of plant protein in all centres. The combined intake of legumes, vegetables and fruit contributed to a greater proportion of plant protein in the southern than in the northern centres. Total meat intake (with some heterogeneity across subtypes of meat) was, with few exceptions, the most important contributor to animal protein in all centres, followed by dairy and fish products. This study shows that intake of protein, especially of animal origin, differs across the 10 European countries, and also shows some differences in food sources of protein across Europe.

  2. 2D-electrophoresis and the urine proteome map: where do we stand?

    PubMed

    Candiano, Giovanni; Santucci, Laura; Petretto, Andrea; Bruschi, Maurizio; Dimuccio, Veronica; Urbani, Andrea; Bagnasco, Serena; Ghiggeri, Gian Marco

    2010-03-10

    The discovery of urinary biomarkers is a main topic in clinical medicine. The development of proteomics has rapidly changed the knowledge on urine protein composition and probably will modify it again. Two-dimensional electrophoresis (2D-PAGE) coupled with mass spectrometry has represented for years the technique of choice for the analysis of urine proteins and it is time to draw some conclusions. This review will focus on major methodological aspects related to urine sample collection, storage and analysis by 2D-PAGE and attempt to define an advanced normal urine protein map. Overall, 1118 spots were reproducibly found in normal urine samples but only 275 were characterized as isoforms of 82 proteins. One-hundred height spots belonging to 30 proteins were also detected in plasma and corresponded to typical plasma components. The identity of most of the proteins found in normal urine by 2D-PAGE remains to be determined, the majority being low-molecular weight proteins (<30 kDa). Equalization procedures would also enhance sensitivity of the analysis and allow low abundance proteins to be characterized. Therefore, we are still on the way to define the normal urine composition. Technology advancements in concentrating procedure will improve sensitivity and give the possibility to purify proteins for mass spectrometry. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  3. Consideration of the degree of increase in urine metadrenalines provides superior specificity in the diagnosis of phaeochromocytoma than additional urine catecholamine measurement.

    PubMed

    Scargill, J J; Reed, P; Kane, J

    2013-01-01

    Measurement of fractionated plasma or urine metadrenalines is the recommended screening test in the diagnosis of phaeochromocytoma, with clinical cut-offs geared towards diagnostic sensitivity. Current practice at Salford Royal Hospital is to add urine catecholamines onto samples with raised urine metadrenalines, with the aim of adding specificity to a diagnosis of phaeochromocytoma. This practice was reviewed by identifying a series of patients with raised urine metadrenalines who had catecholamines reflectively added. A total of 358 samples were identified from 242 patients, of which 228 had urine catecholamines measured. A diagnosis of 'phaeochromocytoma' (n = 41) or 'no phaeochromocytoma' (n = 90) was obtained in 131 of 228 patients, giving raised urine metadrenalines a positive predictive value for phaeochromocytoma of 31%. The finding of increased urine catecholamines in samples with raised urine metadrenalines increased specificity for phaeochromocytoma to 70%. However, 95% diagnostic specificity for phaeochromocytoma could be achieved by the introduction of a second cut-off for urine metadrenalines geared towards maximizing specificity. Consideration of the degree of increase in urine metadrenalines is a superior method of determining the likelihood of phaeochromocytoma than measurement of urine catecholamines.

  4. Examination of the relation between periodontal health status and cardiovascular risk factors: serum total and high density lipoprotein cholesterol, C-reactive protein, and plasma fibrinogen.

    PubMed

    Wu, T; Trevisan, M; Genco, R J; Falkner, K L; Dorn, J P; Sempos, C T

    2000-02-01

    Using data from the Third National Health and Nutrition Examination Survey (1988-1994), the authors examined the relation between periodontal health and cardiovascular risk factors: serum total and high density lipoprotein cholesterol, C-reactive protein, and plasma fibrinogen. A total of 10,146 participants were included in the analyses of cholesterol and C-reactive protein and 4,461 in the analyses of fibrinogen. Periodontal health indicators included the gingival bleeding index, calculus index, and periodontal disease status (defined by pocket depth and attachment loss). While cholesterol and fibrinogen were analyzed as continuous variables, C-reactive protein was dichotomized into two levels. The results show a significant relation between indicators of poor periodontal status and increased C-reactive protein and fibrinogen. The association between periodontal status and total cholesterol level is much weaker. No consistent association between periodontal status and high density lipoprotein cholesterol was detectable. Similar patterns of association were observed for participants aged 17-54 years and those 55 years and older. In conclusion, this study suggests that total cholesterol, C-reactive protein, and fibrinogen are possible intermediate factors that may link periodontal disease to elevated cardiovascular risk.

  5. Quantitation of yeast total proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for uniform loading.

    PubMed

    Sheen, Hyukho

    2016-04-01

    Proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer are difficult to quantitate due to SDS and reducing agents being in the buffer. Although acetone precipitation has long been used to clean up proteins from detergents and salts, previous studies showed that protein recovery from acetone precipitation varies from 50 to 100% depending on the samples tested. Here, this article shows that acetone precipitates proteins highly efficiently from SDS-PAGE sample buffer and that quantitative recovery is achieved in 5 min at room temperature. Moreover, precipitated proteins are resolubilized with urea/guanidine, rather than with SDS. Thus, the resolubilized samples are readily quantifiable with Bradford reagent without using SDS-compatible assays. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Decomposition of total solvation energy into core, side-chains and water contributions: Role of cross correlations and protein conformational fluctuations in dynamics of hydration layer

    NASA Astrophysics Data System (ADS)

    Mondal, Sayantan; Mukherjee, Saumyak; Bagchi, Biman

    2017-09-01

    Dynamical coupling between water and amino acid side-chain residues in solvation dynamics is investigated by selecting residues often used as natural probes, namely tryptophan, tyrosine and histidine, located at different positions on protein surface. Such differently placed residues are found to exhibit different timescales of relaxation. The total solvation response measured by the probe is decomposed in terms of its interactions with (i) protein core, (ii) side-chain and (iii) water. Significant anti cross-correlation among these contributions are observed. When the motion of the protein side-chains is quenched, solvation either becomes faster or slower depending on the location of the probe.

  7. A comparison of two colorimetric assays, based upon Lowry and Bradford techniques, to estimate total protein in soil extracts.

    PubMed

    Redmile-Gordon, M A; Armenise, E; White, R P; Hirsch, P R; Goulding, K W T

    2013-12-01

    Soil extracts usually contain large quantities of dissolved humified organic material, typically reflected by high polyphenolic content. Since polyphenols seriously confound quantification of extracted protein, minimising this interference is important to ensure measurements are representative. Although the Bradford colorimetric assay is used routinely in soil science for rapid quantification protein in soil-extracts, it has several limitations. We therefore investigated an alternative colorimetric technique based on the Lowry assay (frequently used to measure protein and humic substances as distinct pools in microbial biofilms). The accuracies of both the Bradford assay and a modified Lowry microplate method were compared in factorial combination. Protein was quantified in soil-extracts (extracted with citrate), including standard additions of model protein (BSA) and polyphenol (Sigma H1675-2). Using the Lowry microplate assay described, no interfering effects of citrate were detected even with concentrations up to 5 times greater than are typically used to extract soil protein. Moreover, the Bradford assay was found to be highly susceptible to two simultaneous and confounding artefacts: 1) the colour development due to added protein was greatly inhibited by polyphenol concentration, and 2) substantial colour development was caused directly by the polyphenol addition. In contrast, the Lowry method enabled distinction between colour development from protein and non-protein origin, providing a more accurate quantitative analysis. These results suggest that the modified-Lowry method is a more suitable measure of extract protein (defined by standard equivalents) because it is less confounded by the high polyphenolic content which is so typical of soil extracts.

  8. Dipstick Spot urine pH does not accurately represent 24 hour urine PH measured by an electrode.

    PubMed

    Omar, Mohamed; Sarkissian, Carl; Jianbo, Li; Calle, Juan; Monga, Manoj

    2016-01-01

    To determine whether spot urine pH measured by dipstick is an accurate representation of 24 hours urine pH measured by an electrode. We retrospectively reviewed urine pH results of patients who presented to the urology stone clinic. For each patient we recorded the most recente pH result measured by dipstick from a spot urine sample that preceded the result of a 24-hour urine pH measured by the use of a pH electrode. Patients were excluded if there was a change in medications or dietary recommendations or if the two samples were more than 4 months apart. A difference of more than 0.5 pH was considered na inaccurate result. A total 600 patients were retrospectively reviewed for the pH results. The mean difference in pH between spot urine value and the 24 hours collection values was 0.52±0.45 pH. Higher pH was associated with lower accuracy (p<0.001). The accuracy of spot urine samples to predict 24-hour pH values of <5.5 was 68.9%, 68.2% for 5.5 to 6.5 and 35% for >6.5. Samples taken more than 75 days apart had only 49% the accuracy of more recent samples (p<0.002). The overall accuracy is lower than 80% (p<0.001). Influence of diurnal variation was not significant (p=0.588). Spot urine pH by dipstick is not an accurate method for evaluation of the patients with urolithiasis. Patients with alkaline urine are more prone to error with reliance on spot urine pH.

  9. Advantage of multiple spot urine collections for estimating daily sodium excretion: comparison with two 24-h urine collections as reference.

    PubMed

    Uechi, Ken; Asakura, Keiko; Ri, Yui; Masayasu, Shizuko; Sasaki, Satoshi

    2016-02-01

    Several estimation methods for 24-h sodium excretion using spot urine sample have been reported, but accurate estimation at the individual level remains difficult. We aimed to clarify the most accurate method of estimating 24-h sodium excretion with different numbers of available spot urine samples. A total of 370 participants from throughout Japan collected multiple 24-h urine and spot urine samples independently. Participants were allocated randomly into a development and a validation dataset. Two estimation methods were established in the development dataset using the two 24-h sodium excretion samples as reference: the 'simple mean method' estimated by multiplying the sodium-creatinine ratio by predicted 24-h creatinine excretion, whereas the 'regression method' employed linear regression analysis. The accuracy of the two methods was examined by comparing the estimated means and concordance correlation coefficients (CCC) in the validation dataset. Mean sodium excretion by the simple mean method with three spot urine samples was closest to that by 24-h collection (difference: -1.62  mmol/day). CCC with the simple mean method increased with an increased number of spot urine samples at 0.20, 0.31, and 0.42 using one, two, and three samples, respectively. This method with three spot urine samples yielded higher CCC than the regression method (0.40). When only one spot urine sample was available for each study participant, CCC was higher with the regression method (0.36). The simple mean method with three spot urine samples yielded the most accurate estimates of sodium excretion. When only one spot urine sample was available, the regression method was preferable.

  10. Performance assessment of urine flow cytometry (UFC) to screen urines to reflex to culture in immunocompetent and immunosuppressed hosts.

    PubMed

    Stefanovic, Aleksandra; Roscoe, Diane; Ranasinghe, Romali; Wong, Titus; Bryce, Elizabeth; Porter, Charlene; Lim, Adelina; Grant, Jennifer; Ng, Karen; Pudek, Morris

    2017-09-01

    Urine flow cytometry (UFC) is an automated method to quantify bacterial and white blood cell (WBC) counts. We aimed to determine whether a threshold for these parameters can be set to use UFC as a sensitive screen to predict which urine samples will subsequently grow in culture. Urines submitted to our microbiology laboratory at a tertiary care centre from 22 July 2015-17 February 2016 underwent UFC (Sysmex UF-1000i) analysis, regular urinalysis and urine culture. Positive urine cultures were defined as growth ≥104 c.f.u. ml-1 of organisms associated with urinary tract infections. The correlation of UFC bacterial and WBC counts with urine culture was assessed using receiver operating characteristics curves. The sensitivity (SN), specificity (SP), negative predictive values (NPVs), positive predictive values (PPVs) and false negative rate (FNR) were calculated at various thresholds in immunocompetent and immunosuppressed patients. A total of 15 046 urine specimens were submitted, of which 14 908 were analysable in the study. The average time to UFC result from receipt in the laboratory was 0.76 h (+/-1.04). The test performance at a set threshold of UFC bacteria ≥20 or WBC >5 was: SN=96.0 %, SP=39.2 %, PPV=47.0 %, NPV=94.5 % and FNR=4.0 %. This threshold eliminates 26 % of urine cultures. Immunosuppressed hosts had a lower sensitivity of 90.6 % and a higher FNR of 9.4 %. UFC is a rapid and sensitive method to screen out urine samples that will subsequently be negative and to reflex urines to culture that will subsequently grow. UFC results are available within 1 h from receipt and enable the elimination of culture when the set threshold is not met.

  11. Development of online NIR urine analyzing system based on AOTF

    NASA Astrophysics Data System (ADS)

    Wan, Feng; Sun, Zhendong; Li, Xiaoxia

    2006-09-01

    In this paper, some key techniques on development of on-line MR urine analyzing system based on AOTF (Acousto - Optics Tunable Filter) are introduced. Problems about designing the optical system including collimation of incident light and working distance (the shortest distance for separating incident light and diffracted light) are analyzed and researched. DDS (Direct Digital Synthesizer) controlled by microprocessor is used to realize the wavelength scan. The experiment results show that this MR urine analyzing system based on. AOTF has 10000 - 4000cm -1 wavelength range and O.3ms wavelength transfer rate. Compare with the conventional Fourier Transform NIP. spectrophotometer for analyzing multi-components in urine, this system features low cost, small volume and on-line measurement function. Unscrambler software (multivariate statistical software by CAMO Inc. Norway) is selected as the software for processing the data. This system can realize on line quantitative analysis of protein, urea and creatinine in urine.

  12. Total insulinlike growth factor 1 and insulinlike growth factor binding protein levels, functional status, and mortality in older adults.

    PubMed

    Kaplan, Robert C; McGinn, Aileen P; Pollak, Michael N; Kuller, Lewis; Strickler, Howard D; Rohan, Thomas E; Xue, XiaoNan; Kritchevsky, Stephen B; Newman, Anne B; Psaty, Bruce M

    2008-04-01

    To assess the association between total insulinlike growth factor (IGF)-1, IGF binding protein-1 (IGFBP-1), and IGFBP-3 levels and functioning and mortality in older adults. Cohort study. One thousand one hundred twenty-two individuals aged 65 and older without prior cardiovascular disease events participating in the Cardiovascular Health Study. Baseline fasting plasma levels of IGF-1, IGFBP-1, and IGFBP-3 (defined as tertiles, T1-T3) were examined in relationship to handgrip strength, time to walk 15 feet, development of new difficulties with activities of daily living (ADLs), and mortality. Higher IGFBP-1 predicted worse handgrip strength (P-trend(T1-T3)<.01) and slower walking speed (P-trend(T1-T3)=.03), lower IGF-1 had a borderline significant association with worse handgrip strength (P-trend(T1-T3)=.06), and better grip strength was observed in the middle IGFBP-3 tertile than in the low or high tertiles (P=.03). Adjusted for age, sex, and race, high IGFBP-1 predicted greater mortality (P-trend(T1-T3)<.001, hazard ratio (HR)(T3vsT1)=1.48, 95% confidence interval (CI)=1.15-1.90); this association was borderline significant after additional confounder adjustment (P-trend(T1-T3)=.05, HR(T3vsT1)=1.35, 95% CI=0.98-1.87). High IGFBP-1 was associated with greater risk of incident ADL difficulties after adjustment for age, sex, race, and other confounders (P-trend(T1-T3)=.04, HR(T3vsT1)=1.40, CI=1.01-1.94). Neither IGF-1 nor IGFBP-3 level predicted mortality or incident ADL difficulties. In adults aged 65 and older, high IGFBP-1 levels were associated with greater risk of mortality and poorer functional ability, whereas IGF-1 and IGFBP-3 had little association with these outcomes.

  13. Sample handling for mass spectrometric proteomic investigations of human urine.

    PubMed

    Petri, Anette Lykke; Høgdall, Claus; Christensen, Ib Jarle; Simonsen, Anja Hviid; T'jampens, Davy; Hellmann, Marja-Leena; Kjaer, Susanne Krüger; Fung, Eric T; Høgdall, Estrid

    2008-09-01

    Because of its non-invasive sample collection method, human urine is an attractive biological material both for discovering biomarkers and for use in future screening trials for different diseases. Before urine can be used for these applications, standardized protocols for sample handling that optimize protein stability are required. In this explorative study, we examine the influence of different urine collection methods, storage temperatures, storage times, and repetitive freeze-thaw procedures on the protein profiles obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Prospectively collected urine samples from 11 women were collected as either morning or midday specimens. The effects of storage temperature, time to freezing, and freeze-thaw cycles were assessed by calculating the number, intensity, and reproducibility of peaks visualized by SELDI-TOF-MS. On the CM10 array, 122 peaks were detected and 28 peaks were found to be significantly different between urine types, storage temperature and time to freezing. On the IMAC-Cu array, 65 peaks were detected and 1 peak was found to be significantly different according to time to freezing. No significant differences were demonstrated for freeze-thaw cycles. Optimal handling and storage conditions are necessary in clinical urine proteomic investigations. Collection of urine with a single and consistently performed protocol is needed to reduce analytical bias. Collecting only one urine type, which is stored for a limited period at 4°C until freezing at -80°C prior to analysis will provide the most stable profiles. Copyright © 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Is specific gravity a good estimate of urine osmolality?

    PubMed

    Imran, Sethi; Eva, Goldwater; Christopher, Shutty; Flynn, Ethan; Henner, David

    2010-01-01

    Urine specific gravity (USG) is often used by clinicians to estimate urine osmolality. USG is measured either by refractometry or by reagent strip. We studied the correlation of USG obtained by either method with a concurrently obtained osmolality. Using our laboratory's records, we retrospectively gathered data on 504 urine specimens on patients on whom a simultaneously drawn USG and an osmolality were available. Out of these, 253 USG's were measured by automated refractometry and 251 USG's were measured by reagent strip. Urinalysis data on these subjects were used to determine the correlation between USG and osmolality, adjusting for other variables that may impact the relationship. The other variables considered were pH, protein, glucose, ketones, nitrates, bilirubin, urobilinogen, hemoglobin, and leukocyte esterase. The relationships were analyzed by linear regression. This study demonstrated that USG obtained by both reagent strip and refractometry had a correlation of approximately 0.75 with urine osmolality. The variables affecting the correlation included pH, ketones, bilirubin, urobilinogen, glucose, and protein for the reagent strip and ketones, bilirubin, and hemoglobin for the refractometry method. At a pH of 7 and with an USG of 1.010 predicted osmolality is approximately 300  mosm/kg/H(2)O for either method. For an increase in SG of 0.010, predicted osmolality increases by 182  mosm/kg/H(2) O for the reagent strip and 203  mosm/kg/H(2)O for refractometry. Pathological urines had significantly poorer correlation between USG and osmolality than "clean" urines. In pathological urines, direct measurement of urine osmolality should be used. © 2010 Wiley-Liss, Inc.

  15. Development of a Targeted Urine Proteome Assay for kidney diseases.

    PubMed

    Cantley, Lloyd G; Colangelo, Christopher M; Stone, Kathryn L; Chung, Lisa; Belcher, Justin; Abbott, Thomas; Cantley, Jennifer L; Williams, Kenneth R; Parikh, Chirag R

    2016-01-01

    Since human urine is the most readily available biofluid whose proteome changes in response to disease, it is a logical sample for identifying protein biomarkers for kidney diseases. Potential biomarkers were identified by using a multiproteomics workflow to compare urine proteomes of kidney transplant patients with immediate and delayed graft function. Differentially expressed proteins were identified, and corresponding stable isotope labeled internal peptide standards were synthesized for scheduled MRM. The Targeted Urine Proteome Assay (TUPA) was then developed by identifying those peptides for which there were at least two transitions for which interference in a urine matrix across 156 MRM runs was <30%. This resulted in an assay that monitors 224 peptides from 167 quantifiable proteins. TUPA opens the way for using a robust mass spectrometric technology, MRM, for quantifying and validating biomarkers from among 167 urinary proteins. This approach, while developed using differentially expressed urinary proteins from patients with delayed versus immediate graft function after kidney transplant, can be expanded to include differentially expressed urinary proteins in multiple kidney diseases. Thus, TUPA could provide a single assay to help diagnose, prognose, and manage many kidney diseases. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Stabilization of source-separated human urine by chemical oxidation.

    PubMed

    Zhang, Yang; Li, Zifu; Zhao, Yuan; Chen, Shuangling; Mahmood, Ibrahim Babatunde

    2013-01-01

    The inhibitory effect of ozone and hydrogen peroxide (HP) on urea hydrolysis in stored urine was investigated and compared. Ozone showed less effect on urea hydrolysis due to the complicated composition of urine (including a large amount of urease-producing bacteria) and bacteria regeneration. Ozone concentration and total heterotrophic bacteria analysis demonstrated that residual ozone concentration decreased by 43% within 15 hr from 13.50 to 7.72 mg/L in the one-time ozonation urine test, and finally completely decomposed within 4 days. In addition, bacteria regenerated quickly after ozone completely decomposed. However, HP showed a significant effect on inhibiting urea hydrolysis not only in stored urine but also in fecal-contaminated urine. The suitable doses of applied HP to inhibit urea hydrolysis in stored urine, concentrations of 0.5 and 1.0 g feces per liter of fecal-contaminated urine, were 0.03, 0.16 and 0.23 mol/L, respectively. The urea concentrations after 2 months stored were 7,145, 7,109 and 7,234 mg/L, respectively.

  17. Validation of a urine color scale for assessment of urine osmolality in healthy children.

    PubMed

    Kavouras, Stavros A; Johnson, Evan C; Bougatsas, Dimitris; Arnaoutis, Giannis; Panagiotakos, Demosthenes B; Perrier, Erica; Klein, Alexis

    2016-04-01

    Urine color (UC) is a practical tool for hydration assessment. The technique has been validated in adults, but has not been tested in children. The purpose of the study was to test the validity of the urine color scale in young, healthy boys and girls, as a marker of urine concentration, investigate its diagnostic ability of detecting hypohydration and examine the ability of children to self-assess UC. A total of 210 children participated (age: 8-14 years, body mass: 43.4 ± 12.6 kg, height: 1.49 ± 0.13 m, body fat: 25.2 ± 7.8 %). Data collection included: two single urine samples (first morning and before lunch) and 24-h sampling. Hydration status was assessed via urine osmolality (UOsmo) and UC via the eight-point color scale. Mean UC was 3 ± 1 and UOsmo 686 ± 223 mmol kg(-1). UC displayed a positive relationship as a predictor of UOsmo (R (2): 0.45, P < 0.001). Based on the receiver operating curve, UC has good overall classification ability for the three samples (area under the curve 85-92 %), with good sensitivity (92-98 %) and specificity (55-68 %) for detecting hypohydration. The overall accuracy of the self-assessment of UC in the morning or the noon samples ranged from 67 to 78 %. Further threshold analysis indicated that the optimal self-assessed UC threshold for hypohydration was ≥4. The classical eight-point urine color scale is a valid method to assess hydration in children of age 8-14 years, either by researchers or self-assessment.

  18. Evaluation of plant and animal protein sources as partial or total replacement of fish meal in diets for juvenile Nile tilapia

    USDA-ARS?s Scientific Manuscript database

    A feeding trial was conducted in a closed system with Nile tilapia (Oreochromis niloticus) juveniles (mean weight, 2.84 g) to examine the effects of total replacement of fish meal (FM), with and without supplementation of DL-methionine (Met) and L-lysine (Lys), by plant protein sources. Fish were f...

  19. Urine 24-hour volume

    MedlinePlus

    ... a day, such as: Creatinine Sodium Potassium Urea nitrogen Protein This test may also be done if ... 2016:chap 114. Verbalis JG. Disorders of water balance. In: Skorecki K, Chertow GM, Marsden PA, Taal ...

  20. The effects of urine concentration, and cushion centrifugation to remove urine, on the quality of cool-stored stallion sperm.

