Proteins In Urine Of Patients Research Articles

Mizoribine combined with steroids and dietary sodium restriction on the treatment of primary membranous nephropathy: a prospective study.

We aimed to initially explore the efficiency and safety of mizoribine (MZR) combined with steroids and dietary sodium restriction on the treatment of primary membranous nephropathy (MN) compared with cyclophosphamide (CPM)-based steroids. Patients with primary MN were enrolled. According to the therapy, they were divided into theMZR combined with steroids and dietary sodium restriction group (N = 30) and CPM-based steroids group (N = 30). Both groups were followed up for 1year to monitor safety and efficacy. Compared with the CPM-based steroids group, theMZR combined with steroids and dietary sodium restriction group had significantly lower daily sodium intake, serum sodium, blood pressure (BP), and 24h urine protein (all P < 0.05). Conversely, plasma albumin and complete remission rate in the MZR group were higher at the 12th follow-up (40.39 ± 5.14g/L vs. 37.63 ± 5.40g/L; 86.67% vs. 66.67%; all P < 0.05). These two groups showed similar adverse events rates (20.00% vs. 26.67%, P = 0.54). This study demonstrates that MZR combined with steroids and dietary sodium restriction is superior to CPM-based steroids in terms of completeremission and 24h urine protein in patients with primary MN.

Urine protein in patients with type I hypersensitivity is indicative of reversible renal tube injury

AimsIn our clinical work, some patients with type I hypersensitivity could be detected protein in their urine. This study focused on the early renal injury in patients with type I hypersensitivity. Main methodsFrom 43 type I hypersensitivity patients with proteinuria, 10 patients were randomly selected for mass spectrometry analysis of 24-h urine together with 5 healthy volunteers. Mice were vaccinated with Dermatophagoides farina (Der f) and ovalbumin (OVA) were used as antigen to establish the type I hypersensitivity animal models. Key findingsThe urine protein of hypersensitivity patients was significantly increased in the alpha-1-microglobulin/ bikunin precursor (Protein AMBP) (t = 3.140, P = 0.008), retinol binding protein 4 (RBP4) (t = 2.426, P = 0.031), kininogen-1 (t = 2.501, P = 0.027), and transferrin appeared only in patients' urine. After immunizing mice with antigens, significant increases of the total serum immunoglobulin E (IgE) were observed in both Der f (86.92 ± 36.01 U/mL, t = 5.231, P = 0.0004) and OVA group (34.65 ± 24.72 U/mL, t = 2.891, P = 0.0161) compared with the negative control group (2.68 ± 0.47 U/mL). Meanwhile, definite eosinophil infiltration around the impaired renal tubules as well as the bronchus in Der f mice were observed, and urine protein appeared. After stopping the allergen stimulation, proteinuria disappeared. Instead, when the mice were treated with the antigen again, proteinuria reappeared. SignificanceOur findings suggest that renal tubular damage in patients with type I hypersensitivity is reversible, and proteinuria disappears with allergy symptoms remission.

Nephroprotective effect of vitamin E added to angiotensin receptor blocker in patients with diabetic nephropathy