    PubMed

    Voge, Jared; Varner, Dickson D; Blanchard, Terry L; Meschini, Marika; Turner, Carly; Teague, Sheila R; Brinsko, Steven P; Love, Charles C

    2016-09-15

    Urine-contaminated stallion semen is a clinical problem due to a variety of causes. The effect of the level of urine contamination on the longevity of sperm quality has not been evaluated. The aim of this study was to determine the effects of urine concentration level (0%, 10%, 20%, 30%, and 40%) and cushioned centrifugation and resuspension of the sperm pellet in fresh extender, on measures of sperm quality, immediately after semen collection (T0), after 1 hour of storage at room temperature (T1), and after 24 hours of cooled storage (T24). In general, most sperm quality measures declined with increasing urine concentration starting at T0. Cushioned centrifugation (CC), but not simple dilution, generally maintained sperm quality at T24 as compared with T1. At T24, total sperm motility was higher in all urine-contaminated CC samples compared with uncentrifuged samples (P < 0.05); sperm viability was lower in CC than uncentrifuged at a urine concentration of 20%, but higher at 30% and 40% (P < 0.05); and DNA quality was decreased (higher % cells outside the main population) in all urine concentrations (P < 0.05). Immediate extension in semen extender, followed by cushioned centrifugation and resuspension of the sperm pellet in fresh extender, provided the best option for preserving sperm quality of urospermic semen. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Triiodothyronine and thyroxine in urine. I. Measurement and application.

    PubMed

    Shakespear, R A; Burke, C W

    1976-03-01

    Urinary triiodothyronine (T3) and thyroxine (T4) were measured by RIA, and T4 was also measured by competitive protein binding (CPB). pH 1-hydrolysable conjugates were 48% of total urinary T3, and enzyme- or pH 1-hydrolysable conjugates were 55% and 61% of total urinary T4. The mean unconjugated T3 excretion was 34.3 ng/h (0.99 mug T3/g creatinine) in normal subjects (no day-night rhythm found), 1.56 mug/g in late pregnancy, 0.82 mug/g in neonates (1-12 days), and was also unchanged in persons with high or low thyroxine-binding globulin (TBG). In thyrotoxicosis, mean T3 excretion was 281 ng/h, no values being in the normal range. In primary hypothyroidism it was 18.3 ng/h, but over half the values were in the normal range. The mean urinary unconjugated T4 was 82.2 ng/h (1.37 mug T4/g creatinine) in normal subjects, 1.6 mug/g in neonates, and unchanged in persons with high or low TBG, except that in pregnancy high values were compatible with increases protein excretion. Apparently increased day-time T4 excretion compared with night-time excretion may also be due to changes in protein excretion rate. The mean T4 in thyrotoxicosis was 337 ng/h (12% of values in the normal range) and 32.8 ng/h in primary hypothyroidism (over half the normal range). All the assays, especially that of T4 by CPB gave readings which were incorrect with protein concentrations above 100 mg/l. Urinary T3 and T4 assays for clinical purposes have few practical advantages over serum assays, despite the relationship of urine T3 and T4 to serum unbound levels.

  2. Estimation of Serum Triglycerides, Serum Cholesterol, Total Protein, IgG Levels in Chronic Periodontitis Affected Elderly Patients: A Cross-Sectional Study

    PubMed Central

    Saravanan, A. V.; Ravishankar, P. L.; Kumar, Pradeep; Rajapandian, K.; Kalaivani, V.; Rajula, M. Prem Blaisie

    2017-01-01

    Aim: The present study was conducted to evaluate the serum triglycerides, serum cholesterol, total protein, and IgG levels in elderly patients who were affected by periodontal disease. Materials and Methods: This study was conducted at the Rajah Muthiah Dental College and Hospital in the periodontics division. The study was conducted for a period of 3 months. This study is a prospective analytical study. Sixty individuals who were systemically healthy in the age group of 50 and above were included in this study. Control and experimental groups of 30 participants each were included. Plaque index, gingival index, probing pocket depth, and clinical attachment loss were recorded. Biochemical parameters such as serum cholesterol, serum triglycerides, total protein, and IgG levels were also evaluated and correlated with the periodontal parameters. Data was analyzed using SPSS version 16.0 (IBM Corp., Armonk, NY). The relationship between periodontal status and the biochemical parameters such as serum cholesterol, serum triglycerides, total protein, and IgG levels were evaluated by Student's t-test. Results: There was no significant difference in the plaque and gingival scores between the experimental and control group. It was observed that serum cholesterol level and total protein level was lower in participants suffering from chronic periodontitis. Triglycerides level was significantly elevated in the experimental group. IgG, a level which is not significant, concluded that there is no difference in control and experimental group. Conclusion: It was concluded from the results obtained from the study that there is an association between serum triglycerides, serum cholesterol, total protein, and periodontal disease. However, further longitudinal and well-controlled studies are required to evaluate the relationship between these biochemical parameters and periodontal disease. PMID:28462181

  3. Estimation of Serum Triglycerides, Serum Cholesterol, Total Protein, IgG Levels in Chronic Periodontitis Affected Elderly Patients: A Cross-Sectional Study.

    PubMed

    Saravanan, A V; Ravishankar, P L; Kumar, Pradeep; Rajapandian, K; Kalaivani, V; Rajula, M Prem Blaisie

    2017-01-01

    The present study was conducted to evaluate the serum triglycerides, serum cholesterol, total protein, and IgG levels in elderly patients who were affected by periodontal disease. This study was conducted at the Rajah Muthiah Dental College and Hospital in the periodontics division. The study was conducted for a period of 3 months. This study is a prospective analytical study. Sixty individuals who were systemically healthy in the age group of 50 and above were included in this study. Control and experimental groups of 30 participants each were included. Plaque index, gingival index, probing pocket depth, and clinical attachment loss were recorded. Biochemical parameters such as serum cholesterol, serum triglycerides, total protein, and IgG levels were also evaluated and correlated with the periodontal parameters. Data was analyzed using SPSS version 16.0 (IBM Corp., Armonk, NY). The relationship between periodontal status and the biochemical parameters such as serum cholesterol, serum triglycerides, total protein, and IgG levels were evaluated by Student's t-test. There was no significant difference in the plaque and gingival scores between the experimental and control group. It was observed that serum cholesterol level and total protein level was lower in participants suffering from chronic periodontitis. Triglycerides level was significantly elevated in the experimental group. IgG, a level which is not significant, concluded that there is no difference in control and experimental group. It was concluded from the results obtained from the study that there is an association between serum triglycerides, serum cholesterol, total protein, and periodontal disease. However, further longitudinal and well-controlled studies are required to evaluate the relationship between these biochemical parameters and periodontal disease.

  4. Serum Markers of Neurodegeneration in Maple Syrup Urine Disease.

    PubMed

    Scaini, Giselli; Tonon, Tássia; de Souza, Carolina F Moura; Schuk, Patricia F; Ferreira, Gustavo C; Neto, Joao Seda; Amorin, Tatiana; Schwartz, Ida Vanessa D; Streck, Emilio L

    2017-09-01

    Maple syrup urine disease (MSUD) is an inherited disorder caused by deficient activity of the branched-chain α-keto acid dehydrogenase complex involved in the degradation pathway of branched-chain amino acids (BCAAs) and their respective α-keto-acids. Patients affected by MSUD present severe neurological symptoms and brain abnormalities, whose pathophysiology is poorly known. However, preclinical studies have suggested alterations in markers involved with neurodegeneration. Because there are no studies in the literature that report the neurodegenerative markers in MSUD patients, the present study evaluated neurodegenerative markers (brain-derived neurotrophic factor (BDNF), cathepsin D, neural cell adhesion molecule (NCAM), plasminogen activator inhibitor-1 total (PAI-1 (total)), platelet-derived growth factor AA (PDGF-AA), PDGF-AB/BB) in plasma from 10 MSUD patients during dietary treatment. Our results showed a significant decrease in BDNF and PDGF-AA levels in MSUD patients. On the other hand, NCAM and cathepsin D levels were significantly greater in MSUD patients compared to the control group, while no significant changes were observed in the levels of PAI-1 (total) and PDGF-AB/BB between the control and MSUD groups. Our data show that MSUD patients present alterations in proteins involved in the neurodegenerative process. Thus, the present findings corroborate previous studies that demonstrated that neurotrophic factors and lysosomal proteases may contribute, along with other mechanisms, to the intellectual deficit and neurodegeneration observed in MSUD.

  5. Treating urine by Spirulina platensis

    NASA Astrophysics Data System (ADS)

    Yang, Chenliang; Liu, Hong; Li, Ming; Yu, Chengying; Yu, Gurevich

    In this paper Spirulina platensis with relatively high nutrition was cultivated to treat human urine. Batch culture showed that the consumption of N in human urine could reach to 99%, and the consumption of P was more than 99.9%, and 1.05 g biomass was obtained by treating 12.5 ml synthetic human urine; continuous culture showed that S. platensis could consume N, Cl, K and S in human urine effectively, and the consumption could reach to 99.9%, 75.0%, 83.7% and 96.0%, respectively, and the consumption of P was over 99.9%, which is very important to increase the closure and safety of the bioregenerative life support system (BLSS).

  6. Urine collection apparatus. [feminine hygiene

    NASA Technical Reports Server (NTRS)

    Michaud, R. B. (Inventor)

    1981-01-01

    A urine collection device for females comprises an interface body with an interface surface for engagement with the user's body. The interface body comprises a forward portion defining a urine-receiving bore which has an inlet in the interface surface adapted to be disposed in surrounding relation to the urethral opening of the user. The interface body also has a rear portion integrally adjoining the forward portion and a non-invasive vaginal seal on the interface surface for sealing the vagina of the user from communication with the urine-receiving bore. An absorbent pad is removably supported on the interface body and extends laterally therefrom. A garment for supporting the urine collection is also disclosed.

  7. Optimizing total reflection X-ray fluorescence for direct trace element quantification in proteins I: Influence of sample homogeneity and reflector type

    NASA Astrophysics Data System (ADS)

    Wellenreuther, G.; Fittschen, U. E. A.; Achard, M. E. S.; Faust, A.; Kreplin, X.; Meyer-Klaucke, W.

    2008-12-01

    Total reflection X-ray fluorescence (TXRF) is a very promising method for the direct, quick and reliable multi-elemental quantification of trace elements in protein samples. With the introduction of an internal standard consisting of two reference elements, scandium and gallium, a wide range of proteins can be analyzed, regardless of their salt content, buffer composition, additives and amino acid composition. This strategy also enables quantification of matrix effects. Two potential issues associated with drying have been considered in this study: (1) Formation of heterogeneous residues of varying thickness and/or density; and (2) separation of the internal standard and protein during drying (which has to be prevented to allow accurate quantification). These issues were investigated by microbeam X-ray fluorescence (μXRF) with special emphasis on (I) the influence of sample support and (II) the protein / buffer system used. In the first part, a model protein was studied on well established sample supports used in TXRF, PIXE and XRF (Mylar, siliconized quartz, Plexiglas and silicon). In the second part we imaged proteins of different molecular weight, oligomerization state, bound metals and solubility. A partial separation of protein and internal standard was only observed with untreated silicon, suggesting it may not be an adequate support material. Siliconized quartz proved to be the least prone to heterogeneous drying of the sample and yielded the most reliable results.

  8. A urine volume measurement system

    NASA Technical Reports Server (NTRS)

    Poppendiek, H. F.; Mouritzen, G.; Sabin, C. M.

    1972-01-01

    An improved urine volume measurement system for use in the unusual environment of manned space flight is reported. The system utilizes a low time-constant thermal flowmeter. The time integral of the transient response of the flowmeter gives the urine volume during a void as it occurs. In addition, the two phase flows through the flowmeter present no problem. Developments of the thermal flowmeter and a verification of the predicted performance characteristics are summarized.

  9. Uncertainties of Mayak urine data

    SciTech Connect

    Miller, Guthrie; Vostrotin, Vadim; Vvdensky, Vladimir

    2008-01-01

    For internal dose calculations for the Mayak worker epidemiological study, quantitative estimates of uncertainty of the urine measurements are necessary. Some of the data consist of measurements of 24h urine excretion on successive days (e.g. 3 or 4 days). In a recent publication, dose calculations were done where the uncertainty of the urine measurements was estimated starting from the statistical standard deviation of these replicate mesurements. This approach is straightforward and accurate when the number of replicate measurements is large, however, a Monte Carlo study showed it to be problematic for the actual number of replicate measurements (median from 3more » to 4). Also, it is sometimes important to characterize the uncertainty of a single urine measurement. Therefore this alternate method has been developed. A method of parameterizing the uncertainty of Mayak urine bioassay measmements is described. The Poisson lognormal model is assumed and data from 63 cases (1099 urine measurements in all) are used to empirically determine the lognormal normalization uncertainty, given the measurement uncertainties obtained from count quantities. The natural logarithm of the geometric standard deviation of the normalization uncertainty is found to be in the range 0.31 to 0.35 including a measurement component estimated to be 0.2.« less

  10. Using human urine as food for cyanobacteria in LSS

    NASA Astrophysics Data System (ADS)

    Kalacheva, Galina; Gribovskaya, Iliada; Kolmakova, Angela

    In biological LSS: human, higher plants, algae, united by common cycle of matter, native human urine is the most problematic substance for using in inter-link exchange. It contains urea, ammonium compounds and up to 10 g/l of NaCl. Each of the mentioned components is toxic for growing higher plants. As for inferior plants, experiments showed that cyanobacteria of genus Spirulina platensis and similar genus Oscillatoria deflexa can grow at NaCl concentrations up to 20 g/l and NH4Cl concentrations up to 800 mg/l. These cyanobacteria can be used in LSS as a photosynthesizing link. Besides, S. platensis is edible for humans and fish. To use urine as food for algae, it is necessary to remove urea and organics. All previously used methods for urine treatment aimed at urea destruction: heating to 300oC, ultraviolet exposure, freezing, oxidation on reactor with hydrogen peroxide, had no effect. We used the following method of urine treatment: urine evaporation till dry residue, subsequent combustion in muffle furnace at 450-500oC and creation of ash water extract of the same volume as the initial urine. Comparison of standard Zarrouk's solution for S. platensis and O. deflexa with the water extract of urine ash showed that the concentrations of K, Ca, Mg, P, S were similar. Successful experiments were made with O. deflexa that were grown on nutrient solution made of the water extract of urine ash with 10 g/l of NaHCO3 and 2 g/l of NaNO3. The sources of intersystem production of HCO3 and NO3 were shown, and the biochemical composition of the investigated algae species, including mineral composition, protein, carbohydrate, amino acid, lipid and vitamin content were studied.

  11. A new mutation in the AFP gene responsible for a total absence of alpha feto-protein on second trimester maternal serum screening for Down syndrome

    PubMed Central

    Petit, François M; Hébert, Marylise; Picone, Olivier; Brisset, Sophie; Maurin, Marie-Laure; Parisot, Frédéric; Capel, Liliane; Benattar, Clarisse; Sénat, Marie-Victoire; Tachdjian, Gérard; Labrune, Philippe

    2009-01-01

    Alpha feto-protein (AFP) is a major plasma protein produced by the yolk sac and the liver during the fetal period. During the second trimester of pregnancy, APF and βhCG serum concentrations are commonly used for screening Down syndrome. AFP deficiency is rare (estimated to be 1/105 000 newborns) and only one sequence alteration has previously been reported in the AFP gene. We report a new mutation in exon 5 of the AFP gene, leading to a total absence of AFP on 2nd-trimester maternal serum screening for Down syndrome, confirmed on the amniotic fluid. Despite this, fetal development and birth were normal. After PCR-amplification, the whole AFP gene was sequenced. The new mutation was a guanine to adenine transition in position 543 creating a premature stop codon in position 181. In order to search for eventual modifications of the amniotic fluid profile, proteins were separated by electrophoresis and compared with 10 normal amniotic fluids sampled at the same developmental age (18 weeks). In the amniotic fluid of our patient albumin rate was reduced whereas alpha1 and beta protein fractions were increased, suggesting that AFP deficiency may modify the distribution of protein fractions. This observation emphasizes the complex molecular mechanisms of compensation of serum protein deficiency. Studies on other families with AFP deficiency are necessary to confirm this observation. PMID:18854864

  12. Estimating residual kidney function in dialysis patients without urine collection

    PubMed Central

    Shafi, Tariq; Michels, Wieneke M.; Levey, Andrew S.; Inker, Lesley A.; Dekker, Friedo W.; Krediet, Raymond T.; Hoekstra, Tiny; Schwartz, George J.; Eckfeldt, John H.; Coresh, Josef

    2016-01-01

    Residual kidney function contributes substantially to solute clearance in dialysis patients but cannot be assessed without urine collection. We used serum filtration markers to develop dialysis-specific equations to estimate urinary urea clearance without the need for urine collection. In our development cohort, we measured 24-hour urine clearances under close supervision in 44 patients and validated these equations in 826 patients from the Netherlands Cooperative Study on the Adequacy of Dialysis. For the development and validation cohorts, median urinary urea clearance was 2.6 and 2.4 mL/min, respectively. During the 24-hour visit in the development cohort, serum β-trace protein concentrations remained in steady state but concentrations of all other markers increased. In the validation cohort, bias (median measured minus estimated clearance) was low for all equations. Precision was significantly better for β-trace protein and β2-microglobulin equations and the accuracy was significantly greater for β-trace protein, β2-microglobulin and cystatin C equations, compared with the urea plus creatinine equation. Area under the receiver operator characteristic curve for detecting measured urinary urea clearance by equation-estimated urinary urea clearance (both 2 mL/min or more) were 0.821, 0.850 and 0.796 for β-trace protein, β2-microglobulin and cystatin C equations, respectively; significantly greater than the 0.663 for the urea plus creatinine equation. Thus, residual renal function can be estimated in dialysis patients without urine collections. PMID:26924062

  13. Comparison of urine analysis using manual and sedimentation methods.

    PubMed

    Kurup, R; Leich, M

    2012-06-01

    Microscopic examination of urine sediment is an essential part in the evaluation of renal and urinary tract diseases. Traditionally, urine sediments are assessed by microscopic examination of centrifuged urine. However the current method used by the Georgetown Public Hospital Corporation Medical Laboratory involves uncentrifuged urine. To encourage high level of care, the results provided to the physician must be accurate and reliable for proper diagnosis. The aim of this study is to determine whether the centrifuge method is more clinically significant than the uncentrifuged method. In this study, a comparison between the results obtained from centrifuged and uncentrifuged methods were performed. A total of 167 urine samples were randomly collected and analysed during the period April-May 2010 at the Medical Laboratory, Georgetown Public Hospital Corporation. The urine samples were first analysed microscopically by the uncentrifuged, and then by the centrifuged method. The results obtained from both methods were recorded in a log book. These results were then entered into a database created in Microsoft Excel, and analysed for differences and similarities using this application. Analysis was further done in SPSS software to compare the results using Pearson ' correlation. When compared using Pearson's correlation coefficient analysis, both methods showed a good correlation between urinary sediments with the exception of white bloods cells. The centrifuged method had a slightly higher identification rate for all of the parameters. There is substantial agreement between the centrifuged and uncentrifuged methods. However the uncentrifuged method provides for a rapid turnaround time.

  14. A comparison of the effects of 2 doses of soy protein or casein on serum lipids, serum lipoproteins, and plasma total homocysteine in hypercholesterolemic subjects.

    PubMed

    Tonstad, Serena; Smerud, Knut; Høie, Lars

    2002-07-01

    Studies have shown that soy protein reduces some atherogenic lipid and lipoprotein concentrations, although lipoprotein(a) concentrations may be increased. The dose response of soy protein has not been established; neither has its effect on plasma total homocysteine. Our objective was to evaluate the effect of 2 doses of soy protein on lipid, lipoprotein, and homocysteine concentrations. Four to 24 wk after being instructed to consume a lipid-lowering diet, 130 men and women with LDL-cholesterol concentrations > or = 4 mmol/L were studied during a parallel group trial in which 4 interventions were assigned randomly. Thirty grams isolated soy protein (ISP) and 10 g cotyledon fiber or 50 g ISP and 16.6 g cotyledon fiber or equivalent doses of casein and cellulose were consumed daily as a beverage for 16 wk. When the 2 groups who consumed ISP were compared with the 2 groups who consumed casein, the differences in the net changes from baseline to week 16 in the concentrations of LDL cholesterol and plasma total homocysteine were -0.26 mmol/L (95% CI: -0.43, -0.09 mmol/L; P = 0.01) and -0.8 micromol/L (-1.4, -0.2 micromol/L; P = 0.005), respectively. The effect of the ISP dose was not significant. There were no significant differences between the 2 ISP and the 2 casein groups in changes in lipoprotein(a), HDL-cholesterol, or triacylglycerol concentrations. Adding 30-50 g soy protein/d to a lipid-lowering diet significantly reduced LDL-cholesterol concentrations without increasing lipoprotein(a) concentrations. Plasma total homocysteine concentrations also decreased, suggesting a novel, possibly antiatherosclerotic effect.

  15. Cleaved Form of Osteopontin in Urine as a Clinical Marker of Lupus Nephritis

    PubMed Central

    Kitagori, Koji; Yoshifuji, Hajime; Oku, Takuma; Sasaki, Chiyomi; Miyata, Hitomi; Mori, Keita P.; Nakajima, Toshiki; Ohmura, Koichiro; Kawabata, Daisuke; Yukawa, Naoichiro; Imura, Yoshitaka; Murakami, Kosaku; Nakashima, Ran; Usui, Takashi; Fujii, Takao; Sakai, Kaoru; Yanagita, Motoko; Hirayama, Yoshitaka; Mimori, Tsuneyo

    2016-01-01

    We assessed the utility of two forms of osteopontin (OPN), OPN full and its cleaved form (OPN N-half), in plasma and urine as markers of disease activity in lupus nephritis (LN). Samples were collected from patients with systemic lupus erythematosus (SLE) (LN: N = 29, non-LN: N = 27), IgA nephropathy (IgAN) (N = 14), minimal change nephrotic syndrome (MCNS) (N = 5), diabetic nephropathy (DN) (N = 14) and healthy volunteers (HC) (N = 17). While there was no significant difference in urine OPN full concentration between groups, urine OPN N-half concentration was significantly higher in patients with LN than HC (p < 0.05). Moreover, urine OPN N-half was higher in LN patients with overt proteinuria (urine protein/creatinine ratio: P/C > 0.5) than LN patients with minimal proteinuria (P/C < 0.5, p < 0.0001), and also higher than in DN patients with overt proteinuria (P/C > 0.5, p < 0.01). Urine thrombin activity correlated with urine OPN N-half concentration (p < 0.0001), but not with urine OPN full concentration. These results suggest that urine OPN N-half concentration reflects renal inflammation. Thus, urine OPN N-half may be a novel disease activity marker for LN. PMID:27992535

  16. Current state of the art for enhancing urine biomarker discovery

    PubMed Central

    Harpole, Michael; Davis, Justin; Espina, Virginia

    2016-01-01

    Urine is a highly desirable biospecimen for biomarker analysis because it can be collected recurrently by non-invasive techniques, in relatively large volumes. Urine contains cellular elements, biochemicals, and proteins derived from glomerular filtration of plasma, renal tubule excretion, and urogenital tract secretions that reflect, at a given time point, an individual's metabolic and pathophysiologic state. High-resolution mass spectrometry, coupled with state of the art fractionation systems are revealing the plethora of diagnostic/prognostic proteomic information existing within urinary exosomes, glycoproteins, and proteins. Affinity capture pre-processing techniques such as combinatorial peptide ligand libraries and biomarker harvesting hydrogel nanoparticles are enabling measurement/identification of previously undetectable urinary proteins. Future challenges in the urinary proteomics field include a) defining either single or multiple, universally applicable data normalization methods for comparing results within and between individual patients/data sets, and b) defining expected urinary protein levels in healthy individuals. PMID:27232439

  17. Metabolomics Profiling of Serum and Urine in Three Beef Cattle Breeds Revealed Different Levels of Tolerance to Heat Stress.

    PubMed

    Liao, Yupeng; Hu, Rui; Wang, Zhisheng; Peng, Quanhui; Dong, Xianwen; Zhang, Xiangfei; Zou, Huawei; Pu, Qijian; Xue, Bai; Wang, Lizhi

    2018-06-25

    This study was to determine differences in the global metabolic profiles of serum and urine of Xuanhan yellow cattle, Simmental crossbred cattle (Simmental × Xuanhan yellow cattle), and cattle-yaks (Jersey × Maiwa yak) under heat stress (temperature-humidity index remained above 80 for 1 week). A total of 55 different metabolites associated with the three breeds were identified in the serum and urine samples by gas chromatography-mass spectrometry. The metabolic adaptations to heat stress are heterogeneous. Cattle-yaks mobilize a greater amount of body protein to release glucogenic amino acids to supply energy, whereas the tricarboxylic acid cycle is inhibited. Simmental crossbred cattle mobilize a greater amount of body fat to use free fatty acids as an energy source. In comparison with Simmental crossbred cattle and cattle-yaks, Xuanhan yellow cattle have higher glycolytic activity and possess a stronger antioxidant defense system and are, in conclusion, more adapted to hot and humid environments.

  18. Urine therapy through the centuries.

    PubMed

    Savica, Vincenzo; Calò, Lorenzo A; Santoro, Domenico; Monardo, Paolo; Mallamace, Agostino; Bellinghieri, Guido

    2011-01-01

    Urine has always interested and attracted the attention of people. It was in fact never considered a waste product of the body but rather as a distilled product selected from the blood and containing useful substances for the care of the body. It was referred to as the "gold of the blood" and "elixir of long life," indicating its therapeutic potential. This paper reports on the practice of urine therapy since its origin attributed to the Indian culture, and briefly reviews its use through the centuries and different cultures and traditions. Records from the Egyptians to Jews, Greeks, Romans and from the Middle Ages and the Renaissance testify to the practice of urine therapy--a practice that continues to be found in more recent times, from the 18th century to the present. Experiences with the practice of urine therapy have even been discussed and shared recently in 2 different conferences: in 1996 in India and in 1999 in Germany, where people from different countries shared and presented their own research on urine therapy.

  19. Creating a urine black hole

    NASA Astrophysics Data System (ADS)

    Hurd, Randy; Pan, Zhao; Meritt, Andrew; Belden, Jesse; Truscott, Tadd

    2015-11-01

    Since the mid-nineteenth century, both enlisted and fashion-conscious owners of khaki trousers have been plagued by undesired speckle patterns resulting from splash-back while urinating. In recent years, industrial designers and hygiene-driven entrepreneurs have sought to limit this splashing by creating urinal inserts, with the effectiveness of their inventions varying drastically. From this large assortment of inserts, designs consisting of macroscopic pillar arrays seem to be the most effective splash suppressers. Interestingly this design partially mimics the geometry of the water capturing moss Syntrichia caninervis, which exhibits a notable ability to suppress splash and quickly absorb water from impacting rain droplets. With this natural splash suppressor in mind, we search for the ideal urine black hole by performing experiments of simulated urine streams (water droplet streams) impacting macroscopic pillar arrays with varying parameters including pillar height and spacing, draining and material properties. We propose improved urinal insert designs based on our experimental data in hopes of reducing potential embarrassment inherent in wearing khakis.

  20. Screening for urinary tract infection with the Sysmex UF-1000i urine flow cytometer.

    PubMed

    Broeren, Maarten A C; Bahçeci, Semiha; Vader, Huib L; Arents, Niek L A

    2011-03-01

    The diagnosis of urinary tract infection (UTI) by urine culture is time-consuming and can produce up to 60 to 80% negative results. Fast screening methods that can reduce the necessity for urine cultures will have a large impact on overall turnaround time and laboratory economics. We have evaluated the detection of bacteria and leukocytes by a new urine analyzer, the UF-1000i, to identify negative urine samples that can be excluded from urine culture. In total, 1,577 urine samples were analyzed and compared to urine culture. Urine culture showed growth of ≥10(3) CFU/ml in 939 samples (60%). Receiver operating characteristics (ROC) curves and ROC decision plots were been prepared at three different gold standard definitions of a negative urine culture: no growth, growth of bacteria at <10(4) CFU/ml, and growth of bacteria at <10(5) CFU/ml. Also, the reduction in urine cultures and the percentage of false negatives were calculated. At the most stringent gold standard definition of no growth, a chosen sensitivity of 95% resulted in a cutoff value of 26 bacteria/μl, a specificity of 43% and a reduction in urine cultures of only 20%, of which 14% were false negatives. However, at a gold standard definition of <10(5) CFU/ml and a sensitivity of 95%, the UF-1000i cutoff value was 230 bacteria/μl, the specificity was 80%, and the reduction in urine cultures was 52%, of which 0.3% were false negatives. The applicability of the UF-1000i to screen for negative urine samples strongly depends on population characteristics and the definition of a negative urine culture. In our setting, however, the low workload savings and the high percentage of false-negative results do not warrant the UF-1000i to be used as a screening analyzer.

  1. Estimating population salt intake in India using spot urine samples.

    PubMed

    Petersen, Kristina S; Johnson, Claire; Mohan, Sailesh; Rogers, Kris; Shivashankar, Roopa; Thout, Sudhir Raj; Gupta, Priti; He, Feng J; MacGregor, Graham A; Webster, Jacqui; Santos, Joseph Alvin; Krishnan, Anand; Maulik, Pallab K; Reddy, K Srinath; Gupta, Ruby; Prabhakaran, Dorairaj; Neal, Bruce

    2017-11-01

    To compare estimates of mean population salt intake in North and South India derived from spot urine samples versus 24-h urine collections. In a cross-sectional survey, participants were sampled from slum, urban and rural communities in North and in South India. Participants provided 24-h urine collections, and random morning spot urine samples. Salt intake was estimated from the spot urine samples using a series of established estimating equations. Salt intake data from the 24-h urine collections and spot urine equations were weighted to provide estimates of salt intake for Delhi and Haryana, and Andhra Pradesh. A total of 957 individuals provided a complete 24-h urine collection and a spot urine sample. Weighted mean salt intake based on the 24-h urine collection, was 8.59 (95% confidence interval 7.73-9.45) and 9.46 g/day (8.95-9.96) in Delhi and Haryana, and Andhra Pradesh, respectively. Corresponding estimates based on the Tanaka equation [9.04 (8.63-9.45) and 9.79 g/day (9.62-9.96) for Delhi and Haryana, and Andhra Pradesh, respectively], the Mage equation [8.80 (7.67-9.94) and 10.19 g/day (95% CI 9.59-10.79)], the INTERSALT equation [7.99 (7.61-8.37) and 8.64 g/day (8.04-9.23)] and the INTERSALT equation with potassium [8.13 (7.74-8.52) and 8.81 g/day (8.16-9.46)] were all within 1 g/day of the estimate based upon 24-h collections. For the Toft equation, estimates were 1-2 g/day higher [9.94 (9.24-10.64) and 10.69 g/day (9.44-11.93)] and for the Kawasaki equation they were 3-4 g/day higher [12.14 (11.30-12.97) and 13.64 g/day (13.15-14.12)]. In urban and rural areas in North and South India, most spot urine-based equations provided reasonable estimates of mean population salt intake. Equations that did not provide good estimates may have failed because specimen collection was not aligned with the original method.

  2. Analyte variations in consecutive 24-hour urine collections in children.

    PubMed

    Ellison, Jonathan S; Hollingsworth, John M; Langman, Craig B; Asplin, John R; Schwaderer, Andrew L; Yan, Phyllis; Bierlein, Maggie; Barraza, Mark A; Defoor, William R; Figueroa, T Ernesto; Jackson, Elizabeth C; Jayanthi, Venkata R; Johnson, Emilie K; Joseph, David B; Shnorhavorian, Margarett

    2017-12-01

    The metabolic evaluation of children with nephrolithiasis begins with a 24-h urine collection. For adults, the diagnostic yield increases with consecutive collections; however, little is known regarding the variability of multiple 24-h studies in the pediatric population. We sought to evaluate the variability of consecutive 24-h urine collection in children through a multi-institutional study hypothesizing that compared with a single collection, consecutive 24-h urine collections would reveal a greater degree of clinically useful information in the evaluation of children at risk for nephrolithiasis. Including data from six institutions, we identified children less than 18 years of age considered at risk for recurrent nephrolithiasis, undergoing metabolic evaluation. We evaluated a subset of patients performing two collections with urine creatinine varying by 10% or less during a 7-day period. Discordance between repeat collections based on normative urine chemistry values was evaluated. A total of 733 children met inclusion criteria, and in over a third both urine calcium and urine volume differed by 30% or more between samples. Urine oxalate demonstrated greater variation between collections in children <5 years than among older children (p = 0.030) while variation in other parameters did not differ by age. Discordance between repeat samples based on normative values was most common for urine oxalate (22.5%) and the derived relative supersaturation ratios for both calcium phosphate (25.1%) and calcium oxalate (20.5%). The proportion of discordant samples, based on normative thresholds, as well as variability greater ≥30% and 50%, respectively, are shown in the table. Our analysis indicates that stone risk in as many as one in four children may be misclassified if normative values of only a single 24-h urine are used. In light of these findings, repeat 24-h urine collections prior to targeted intervention to modify stone risk are advised to increase

  3. Asymptomatic bacteriuria in pregnant women attending Boo-Ali Hospital Tehran Iran: Urine analysis vs. urine culture.

    PubMed

    Etminan-Bakhsh, Mina; Tadi, Sima; Darabi, Roksana

    2017-11-01

    Asymptomatic bacteriuria is one of the common problems in pregnancy. Asymptomatic bacteriuria is associated with pyelonephritis, preterm labor and low birth weight infants. The physiological and anatomical changes in pregnancy facilitate urinary tract infection (UTI) during pregnancy. Several tests are available for diagnosis of asymptomatic bacteriuria. The urine culture is a gold standard diagnostic test for asymptomatic bacteriuria but it is expensive and time-consuming. Screening methods may be useful in detecting high-risk pregnant women for asymptomatic bacteriuria. The aim of the present study was to compare urine analysis as a rapid screening test to urine culture in diagnosis of asymptomatic bacteriuria. A total of 123 pregnant women attending the obstetrics clinic of Boo-Ali hospital in Tehran, Iran from March 2013 to September 2014 were included in the present diagnostic cross-sectional study. One hundred twenty three mid-stream urine samples were inoculated into cultures and were processed by dipstick (nitrite test and leucocyte esterase test) and microscopic pus cell count. The sensitivity, specificity, positive predictive value and negative predictive value of nitrite test, leucocyte esterase test and microscopic pus cell count were compared with urine culture in diagnosis of asymptomatic bacteriuria by using SPSS version 19. Of 123 urine samples, significant asymptomatic bacteriuria (≥10 4 cfu/Ml) was detected in 8 (6.5%) subjects. The sensitivity and specificity of nitrite test were 37% and 100% respectively. The sensitivity of pus cell count alone and leucocyte esterase test alone were 100% but the specificity of them were 64% and 65% respectively. We found high negative predictive value by Pus cell count and the leucocyte esterase test (100%) and low positive predictive value by them (16% and 17% respectively). Urine culture is the most useful test for diagnosis of asymptomatic bacteriuria. None of our screening tests had a sensitivity and

  4. Two-Dimensional Differential In-Gel Electrophoresis Proteomic Approaches Reveal Urine Candidate Biomarkers in Pediatric Obstructive Sleep Apnea

    PubMed Central

    Gozal, David; Jortani, Saeed; Snow, Ayelet B.; Kheirandish-Gozal, Leila; Bhattacharjee, Rakesh; Kim, Jinkwan; Capdevila, Oscar Sans

    2009-01-01

    Rationale: Sleep studies are laborious, expensive, inaccessible, and inconvenient for diagnosing obstructive sleep apnea (OSA) in children. Objectives: To examine whether the urinary proteome uncovers specific clusters that are differentially expressed in the urine of children with OSA. Methods: Two-dimensional differential in-gel electrophoresis (2D-DIGE) and mass spectrometry proteomics followed by validation with western blot of ELISA. Measurements and Main Results: Morning urine proteins from 60 children with polysomnographically confirmed OSA and from matched children with primary snoring (n = 30) and control subjects (n = 30) were assessed. A total of 16 proteins that are differentially expressed in OSA were identified, and 7 were confirmed by either immunoblots or ELISA. Among the latter, receiver–operator curve analyses of urinary concentrations of uromodulin, urocortin-3, orosomucoid-1, and kallikrein assigned favorable predictive properties to these proteins. Furthermore, combinatorial approaches indicated that the presence of values beyond the calculated cutoff concentrations for three or more of the proteins yielded a sensitivity of 95% and a specificity of 100%. Conclusions: Proteomic approaches reveal that pediatric OSA is associated with specific and consistent alterations in urinary concentrations of specific protein clusters. Future studies aiming to validate this approach as a screening method of habitually snoring children appears warranted. PMID:19797158

  5. Generation of Mesenchymal-Like Stem Cells From Urine in Pediatric Patients.

    PubMed

    He, W; Zhu, W; Cao, Q; Shen, Y; Zhou, Q; Yu, P; Liu, X; Ma, J; Li, Y; Hong, K

    2016-01-01

    Mesenchymal stem cells (MSCs) have been widely used for regenerative medicine. Traditionally, the procedures of MSC isolation are usually invasive and time-consuming. Urine is merely a body waste, and recent studies have suggested that urine represents an alternative source of stem cells. We, therefore, determined whether the possibility of isolating mesenchymal-like stem cells was practical from human urine. A total of 16 urine samples were collected from pediatric patients. Urine-derived cells were isolated, expanded, and identified for specific cell surface markers using flow cytometry. Cell morphology was observed by microscopy. Osteogenic and adipogenic differentiation potential were determinded by culturing cells in specific induction medium, and assessed by alkaline phosphatase and oil red O stainings, respectively. Clones were established and passaged successfully from primary cultures of urine cells. Cultured urine-derived cells at passage 3 were fusiform and arranged with certain directionality. Urine-derived cells at passage 5 displayed expressions of cell surface markers (CD29, CD105, CD166, CD90, and CD13). There was no expression of the general hematopoietic cell markers (CD45, CD34, and HLA-DR). Under in vitro induction conditions, urine-derived cells at passage 5 were able to differentiate into osteoblasts, but not adipocytes. Urine may be a noninvasive source for mesenchymal-like stem cells. These cells could potentially provide a new source of autologous stem cells for regenerative medicine and cell therapy. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Uranium Associations with Kidney Outcomes Vary by Urine Concentration Adjustment Method

    PubMed Central

    Shelley, Rebecca; Kim, Nam-Soo; Parsons, Patrick J.; Lee, Byung-Kook; Agnew, Jacqueline; Jaar, Bernard G.; Steuerwald, Amy J.; Matanoski, Genevieve; Fadrowski, Jeffrey; Schwartz, Brian S.; Todd, Andrew C.; Simon, David; Weaver, Virginia M.

    2017-01-01

    Uranium is a ubiquitous metal that is nephrotoxic at high doses. Few epidemiologic studies have examined the kidney filtration impact of chronic environmental exposure. In 684 lead workers environmentally exposed to uranium, multiple linear regression was used to examine associations of uranium measured in a four-hour urine collection with measured creatinine clearance, serum creatinine- and cystatin-C-based estimated glomerular filtration rates, and N-acetyl-β-D-glucosaminidase (NAG). Three methods were utilized, in separate models, to adjust uranium levels for urine concentration - μg uranium/g creatinine; μg uranium/L and urine creatinine as separate covariates; and μg uranium/4 hr. Median urine uranium levels were 0.07 μg/g creatinine and 0.02 μg/4 hr and were highly correlated (rs =0.95). After adjustment, higher ln-urine uranium was associated with lower measured creatinine clearance and higher NAG in models that used urine creatinine to adjust for urine concentration but not in models that used total uranium excreted (μg/4 hr). These results suggest that, in some instances, associations between urine toxicants and kidney outcomes may be statistical, due to the use of urine creatinine in both exposure and outcome metrics, rather than nephrotoxic. These findings support consideration of non-creatinine-based methods of adjustment for urine concentration in nephrotoxicant research. PMID:23591699

  7. Microanalyzer for Biomonitoring of Lead (Pb) in Blood and Urine

    SciTech Connect

    Yantasee, Wassana; Timchalk, Chuck; Lin, Yuehe

    2007-01-01

    Biomonitoring of lead (Pb) in blood and urine enables quantitative evaluation of human occupational and environmental exposures to Pb. The state-of-the-art ICP-MS instruments analyze metals in laboratories, resulting in lengthy turn around time, and are expensive. In response to the growing need for metal analyzer for on-site, real-time monitoring of trace metals in individuals, we developed a portable microanalyzer based on flow-injection/adsorptive stripping voltammetry and used it to analyze Pb in rat blood and urine. Fouling of electrodes by proteins often prevents the effective use of electrochemical sensors in biological matrices. Minimization of such fouling was accomplished with the suitablemore » sample pretreatment and the turbulent flowing of Pb contained blood and urine onto the glassy electrode inside the microanalyzer, which resulted in no apparent electrode fouling even when the samples contained 50% urine or 10% blood by volume. There was no matrix effect on the voltammetric Pb signals even when the samples contained 10% blood or 10% urine. The microanalyzer offered linear concentration range relevant to Pb exposure levels in human (0-20 ppb in 10%-blood samples, 0-50 ppb in 50%-urine samples). The device had excellent sensitivity and reproducibility; Pb detection limits were 0.54 ppb and 0.42 ppb, and % RSDs were 4.9 and 2.4 in 50%-urine and 10%-blood samples, respectively. It offered a high throughput (3 min per sample) and had economical use of samples (60 ?L per measurement), making the collection of blood being less invasive especially to children, and had low reagent consumption (1 ?g of Hg per measurement), thus minimizing the health concerns of mercury use. Being miniaturized in size, the microanalyzer is portable and field-deployable. Thus, it has a great potential to be the next-generation analyzer for biomonitoring of toxic metals.« less

  8. Dipping your feet in the water: podocytes in urine.

    PubMed

    Sir Elkhatim, Rashid; Li, Jordan Y Z; Yong, Tuck Y; Gleadle, Jonathan M

    2014-05-01

    Podocyte injury and loss plays an important role in the pathogenesis and progression of many kidney diseases. Studies have shown that podocyte-related markers and products can be detected in the urine of patients with glomerular diseases such as focal segmental glomerulosclerosis, IgA nephropathy, lupus nephritis, diabetic nephropathy and pre-eclampsia. Therefore, detecting the loss of podocytes in the urine provides a useful noninvasive technique of gathering information about the disease type and/or activity of glomerular diseases. Currently, urine podocyte-related protein markers, mRNA, microRNA and exosomes have been used with varying degrees of success to study glomerular diseases. The determination of urinary podocyte loss may become an important noninvasive tool in the evaluation of glomerular diseases.

  9. Determination of sulfur in human hair using high resolution continuum source graphite furnace molecular absorption spectrometry and its correlation with total protein and albumin

    NASA Astrophysics Data System (ADS)

    Ozbek, Nil; Baysal, Asli

    2017-04-01

    Human hair is a valuable contributor for biological monitoring. It is an information storage point to assess the effects of environmental, nutritional or occupational sources on the body. Human proteins, amino acids or other compounds are among the key components to find the sources of different effects or disorders in the human body. Sulfur is a significant one of these compounds, and it has great affinity to some metals and compounds. This property of the sulfur affects the human health positively or negatively. In this manuscript, sulfur was determined in hair samples of autistic and age-match control group children via molecular absorption of CS using a high-resolution continuum source graphite furnace atomic absorption spectrometer. For this purpose, hair samples were appropriately washed and dried at 75 °C. Then samples were dissolved in microwave digestion using HNO3 for sulfur determination. Extraction was performed with HCl hydrolysation by incubation for 24 h at 110 °C for total protein and albumin determination. The validity of the method for the sulfur determination was tested using hair standard reference materials. The results were in the uncertainty limits of the certified values at 95% confidence level. Finally correlation of sulfur levels of autistic children's hair with their total protein and albumin levels were done.

  10. Gender-specific association between dietary acid load and total lean body mass and its dependency on protein intake in seniors.

    PubMed

    Faure, A M; Fischer, K; Dawson-Hughes, B; Egli, A; Bischoff-Ferrari, H A

    2017-12-01

    Diet-related mild metabolic acidosis may play a role in the development of sarcopenia. We investigated the relationship between dietary acid load and total lean body mass in male and female seniors age ≥ 60 years. We found that a more alkaline diet was associated with a higher %TLM only among senior women. The aim of this study was to determine if dietary acid load is associated with total lean body mass in male and female seniors age ≥ 60 years. We investigated 243 seniors (mean age 70.3 ± 6.3; 53% women) age ≥ 60 years who participated in the baseline assessment of a clinical trial on vitamin D treatment and rehabilitation after unilateral knee replacement due to severe knee osteoarthritis. The potential renal acid load (PRAL) was assessed based on individual nutrient intakes derived from a food frequency questionnaire. Body composition including percentage of total lean body mass (%TLM) was determined using dual-energy X-ray absorptiometry. Cross-sectional analyses were performed for men and women separately using multivariable regression models controlling for age, physical activity, smoking status, protein intake (g/kg BW per day), energy intake (kcal), and serum 25-hydroxyvitamin D concentration. We included a pre-defined subgroup analysis by protein intake (< 1 g/kg BW day, > 1 g/kg BW day) and by age group (< 70 years, ≥ 70 years). Adjusted %TLM decreased significantly across PRAL quartiles only among women (P trend  = 0.004). Moreover, in subgroup analysis, the negative association between the PRAL and %TLM was most pronounced among women with low protein intake (< 1 g/kg BW per day) and age below 70 years (P = 0.002). Among men, there was no association between the PRAL and %TLM. The association between dietary acid load and %TLM seems to be gender-specific, with a negative impact on total lean mass only among senior women. Therefore, an alkaline diet may be beneficial for preserving total lean mass in senior women

  11. MELFI Urine Sample First Insertion

    NASA Image and Video Library

    2009-04-11

    ISS019-E-005715 (11 April 2009) --- Astronaut Michael Barratt, Expedition 19/20 flight engineer, performs an insertion of urine samples into the Minus Eighty Degree Laboratory Freezer for ISS (MELFI) as part of the Nutritional Status Assessment (NUTRITION) study in the Japanese Kibo laboratory of the International Space Station.

  12. Chemical measurement of urine volume

    NASA Technical Reports Server (NTRS)

    Sauer, R. L.

    1978-01-01

    Chemical method of measuring volume of urine samples using lithium chloride dilution technique, does not interfere with analysis, is faster, and more accurate than standard volumetric of specific gravity/weight techniques. Adaptation of procedure to urinalysis could prove generally practical for hospital mineral balance and catechoamine determinations.