Background: Diabetes mellitus is a condition associated with increased oxidative stress as a consequence of hyperglycemia. Therefore, the use of antioxidants in people with diabetes has been advocated. Vitamin E is the most prevalent naturally occurring anti-oxidant. Aim: To evaluate the effect of the combination of angiotensin receptor blocker (ARB) and vitamin E on urine protein in patients with diabetic nephropathy. Methods: One hundred and six patients with diabetic nephropathy, who visited the Alsader medical city in Najaf governorate and Al-Furat Al-Awsat general hospital in Kufa, were investigated, 39 of them are type 1diabetics, and 67 of them are type 2 diabetics. Each patient was followed up for 3 months with valsartan (160mg/day), then for 3 months with addition of tocopherol (400mg). At the screening visits, proteinuria was determined in two 24 h urine sample, arterial blood pressure was measured twice at ten minutes rest with 2 minutes interval and serum level of potassium, sodium &amp; creatinine were determined. Results: Proteinuria was significantly lowered when tocopherol 400mg was added to valsartan, as compared with valsartan without tocopherol. Valsartan alone caused a mean reduction of 24 h urine protein of 27% from the baseline, while the addition of tocopherol caused a mean reduction of 24 hour proteinuria of 30% from result of valsartan. That is mean the combination therapy of valsartan and tocopherol caused a reduction in proteinuria of 44% from baseline. Conclusion: The current study suggests that combination therapy of ARB and vitamin E offers an additional antiproteinuric and nephroprotective effect. Recommendation: Antioxidants, in particular vitamin E have promising nephroprotective effects. However, studies on other antioxidants like vitamin C are needed in the efforts of prevention of diabetic nephropathy

Optimized Preparation of Urine Samples from Acute Melioidosis Patients for In-Solution Proteomic Studies using LCMS QTOF or MALDI TOF MS

Introduction: Investigation of urine proteome in patients with acute melioidosis may reveal potential disease markers, from either bacterial or human proteins. We used an in-solution gel-free method instead of 2-DE to detect human and Burkholderia pseudomallei proteins in urine of patients with acute melioidosis. Here, we propose a simpler, economical method for preparing urine samples directly from melioidosis patients, for in-solution proteomic analysis using LCMS-QTOF MS/MS or MALDI-TOF MS/MS. Material and Methods: We adapted an acetone-TCA based protein precipitation method with LCMS-QTOF MS to detect the B. pseudomallei proteins directly from urine of acute melioidosis patients (culture positive and negative). This process involves protein precipitation, desalting, trypsin digestion, and optimization for the mass spectrometry. Results: A total of 3,866 human peptides were detected across 11 urine samples from clinically suspected acute melioidosis patients. Among them were three Burkholderia specific proteins detected in 75% of culture positive samples. Large amounts of acute phase proteins, cell mediated immunity proteins, complement pathway proteins and inflammatory mediators were seen upon gene ontology (GO) annotation and GO enrichment analysis. Conclusions: This simple in-solution sample preparation method can be replicated easily for LCMS/MS-QTOF and MALDI-TOF proteomic analyses, avoiding tedious optimization steps in 2-DE. This method is cost effective and can be done in centres without specialized 2-DE or MS equipment and elutes can be easily transported for analysis and bioinformatics. This is the first study to analyse urine samples directly for B. pseudomallei proteins. Discovery of the entire proteome as a whole is important in leading to biomarker discovery.

Urine Protein in Patients with Type I Hypersensitivity is Indicative of Reversible Renal Tube Injury

Urine Protein in Patients with Type I Hypersensitivity is Indicative of Reversible Renal Tube Injury

Expression of urotensin II is positively correlated with pyroptosis-related molecules in patients with severe preeclampsia

ABSTRACT Purpose: We studied the expression of urotensin II (UII) and its relationships with markers of pyroptosis in preeclampsia. Methods: 48 pregnant subjects were recruited consisting of 28 severe preeclampsia pregnancies (SPE) and 20 healthy pregnancies. We detected expressions of UII and markers of pyroptosis such as NLR-family pyrin domain (PYD)-containing 3 (NLRP-3), caspase-1/4/5, interleukin-1β (IL-1β), and gasdermin D (GSDMD) in placentas of patients with SPE and healthy pregnancies. Results: SPE group have higher expression of UII and NLRP-3, caspase-1, interleukin-1β (IL-1β), and GSDMD than that normal controls by IHC, real-time PCR, and western blot. IHC analysis manifests that the expressions of UII and pyroptosis-related molecules are mainly located in the placental cytotrophoblasts. Expressions of UII mRNA and protein are significantly positively correlated with pyroptosis marker such as NLRP3, caspase-1, GSDMD mRNA and protein by Pearson correlation analysis. Moreover, UII, NLRP-3, caspase-1, interleukin-1β (IL-1β), and GSDMD are positively related with systolic blood pressure, meanwhile caspase-1 and GSDMD are positively correlated with urine protein in SPE patients. We firstly verify that UII has a positive correlation with pyroptosis markers in placentas of preeclampsia patients; besides, pyroptosis-related proteins are positively correlated with systolic blood pressure and urine protein in patients with severe preeclampsia.