  13. Measurement of thioridazine in blood and urine.

    PubMed Central

    Ng, C H; Crammer, J L

    1977-01-01

    1 Thioridazine can be specifically, simply, and reliably measured in plasma and urine by gas chromatography using hexane extraction and prochlorperazine as internal standard; fluorimetry is non-specific. 2 The method can also measure thioridazine ring sulphoxide, and mesoridazine-plus-sulphoridazine (M/S). 3 After single doses plasma sometimes shows M/S in addition to thioridazine itself; it always does so on continued treatment. There is great individual variation in both components, and evidence of changes in metabolism during the early weeks. 4 Urinary excretion may be influenced by pH, but between pH 6.0-7.0 about 1% of the daily dose appears in 24 h urine as the following: free thioridazine in microng quantities, M/S and ring sulphoxide each in mg amounts. 5 Patients attain steady state conditions, although plasma levels rise considerably after each dose and settle again in about 10 h. After chronic treatment is stopped to half-life is at about 30 h. 6 Plasma levels cannot be related to therapeutic response when this is slow, as in schizophrenia, but interpretations are complicated by the production of clinically active metabolites, and by plasma protein binding. PMID:16633

  14. Antibiotic Screening of Urine Culture for Internal Quality Audit at Amrita Hospital, Kochi.

    PubMed

    Suresh, Aswathy; Gopinathan, Anusha; Dinesh, Kavitha R; Kumar, Anil

    2017-07-01

    Urine antimicrobial activity is a seldom analysed laboratory test which greatly impacts the quantification of urine specimens. Presence of antimicrobial activity in the urine reduces the bacterial load in these specimens. Hence, the chances of erroneously reporting insignificant bacteriuria can be reduced on analysis of the antimicrobial activity in urine. The aim of the study was to measure the antimicrobial activity of urine samples obtained from patients in a tertiary care hospital. A total of 100 urine specimens were collected from the study group. Tests like wet mount, Gram staining and culture were performed. Antimicrobial susceptibility testing was done on the bacteria isolated from each specimen. The urine specimens were reported as significant bacteriuria (>105 Colony Forming Unit (CFU)/ml) and insignificant bacteriuria (<105 CFU/ml - clean catch midstream urine; <102 CFU/ml - catheterized urine sample) according to the CFU/ml. Staphylococcus aureus ATCC ® 25923 ™ and Escherichia coli ATCC ® 25922 ™ were used to identify the presence of antimicrobial activity in the urine sample by Urine Anti-Bacterial substance Assay (UABA). McNemar test was used for statistical analysis using Statistical Package for the Social Sciences (SPSS) version 21.0. On analysis of the antimicrobial activity of urine sample with the prior antibiotic history of the patients, 17 were true positives and 43 were true negatives. Twenty six of samples with UABA positivity were culture negative and 28 samples with UABA positivity were culture positive. Sensitivity and specificity of the test was 85% and 53.8% respectively. Accuracy of the test was 60%. The p-value of UABA was <0.001. Enterobacteriaceae was the most common bacterial family isolated from the urine specimens. A total of 85% patients responded to treatment. Presence of antimicrobial activity in urine has a great impact on the interpretation of urine culture reports. Identification of urine antimicrobial activity helps

  15. Intake of branched-chain amino acids influences the levels of MAFbx mRNA and MuRF-1 total protein in resting and exercising human muscle.

    PubMed

    Borgenvik, Marcus; Apró, William; Blomstrand, Eva

    2012-03-01

    Resistance exercise and amino acids are two major factors that influence muscle protein turnover. Here, we examined the effects of resistance exercise and branched-chain amino acids (BCAA), individually and in combination, on the expression of anabolic and catabolic genes in human skeletal muscle. Seven subjects performed two sessions of unilateral leg press exercise with randomized supplementation with BCAA or flavored water. Biopsies were collected from the vastus lateralis muscle of both the resting and exercising legs before and repeatedly after exercise to determine levels of mRNA, protein phosphorylation, and amino acid concentrations. Intake of BCAA reduced (P < 0.05) MAFbx mRNA by 30 and 50% in the resting and exercising legs, respectively. The level of MuRF-1 mRNA was elevated (P < 0.05) in the exercising leg two- and threefold under the placebo and BCAA conditions, respectively, whereas MuRF-1 total protein increased by 20% (P < 0.05) only in the placebo condition. Phosphorylation of p70(S6k) increased to a larger extent (∼2-fold; P < 0.05) in the early recovery period with BCAA supplementation, whereas the expression of genes regulating mTOR activity was not influenced by BCAA. Muscle levels of phenylalanine and tyrosine were reduced (13-17%) throughout recovery (P < 0.05) in the placebo condition and to a greater extent (32-43%; P < 0.05) following BCAA supplementation in both resting and exercising muscle. In conclusion, BCAA ingestion reduced MAFbx mRNA and prevented the exercise-induced increase in MuRF-1 total protein in both resting and exercising leg. Further-more, resistance exercise differently influenced MAFbx and MuRF-1 mRNA expression, suggesting both common and divergent regulation of these two ubiquitin ligases.

  16. Influence of dietary protein and fructooligosaccharides on fecal fermentative end-products, fecal bacterial populations and apparent total tract digestibility in dogs.

    PubMed

    Pinna, Carlo; Vecchiato, Carla Giuditta; Bolduan, Carmen; Grandi, Monica; Stefanelli, Claudio; Windisch, Wilhelm; Zaghini, Giuliano; Biagi, Giacomo

    2018-03-20

    Feeding dogs with diets rich in protein may favor putrefactive fermentations in the hindgut, negatively affecting the animal's intestinal environment. Conversely, prebiotics may improve the activity of health-promoting bacteria and prevent bacterial proteolysis in the colon. The aim of this study was to evaluate the effects of dietary supplementation with fructooligosaccharides (FOS) on fecal microbiota and apparent total tract digestibility (ATTD) in dogs fed kibbles differing in protein content. Twelve healthy adult dogs were used in a 4 × 4 replicated Latin Square design to determine the effects of four diets: 1) Low protein diet (LP, crude protein (CP) 229 g/kg dry matter (DM)); 2) High protein diet (HP, CP 304 g/kg DM); 3) Diet 1 + 1.5 g of FOS/kg; 4) Diet 2 + 1.5 g of FOS/kg. The diets contained silica at 5 g/kg as a digestion marker. Differences in protein content were obtained using different amounts of a highly digestible swine greaves meal. Each feeding period lasted 28 d, with a 12 d wash-out in between periods. Fecal samples were collected from dogs at 0, 21 and 28 d of each feeding period. Feces excreted during the last five days of each feeding period were collected and pooled in order to evaluate ATTD. Higher fecal ammonia concentrations were observed both when dogs received the HP diets (p < 0.001) and the supplementation with FOS (p < 0.05). The diets containing FOS resulted in greater ATTD of DM, Ca, Mg, Na, Zn, and Fe (p < 0.05) while HP diets were characterized by lower crude ash ATTD (p < 0.05). Significant interactions were observed between FOS and protein concentration in regards to fecal pH (p < 0.05), propionic acid (p < 0.05), acetic to propionic acid and acetic + n-butyric to propionic acid ratios (p < 0.01), bifidobacteria (p < 0.05) and ATTD of CP (p < 0.05) and Mn (p < 0.001). A relatively moderate increase of dietary protein resulted in higher concentrations of ammonia in

  17. Residual urine output and postoperative mortality in maintenance hemodialysis patients.

    PubMed

    Lin, Yu-Feng; Wu, Vin-Cent; Ko, Wen-Je; Chen, Yih-Sharng; Chen, Yung-Ming; Li, Wen-Yi; Chou, Nai-Kuan; Chao, Anne; Huang, Tao-Min; Chang, Fan-Chi; Chen, Shih-I; Shiao, Chih-Chung; Wang, Wei-Jie; Tsai, Hung-Bin; Tsai, Pi-Ru; Hu, Fu-Chang; Wu, Kwan-Dun

    2009-09-01

    The relationship between residual urine output and postoperative survival in maintenance hemodialysis patients is unknown. To explore the relationship between amount of urine before surgery and postoperative mortality and differences between postoperative nonanuria and anuria in maintenance hemodialysis patients. A total of 109 maintenance hemodialysis patients underwent major operations. Anuria was defined as urine output <30 mL in the 8 hours before the first session of postoperative dialysis. Propensity scores for postoperative anuria were developed. Postoperative residual urine output was 159.2 mL/8 h (SD, 115.1) in 33 patients; 76 patients were anuric. Preoperative residual urine output and adequate perioperative blood transfusion were positively related to postoperative urine output. Propensity-adjusted 30-day mortality was associated with postoperative anuria (odds ratio [OR], 4.56; 95% confidence interval [CI], 1.16-17.96; P = .03), prior stroke (OR, 4.46; 95% CI, 1.43-13.89; P = .01) and higher disease severity (OR, 1.10; 95% CI, 1.00-1.21; P = .049) at the first postoperative dialysis. OR of 30-day mortality was 5.38 for nonanuria to anuria vs nonanuria to nonanuria (P = .03) and 5.13 for preoperative anuria vs nonanuria to nonanuria (P = .01). By Kaplan-Meier analysis, 30-day mortality differed significantly among patients for nonanuria to nonanuria, anuria, and nonanuria to anuria (log rank, P = .045). Patients with preoperative nonanuria and postoperative anuria had higher mortality than did patients with no anuria before and after surgery and patients with anuria before surgery. Postoperative residual urine output is an important surrogate marker for disease severity.

  18. Validation of Surrogates of Urine Osmolality in Population Studies.

    PubMed

    Youhanna, Sonia; Bankir, Lise; Jungers, Paul; Porteous, David; Polasek, Ozren; Bochud, Murielle; Hayward, Caroline; Devuyst, Olivier

    2017-01-01

    The importance of vasopressin and/or urine concentration in various kidney, cardiovascular, and metabolic diseases has been emphasized recently. Due to technical constraints, urine osmolality (Uosm), a direct reflect of urinary concentrating activity, is rarely measured in epidemiologic studies. We analyzed 2 possible surrogates of Uosm in 4 large population-based cohorts (total n = 4,247) and in patients with chronic kidney disease (CKD, n = 146). An estimated Uosm (eUosm) based on the concentrations of sodium, potassium, and urea, and a urine concentrating index (UCI) based on the ratio of creatinine concentrations in urine and plasma were compared to the measured Uosm (mUosm). eUosm is an excellent surrogate of mUosm, with a highly significant linear relationship and values within 5% of mUosm (r = 0.99 or 0.98 in each population cohort). Bland-Altman plots show a good agreement between eUosm and mUosm with mean differences between the 2 variables within ±24 mmol/L. This was verified in men and women, in day and night urine samples, and in CKD patients. The relationship of UCI with mUosm is also significant but is not linear and exhibits more dispersed values. Moreover, the latter index is no longer representative of mUosm in patients with CKD as it declines much more quickly with declining glomerular filtration rate than mUosm. The eUosm is a valid marker of urine concentration in population-based and CKD cohorts. The UCI can provide an estimate of urine concentration when no other measurement is available, but should be used only in subjects with normal renal function. © 2017 S. Karger AG, Basel.

  19. Evaluation of five commercially available assays and measurement of serum total protein concentration via refractometry for the diagnosis of failure of passive transfer of immunity in foals.

    PubMed

    Davis, Rachel; Giguère, Steeve

    2005-11-15

    To determine and compare sensitivity, specificity, accuracy, and predictive values of measurement of serum total protein concentration by refractometry as well as 5 commercially available kits for the diagnosis of failure of passive transfer (FPT) of immunity in foals. Prospective study. 65 foals with various medical problems and 35 clinically normal foals. IgG concentration in serum was assessed by use of zinc sulfate turbidity (assay C), glutaraldehyde coagulation (assay D), 2 semiquantitative immunoassays (assays F and G), and a quantitative immunoassay (assay H). Serum total protein concentration was assessed by refractometry. Radial immunodiffusion (assays A and B) was used as the reference method. For detection of IgG < 400 mg/dL, sensitivity of assay H (100%) was not significantly different from that of assays C, E, and G (88.9%). Specificity of assays H (96.0%) and G (95.8%) was significantly higher than that of assays C (79.4%) and E (78.1 %). For detection of IgG < 800 mg/dL, sensitivities of assays H (976%), D (92.9%), C (81.0%), and G (81.0%) were significantly higher than that of assay F (52.4%). Specificity of assays F (100%), G (94.7%), and H (82.8%) was significantly higher than that of assays C (56.9%) and D (58.6%). Serum total protein concentration < or = 4.5 g/dL was suggestive of FPT, whereas values > or = 6.0 g/dL indicated adequate IgG concentrations. Most assays were adequate as initial screening tests. However, their use as a definitive test would result in unnecessary treatment of foals with adequate IgG concentrations.

  20. Apparent total tract macronutrient digestibility, fecal characteristics, and fecal fermentative end-product concentrations of healthy adult dogs fed bioprocessed soy protein.

    PubMed

    Beloshapka, A N; de Godoy, M R C; Detweiler, K B; Newcomb, M; Ellegård, K H; Fahey, G C; Swanson, K S

    2016-09-01

    Animal proteins are commonly used in extruded dog foods. Plant-based proteins have a more consistent nutrient profile than animal sources but may contain antinutritional factors, including trypsin inhibitors and oligosaccharides. Bioprocessed soy protein (SP; HP-300; Hamlet Protein, Inc., Findlay, OH) is a processed soy-based product with low antinutritional factor concentrations and high protein quality. The objective was to evaluate the effects of SP on apparent total tract macronutrient digestibility, fecal characteristics, and fecal fermentative end products. Furthermore, this study aimed to identify if SP can be a replacement for poultry byproduct meal (PBPM) in dog food and determine if there are practical limits to its use. Three palatability experiments were conducted to evaluate 1) 0 vs. 12% SP, 2) 0 vs. 48% SP, and 3) 12 vs. 48% SP. For digestibility, 48 healthy adult Beagle dogs (20 females and 28 males; 3.4 yr mean age and 10.0 kg mean BW) were randomly allotted to 1 of 6 dietary treatments, 0 (control), 4, 8, 12, 24, and 48% SP, in a completely randomized design. All diets were formulated to meet Association of American Feed Control Officials nutrient profiles and contained approximately 30% CP and 16% fat. The treatment period consisted of a 10-d diet adaptation phase followed by a 4-d fresh and total fecal collection phase. The palatability results suggest that of the 3 inclusion levels tested (0, 12, or 48% SP), the best inclusion of SP is 12%, which was preferred over 0 and 48% SP. Digestibility and fecal data were evaluated for linear and quadratic effects using SAS. Stool output (on both an as-is and a DM basis) did not differ from the control except for the 48% SP treatment ( < 0.01). Fecal output per unit food intake differed ( < 0.01) from the control only at the 24 and 48% SP inclusion rates. No significant effects of feeding SP were found on stool consistency scores. Digestibility of DM, OM, and energy did not differ from the control at any

  1. Hemoglobinuria Misidentified as Hematuria: Review of Discolored Urine and Paroxysmal Nocturnal Hemoglobinuria

    PubMed Central

    Veerreddy, Prashant

    2013-01-01

    Discolored urine is a common reason for office visits to a primary care physician and urology referral. Early differentiation of the type or cause of discolored urine is necessary for accurate diagnosis and prompt management. Paroxysmal nocturnal hemoglobinuria is a clonal disorder caused by acquired somatic mutations in the PIG-A gene on the X- chromosome of hemopoietic stem cells and leads to deficiency of surface membrane anchor proteins. The deficiency of these proteins leads to an increased risk of hemolysis of erythrocytes and structural damage of platelets, resulting in a clinical syndrome characterized by complement-mediated intravascular hemolytic anemia, bone marrow failure, and venous thrombosis. Patients with this clinical syndrome present with paroxysms of hemolysis, causing hemoglobinuria manifesting as discolored urine. This can be easily confused with other common causes of discolored urine and result in extensive urologic work-up. Three commonly confused entities of discolored urine include hematuria, hemoglobinuria, and myoglobinuria. Specific characteristics in a dipstick test or urinalysis can guide differentiation of these three causes of discolored urine. This article begins with a case summary of a woman presenting with cranberry-colored urine and a final delayed diagnosis of paryxysmal nocturnal hemoglobinuria. Her hemoglobinuria was misdiagnosed as hematuria, leading to extensive urologic work-up. The article also gives an overview of the approach to diagnosing and treating discolored urine. PMID:25512715

  2. Fermented dairy products consumption is associated with attenuated cortical bone loss independently of total calcium, protein, and energy intakes in healthy postmenopausal women.

    PubMed

    Biver, E; Durosier-Izart, C; Merminod, F; Chevalley, T; van Rietbergen, B; Ferrari, S L; Rizzoli, R

    2018-05-03

    A longitudinal analysis of bone microstructure in postmenopausal women of the Geneva Retirees Cohort indicates that age-related cortical bone loss is attenuated at non-bearing bone sites in fermented dairy products consumers, not in milk or ripened cheese consumers, independently of total energy, calcium, or protein intakes. Fermented dairy products (FDP), including yogurts, provide calcium, phosphorus, and proteins together with prebiotics and probiotics, all being potentially beneficial for bone. In this prospective cohort study, we investigated whether FDP, milk, or ripened cheese consumptions influence age-related changes of bone mineral density (BMD) and microstructure. Dietary intakes were assessed at baseline and after 3.0 ± 0.5 years with a food frequency questionnaire in 482 postmenopausal women enrolled in the Geneva Retirees Cohort. Cortical (Ct) and trabecular (Tb) volumetric (v) BMD and microstructure at the distal radius and tibia were assessed by high-resolution peripheral quantitative computerized tomography, in addition to areal (a) BMD and body composition by dual-energy X-ray absorptiometry, at the same time points. At baseline, FDP consumers had lower abdominal fat mass and larger bone size at the radius and tibia. Parathyroid hormone and β-carboxyterminal cross-linked telopeptide of type I collagen levels were inversely correlated with FDP consumption. In the longitudinal analysis, FDP consumption (mean of the two assessments) was associated with attenuated loss of radius total vBMD and of Ct vBMD, area, and thickness. There was no difference in aBMD and at the tibia. These associations were independent of total energy, calcium, or protein intakes. For other dairy products categories, only milk consumption was associated with lower decrease of aBMD and of failure load at the radius. In this prospective cohort of healthy postmenopausal women, age-related Ct bone loss was attenuated at non-bearing bone sites in FDP consumers, not in milk

  3. Clonorchis sinensis-derived total protein attenuates airway inflammation in murine asthma model by inducing regulatory T cells and modulating dendritic cell functions

    SciTech Connect

    Jeong, Young-Il; Kim, Seung Hyun; Ju, Jung Won

    Highlights: {yields} Treatment with Clonorchis sinensis-derived total protein attenuates OVA-induced airway inflammation and AHR to methacholine. {yields} Induction of CD4{sup +}CD25{sup +}Foxp3{sup +} T cells and IL-10 along with suppression of splenocyte proliferation by C. sinensis-derived total protein. {yields} C. sinensis-derived total protein interferes with the expression of co-stimulatory molecules in DCs. -- Abstract: Asthma is characterized by Th2-mediated inflammation, resulting in airway hyperresponsiveness (AHR) through airway remodeling. Recent epidemiological and experimental reports have suggested an inverse relationship between the development of allergy and helminth infections. Infection by Clonorchis sinensis, a liver fluke that resides in the bile duct ofmore » humans, is endemic predominantly in Asia including Korea and China. Using a murine model for asthma, we investigated the effects of C. sinensis-derived total protein (Cs-TP) on allergen-induced airway inflammation and the mechanism underlying the protective effects of Cs-TP administration on asthma. Treatment with Cs-TP attenuated OVA-induced airway inflammation and methacholine-induced AHR, as well as eosinophilia development, lymphocyte infiltration into the lung, and goblet cell metaplasia. This protective effect of Cs-TP is associated with markedly reduced OVA-specific IgE and Th1/Th2 cytokine production. Moreover, Cs-TP increased the number of CD4{sup +}CD25{sup +}Foxp3{sup +} regulatory T (Treg) cells as well as their suppressive activity. In fact, proliferation of OVA-restimulated splenocytes was suppressed significantly. Cs-TP also inhibited the expression of such co-stimulatory molecules as CD80, CD86, and CD40 in LPS- or OVA-stimulated dendritic cells (DCs), suggesting that Cs-TP could interfere with the capacity of airway DCs to prime naive T cells. These data demonstrate the capacity of C. sinensis to ameliorate allergic asthma and broaden our understanding of the

  4. Importance of Urine Dipstick in Evaluation of Young Febrile Infants With Positive Urine Culture: A Spanish Pediatric Emergency Research Group Study.

    PubMed

    Velasco, Roberto; Benito, Helvia; Mozun, Rebeca; Trujillo, Juan E; Merino, Pedro A; de la Torre, Mercedes; Gomez, Borja; Mintegi, Santiago

    2016-12-01

    Guidelines from the American Academy of Pediatrics define urinary tract infection (UTI) as the growth of greater than 50,000 ufc/mL of a single bacterium in a urine culture with a positive urine dipstick or with a urinalysis associated. Our objective was to evaluate the adequacy of this cutoff point for the diagnosis of UTI in young febrile infants. Subanalysis of a prospective multicenter study developed in RISeuP-SPERG Network between October 11 and September 13. To carry out the study, it was performed a comparison of analytical and microbiological characteristics of patients younger than 90 days with fever without focus, taking into account the results of urine dipstick and urine culture. Of a total of 3333 infants younger than 90 days with fever without focus which were included in the study, 538 were classified as UTI in accordance with American Academy of Pediatrics' guidelines. These patients were similar to those who had a positive urine dipstick and a urine culture yielding of 10,000 to 50,000 ufc/mL, and they were different from those who had a normal urine dipstick and a urine culture >50,000 ufc/mL, being focused on the isolated bacteria and blood biomarkers values. Forty-five invasive bacterial infections were diagnosed (5.9% of the 756 with a urine culture >10,000 ufc/mL). Half of the infants with a normal urine dipstick diagnosed with invasive bacterial infections were younger than 15 days. It might be inadequate to use a threshold of 50,000 cfu/mL to consider a urine culture as positive in young febrile infants given the fact that it would misdiagnose several UTIs.

  5. Urine interleukin-6, interleukin-8 and transforming growth factor β1 in infants with urinary tract infection and asymptomatic bacteriuria

    PubMed Central

    Krzemień, Grażyna; Turczyn, Agnieszka; Pańczyk-Tomaszewska, Małgorzata

    2016-01-01

    Introduction Urinary tract infection (UTI) occurs in 1.1% of girls and 1.4% of boys during the first year of life. Asymptomatic bacteriuria (ABU) is usually detected incidentally in 0.9% of girls and 2.5% of boys at this age. The aim of the study was to assess the usefulness of measurement of pro-inflammatory urine interleukin (IL)-6 and IL-8 concentrations and anti-inflammatory transforming growth factor β1 (TGF-β1) level in infants with febrile UTI, non-febrile UTI and ABU. Material and methods A total of 35 children, mean age 6.14 ±3.47 months, were divided into three groups: group I – febrile UTI (n = 13), group II – non-febrile UTI (n = 13) and group III – ABU (n = 9). At the time of enrollment urine IL-6, IL-8, TGF-β1 and serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell count (WBC) were measured. Renal ultrasound was performed in all children, 99mTc-dimercaptosuccinic acid scintigraphy (DMSA) and voiding cystourethrography in children with UTI. Results Urine concentrations of IL-6 and IL-8 were significantly higher in febrile UTI compared to those with non-febrile UTI and ABU (p < 0.5, p < 0.01) and positively correlated with CRP, ESR and WBC (p < 0.01). Urine levels of TGF-β1 were significantly higher in children with febrile UTI compared to those with ABU (p < 0.05) and positively correlated with WBC (p < 0.01). Inflammatory changes in the DMSA scan were detected in 66.6% of children with UTI. No significant difference in frequency of an abnormal DMSA scan compared to a normal scan was found in groups with febrile and non-febrile UTI. No relations between urine cytokines, systemic inflammatory markers and changes in DMSA scan were observed. The cutoff value for detection of inflammatory changes in the DMSA scan for IL-8 was 120 pg/mg creatinine (Cr) and 40 pg/mg Cr for TGF-β1. Based on this value, the sensitivity for IL-8 was 58.3%, specificity 100% and for TGF-β1 66.7% and 83.7%, respectively

  6. Urine Potassium Excretion, Kidney Failure, and Mortality in CKD.

    PubMed

    Leonberg-Yoo, Amanda K; Tighiouart, Hocine; Levey, Andrew S; Beck, Gerald J; Sarnak, Mark J

    2017-03-01

    Low urine potassium excretion, as a surrogate for dietary potassium intake, is associated with higher risk for hypertension and cardiovascular disease in a general population. Few studies have investigated the relationship of urine potassium with clinical outcomes in chronic kidney disease (CKD). Longitudinal cohort study. The MDRD (Modification of Diet in Renal Disease) Study was a randomized controlled trial (N = 840) conducted in 1989 to 1993 to examine the effects of blood pressure control and dietary protein restriction on kidney disease progression in adults aged 18 to 70 years with CKD stages 2 to 4. This post hoc analysis included 812 participants. The primary predictor variable was 24-hour urine potassium excretion, measured at baseline and at multiple time points (presented as time-updated average urine potassium excretion). Kidney failure, defined as initiation of dialysis therapy or transplantation, was determined from US Renal Data System data. All-cause mortality was assessed using the National Death Index. Median follow-up for kidney failure was 6.1 (IQR, 3.5-11.7) years, with 9 events/100 patient-years. Median all-cause mortality follow-up was 19.2 (IQR, 10.8-20.6) years, with 3 deaths/100 patient-years. Baseline mean urine potassium excretion was 2.39±0.89 (SD) g/d. Each 1-SD higher baseline urine potassium level was associated with an adjusted HR of 0.95 (95% CI, 0.87-1.04) for kidney failure and 0.83 (95% CI, 0.74-0.94) for all-cause mortality. Results were consistent using time-updated average urine potassium measurements. Analyses were performed using urine potassium excretion as a surrogate for dietary potassium intake. Results are obtained from a primarily young, nondiabetic, and advanced CKD population and may not be generalizable to the general CKD population. Higher urine potassium excretion was associated with lower risk for all-cause mortality, but not kidney failure. Copyright © 2016 National Kidney Foundation, Inc. Published by

  7. Genital region cleansing wipes: Effects on urine culture contamination.

    PubMed

    Selek, Mehmet Burak; Bektöre, Bayhan; Sezer, Ogün; Kula Atik, Tuğba; Baylan, Orhan; Özyurt, Mustafa

    2017-01-30

    Urine culture is the gold standard test for revealing the microbial agent causing urinary tract infection (UTI). Culture results are affected by sampling techniques; improper sampling leads to contamination of urine and thus contamination of the culture with urogenital flora. We aimed to evaluate the effect of urogenital cleansing, performed with chlorhexidine-containing genital region cleansing wipes (GRCW) on contamination rates. A total of 2,665 patients with UTI-related complaints and with urine culture requests from various outpatient clinics were enrolled in the study. Of the patients, 1,609 in the experimental group used GRCW before sampling, while 1,046 in the control group did not use any wipes. The contamination rate in the experimental group patients was 7.7%, while it was 15.8% in the control group. Contamination rates were significantly higher in the control group than in the experimental group for both women and men. Contamination rates for children and adults were also significantly lower in the experimental group than in the control group. Our study, conducted in a large population, showed that the use of chlorhexidine-containing cleansing wipes significantly reduced urine culture contamination rates in both genders, in both child and adult age groups. Using GRCW, collection of urine after urogenital area cleansing will decrease the contamination problem.

  8. [Effect of remifentanil on urine output during gynecological laparoscopic surgery].

    PubMed

    Yago, Yasuko; Tajiri, Osamu; Ito, Hiroyuki; Kanazawa, Masashi; Tateda, Takeshi

    2009-05-01

    We retrospectively examined the effect of remifentanil on urine output during gynecological laparoscopic surgery under general anesthesia performed from April 2006 to July 2007. Forty six patients undergoing gynecological laparoscopic surgery under general anesthesia were divided into 2 groups. In group C (n=23), anesthesia was performed using sevoflurane and/or propofol with intermittent fentanyl. In group R (n=23), remifentanil was additionally used with the method of group C. Patient's demography was not different between the two groups. Intraoperative conditions were compatible in both groups. In group R, total dose of fentanyl is significantly lower than group C. BP and HR measured at 20 min after pneumoperitoneum were significantly lower in group R. Intraoperative urine output was significantly greater in group R than group C. A decrease in urine output is commonly seen particularly in laparoscopic surgery. Increased stress hormonal responses due to pneumoperitoneum have been explained as one of the causes of this phenomenon. Remifentanil has been reported to maintain urine output as well as to blunt hormonal responses in CABG surgery. Although we did not measure hormonal responses in the present study, increased urine output could be attributed to decreased catecholamine levels by remifentanil.

  9. 28 CFR 550.41 - Urine surveillance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Urine surveillance. 550.41 Section 550.41 Judicial Administration BUREAU OF PRISONS, DEPARTMENT OF JUSTICE INSTITUTIONAL MANAGEMENT DRUG PROGRAMS Drug Services (Urine Surveillance and Counseling for Sentenced Inmates in Contract CTCs) § 550.41 Urine...

  10. 28 CFR 550.41 - Urine surveillance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Urine surveillance. 550.41 Section 550.41 Judicial Administration BUREAU OF PRISONS, DEPARTMENT OF JUSTICE INSTITUTIONAL MANAGEMENT DRUG PROGRAMS Drug Services (Urine Surveillance and Counseling for Sentenced Inmates in Contract CTCs) § 550.41 Urine...

  11. 28 CFR 550.41 - Urine surveillance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Urine surveillance. 550.41 Section 550.41 Judicial Administration BUREAU OF PRISONS, DEPARTMENT OF JUSTICE INSTITUTIONAL MANAGEMENT DRUG PROGRAMS Drug Services (Urine Surveillance and Counseling for Sentenced Inmates in Contract CTCs) § 550.41 Urine...

  12. 28 CFR 550.41 - Urine surveillance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Urine surveillance. 550.41 Section 550.41 Judicial Administration BUREAU OF PRISONS, DEPARTMENT OF JUSTICE INSTITUTIONAL MANAGEMENT DRUG PROGRAMS Drug Services (Urine Surveillance and Counseling for Sentenced Inmates in Contract CTCs) § 550.41 Urine...

  13. 28 CFR 550.41 - Urine surveillance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 2 2011-07-01 2011-07-01 false Urine surveillance. 550.41 Section 550.41 Judicial Administration BUREAU OF PRISONS, DEPARTMENT OF JUSTICE INSTITUTIONAL MANAGEMENT DRUG PROGRAMS Drug Services (Urine Surveillance and Counseling for Sentenced Inmates in Contract CTCs) § 550.41 Urine...

  14. Determination of free and total (free plus protein-bound) melatonin in plasma and cerebrospinal fluid by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Rizzo, Vittoria; Porta, Camillo; Moroni, Mauro; Scoglio, Enrico; Moratti, Remigio

    2002-07-05

    A simple, sensitive and accurate method for the estimation of free and total (free plus protein-bound) melatonin (MLT) in human plasma and cerebrospinal fluid (CSF) is described. Via Chem-Elut cartridges, free and total MLT (the latter obtained after a deproteinization step) were quantified in dichloromethane-extracted samples and analyzed in one chromatographic run by high-performance liquid chromatography (HPLC) with fluorimetric detection. The column used was an Extrasil ODS-2 (3 microm, 150 x 4.6 mm I.D.), while the mobile phase consisted of 75 mM sodium acetate-acetonitrile (72:28, v/v) (pH 5.0). Repeatability and reproducibility of the method were 3.24 and 9.4%, respectively. The recovery of melatonin from plasma and CSF was 99.9+/-4.0% for non-deproteinized samples and 93.2+/-4.8% for deproteinized samples. The detection limit of the assay was 0.5 pg/ml. In human plasma, the mean+/-SD concentrations in the darkness period were 23.18+/-7.44 pg/ml for free melatonin and 82.5+/-36.48 pg/ml for total melatonin, while the lowest concentrations detected during daytime were 2.23+/-2.22 and 7.40+/-5.68 pg/ml, respectively. Detection of MLT in CSF was 5.01+/-2.31 and 28.55+/-6.95 pg/ml for the free and total fraction, respectively.

  15. Determination of uromodulin in human urine: influence of storage and processing.

    PubMed

    Youhanna, Sonia; Weber, Julien; Beaujean, Viviane; Glaudemans, Bob; Sobek, Jens; Devuyst, Olivier

    2014-01-01

    Uromodulin (Tamm-Horsfall protein) is the most abundant protein excreted in the urine under physiological conditions. It is exclusively produced in the kidney and secreted into the urine via proteolytic cleavage. The involvement of UMOD, the gene that encodes uromodulin, in rare autosomal dominant diseases, and its robust genome-wide association with the risk of chronic kidney disease suggest that the level of uromodulin in urine could represent a critical biomarker for kidney function. The structure of uromodulin is complex, with multiple disulfide bonds and typical domains of extracellular proteins. Thus far, the conditions influencing stability and measurement of uromodulin in human urine have not been systematically investigated, giving inconsistent results. In this study, we used a robust, in-house ELISA to characterize the conditions of sampling and storage necessary to provide a faithful dosage of uromodulin in the urine. The levels of uromodulin in human urine were significantly affected by centrifugation and vortexing, as well as by the conditions and duration of storage. These results validate a simple, low-cost ELISA and document the optimal conditions of processing and storage for measuring uromodulin in human urine.

  16. Building global models for fat and total protein content in raw milk based on historical spectroscopic data in the visible and short-wave near infrared range.

    PubMed

    Melenteva, Anastasiia; Galyanin, Vladislav; Savenkova, Elena; Bogomolov, Andrey

    2016-07-15

    A large set of fresh cow milk samples collected from many suppliers over a large geographical area in Russia during a year has been analyzed by optical spectroscopy in the range 400-1100 nm in accordance with previously developed scatter-based technique. The global (i.e. resistant to seasonal, genetic, regional and other variations of the milk composition) models for fat and total protein content, which were built using partial least-squares (PLS) regression, exhibit satisfactory prediction performances enabling their practical application in the dairy. The root mean-square errors of prediction (RMSEP) were 0.09 and 0.10 for fat and total protein content, respectively. The issues of raw milk analysis and multivariate modelling based on the historical spectroscopic data have been considered and approaches to the creation of global models and their transfer between the instruments have been proposed. Availability of global models should significantly facilitate the dissemination of optical spectroscopic methods for the laboratory and in-line quantitative milk analysis. Copyright © 2016. Published by Elsevier Ltd.

  17. Pragmatic and evidence-based approach to paediatric cerebrospinal fluid reference limits for white cell count and concentrations of total protein and glucose.

    PubMed

    Josman, Nicky; Tee, Nancy W S; Maiwald, Matthias; Loo, Liat Hui; Ho, Clement K M

    2018-06-15

    It is often impractical for each laboratory to establish its own paediatric reference intervals. This is particularly true for specimen types collected using invasive procedures, for example, cerebrospinal fluid (CSF). Published CSF reference intervals for white cell count, and concentrations of total protein and glucose were reviewed by stakeholders in a paediatric hospital. Consensus reference intervals for the three CSF parameters were then subjected to verification using guidelines from the Clinical Laboratory Standards Institute and residual CSF specimens. Consensus paediatric reference intervals adapted from published studies with minor modifications were locally verified as follows. White cell count (x10 6 cells/L): 0-20 (<1 month); 0-10 (1-2 months); 0-5 (>2 months). Total protein (g/L): 0.3-1.2 (<1 month); 0.2-0.6 (1-3 months); 0.1-0.4 (>3 months). Glucose (mmol/L): 2.0-5.6 (<6 months); 2.4-4.3 (6 months or older). © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  18. Bisphenol A concentrations in maternal breast milk and infant urine

    PubMed Central

    Mendonca, K.; Hauser, R.; Calafat, A.M.; Arbuckle, T.E.; Duty, S.M.

    2013-01-01

    Purpose The present report describes the distribution of breast milk and urinary free and total bisphenol A (BPA) concentrations, from 27 post-partum women and their 31 infants, and explores the influence of age, sex, and nutritional source on infant BPA urinary concentration. Methods Both free (unconjugated) and total (free plus conjugated) BPA concentrations from women’s breast milk samples and infants’ urine samples were measured by online solid-phase extraction coupled to high-performance liquid chromatography–isotope dilution tandem mass spectrometry. Descriptive statistics and non-parametric tests of group comparisons were conducted. Results Total BPA was detected in 93% of urine samples in this healthy infant population aged 3–15 months who were without known environmental exposure to BPA (interquartile range [IQR]=1.2 – 4.4 μg/L). Similarly, 75% of the mothers’ breast milk samples had detectable concentrations of total BPA (IQR=0.4 – 1.4 μg/L). The magnitude and frequency of detection of free BPA in the children’s urine and the mothers’ breast milk were much lower than the total concentrations. Conclusions Total BPA was detected in 93% of this healthy infant population aged 3–15 months who are without known environmental exposure to BPA. Neither free nor total BPA urinary concentrations differed significantly by infant’s sex or by nutritional source (breast milk and/or formula) while age group was of borderline significance. There were no significant correlations between free or total BPA concentrations in mothers’ breast milk and their infants’ urine. PMID:23212895

  19. On-Demand Urine Analyzer

    NASA Technical Reports Server (NTRS)

    Farquharson, Stuart; Inscore, Frank; Shende, Chetan

    2010-01-01

    A lab-on-a-chip was developed that is capable of extracting biochemical indicators from urine samples and generating their surface-enhanced Raman spectra (SERS) so that the indicators can be quantified and identified. The development was motivated by the need to monitor and assess the effects of extended weightlessness, which include space motion sickness and loss of bone and muscle mass. The results may lead to developments of effective exercise programs and drug regimes that would maintain astronaut health. The analyzer containing the lab-on-a- chip includes materials to extract 3- methylhistidine (a muscle-loss indicator) and Risedronate (a bone-loss indicator) from the urine sample and detect them at the required concentrations using a Raman analyzer. The lab-on- a-chip has both an extractive material and a SERS-active material. The analyzer could be used to monitor the onset of diseases, such as osteoporosis.

  20. Urine Bacterial Community Convergence through Fertilizer Production: Storage, Pasteurization, and Struvite Precipitation.

    PubMed

    Lahr, Rebecca H; Goetsch, Heather E; Haig, Sarah J; Noe-Hays, Abraham; Love, Nancy G; Aga, Diana S; Bott, Charles B; Foxman, Betsy; Jimenez, Jose; Luo, Ting; Nace, Kim; Ramadugu, Kirtana; Wigginton, Krista R

    2016-11-01

    Source-separated human urine was collected from six public events to study the impact of urine processing and storage on bacterial community composition and viability. Illumina 16S rRNA gene sequencing revealed a complex community of bacteria in fresh urine that differed across collection events. Despite the harsh chemical conditions of stored urine (pH > 9 and total ammonia nitrogen > 4000 mg N/L), bacteria consistently grew to 5 ± 2 × 10 8 cells/mL. Storing hydrolyzed urine for any amount of time significantly reduced the number of operational taxonomic units (OTUs) to 130 ± 70, increased Pielou evenness to 0.60 ± 0.06, and produced communities dominated by Clostridiales and Lactobacillales. After 80 days of storage, all six urine samples from different starting materials converged to these characteristics. Urine pasteurization or struvite precipitation did not change the microbial community, even when pasteurized urine was stored for an additional 70 days. Pasteurization decreased metabolic activity by 50 ± 10% and additional storage after pasteurization did not lead to recovery of metabolic activity. Urine-derived fertilizers consistently contained 16S rRNA genes belonging to Tissierella, Erysipelothrix, Atopostipes, Bacteroides, and many Clostridiales OTUs; additional experiments must determine whether pathogenic species are present, responsible for observed metabolic activity, or regrow when applied.

  1. Dynamic detection of non-protein-bound strychnine and brucine in rabbit muscle and synovial fluid after topical application of total Strychnos alkaloid patches.

    PubMed

    Tang, Huaibo; Yan, Miao; Li, Huande; Xun, Tianrong; Deng, Yang; Zhao, Yeye; Deng, Long

    2014-04-01

    Semen Strychni, a known toxic drug in Chinese pharmacopoeia, is notable for its therapeutic effects on local muscle and joint pain. However, oral administration can be risky. Topically administered drugs accumulate in the topical muscles and knee joints without any major increase in plasma levels; only non-protein-bound drugs in the biological fluids of target tissues are effective for therapeutic effects. A sensitive and rapid ultra performance liquid chromatography - mass spectrometry (UPLC-MS) method coupled with a microdialysis technique was developed to determine the non-protein-bound strychnine (Str) and brucine (Bru) in rabbit muscle and synovial fluid microdialysate. The UPLC separation was carried out using a 1.7μm BEH C18 column (50 mm × 2.1 mm) with a mobile phase consisting of methanol: water (29.5:70.5, v/v) with 0.1% formic acid and 20 mM ammonium acetate in water. The method was validated at concentrations ranging from 0.58 ng/ml to 467.20 ng/ml for Str and from 0.42 ng/ml to 422.40 ng/ml for Bru. Intra-day and inter-day accuracy ranged from 99.1% to 103.2% for Str and from 95.8% to 108.8% for Bru with intra-day and inter-day precision within 9.7%. The proposed method was successfully applied to determine non-protein-bound Str and Bru, and the analysates concentration remained stable in rabbit muscle and synovial fluid after topical application of total Strychnos alkaloid patches, which indicated that total Strychnos alkaloid patches could substitute for the traditional oral administration of Semen Strychni. Copyright © 2013 John Wiley & Sons, Ltd.

  2. Doping control container for urine stabilization: a pilot study.

    PubMed

    Tsivou, Maria; Giannadaki, Evangelia; Hooghe, Fiona; Roels, Kris; Van Gansbeke, Wim; Garribba, Flaminia; Lyris, Emmanouil; Deventer, Koen; Mazzarino, Monica; Donati, Francesco; Georgakopoulos, Dimitrios G; Van Eenoo, Peter; Georgakopoulos, Costas G; de la Torre, Xavier; Botrè, Francesco

    2017-05-01

    Urine collection containers used in the doping control collection procedure do not provide a protective environment for urine, against degradation by microorganisms and proteolytic enzymes. An in-house chemical stabilization mixture was developed to tackle urine degradation problems encountered in human sport samples, in cases of microbial contamination or proteolytic activity. The mixture consists of antimicrobial substances and protease inhibitors for the simultaneous inactivation of a wide range of proteolytic enzymes. It has already been tested in lab-scale, as part of World Anti-Doping Agency's (WADA) funded research project, in terms of efficiency against microbial and proteolytic activity. The present work, funded also by WADA, is a follow-up study on the improvement of chemical stabilization mixture composition, application mode and limitation of interferences, using pilot urine collection containers, spray-coated in their internal surface with the chemical stabilization mixture. Urine in plastic stabilized collection containers have been gone through various incubation cycles to test for stabilization efficiency and analytical matrix interferences by three WADA accredited Laboratories (Athens, Ghent, and Rome). The spray-coated chemical stabilization mixture was tested against microorganism elimination and steroid glucuronide degradation, as well as enzymatic breakdown of proteins, such as intact hCG, recombinant erythropoietin and small peptides (GHRPs, ipamorelin), induced by proteolytic enzymes. Potential analytical interferences, observed in the presence of spray-coated chemical stabilization mixture, were recorded using routine screening procedures. The results of the current study support the application of the spray-coated plastic urine container, in the doping control collection procedure. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Urine Anion Gap to Predict Urine Ammonium and Related Outcomes in Kidney Disease.

    PubMed

    Raphael, Kalani L; Gilligan, Sarah; Ix, Joachim H

    2018-02-07

    Low urine ammonium excretion is associated with ESRD in CKD. Few laboratories measure urine ammonium, limiting clinical application. We determined correlations between urine ammonium, the standard urine anion gap, and a modified urine anion gap that includes sulfate and phosphate and compared risks of ESRD or death between these ammonium estimates and directly measured ammonium. We measured ammonium, sodium, potassium, chloride, phosphate, and sulfate from baseline 24-hour urine collections in 1044 African-American Study of Kidney Disease and Hypertension participants. We evaluated the cross-sectional correlations between urine ammonium, the standard urine anion gap (sodium + potassium - chloride), and a modified urine anion gap that includes urine phosphate and sulfate in the calculation. Multivariable-adjusted Cox models determined the associations of the standard urine anion gap and the modified urine anion gap with the composite end point of death or ESRD; these results were compared with results using urine ammonium as the predictor of interest. The standard urine anion gap had a weak and direct correlation with urine ammonium ( r =0.18), whereas the modified urine anion gap had a modest inverse relationship with urine ammonium ( r =-0.58). Compared with the highest tertile of urine ammonium, those in the lowest urine ammonium tertile had higher risk of ESRD or death (hazard ratio, 1.46; 95% confidence interval, 1.13 to 1.87) after adjusting for demographics, GFR, proteinuria, and other confounders. In comparison, participants in the corresponding standard urine anion gap tertile did not have higher risk of ESRD or death (hazard ratio, 0.82; 95% confidence interval, 0.64 to 1.07), whereas the risk for those in the corresponding modified urine anion gap tertile (hazard ratio, 1.32; 95% confidence interval, 1.03 to 1.68) approximated that of directly measured urine ammonium. Urine anion gap is a poor surrogate of urine ammonium in CKD unless phosphate and

  4. ELKS, a Protein Structurally Related to the Active Zone-associated Protein CAST, Is Expressed in Pancreatic β Cells and Functions in Insulin Exocytosis: Interaction of ELKS with Exocytotic Machinery Analyzed by Total Internal Reflection Fluorescence MicroscopyV⃞

    PubMed Central

    Ohara-Imaizumi, Mica; Ohtsuka, Toshihisa; Matsushima, Satsuki; Akimoto, Yoshihiro; Nishiwaki, Chiyono; Nakamichi, Yoko; Kikuta, Toshiteru; Nagai, Shintaro; Kawakami, Hayato; Watanabe, Takashi; Nagamatsu, Shinya

    2005-01-01

    The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitters. Here, we demonstrate the expression and function of ELKS, a protein structurally related to the CAZ protein CAST, in insulin exocytosis. The results of confocal and immunoelectron microscopic analysis showed that ELKS is present in pancreatic β cells and is localized close to insulin granules docked on the plasma membrane-facing blood vessels. Total internal reflection fluorescence microscopy imaging in insulin-producing clonal cells revealed that the ELKS clusters are less dense and unevenly distributed than syntaxin 1 clusters, which are enriched in the plasma membrane. Most of the ELKS clusters were on the docking sites of insulin granules that were colocalized with syntaxin 1 clusters. Total internal reflection fluorescence images of single-granule motion showed that the fusion events of insulin granules mostly occurred on the ELKS cluster, where repeated fusion was sometimes observed. When the Bassoon-binding region of ELKS was introduced into the cells, the docking and fusion of insulin granules were markedly reduced. Moreover, attenuation of ELKS expression by small interfering RNA reduced the glucose-evoked insulin release. These data suggest that the CAZ-related protein ELKS functions in insulin exocytosis from pancreatic β cells. PMID:15888548

  5. Surface Glycosylation Profiles of Urine Extracellular Vesicles

    PubMed Central

    Gerlach, Jared Q.; Krüger, Anja; Gallogly, Susan; Hanley, Shirley A.; Hogan, Marie C.; Ward, Christopher J.