The effect of ginger on blood sugar and urine protein in patients with type 2 diabetes mellitus; a clinical trial

Introduction: The prevalence of diabetes mellitus has been increasing exponentially and its complications are a major cause of morbidity and mortality. Ginger is one of the herbal remedies which has preventive effects on nephropathy in animal models in some studies. Objectives: We aimed to investigate the effect of ginger on urine protein analysis and blood glucose levels in diabetic patients. Patients and Methods: In a clinical trial, 98 patients with type 2 diabetes were randomly divided into two groups; the intervention group received 500 mg of Zingiber as capsule, daily in addition to routine therapies, since control group received only conventional treatments for diabetes. Before and after intervention, blood glucose, urine microalbumin and hemoglobin A1c (HbA1c) were measured. Results: The findings showed that Zingiber decreased HbA1c levels in diabetic patients (P=0.006), however, it did not have a significant effect on fasting blood glucose (P=0.179), 2 hours post-prandial glucose levels (P= 0.272), and the microalbuminuria quantity in urine (P=0.109). Conclusion: In our study, the hypoglycemic short-term effects of ginger observed as a reduction of HbA1c, however its effect on reducing blood glucose levels [FBS and 2 hours postprandial glucose (2hpp)] was not significant, therefore ginger can be considered as a supplementary agent in the treatment of diabetes. However it is recommended to conduct more studies to obtain results with more reliability. Trial registration: The trial protocol was approved by the Iranian Registry of Clinical Trial (identifier: IRCT2017103025732N27; https://www.irct.ir/trial/21503, ethical code; IR.SEMUMS.REC.1395.207).

Efficacy of Bailing Capsule (Traditional Chinese Medicine) in the Treatment of Nephrotic Syndrome: A Meta-analysis

Meta-analysis of the existing literature on the treatment of nephrotic syndrome with Bailing capsule in adult patients was carried out to provide evidences for clinical practice. Relevant articles describing the effect were included. A total of 5 documents were selected. The meta-analysis showed that Bailing capsule reduced 24 h urine protein in patients with nephrotic syndrome (standardized mean difference=2.35 (3.72, 0.97), p<0.01), increased the level of serum albumin (standardized mean difference=0.94 (0.37, 1.52), p=0.01), but had no improvement on serum creatinine (standardized mean difference=0.50 (1.25, 0.26), p=0.20) as well as blood urea nitrogen (standardized mean difference=0.10 (0.36,0.17), p=0.48). The adjuvant treatment of nephrotic syndrome with Bailing capsule could reduce proteinuria, enhance plasma albumin and decrease blood lipids in patients. However, these conclusions need to be confirmed by further clinical trials.

STIM promotes the epithelial-mesenchymal transition of podocytes through regulation of FcγRII activity in diabetic nephropathy.