    2013-01-01

    Urinary extracellular vesicles (uEVs) are released by cells throughout the nephron and contain biomolecules from their cells of origin. Although uEV-associated proteins and RNA have been studied in detail, little information exists regarding uEV glycosylation characteristics. Surface glycosylation profiling by flow cytometry and lectin microarray was applied to uEVs enriched from urine of healthy adults by ultracentrifugation and centrifugal filtration. The carbohydrate specificity of lectin microarray profiles was confirmed by competitive sugar inhibition and carbohydrate-specific enzyme hydrolysis. Glycosylation profiles of uEVs and purified Tamm Horsfall protein were compared. In both flow cytometry and lectin microarray assays, uEVs demonstrated surface binding, at low to moderate intensities, of a broad range of lectins whether prepared by ultracentrifugation or centrifugal filtration. In general, ultracentrifugation-prepared uEVs demonstrated higher lectin binding intensities than centrifugal filtration-prepared uEVs consistent with lesser amounts of co-purified non-vesicular proteins. The surface glycosylation profiles of uEVs showed little inter-individual variation and were distinct from those of Tamm Horsfall protein, which bound a limited number of lectins. In a pilot study, lectin microarray was used to compare uEVs from individuals with autosomal dominant polycystic kidney disease to those of age-matched controls. The lectin microarray profiles of polycystic kidney disease and healthy uEVs showed differences in binding intensity of 6/43 lectins. Our results reveal a complex surface glycosylation profile of uEVs that is accessible to lectin-based analysis following multiple uEV enrichment techniques, is distinct from co-purified Tamm Horsfall protein and may demonstrate disease-specific modifications. PMID:24069349

  6. Comparison of Plasma and Urine Biomarker Performance in Acute Kidney Injury

    PubMed Central

    Schley, Gunnar; Köberle, Carmen; Manuilova, Ekaterina; Rutz, Sandra; Forster, Christian; Weyand, Michael; Formentini, Ivan; Kientsch-Engel, Rosemarie; Eckardt, Kai-Uwe; Willam, Carsten

    2015-01-01

    Background New renal biomarkers measured in urine promise to increase specificity for risk stratification and early diagnosis of acute kidney injury (AKI) but concomitantly may be altered by urine concentration effects and chronic renal insufficiency. This study therefore directly compared the performance of AKI biomarkers in urine and plasma. Methods This single-center, prospective cohort study included 110 unselected adults undergoing cardiac surgery with cardiopulmonary bypass between 2009 and 2010. Plasma and/or urine concentrations of creatinine, cystatin C, neutrophil gelatinase-associated lipocalin (NGAL), liver fatty acid-binding protein (L-FABP), kidney injury molecule 1 (KIM1), and albumin as well as 15 additional biomarkers in plasma and urine were measured during the perioperative period. The primary outcome was AKI defined by AKIN serum creatinine criteria within 72 hours after surgery. Results Biomarkers in plasma showed markedly better discriminative performance for preoperative risk stratification and early postoperative (within 24h after surgery) detection of AKI than urine biomarkers. Discriminative power of urine biomarkers improved when concentrations were normalized to urinary creatinine, but urine biomarkers had still lower AUC values than plasma biomarkers. Best diagnostic performance 4h after surgery had plasma NGAL (AUC 0.83), cystatin C (0.76), MIG (0.74), and L-FAPB (0.73). Combinations of multiple biomarkers did not improve their diagnostic power. Preoperative clinical scoring systems (EuroSCORE and Cleveland Clinic Foundation Score) predicted the risk for AKI (AUC 0.76 and 0.71) and were not inferior to biomarkers. Preexisting chronic kidney disease limited the diagnostic performance of both plasma and urine biomarkers. Conclusions In our cohort plasma biomarkers had higher discriminative power for risk stratification and early diagnosis of AKI than urine biomarkers. For preoperative risk stratification of AKI clinical models showed

  7. Estimating residual kidney function in dialysis patients without urine collection.

    PubMed

    Shafi, Tariq; Michels, Wieneke M; Levey, Andrew S; Inker, Lesley A; Dekker, Friedo W; Krediet, Raymond T; Hoekstra, Tiny; Schwartz, George J; Eckfeldt, John H; Coresh, Josef

    2016-05-01

    Residual kidney function contributes substantially to solute clearance in dialysis patients but cannot be assessed without urine collection. We used serum filtration markers to develop dialysis-specific equations to estimate urinary urea clearance without the need for urine collection. In our development cohort, we measured 24-hour urine clearances under close supervision in 44 patients and validated these equations in 826 patients from the Netherlands Cooperative Study on the Adequacy of Dialysis. For the development and validation cohorts, median urinary urea clearance was 2.6 and 2.4 ml/min, respectively. During the 24-hour visit in the development cohort, serum β-trace protein concentrations remained in steady state but concentrations of all other markers increased. In the validation cohort, bias (median measured minus estimated clearance) was low for all equations. Precision was significantly better for β-trace protein and β2-microglobulin equations and the accuracy was significantly greater for β-trace protein, β2-microglobulin, and cystatin C equations, compared with the urea plus creatinine equation. Area under the receiver operator characteristic curve for detecting measured urinary urea clearance by equation-estimated urinary urea clearance (both 2 ml/min or more) were 0.821, 0.850, and 0.796 for β-trace protein, β2-microglobulin, and cystatin C equations, respectively; significantly greater than the 0.663 for the urea plus creatinine equation. Thus, residual renal function can be estimated in dialysis patients without urine collections. Copyright © 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

  8. Proteomic Investigation of the Response of Enterococcus faecalis V583 when Cultivated in Urine

    PubMed Central

    Arntzen, Magnus Øverlie; Karlskås, Ingrid Lea; Skaugen, Morten; Eijsink, Vincent G. H.; Mathiesen, Geir

    2015-01-01

    Enterococcus faecalis is a robust bacterium, which is able to survive in and adapt to hostile environments such as the urinary tract and bladder. In this label-free quantitative proteomic study based on MaxQuant LFQ algorithms, we identified 127 proteins present in the secretome of the clinical vancomycin-resistant isolate E. faecalis V583 and we compared proteins secreted in the initial phase of cultivation in urine with the secretome during cultivation in standard laboratory medium, 2xYT. Of the 54 identified proteins predicted to be secreted, six were exclusively found after cultivation in urine including the virulence factor EfaA (“endocarditis specific antigen”) and its homologue EF0577 (“adhesion lipoprotein”). These two proteins are both involved in manganese transport, known to be an important determinant of colonization and infection, and may additionally function as adhesins. Other detected urine-specific proteins are involved in peptide transport (EF0063 and EF3106) and protease inhibition (EF3054). In addition, we found an uncharacterized protein (EF0764), which had not previously been linked to the adaptation of V583 to a urine environment, and which is unique to E. faecalis. Proteins found in both environments included a histone-like protein, EF1550, that was up-regulated during cultivation in urine and that has a homologue in streptococci (HlpA) known to be involved in bacterial adhesion to host cells. Up-regulated secreted proteins included autolysins. These results from secretome analyses are largely compatible with previously published data from transcriptomics studies. All in all, the present data indicate that transport, in particular metal transport, adhesion, cell wall remodelling and the unknown function carried out by the unique EF0764 are important for enterococcal adaptation to the urine environment. These results provide a basis for a more targeted exploration of novel proteins involved in the adaptability and pathogenicity of E

  9. Proteomic Investigation of the Response of Enterococcus faecalis V583 when Cultivated in Urine.

    PubMed

    Arntzen, Magnus Øverlie; Karlskås, Ingrid Lea; Skaugen, Morten; Eijsink, Vincent G H; Mathiesen, Geir

    2015-01-01

    Enterococcus faecalis is a robust bacterium, which is able to survive in and adapt to hostile environments such as the urinary tract and bladder. In this label-free quantitative proteomic study based on MaxQuant LFQ algorithms, we identified 127 proteins present in the secretome of the clinical vancomycin-resistant isolate E. faecalis V583 and we compared proteins secreted in the initial phase of cultivation in urine with the secretome during cultivation in standard laboratory medium, 2xYT. Of the 54 identified proteins predicted to be secreted, six were exclusively found after cultivation in urine including the virulence factor EfaA ("endocarditis specific antigen") and its homologue EF0577 ("adhesion lipoprotein"). These two proteins are both involved in manganese transport, known to be an important determinant of colonization and infection, and may additionally function as adhesins. Other detected urine-specific proteins are involved in peptide transport (EF0063 and EF3106) and protease inhibition (EF3054). In addition, we found an uncharacterized protein (EF0764), which had not previously been linked to the adaptation of V583 to a urine environment, and which is unique to E. faecalis. Proteins found in both environments included a histone-like protein, EF1550, that was up-regulated during cultivation in urine and that has a homologue in streptococci (HlpA) known to be involved in bacterial adhesion to host cells. Up-regulated secreted proteins included autolysins. These results from secretome analyses are largely compatible with previously published data from transcriptomics studies. All in all, the present data indicate that transport, in particular metal transport, adhesion, cell wall remodelling and the unknown function carried out by the unique EF0764 are important for enterococcal adaptation to the urine environment. These results provide a basis for a more targeted exploration of novel proteins involved in the adaptability and pathogenicity of E. faecalis.

  10. Optimization for Peptide Sample Preparation for Urine Peptidomics

    SciTech Connect

    Sigdel, Tara K.; Nicora, Carrie D.; Hsieh, Szu-Chuan

    2014-02-25

    proteins when utilizing the conventional SPE method. In conclusion, the mSPE method was found to be superior to the conventional, standard SPE method for urine peptide sample preparation when applying LC-MS peptidomics analysis due to the optimized sample clean up that provided improved experimental inference from the confidently identified peptides.« less

  11. Associations between IVF outcomes and essential trace elements measured in follicular fluid and urine: a pilot study.

    PubMed

    Ingle, Mary E; Bloom, Michael S; Parsons, Patrick J; Steuerwald, Amy J; Kruger, Pamela; Fujimoto, Victor Y

    2017-02-01

    A hypothesis-generating pilot study exploring associations between essential trace elements measured in follicular fluid (FF) and urine and in vitro fertilization (IVF) endpoints. We recruited 58 women undergoing IVF between 2007 and 2008, and measured cobalt, chromium, copper, manganese, molybdenum, and zinc in FF (n = 46) and urine (n = 45) by inductively coupled plasma mass spectrometry (ICP-MS). We used multivariable regression models to assess the impact of FF and urine trace elements on IVF outcomes, adjusted for age, body mass index, race, and cigarette smoking. Trace elements were mostly present at lower concentrations in FF than in urine. The average number of oocytes retrieved was positively associated with higher urine cobalt, chromium, copper, and molybdenum concentrations. FF chromium and manganese were negatively associated with the proportion of mature oocytes, yet urine manganese had a positive association. FF zinc was inversely associated with average oocyte fertilization. Urine trace elements were significant positive predictors for the total number of embryos generated. FF copper predicted lower embryo fragmentation while urine copper was associated with higher embryo cell number and urine manganese with higher embryo fragmentation. No associations were detected for implantation, pregnancy, or live birth. Our results suggest the importance of trace elements in both FF and urine for intermediate, although not necessarily clinical, IVF endpoints. The results differed using FF or urine biomarkers of exposure, which may have implications for the design of clinical and epidemiologic investigations. These initial findings will form the basis of a more definitive future study.

  12. Association of total mixed ration particle fractions retained on the Penn State Particle Separator with milk, fat, and protein yield lactation curves at the cow level.

    PubMed

    Caccamo, M; Ferguson, J D; Veerkamp, R F; Schadt, I; Petriglieri, R; Azzaro, G; Pozzebon, A; Licitra, G

    2014-01-01

    As part of a larger project aiming to develop management evaluation tools based on results from test-day (TD) models, the objective of this study was to examine the effect of physical composition of total mixed rations (TMR) tested quarterly from March 2006 through December 2008 on milk, fat, and protein yield curves for 25 herds in Ragusa, Sicily. A random regression sire-maternal grandsire model was used to estimate variance components for milk, fat, and protein yields fitted on a full data set, including 241,153 TD records from 9,809 animals in 42 herds recorded from 1995 through 2008. The model included parity, age at calving, year at calving, and stage of pregnancy as fixed effects. Random effects were herd × test date, sire and maternal grandsire additive genetic effect, and permanent environmental effect modeled using third-order Legendre polynomials. Model fitting was carried out using ASREML. Afterward, for the 25 herds involved in the study, 9 particle size classes were defined based on the proportions of TMR particles on the top (19-mm) and middle (8-mm) screen of the Penn State Particle Separator. Subsequently, the model with estimated variance components was used to examine the influence of TMR particle size class on milk, fat, and protein yield curves. An interaction was included with the particle size class and days in milk. The effect of the TMR particle size class was modeled using a ninth-order Legendre polynomial. Lactation curves were predicted from the model while controlling for TMR chemical composition (crude protein content of 15.5%, neutral detergent fiber of 40.7%, and starch of 19.7% for all classes), to have pure estimates of particle distribution not confounded by nutrient content of TMR. We found little effect of class of particle proportions on milk yield and fat yield curves. Protein yield was greater for sieve classes with 10.4 to 17.4% of TMR particles retained on the top (19-mm) sieve. Optimal distributions different from those

  13. Spot Urine-guided Salt Reduction in Chronic Kidney Disease Patients.

    PubMed

    Uchiyama, Kiyotaka; Yanai, Akane; Ishibashi, Yoshitaka

    2017-09-01

    Dietary salt restriction is important in patients with chronic kidney disease (CKD) to reduce hypertension, cardiovascular events, progression of CKD, and mortality. However, recommending salt reduction for patients is difficult without knowing their actual sodium intake. This study evaluated the effectiveness of spot urine-guided salt reduction in CKD outpatients. A prospective cohort study was used. This study included a total of 127 adult outpatients (aged 60 ± 18 years, 80 males) with CKD. Their baseline estimated glomerular filtration rate was 51.4 ± 25.1 (mL/minute/1.73 m 2 ), and 64 (50%) of them were with CKD stage 3a or 3b (both 32 [25%]). We informed the patients of their individual spot urine-estimated salt intake every time they visited the outpatient clinic. Based on the data, the nephrologist encouraged the patients to achieve their salt restriction goal. The primary outcome was the estimated salt excretion, and the secondary outcome was the urinary protein-to-Cr ratio (UPCR). Multiple regression analyses were performed to clarify the contributing factors of changes in both outcomes. Over a follow-up of 12 months, the median number of patients' visits was 7 (5-8). The estimated salt intake was significantly reduced from 7.98 ± 2.49 g/day to 6.77 ± 1.77 g/day (P < .0001). The median UPCR was also reduced from 0.20 (0.10-0.80) to 0.10 (0.10-0.48) (P < .0001). On multiple regression analysis, a reduction in UPCR was positively associated with the baseline UPCR and a reduction in systolic blood pressure significantly (P < .0001 and P < .01, respectively) as well as positively correlated with a reduction in the estimated salt intake, with borderline significance (P = .08). Providing spot urine-estimated salt intake feedback effectively motivated CKD patients to reduce their salt intake. Spot urine-guided salt reduction may slow CKD progression through decreased urinary protein excretion. Copyright © 2017 National Kidney

  14. The Kjeldahl method as a primary reference procedure for total protein in certified reference materials used in clinical chemistry. II. Selection of direct Kjeldahl analysis and its preliminary performance parameters.

    PubMed

    Vinklárková, Bára; Chromý, Vratislav; Šprongl, Luděk; Bittová, Miroslava; Rikanová, Milena; Ohnútková, Ivana; Žaludová, Lenka

    2015-01-01

    To select a Kjeldahl procedure suitable for the determination of total protein in reference materials used in laboratory medicine, we reviewed in our previous article Kjeldahl methods adopted by clinical chemistry and found an indirect two-step analysis by total Kjeldahl nitrogen corrected for its nonprotein nitrogen and a direct analysis made on isolated protein precipitates. In this article, we compare both procedures on various reference materials. An indirect Kjeldahl method gave falsely lower results than a direct analysis. Preliminary performance parameters qualify the direct Kjeldahl analysis as a suitable primary reference procedure for the certification of total protein in reference laboratories.

  15. Amino acid profiles of rumen undegradable protein: a comparison between forages including cereal straws and alfalfa and their respective total mixed rations.

    PubMed

    Wang, B; Jiang, L S; Liu, J X

    2018-06-01

    Optimizing the amino acid (AA) profile of rumen undegradable protein (RUP) can positively affect the amount of milk protein. This study was conducted to improve knowledge regarding the AA profile of rumen undegradable protein from corn stover, rice straw and alfalfa hay as well as the total mixed ratio diets (TMR) based on one of them as forage source [forage-to-concentrate ratio of 45:55 (30% of corn stover (CS), 30% of rice straw (RS), 23% of alfalfa hay (AH) and dry matter basis)]. The other ingredients in the three TMR diets were similar. The RUP of all the forages and diets was estimated by incubation for 16 hr in the rumen of three ruminally cannulated lactating cows. All residues were corrected for microbial colonization, which was necessary in determining the AA composition of RUP from feed samples using in situ method. Compared with their original AA composition, the AA pattern of forages and forage-based diets changed drastically after rumen exposure. In addition, the extent of ruminal degradation of analysed AA was not constant among the forages. The greatest individual AA degradability of alfalfa hay and corn stover was Pro, but was His of rice straw. A remarkable difference was observed between microbial attachment corrected and uncorrected AA profiles of RUP, except for alfalfa hay and His in the three forages and TMR diets. The ruminal AA degradability of cereal straws was altered compared with alfalfa hay but not for the TMR diets. In summary, the AA composition of forages and TMR-based diets changed significantly after ruminal exposure, indicating that the original AA profiles of the feed cannot represent its AA composition of RUP. The AA profile of RUP and ruminal AA degradability for corn stover and rice straw contributed to missing information in the field. © 2017 Blackwell Verlag GmbH.

  16. Master Amino acid Pattern as sole and total substitute for dietary proteins during a weight-loss diet to achieve the body's nitrogen balance equilibrium.

    PubMed

    Lucà-Moretti, M; Grandi, A; Lucà, E; Muratori, G; Nofroni, M G; Mucci, M P; Gambetta, P; Stimolo, R; Drago, P; Giudice, G; Tamburlin, N; Karbalai, M; Valente, C; Moras, G

    2003-01-01

    Results of this multicentric study have shown that by giving Master Amino acid Pattern (MAP) as a sole and total substitute of dietary proteins to 500 overweight participants undergoing the American Nutrition Clinics/Overweight Management Program (ANC/OMP), the participants' body nitrogen balance could be maintained in equilibrium with essentially no calories (MAP 1 g=0.04 kcal), thereby preserving the body's structural and functional proteins, eliminating excessive water retention from the interstitial compartment, and preventing the sudden weight increase after study conclusion commonly known as the yo-yo effect. Study results have shown that the use of MAP, in conjunction with the ANC/OMP regimen, has proven to be safe and effective by preventing those adverse effects associated with a negative nitrogen balance, such as oversized or flabby tissue, stretch marks, the sagging of breast tissue, increased hair loss, faded hair color, and fragile or brittle nails. Also prevented were those anomalies commonly associated with weight-loss diets, such as hunger, weakness, headache caused by ketosis, constipation, and decreased libido. The use of MAP in conjunction with the ANC/OMP also allowed for mean weight loss of 2.5 kg (5.5 lb) per week, achieved through reduction of excessive fat tissue and elimination of excessive water retention from the interstitial compartment.

  17. Plasma-deposited tetraglyme surfaces greatly reduce total blood protein adsorption, contact activation, platelet adhesion, platelet procoagulant activity, and in vitro thrombus deposition.

    PubMed

    Cao, Lan; Chang, Mark; Lee, Chi-Ying; Castner, David G; Sukavaneshvar, Sivaprasad; Ratner, Buddy D; Horbett, Thomas A

    2007-06-15

    The ability of tetraethylene glycol dimethyl ether (tetraglyme) plasma deposited coatings exhibiting ultralow fibrinogen adsorption to reduce blood activation was studied with six in vitro methods, namely fibrinogen and von Willebrand's factor adsorption, total protein adsorption, clotting time in recalcified plasma, platelet adhesion and procoagulant activity, and whole blood thrombosis in a disturbed flow catheter model. Surface plasmon resonance results showed that tetraglyme surfaces strongly resisted the adsorption of all proteins from human plasma. The clotting time in the presence of tetraglyme surfaces was lengthened compared with controls, indicating a lower activation of the intrinsic coagulation cascade. Platelet adhesion and thrombin generation by adherent platelets were greatly reduced on tetraglyme-coated materials, compared with uncoated and Biospan-coated glass slides. In the in vitro disturbed blood flow model, tetraglyme plasma coated catheters had 50% less thrombus than did the uncoated catheters. Tetraglyme-coated materials thus had greatly reduced blood interactions as measured with all six methods. The improved blood compatibility of plasma-deposited tetraglyme is thus not only due to their reduced platelet adhesion and activation, but also to a generalized reduction in blood interactions. (c) 2007 Wiley Periodicals, Inc.

  18. The method of urine sampling is not a valid predictor for vesicoureteral reflux in children after febrile urinary tract infections.

    PubMed

    Haid, Bernhard; Roesch, Judith; Strasser, Christa; Oswald, Josef

    2017-10-01

    The likelihood of detecting vesicoureteral reflux (VUR) after febrile urinary tract infections (UTI) in children logically should correlate with the correct diagnosis of the UTI. Beneath the unspecific symptoms of fever urine analysis is the main diagnostic criterion for the exact diagnosis of febrile UTIs in children. Use of inadequate urine sampling techniques during diagnosis may lead to impaired accuracy in UTI diagnosis. This could lead to the assumption that children, having diagnosed their UTI by the use of possibly inadequate urine sampling techniques should not be evaluated as consequently compared to those, where the diagnosis relied on sterile urine sampling techniques. We hypothesized that children with possibly contaminated urine samples during the initial diagnosis may show a lower rate of VUR in subsequent VCUGs because of a wrong diagnosis initially compared to children, where accurate urine sampling techniques were used. Between 2009 and 2014, a total of 555 patients underwent a primary VCUG at our department indicated because of febrile UTIs. Patients with urine collection methods other than bag urine and catheter/suprapubic aspiration (SPA) were excluded from this study (mid-stream urine, potty urine, n = 149). We evaluated 402 patients (male/female 131/271, mean age 1.91 years), VUR rates and grades were compared between patients where urine was sampled by the use of a urine bag only at the time of diagnosis (n = 296, 73.6%) and those where sterile urine sampling (catheter, suprapubic puncture) was performed (n = 106, 26.3%). 4 patients were excluded due to equivocal data on urine sampling. VUR rate in children after sterile urine sampling using a catheter or SPA accounted to 31.1%. In those where urine samples acquired by the use of urine bags were used, 33.7% showed VUR on subsequent VCUG (p = 0.718). There were no significant differences as to VUR grades or gender, although VUR was much more commonly diagnosed in female patients (37

  19. The Use of Chlorhexidine/n-Propyl Gallate (CPG) as an Ambient-Temperature Urine Preservative

    NASA Technical Reports Server (NTRS)

    Nillen, Jeannie L.; Smith, Scott M.

    2003-01-01

    A safe, effective ambient temperature urine preservative, chlorhexidine/n-propyl gallate (CPG), has been formulated for use during spacefli ght that reduces the effects of oxidation and bacterial contamination on sample integrity while maintaining urine pH. The ability of this preservative to maintain stability of nine key analytes was evaluated for a period of one year. CPG effectively maintained stability of a mmonia, total nitrogen, 3-methylhistidine, chloride, sodium, potassiu m, and urea; however, creatinine and osmolality were not preserved by CPG. These data indicate that CPG offers prolonged room-temperature storage for multiple urine analytes, reducing the requirements for f rozen urine storage on future spaceflights. Iii medical applications on Earth, this technology can allow urine samples to be collected in remote settings and eliminate the need to ship frozen samples.

  20. Immunodetection and molecular determination of visceral and cutaneous Leishmania infection using patients' urine.

    PubMed

    Mirzaei, Asad; Ahmadipour, Fereshteh; Cannet, Arnaud; Marty, Pierre; Delaunay, Pascal; Perrin, Pascale; Dorkeld, Franck; Sereno, Denis; Akhoundi, Mohammad

    2018-05-27

    The diagnosis of leishmaniasis relies mainly on the use of invasive processes, to collect the biological material for detecting Leishmania parasites. Body fluids, which can be collected by non-invasive process, would greatly facilitate the leishmaniasis diagnosis. In the present study, we investigated the potency of urine immunoblotting to diagnose cutaneous and visceral leishmaniasis and we compared with routine molecular methods. A total of 80 samples, including 40 sera and their 40 corresponding urine samples were collected from 37 suspected patients with cutaneous and visceral leishmaniasis, and 3 healthy individuals (as control), in Ilam and Ardabil provinces of Iran. All sera and urine samples were analyzed, using immunoblotting. The confirmation of leishmaniasis infection was performed, using conventional and quantitative PCRs as well as by sequencing the amplicons. Among 37 suspected patients, 23 patients presented cutaneous lesions (CL) and 14 exhibited clinical symptoms reminiscent of visceral leishmaniasis (L. infantum). Among cutaneous patients, 15 were positive for zoonotic cutaneous leishmaniasis (L. major), and eight for anthroponotic cutaneous leishmaniasis (L. tropica). Molecular quantification of Leishmania parasites was performed on sera, urines and cutaneous biopsies of CL and VL patients, demonstrating that parasite load is lower in urines, compared to sera or biopsy. DNA can be detected in 20 out of 23 (86.9%) CL urine samples and in 13 out of 14 (92.8%) VL urine samples. Immunodetection analysis demonstrates that 22 out of 23 (95.6%) sera from CL patients and all patients suspected with VL are positive. For urine samples, 18 out of 23 (78.2%) urine of CL patients and 13 out of 14 (92.8%) urine of VL patients were positive, using Western blot. Therefore, immunodetection and molecular analysis using urine samples can be used as a diagnostic tool for surveying cutaneous and visceral leishmaniasis. Copyright © 2017. Published by Elsevier B.V.

  1. Determination of Cefalothin and Cefazolin in Human Plasma, Urine and Peritoneal Dialysate by UHPLC-MS/MS: application to a pilot pharmacokinetic study in humans.

    PubMed

    Parker, Suzanne L; Guerra Valero, Yarmarly C; Roberts, Darren M; Lipman, Jeffrey; Roberts, Jason A; Wallis, Steven C

    2016-06-01

    An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the analysis of cefazolin and cefalothin in human plasma (total and unbound), urine and peritoneal dialysate has been developed and validated. Total plasma concentrations are measured following protein precipitation and are suitable for the concentration range of 1-500 µg/mL. Unbound concentrations are measured from ultra-filtered plasma acquired using Centrifree(®) devices and are suitable for the concentration range of 0.1-500 µg/mL for cefazolin and 1-500 µg/mL for cefalothin. The urine method is suitable for a concentration range of 0.1-20 mg/mL for cefazolin and 0.2-20 mg/mL for cefalothin. Peritoneal dialysate concentrations are measured using direct injection, and are suitable for the concentration range of 0.2-100 µg/mL for both cefazolin and cefalothin. The cefazolin and cefalothin plasma (total and unbound), urine and peritoneal dialysate results are reported for recovery, inter-assay precision and accuracy, and the lower limit of quantification, linearity, stability and matrix effects, with all results meeting acceptance criteria. The method was used successfully in a pilot pharmacokinetic study with patients with peritoneal dialysis-associated peritonitis, receiving either intraperitoneal cefazolin or cefalothin. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  2. Effect of sodium chloride intake on urine volume, urinary urea excretion, and milk urea concentration in lactating dairy cattle.

    PubMed

    Spek, J W; Bannink, A; Gort, G; Hendriks, W H; Dijkstra, J

    2012-12-01

    Milk urea nitrogen (MUN; mg of N/dL) has been shown to be related to excretion of urinary urea N (UUN; g of N/d) and total excretion of urinary N (UN; g of N/d) in dairy cows. In the present experiment, it was hypothesized that MUN and the relationship between MUN and UUN or UN is affected by urine volume as a result of dietary sodium chloride intake. Twelve lactating Holstein-Friesian dairy cows (mean ± SD: milk production 28.1±3.23 kg/d and 190±41 d in milk), of which 4 were fitted with catheters in the urine bladder and jugular vein, were randomly assigned to 4 dietary levels of sodium chloride (3, 9, 14, and 19 g of Na/kg of DM) according to a triple 4×4 Latin square design. Cows were fed at 95% of ad libitum intake, excluding salt addition. Milk was analyzed for MUN and protein content; urine was analyzed for total N, urea, and creatinine content; feces were analyzed for total N and DM content; and blood plasma was analyzed for urea and creatinine content. Creatinine clearance rate (CCR; L/min) and renal urea reabsorption ratio were estimated based on plasma concentrations of urea and creatinine, and total excretion of urea and creatinine in urine. Intake of DM and N, milk production, and milk protein content were (mean ± SD), on average, 21.4±1.24 kg/d, 522±32.0 g/d, 25.4±2.53 kg/d, and 3.64±0.186%, respectively. A linear relationship was found between Na intake and urine production [urine (kg/d; mean ± SE)=7.5±4.33+0.136±0.0143 × Na intake (g/d)] and between Na intake and MUN [MUN (mg/dL; mean ± SE)=13.5±0.35-0.0068±0.00104 × Na intake (g/d)]. Despite the decrease in MUN with increased Na intake, UN excretion increased linearly with Na intake. Excretion of UUN was not affected by dietary Na content. A linear plateau relationship was observed between CCR and renal urea reabsorption. An increase in CCR coincided with an increase in calculated renal urea reabsorption until a CCR breakpoint value (mean ± SD) of 1.56±0.063 L/min was reached. We

  3. Urine TREM-1 as a marker of urinary tract infection in children.

    PubMed

    Sierra-Diaz, Erick; Bravo Cuéllar, Alejandro; Ortiz Lazareno, Pablo Cesar; García Gutiérrez, Mariana; Georgina, Hernandez Flores; Anaya Prado, Roberto

    2017-04-01

    Objective Triggering receptor expressed on myeloid cells (TREM)-1 is a receptor that is thought to improve recognition of patients with true infection. In this study, we investigated whether Triggering receptor expressed on myeloid cells (TREM-1) is present in urine samples from children with urinary tract infection (UTI) and in samples from healthy children. Methods A total of 128 samples met the inclusion criteria for the study. Urine samples were processed for culture and urinalysis as a regular protocol for patients with UTI. Samples were classified according to culture and urinalysis results. TREM-1 protein expression was detected with flow cytometry and sTREM-1 was assessed by ELISA. Results Flow cytometry showed detectable expression of TREM-1 in 100% of samples, UTI and non-UTI groups ( p < 0.001). Mean fluorescence intensity of TREM-1 was different between the groups ( p < 0.001). Levels of sTREM-1 were detected in patients with UTI, but not in non-UTI patients. Conclusions All of our patients (healthy and diseased) showed TREM-1 expression. However, TREM-1 levels in patients with UTI tend to be higher and are associated with increased neutrophils and cytokine activity induced by bacteria.

  4. Urine TREM-1 as a marker of urinary tract infection in children

    PubMed Central

    Sierra-Diaz, Erick; Ortiz Lazareno, Pablo Cesar; García Gutiérrez, Mariana; Georgina, Hernandez Flores; Anaya Prado, Roberto

    2017-01-01

    Objective Triggering receptor expressed on myeloid cells (TREM)-1 is a receptor that is thought to improve recognition of patients with true infection. In this study, we investigated whether Triggering receptor expressed on myeloid cells (TREM-1) is present in urine samples from children with urinary tract infection (UTI) and in samples from healthy children. Methods A total of 128 samples met the inclusion criteria for the study. Urine samples were processed for culture and urinalysis as a regular protocol for patients with UTI. Samples were classified according to culture and urinalysis results. TREM-1 protein expression was detected with flow cytometry and sTREM-1 was assessed by ELISA. Results Flow cytometry showed detectable expression of TREM-1 in 100% of samples, UTI and non-UTI groups (p < 0.001). Mean fluorescence intensity of TREM-1 was different between the groups (p < 0.001). Levels of sTREM-1 were detected in patients with UTI, but not in non-UTI patients. Conclusions All of our patients (healthy and diseased) showed TREM-1 expression. However, TREM-1 levels in patients with UTI tend to be higher and are associated with increased neutrophils and cytokine activity induced by bacteria. PMID:28367708

  5. Effects of nonstructural carbohydrates and protein sources on intake, apparent total tract digestibility, and ruminal metabolism in vivo and in vitro with high-concentrate beef cattle diets.

    PubMed

    Rotger, A; Ferret, A; Calsamiglia, S; Manteca, X

    2006-05-01

    To investigate the effects of synchronizing nonstructural carbohydrate (NSC) and protein degradation on intake and rumen microbial fermentation, four ruminally fistulated Holstein heifers (BW = 132.3 +/- 1.61 kg) fed high-concentrate diets were assigned to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments studied in vivo and in vitro with a dual-flow continuous culture system. Two NSC sources (barley and corn) and 2 protein sources [soybean meal (SBM) and sunflower meal (SFM)] differing in their rate and extent of ruminal degradation were combined resulting in a synchronized rapid fermentation diet (barley-SFM), a synchronized slow fermentation diet (corn-SBM), and 2 unsynchronized diets with a rapidly and a slowly fermenting component (barley-SBM, and corn-SFM). In vitro, the fermentation profile was studied at a constant pH of 6.2, and at a variable pH with 12 h at pH 6.4 and 12 h at pH 5.8. Synchronization tended to result in greater true OM digestion (P = 0.072), VFA concentration (P = 0.067), and microbial N flow (P = 0.092) in vitro, but had no effects on in vivo fermentation pattern or on apparent total tract digestibility. The NSC source affected the efficiency of microbial protein synthesis in vitro, tending to be greater (P = 0.07) for barley-based diets, and in vivo, the NSC source tended to affect intake. Dry matter and OM intake tended to be greater (P > or = 0.06) for corn- than barley-based diets. Ammonia N concentration was lower in vitro (P = 0.006) and tended to be lower in vivo (P = 0.07) for corn- than barley-based diets. In vitro, pH could be reduced from 6.4 to 5.8 for 12 h/d without any effect on ruminal fermentation or microbial protein synthesis. In summary, ruminal synchronization seemed to have positive effects on in vitro fermentation, but in vivo recycling of endogenous N or intake differences could compensate for these effects.

  6. Green Urine in Traditional Persian Medicine

    PubMed Central

    Kolouri, Sepideh; Daneshfard, Babak; Jaladat, Amir-Mohammad; Tafazoli, Vahid

    2016-01-01

    The color of urine is an important factor in urine examination, which can help physicians differentiate various diseases. Today, it is known that certain dyes, drug intoxications, and diseases can induce green urine discoloration. In the view of traditional Persian medicine, which is based on humoral medicine, green urine discoloration is generally referred to the dominance of coldness in the body. In fact, it is considered to be a result of a special kind of humoral imbalance and fluid depletion or retention in the human body. Persian scholars believed that green urine could be an indicator of intoxication or a predictor of an imminent spasm or convulsion in pediatric patients. Further investigations could result in finding new diagnostic scales of urine color based on the teachings of traditional Persian medicine. PMID:27103627

  7. Analysis of urine composition in type Ⅱ diabetic mice after intervention therapy using holothurian polypeptides

    NASA Astrophysics Data System (ADS)

    Li, Yanyan; Xu, Jiajie; Su, Xiurong

    2017-07-01

    Hydrolysates and peptide fractions (PF) obtained from sea cucumber with commercial enzyme were studied on the hpyerglycemic and renal protective effects on db/db rats using urine metabolomics. Compared with the control group the polypeptides from the two species could significantly reduce the urine glucose and urea. We also tried to address the compositions of highly expressed urinary proteins using a proteomics approach. They were serum albumins, AMBP proteins, negative trypsin, elastase and urinary protein, GAPDH, a receptor of urokinase-type plasminogen activator (uPAR), and Ig kappa chain C region. We used the electronic nose to quickly detect changes in the volatile substances in mice urine after holothurian polypeptides fed, and the results show it can identify the difference between treatment groups with the control group without overlapping. The protein express mechanism of holothurian polypeptides treating diabetes was discussed, and we suggested these two peptides with the hypoglycemic and renal protective activity might be utilized as nutraceuticals.

  8. The Kjeldahl method as a primary reference procedure for total protein in certified reference materials used in clinical chemistry. I. A review of Kjeldahl methods adopted by laboratory medicine.

    PubMed

    Chromý, Vratislav; Vinklárková, Bára; Šprongl, Luděk; Bittová, Miroslava

    2015-01-01

    We found previously that albumin-calibrated total protein in certified reference materials causes unacceptable positive bias in analysis of human sera. The simplest way to cure this defect is the use of human-based serum/plasma standards calibrated by the Kjeldahl method. Such standards, commutative with serum samples, will compensate for bias caused by lipids and bilirubin in most human sera. To find a suitable primary reference procedure for total protein in reference materials, we reviewed Kjeldahl methods adopted by laboratory medicine. We found two methods recommended for total protein in human samples: an indirect analysis based on total Kjeldahl nitrogen corrected for its nonprotein nitrogen and a direct analysis made on isolated protein precipitates. The methods found will be assessed in a subsequent article.

  9. Glutamyl aminopeptidase in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats.

    PubMed

    Quesada, Andrés; Segarra, Ana Belén; Montoro-Molina, Sebastián; de Gracia, María Del Carmen; Osuna, Antonio; O'Valle, Francisco; Gómez-Guzmán, Manuel; Vargas, Félix; Wangensteen, Rosemary

    2017-01-01

    The aim of this work was to investigate if the content of glutamyl aminopeptidase (GluAp) in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats. Urine samples were collected 24 hours after injection of cisplatin (7 mg/kg, n = 10) or saline serum (n = 10), and they were subjected to differential centrifugation at 1.000, 17.000 and 200.000 g to obtain microvesicular and exosomal fractions. GluAp was measured with a commercial ELISA kit in both fractions. Serum creatinine (SCr) and body weight were measured 15 days after treatment. We analyzed if early excretion of GluAp in microsomal and exosomal fractions was correlated with final SCr and body weight increase. In a second experiment, enzymatic activities of GluAp and alanyl aminopeptidase (AlaAp) in urine, microvesicular and exosomal fractions were measured three days after injection. We analyzed the correlation of both markers with SCr determined at this point. Finally, we studied the expression of GluAp and extracellular vesicles markers Alix and tumor susceptibility gene (TSG101) in both fractions by immunoblotting. GluAp excretion was increased in all fractions of urine after cisplatin treatment, even if data were normalized per mg of creatinine, per body weight or per total protein content of each fraction. We found significant predictive correlations with SCr concentration, and inverse correlations with body weight increase determined 15 days later. Three days after injection, aminopeptidasic activities were markedly increased in all fractions of urine in cisplatin-treated rats. The highest correlation coefficient with SCr was found for GluAp in microvesicular fraction. Increase of GluAp in microvesicular and exosomal fractions from cisplatin-treated rats was confirmed by immunoblotting. Alix and TSG101 showed different patterns of expression in each fraction. Determination of GluAp content or its enzymatic activity in microvesicular and exosomal

  10. Evaluation of the process of recycling and renal parenchymal injury after eswl with metabolites excreted in the urine.

    PubMed

    Ceylan, Cavit; Dogan, Serkan; Saydam, Gulsevim; Kocak, Mehmet Zait; Doluoglu, Omer Gokhan

    2013-01-01

    To show renal parenchymal injury depending on extracorporeal shock wave lithotripsy (ESWL). The patients with one renal stone and in whom ESWL is planned among the patients in whom renal stone was determined. Their 24-h urine samples were collected just before and after the ESWL treatment. Cit (citrate), UrA (uric acid), RBP (retinol-binding protein), NAG (N-acetyl-β-Đ-glucosaminidase), Cr (creatinine), Na (sodium), K (potassium), P (phosphor), Ca (calcium), and Cl (chlorine) metabolites excreted in urine were evaluated after urine samples were taken on the study day. Changes in the metabolites excreted; the number, frequency, and duration of ESWL shock wave; the energy; and the body mass index were recorded. The results for p < 0.05 will be accepted as statistically significant. Two sessions of ESWL were applied to a total of 20 patients. When metabolites excreted in the urine before (B1E) and after (A1E) the first session of ESWL, and before (B2E) and after (A2E) the second session of ESWL, were evaluated, no statistically significant result for Ca and Cl excretion was noted. For NAG and Cr, a significant difference was observed in terms of metabolite excretion between B1E and B2E. For other metabolites, we saw that there is no difference between B1E and B2E. While a significant metabolite change was observed for RBP, NAG, Cr, and Na as long as A1E and A2E ESWL session number increases, other metabolites were not significant. Shock waves induce significant damage to the renal and adjacent tissues as indicated by a significant increase in cell-escaped enzymes and electrolytes and the extent of damage depends on the energy and the number of shock wave exposure.

  11. A method for estimating radioactive cesium concentrations in cattle blood using urine samples.

    PubMed

    Sato, Itaru; Yamagishi, Ryoma; Sasaki, Jun; Satoh, Hiroshi; Miura, Kiyoshi; Kikuchi, Kaoru; Otani, Kumiko; Okada, Keiji

    2017-12-01

    In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the 'difficult-to-return zone'. Urine 137 Cs, specific gravity, electrical conductivity, pH, sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood 137 Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood 137 Cs] = [urinary 137 Cs]/([specific gravity] - 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher 137 Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood 137 Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle. © 2017 Japanese Society of Animal Science.

  12. Comparison of vacuum and non-vacuum urine tubes for urinary sediment analysis.

    PubMed

    Topcuoglu, Canan; Sezer, Sevilay; Kosem, Arzu; Ercan, Mujgan; Turhan, Turan

    2017-12-01

    Urine collection systems with aspiration system for vacuum tubes are becoming increasingly common for urinalysis, especially for microscopic examination of the urine. In this study, we aimed to examine whether vacuum aspiration of the urine sample has any adverse effect on sediment analysis by comparing results from vacuum and non-vacuum urine tubes. The study included totally 213 urine samples obtained from inpatients and outpatients in our hospital. Urine samples were collected to containers with aspiration system for vacuum tubes. Each sample was aliquoted to both vacuum and non-vacuum urine tubes. Urinary sediment analysis was performed using manual microscope. Results were evaluated using chi-square test. Comparison of the sediment analysis results from vacuum and non-vacuum urine tubes showed that results were highly concordant for erythrocyte, leukocyte and epithelial cells (gamma values 1, 0.997, and 0.994, respectively; p < .001). Results were also concordant for urinary casts, crystals and yeast (kappa values 0.815, 0.945 and 1, respectively; p < .001). The results show that in urinary sediment analysis, vacuum aspiration has no adverse effect on the cellular components except on casts.

  13. Neuroradiological findings in maple syrup urine disease

    PubMed Central

    Indiran, Venkatraman; Gunaseelan, R. Emmanuel

    2013-01-01

    Maple syrup urine disease is a rare inborn error of amino acid metabolism involving catabolic pathway of the branched-chain amino acids. This disease, if left untreated, may cause damage to the brain and may even cause death. These patients typically present with distinctive maple syrup odour of sweat and urine. Patients typically present with skin and urine smelling like maple syrup. Here we describe a case with relevant magnetic resonance imaging findings and confirmatory biochemical findings. PMID:23772241

  14. Neuroradiological findings in maple syrup urine disease.

    PubMed

    Indiran, Venkatraman; Gunaseelan, R Emmanuel

    2013-01-01

    Maple syrup urine disease is a rare inborn error of amino acid metabolism involving catabolic pathway of the branched-chain amino acids. This disease, if left untreated, may cause damage to the brain and may even cause death. These patients typically present with distinctive maple syrup odour of sweat and urine. Patients typically present with skin and urine smelling like maple syrup. Here we describe a case with relevant magnetic resonance imaging findings and confirmatory biochemical findings.

  15. Protopine alkaloids in horse urine.

    PubMed

    Wynne, Paul M; Vine, John H; Amiet, R Gary

    2004-11-05

    Protopine was extracted from Fumaria officinalis and purified by column chromatography. Urine samples were collected from horses and a human volunteer that had been administered either F. officinalis or protopine free base. Plant and urine samples were acetylated and analysed by GCMS after solid-phase extraction (SPE). The urinary metabolites of protopine were identified as 4,6,7,13-tetrahydro-9,10-dihydroxy-5-methyl-benzo[e]-l,3-benzodioxolo [4,5-1][2] benzazecin-12(5H)-one, 4,6,7,13-tetrahydro-10-hydroxy-9-methoxy-5-methyl-benzo[e]-1,3-benzodioxolo[4,5-1][2] benzazecin-12(5H)-one and 4,6,7,13-tetrahydro-9-hydroxy-10-methoxy-5-methyl-benzo[e]-1,3-benzodioxolo[4,5-l][2] benzazecin-12(5H)-one, chelianthifoline, isochelianthifoline and 2-O-desmethylchelianthifoline. The metabolic formation of the tetrahydroprotoberberines by closure of the bridge across N5 and C13 is rate limited and protopine-like metabolites accumulate only when the route is overloaded. Metabolism was qualitatively similar in the horse and human.

  16. Age-dependent change in urine proteome of healthy individuals

    NASA Astrophysics Data System (ADS)

    Dobrokhotov, Igor; Liudmila Pastushkova, MRS.; Larina, Irina; Kononikhin, Alexey

    It was analyzed the protein composition of urine samples obtained from twenty Russian cosmonauts and thirty-eight healthy volunteers, that have been selected for the experiments simulating the physiological effects of microgravity. The special sample preparation was performed, followed by liquid chromatography-mass spectrometry. Liquid chromatography-mass spectrometry of the minor proteins was performed on a nano-HPLC Agilent 1100 system (Agilent Technologies Inc., USA) in combination with a LTQ-FT Ultra mass spectrometer (Thermo Electron, Germany). List of masses derived peptides and they fragments have used for search and identification of proteins by database IPI-human (international index of protein) using the program Mascot (MS version 2.0.04 , UK) according to the following criteria: 1 - enzyme-trypsin; 2 - peptide tol. ± 5 ppm; 3 - MS / MS tol. 0.5Da. From list of proteins obtained as a result Mascot-search it was selected only those proteins that were identified based on 2 or more peptides with the rating more than 24. Analysis of the list of proteins was performed using software developed in the laboratory of VA Ivanisenko (ICG SB RAS) Age of healthy individuals was ranged from 18 to 54 years. Depending on the age, the data were divided into three groups: those relating to the group of persons under 25 years (youth and mature age 1), 25-40 years (mature age 2) and 40-54 years (mature age 3) It was detected reliable changes in the number of proteins among groups depending of the age. It was found that the minimum number of different proteins were detected in the urine of the group of young patients (under 25 years old) , and the maximum - was observed in the group of middle-aged persons (25 to 40 years). When the proteins were compared according to their molecular mass it was revealed that in the older group (40-54 years ) there is noticeably smaller percentage of high molecular weight proteins than in groups of young and middle aged persons. Thus

  17. Asymptomatic bacteriuria in pregnant women attending Boo-Ali Hospital Tehran Iran: Urine analysis vs. urine culture

    PubMed Central

    Etminan-Bakhsh, Mina; Tadi, Sima; Darabi, Roksana

    2017-01-01

    Background Asymptomatic bacteriuria is one of the common problems in pregnancy. Asymptomatic bacteriuria is associated with pyelonephritis, preterm labor and low birth weight infants. The physiological and anatomical changes in pregnancy facilitate urinary tract infection (UTI) during pregnancy. Several tests are available for diagnosis of asymptomatic bacteriuria. The urine culture is a gold standard diagnostic test for asymptomatic bacteriuria but it is expensive and time-consuming. Screening methods may be useful in detecting high-risk pregnant women for asymptomatic bacteriuria. Objective The aim of the present study was to compare urine analysis as a rapid screening test to urine culture in diagnosis of asymptomatic bacteriuria. Methods A total of 123 pregnant women attending the obstetrics clinic of Boo-Ali hospital in Tehran, Iran from March 2013 to September 2014 were included in the present diagnostic cross-sectional study. One hundred twenty three mid-stream urine samples were inoculated into cultures and were processed by dipstick (nitrite test and leucocyte esterase test) and microscopic pus cell count. The sensitivity, specificity, positive predictive value and negative predictive value of nitrite test, leucocyte esterase test and microscopic pus cell count were compared with urine culture in diagnosis of asymptomatic bacteriuria by using SPSS version 19. Results Of 123 urine samples, significant asymptomatic bacteriuria (≥104 cfu/Ml) was detected in 8 (6.5%) subjects. The sensitivity and specificity of nitrite test were 37% and 100% respectively. The sensitivity of pus cell count alone and leucocyte esterase test alone were 100% but the specificity of them were 64% and 65% respectively. We found high negative predictive value by Pus cell count and the leucocyte esterase test (100%) and low positive predictive value by them (16% and 17% respectively). Conclusion Urine culture is the most useful test for diagnosis of asymptomatic bacteriuria. None of

  18. Determination of urine ionic composition with potentiometric multisensor system.

    PubMed

    Yaroshenko, Irina; Kirsanov, Dmitry; Kartsova, Lyudmila; Sidorova, Alla; Borisova, Irina; Legin, Andrey

    2015-01-01

    The ionic composition of urine is a good indicator of patient's general condition and allows for diagnostics of certain medical problems such as e.g., urolithiasis. Due to environmental factors and malnutrition the number of registered urinary tract cases continuously increases. Most of the methods currently used for urine analysis are expensive, quite laborious and require skilled personnel. The present work deals with feasibility study of potentiometric multisensor system of 18 ion-selective and cross-sensitive sensors as an analytical tool for determination of urine ionic composition. In total 136 samples from patients of Urolithiasis Laboratory and healthy people were analyzed by the multisensor system as well as by capillary electrophoresis as a reference method. Various chemometric approaches were implemented to relate the data from electrochemical measurements with the reference data. Logistic regression (LR) was applied for classification of samples into healthy and unhealthy producing reasonable misclassification rates. Projection on Latent Structures (PLS) regression was applied for quantitative analysis of ionic composition from potentiometric data. Mean relative errors of simultaneous prediction of sodium, potassium, ammonium, calcium, magnesium, chloride, sulfate, phosphate, urate and creatinine from multisensor system response were in the range 3-13% for independent test sets. This shows a good promise for development of a fast and inexpensive alternative method for urine analysis. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Albuminuria is associated with an increased prostasin in urine while aldosterone has no direct effect on urine and kidney tissue abundance of prostasin.

    PubMed

    Oxlund, Christina; Kurt, Birgül; Schwarzensteiner, Ilona; Hansen, Mie R; Stæhr, Mette; Svenningsen, Per; Jacobsen, Ib A; Hansen, Pernille B; Thuesen, Anne D; Toft, Anja; Hinrichs, Gitte R; Bistrup, Claus; Jensen, Boye L

    2017-06-01

    The proteinase prostasin is a candidate mediator for aldosterone-driven proteolytic activation of the epithelial sodium channel (ENaC). It was hypothesized that the aldosterone-mineralocorticoid receptor (MR) pathway stimulates prostasin abundance in kidney and urine. Prostasin was measured in plasma and urine from type 2 diabetic patients with resistant hypertension (n = 112) randomized to spironolactone/placebo in a clinical trial. Prostasin protein level was assessed by immunoblotting in (1) human and rat urines with/without nephrotic syndrome, (2) human nephrectomy tissue, (3) urine and kidney from aldosterone synthase-deficient (AS -/- ) mice and ANGII- and aldosterone-infused mice, and in (4) kidney from adrenalectomized rats. Serum aldosterone concentration related to prostasin concentration in urine but not in plasma. Plasma prostasin concentration increased significantly after spironolactone compared to control. Urinary prostasin and albumin related directly and were reduced by spironolactone. In patients with nephrotic syndrome, urinary prostasin protein was elevated compared to controls. In rat nephrosis, proteinuria coincided with increased urinary prostasin, unchanged kidney tissue prostasin, and decreased plasma prostasin while plasma aldosterone was suppressed. Prostasin protein abundance in human nephrectomy tissue was similar across gender and ANGII inhibition regimens. Prostasin urine abundance was not different in AS -/- and aldosterone-infused mice. Prostasin kidney level was not different from control in adrenalectomized rats and AS -/- mice. We found no evidence for a direct relationship between mineralocorticoid receptor signaling and kidney and urine prostasin abundance. The reduction of urinary prostasin in spironolactone-treated patients is most likely the result of an improved glomerular filtration barrier function and generally reduced proteinuria.

  20. The Characterization of Feces and Urine: A Review of the Literature to Inform Advanced Treatment Technology

    PubMed Central

    Rose, C.; Parker, A.; Jefferson, B.; Cartmell, E.

    2015-01-01

    generation rates were 1.42 L/cap/day with a dry solids content of 59 g/cap/day. Variation in the volume and composition of urine is caused by differences in physical exertion, environmental conditions, as well as water, salt, and high protein intakes. Urine has a pH 6.2 and contains the largest fractions of nitrogen, phosphorus, and potassium released from the body. The urinary excretion of nitrogen was significant (10.98 g/cap/day) with urea the most predominant constituent making up over 50% of total organic solids. The dietary intake of food and fluid is the major cause of variation in both the fecal and urine composition and these variables should always be considered if the generation rate, physical, and chemical composition of feces and urine is to be accurately predicted. PMID:26246784

  1. Mean population salt intake estimated from 24-h urine samples and spot urine samples: a systematic review and meta-analysis.

    PubMed

    Huang, Liping; Crino, Michelle; Wu, Jason H Y; Woodward, Mark; Barzi, Federica; Land, Mary-Anne; McLean, Rachael; Webster, Jacqui; Enkhtungalag, Batsaikhan; Neal, Bruce

    2016-02-01

    Estimating equations based on spot urine samples have been identified as a possible alternative approach to 24-h urine collections for determining mean population salt intake. This review compares estimates of mean population salt intake based upon spot and 24-h urine samples. We systematically searched for all studies that reported estimates of daily salt intake based upon both spot and 24-h urine samples for the same population. The associations between the two were quantified and compared overall and in subsets of studies. A total of 538 records were identified, 108 were assessed as full text and 29 were included. The included studies involved 10,414 participants from 34 countries and made 71 comparisons available for the primary analysis. Overall average population salt intake estimated from 24-h urine samples was 9.3 g/day compared with 9.0 g/day estimated from the spot urine samples. Estimates based upon spot urine samples had excellent sensitivity (97%) and specificity (100%) at classifying mean population salt intake as above or below the World Health Organization maximum target of 5 g/day. Compared with the 24-h samples, estimates based upon spot urine overestimated intake at lower levels of consumption and underestimated intake at higher levels of consumption. Estimates of mean population salt intake based upon spot urine samples can provide countries with a good indication of mean population salt intake and whether action on salt consumption is required. Published by Oxford University Press on behalf of the International Epidemiological Association 2015. This work is written by US Government employees and is in the public domain in the US.

  2. Evaluation of four commercial biuret reagent kits of serum total protein by the American Association for Clinical Chemistry reference measurement procedure.

    PubMed

    He, Meilin; Zhang, Jie

    2011-06-01

    In China, the traceability of clinical chemistry methods is still immature. Therefore, it is necessary to establish a reference measurement procedure and evaluate commercial reagent kits using such established procedures. We reproduced the reference measurement procedure for serum total protein, as recommended by the American Association for Clinical Chemistry (AACC). We evaluated the performance by Clinical Laboratory Standard Institute (CLSI) guidelines EP15-A and EP6-A. Subsequently, four commercial reagent kits were evaluated by the reproduced reference procedure following CLSI guideline EP9-A2. The performance of the reproduced reference procedure was as follows: CVs ranged from 0.47% to 0.85% at medical decision levels (X(c)) of 45 g/L, 60 g/L and 80 g/L. Linearity was Y=1.0022X-0.2121 (r=0.9999), and recovery ranged from 100.2% to 102.4%. The External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine (RELA) was applied, and the result was within the limit of equivalence. The linear relationships of four commercial reagent kits, Merit Choice, KHB, Leadman, and Olympus, were, respectively: Y=0.9922X+0.5776 (r=0.9961); Y=0.9936X+0.4316 (r=0.9992); Y=0.9949X+0.9129 (r=0.9987) and Y=0.9923X+0.8876 (r=0.9989). KHB showed slight negative bias, and the mean±SD was -0.03±0.60 g/L. Merit Choice, Leadman, and Olympus all showed positive bias, and the mean±SDs were 0.02±0.63 g/L, 0.55±0.77 g/L and 0.34±0.71 g/L, respectively. The correlation and bias of four commercial reagent kits for serum total protein were found to be acceptable. Thus, these reagent kits can be used reliably in China.

  3. Comparative evaluation of serum and salivary immunoglobulin G and A levels with total serum protein in oral submucous fibrosis patients: A case control study.

    PubMed

    Kandasamy, M; Jaisanghar, N; Austin, Ravi David; Srivastava, Kumar Chandan; Anusuya, G Sai; Anisa, N

    2016-10-01

    The objective of this study is to estimate and compare the serum and salivary immunoglobulin G and A (IgG, IgA) levels in various stages of oral submucous fibrosis (OSMF) patients and relate it to total serum protein (TSP) and hemoglobin (Hb) levels. The sample for the present study comprised a total of 20 healthy controls, 20 OSMF patients. About 5 ml of blood and 2 ml of saliva were collected. Quantitative analysis of serum and salivary IgG, IgA was done by turbidometric immunoassay. TSP and Hb were estimated by Biuret and cyanmethemoglobin methods, respectively. Serum and salivary IgA and IgG levels were statistically significantly increased ( P < 0.001) in OSMF patients when compared to controls. Also serum and salivary IgG and IgA levels showed significantly increased ( P < 0.01) in all the three staging of OSMF when compared to control group. Hb levels and TSP levels were significantly decreased ( P < 0.001) in OSMF patients when compared to controls. One-way ANOVA, Pearson's correlation, and unpaired t -test were used for statistical analysis. The elevated levels of IgG and IgA are also in favor of polygammapathy, which are nonspecific and nondiagnostic objective reflections of an underlying disease. Decreased TSP is a result of host response and Hb, acts as an indicator of nutritional status plays an important role. It is also observed from the present study that the severity of OSMF was directly proportional to the estimated elevated levels of the major IgG and IgA. A need is also felt for the knowledge of immunoprofile estimation in etiology and pathogenesis that would prove a great asset in the proper assessment of this condition.

  4. Paper-Plastic Hybrid Microfluidic Device for Smartphone-Based Colorimetric Analysis of Urine.

    PubMed

    Jalal, Uddin M; Jin, Gyeong Jun; Shim, Joon S

    2017-12-19

    In this work, a disposable paper-plastic hybrid microfluidic lab-on-a-chip (LOC) has been developed and successfully applied for the colorimetric measurement of urine by the smartphone-based optical platform using a "UrineAnalysis" Android app. The developed device was cost-effectively implemented as a stand-alone hybrid LOC by incorporating the paper-based conventional reagent test strip inside the plastic-based LOC microchannel. The LOC device quantitatively investigated the small volume (40 μL) of urine analytes for the colorimetric reaction of glucose, protein, pH, and red blood cell (RBC) in integration with the finger-actuating micropump. On the basis of our experiments, the conventional urine strip showed large deviation as the reaction time goes by, because dipping the strip sensor in a bottle of urine could not control the reaction volume. By integrating the strip sensor in the LOC device for urine analysis, our device significantly improves the time-dependent inconstancy of the conventional dipstick-based urine strip, and the smartphone app used for image analysis enhances the visual assessment of the test strip, which is a major user concern for the colorimetric analysis in point-of-care (POC) applications. As a result, the user-friendly LOC, which is successfully implemented in a disposable format with the smartphone-based optical platform, may be applicable as an effective tool for rapid and qualitative POC urinalysis.

  5. CREATININE DETERMINATION IN URINE BY LIQUID CHROMATOGRAPHY-ELECTROSPRAY IONIZATION-TANDEM MASS SPECTROMETRY METHOD.

    PubMed

    Dereziński, Paweł; Klupczyńska, Agnieszka; Sawicki, Wojciech; Kokot, Zenon J

    2016-01-01

    Creatinine determination in urine is used to estimate the completeness of the 24-h urine collection, compensation for variable diuresis and as a preliminary step in protein profiling in urine. Despite the fact that a wide range of methods of measuring creatinine level in biofluids has been developed, many of them are adversely affected by interfering substances. A new liquid chromatography-tandem mass spectrometry method for creatinine determination in urine has been developed. Chromatographic separation was performed by applying C18 column and a gradient elution. Analyses were carried out on a triple quadrupole mass spectrometer equipped with an electrospray ion source. The developed method was fully validated according to the international guidelines. The quantification range of the method was 5-1500 ng/mL, which corresponds to 1-300 mg/dL in urine. Limit of detection and quantitation were 2 and 5 ng/mL, respectively. Additionally, the comparison of creatinine determination by newly developed method to the colorimetric method was performed. The method enables the determination of creatinine in urine samples with a minimal sample preparation, excellent sensitivity and prominent selectivity. Since mass spectrometry allows to measure a number of compounds simultaneously, a future perspective would be to incorporate the determination of other clinically important compounds excreted in urine.

  6. Detection of prostate cancer-specific transcripts in extracellular vesicles isolated from post-DRE urine.

    PubMed

    Pellegrini, Kathryn L; Patil, Dattatraya; Douglas, Kristen J S; Lee, Grace; Wehrmeyer, Kathryn; Torlak, Mersiha; Clark, Jeremy; Cooper, Colin S; Moreno, Carlos S; Sanda, Martin G

    2017-06-01

    The measurement of gene expression in post-digital rectal examination (DRE) urine specimens provides a non-invasive method to determine a patient's risk of prostate cancer. Many currently available assays use whole urine or cell pellets for the analysis of prostate cancer-associated genes, although the use of extracellular vesicles (EVs) has also recently been of interest. We investigated the expression of prostate-, kidney-, and bladder-specific transcripts and known prostate cancer biomarkers in urine EVs. Cell pellets and EVs were recovered from post-DRE urine specimens, with the total RNA yield and quality determined by Bioanalyzer. The levels of prostate, kidney, and bladder-associated transcripts in EVs were assessed by TaqMan qPCR and targeted sequencing. RNA was more consistently recovered from the urine EV specimens, with over 80% of the patients demonstrating higher RNA yields in the EV fraction as compared to urine cell pellets. The median EV RNA yield of 36.4 ng was significantly higher than the median urine cell pellet RNA yield of 4.8 ng. Analysis of the post-DRE urine EVs indicated that prostate-specific transcripts were more abundant than kidney- or bladder-specific transcripts. Additionally, patients with prostate cancer had significantly higher levels of the prostate cancer-associated genes PCA3 and ERG. Post-DRE urine EVs are a viable source of prostate-derived RNAs for biomarker discovery and prostate cancer status can be distinguished from analysis of these specimens. Continued analysis of urine EVs offers the potential discovery of novel biomarkers for pre-biopsy prostate cancer detection. © 2017 Wiley Periodicals, Inc.

  7. Detection of prostate cancer-specific transcripts in extracellular vesicles isolated from post-DRE urine

    PubMed Central

    Pellegrini, Kathryn L.; Patil, Dattatraya; Douglas, Kristen J.S.; Lee, Grace; Wehrmeyer, Kathryn; Torlak, Mersiha; Clark, Jeremy; Cooper, Colin S.; Moreno, Carlos S.; Sanda, Martin G.

    2018-01-01

    Background The measurement of gene expression in post-digital rectal examination (DRE) urine specimens provides a non-invasive method to determine a patient’s risk of prostate cancer. Many currently available assays use whole urine or cell pellets for the analysis of prostate cancer-associated genes, although the use of extracellular vesicles (EVs) has also recently been of interest. We investigated the expression of prostate-, kidney-, and bladder-specific transcripts and known prostate cancer biomarkers in urine EVs. Methods Cell pellets and EVs were recovered from post-DRE urine specimens, with the total RNA yield and quality determined by Bioanalyzer. The levels of prostate, kidney, and bladder-associated transcripts in EVs were assessed by TaqMan qPCR and targeted sequencing. Results RNA was more consistently recovered from the urine EV specimens, with over 80% of the patients demonstrating higher RNA yields in the EV fraction as compared to urine cell pellets. The median EV RNA yield of 36.4 ng was significantly higher than the median urine cell pellet RNA yield of 4.8 ng. Analysis of the post-DRE urine EVs indicated that prostate-specific transcripts were more abundant than kidney- or bladder-specific transcripts. Additionally, patients with prostate cancer had significantly higher levels of the prostate cancer-associated genes PCA3 and ERG. Conclusions Post-DRE urine EVs are a viable source of prostate-derived RNAs for biomarker discovery and prostate cancer status can be distinguished from analysis of these specimens. Continued analysis of urine EVs offers the potential discovery of novel biomarkers for pre-biopsy prostate cancer detection. PMID:28419548

  8. Urine toxicology screening in an urban stroke and TIA population.

    PubMed

    Silver, Brian; Miller, Daniel; Jankowski, Michelle; Murshed, Nawaf; Garcia, Patricia; Penstone, Patricia; Straub, Melissa; Logan, Sean P; Sinha, Anita; Morris, Daniel C; Katramados, Angelos; Russman, Andrew N; Mitsias, Panayiotis D; Schultz, Lonni R

    2013-04-30

    We sought to determine the rate of urine toxicology screening, differences in testing, and outcomes among patients with stroke and TIA presenting to a tertiary care emergency department. In this retrospective cohort study, patients admitted with stroke or TIA to a single tertiary care stroke center between June 2005 and January 2007 were identified through a stroke database. Factors that predicted urine toxicology screening of patients and a positive test, and discharge outcomes of patients based on toxicology result were analyzed. Stroke severity, treatment with tissue plasminogen activator, discharge status, and stroke etiology were compared between toxicology positive and negative patients. A total of 1,024 patients were identified: 704 with ischemic stroke, 133 with intracerebral hemorrhage, and 205 with TIA. Urine toxicology screening was performed in 420 patients (40%); 11% of these studies were positive for cocaine (19% younger than 50 years and 9% 50 years or older). Factors that significantly predicted the performance of a urine toxicology screen were younger age (<50 years) and black race (<0.001). Positive toxicology screens occurred in a broad range of patients. There were no significant differences in admission NIH Stroke Scale score, stroke etiology, and discharge status between toxicology-positive and -negative patients. In this study, patients with stroke and TIA who were young and black were more likely to have urine toxicology screening. Eleven percent of all tested patients (and 9% of patients 50 years or older) were positive for cocaine. To avoid disparities, we suggest that all stroke and TIA patients be tested.

  9. Radioscintigraphic demonstration of unsuspected urine extravasation

    SciTech Connect

    Bocchini, T.; Williams, W.; Patton, D.

    Three cases of unsuspected urine extravasation first detected by radionuclide scintigraphy are presented with subsequent confirmation by CT and, retrograde pyelograms. A renal study done to rule out acute transplant rejection demonstrates gallbladder uptake which was initially thought to be consistent with urine extravasation.

  10. Measurement of Menadione in urine by HPLC

    USDA-ARS?s Scientific Manuscript database

    Menadione may be an important metabolite of vitamin K that is excreted in urine. A high performance liquid chromatography (HPLC) method with a C30 column, fluorescence detection and post-column zinc reduction was developed to measure menadione in urine. The mobile phase was composed of 95% methanol...

  11. Managing Chemotherapy Side Effects: Urination Changes

    MedlinePlus

    ... how important it was to drink lots of water. She told me what changes to call about, such as a fever or ... when you urinate Managing Chemotherapy Side Effects: Urination ... Drink liquids such as water, soup, milkshakes, and cranberry juice. Add extra water ...

  12. Recommendations for provoked challenge urine testing.

    PubMed

    Ruha, Anne-Michelle

    2013-12-01

    "Urine mobilization test," "challenge test," and "provoked urine test" are all terms used to describe the administration of a chelating agent to a person prior to collection of their urine to test for metals. There is no standard, validated challenge test. Despite recommendations by professional and government organizations against the use of provoked urine testing, the tests are still commonly used and recommended by some practitioners. Challenge testing utilizes a variety of chelating agents, including dimercaptosuccinic acid (DMSA), dimercaptopropanesulfonate (DMPS), and ethylenediaminetetraacetic acid (EDTA). The agents are given by a variety of routes of administration, doses used are inconsistent, and urine collection procedures vary. Additional problems with challenge tests include comparison of results to inappropriate reference ranges and creatinine correction of urine obtained within hours of chelator administration. Human volunteer studies demonstrate that mercury is detected in the urine of most people even in the absence of known exposure or chelator administration, and that urinary mercury excretion rises after administration of a chelator, regardless of exposure history and in an unpredictable fashion. Studies also demonstrate that challenge testing fails to reveal a "body burden" of mercury due to remote exposure. Chelating agents have been associated with adverse reactions. Current evidence does not support the use of DMPS, DMSA, or other chelation challenge tests for the diagnosis of metal toxicity. Since there are no established reference ranges for provoked urine samples in healthy subjects, no reliable evidence to support a diagnostic value for the tests, and potential harm, these tests should not be utilized.

  13. Blood in the Urine (Hematuria) (For Parents)

    MedlinePlus

    ... weeks later, just to make sure the urine is free of red blood cells. Hematuria that only happens once won't ... up tests to make sure your child's urine is free of red blood cells. Reviewed by: Robert S. Mathias, MD Date reviewed: ...

  14. Effect of fiber blends, total solids, heat treatment, whey protein concentrate and stage of sugar incorporation on dietary fiber-fortified Kheer.

    PubMed

    Arora, Simran Kaur; Patel, A A

    2017-10-01

    Owing to the proven beneficial role of dietary fiber (DF) on human health, feasibility of incorporating commercially available soluble and insoluble DF preparations into rice-milk-pudding ( kheer, a popular Indian delicacy) was studied through process modification. The novel approach of preparing reduced fat DF-fortified- kheer (DFFK) by developing liquid/cream phase and particulate/rice phase separately, and subsequently blending the two was developed. The major processing variables studied were total solids (TS) in the liquid phase, type of fiber blend, flavor-simulation through heat treatment or added whey protein, and the presence of sugar in water for pre-cooking of rice. Reduced fat DFFK made from three different pre-standardized fiber blends was quite acceptable to the sensory panel (overall rating 7.5). With increasing TS in milk up to 16.5%, sensory acceptability of DFFK increased. There was a small but perceivable improvement in the flavour of DFFK when precooking of rice was carried out in sweetened water. Reduced fat DFFK from different fiber blends was found to be reasonably close to conventional kheer . DFFK prepared from Blend-I provided 3.31 g dietary fiber/100 kcal (suitable for the claim "High-in-Fiber") with 38.71% Reduced-Fat. With the developed process of fortification of kheer with DF, it is possible to reduce fat (and thus calories) to cater to the needs of consumers seeking good health.

  15. [Effect of citric acid stimulation on salivary alpha-amylase, total protein, salivary flow rate and pH value in Pi deficiency children].

    PubMed

    Yang, Ze-min; Chen, Long-hui; Lin, Jing; Zhang, Min; Yang, Xiao-rong; Chen, Wei-wen

    2015-02-01

    To compare the effect of citric acid stimulation on salivary alpha-amylase (sAA), total protein (TP), salivary flow rate, and pH value between Pi deficiency (PD) children and healthy children, thereby providing evidence for Pi controlling saliva theory. Twenty PD children were recruited, and 29 healthy children were also recruited at the same time. Saliva samples from all subjects were collected before and after citric acid stimulation. The sAA activity and amount, TP contents, salivary flow rate, and pH value were determined and compared. (1) Citric acid stimulation was able to significantly increase salivary flow rate, pH value, sAA activities, sAA specific activity and sAA amount (including glycosylated and non-glycosylated sAA amount) in healthy children (P<0.05), while it could markedly increase salivary flow rate, pH value, and glycosylated sAA levels in PD children (P<0.05); (2) Although there was no statistical difference in determined salivary indices between the two groups (P>0.05), saliva