Diabetic nephropathy (DN) is a serious complication in diabetic patients and has been considered as the main cause of end-stage renal disease. However, there are no studies on the role of stromal interaction molecule (STIM) and its two subtypes, STIM1 and STIM2, in the epithelial-to-mesenchymal transition (EMT) of podocytes induced by diabetic kidney disease (DKD). The present study suggests for the first time that STIM inhibition decreases DKD-induced EMT. All DKD patients were diagnosed based on renal biopsies carried out at the Department of Nephrology, Zhejiang Provincial People's Hospital and selected using the Mayo Clinic/Renal Pathology Society Consensus Report on Pathologic Classification, Diagnosis, and Reporting of GN. Images were taken and the number of positive puncta in cells was analyzed using software equipped for immunofluorescence microscopy. STIM1, STIM2, FcγRIIa, FcγRIIb, Nephrin, CTGF, and α-SMA protein levels were detected by Western blotting analysis using the corresponding antibodies. The viability of cells was measured using CCK-8 assays. Absorbance at 450 nm was measured with a Multiskan FC Microplate Reader (Thermo Scientific, USA) and the results were normalized to those of untreated cells. All statistical analyses were performed using SPSS 19.0 software (Stanford University, Stanford, CA, USA). A total of 30 DKD patients and 30 control patients were enrolled in the study. We found that the level of urine protein in patients and db/db diabetic mice is higher than control group and the levels of STIM1 and 2 significantly increased in DKD groups. We also demonstrated that STIM is upregulated during DKD injury. Next, we discovered that DKD-induced podocyte EMT is related to STIM overexpression in vivo and in vitro. Further research demonstrated that STIM siRNA reverses podocytes from DKD-induced injury and EMT and reverses FcγRII activity in HG-treated podocytes. Our study suggests that STIM and FcγRII play an essential role in the regulation of DKD-induced podocyte EMT. STIM is an essential component of FcγR activation and inhibition of STIM-mediated signaling pathway might be a new strategy to treat IgG-dependent renal diseases.

GW29-e0893 Sulodexide effectively alleviated urine protein in patients with hypertension

GW29-e0893 Sulodexide effectively alleviated urine protein in patients with hypertension

Expression of urotensin II is\xa0associated with placental autophagy in patients with severe preeclampsia

The aims of this study are to explore the correlation between the expressions of urotensin II (UII) and autophagic markers (LC3 and P62) in patients with severe preeclampsia (SPE). A total of 64 pregnant subjects were recruited, including 29 healthy pregnancies and 35 preeclamptic patients (7 mild preeclamptic (MPE) patients and 28 SPE patients). UII and autophagic markers expression in placenta specimens was investigated by immunohistochemistry (IHC), RT-qPCR, and western blot. IHC analysis manifested that the expressions of UII and autophagic markers were mainly located in the placental cytotrophoblast and syncytiotrophoblast. Western blot and IHC analysis both indicated that the expression of UII was significantly correlated with autophagic marker LC3II (by western blot) or LC3 (by IHC) (r = 0.495, P = 0.010; r = 0.816, P = 0.007). Moreover, SPE group had higher expression of UII and LC3II, lower expression of P62 than that of normal controls. The expression of LC3II was positively related with systolic blood pressure (SBP) and urinary protein level (SBP (r = 0.501, P = 0.003) and urine protein quantitation (r = 0.509, P = 0.022)), whereas P62 had negative correlation with SBP. We first verify that UII has positive correlation with autophagic marker LC3 in placentas of preeclampsia patients; besides, autophagic levels are positively correlated with SBP and urine protein in patients with SPE.

Association of serum bilirubin with renal outcomes in Han Chinese patients with chronic kidney disease

BackgroundOxidative stress and inflammation play pivotal roles in chronic kidney disease (CKD). Bilirubin is an endogenous anti-inflammatory antioxidant. However, the relationship between serum bilirubin and renal outcomes in CKD is controversial. We explored the association of serum bilirubin levels with renal outcomes in Han Chinese patients with CKD. MethodsClinical and laboratory data were collected from 316 patients with CKD. The primary clinical endpoint was renal replacement therapy or death. The association between serum bilirubin and clinical parameters was assessed by correlation analysis. Multiple Cox regression analysis was used to explore the relationship between serum bilirubin and renal outcomes in patients with CKD. ResultsSerum total and indirect bilirubin were positively correlated with estimated glomerular filtration rate, but negatively correlated with 24-h urine protein in patients with CKD. Serum total and indirect bilirubin were inversely associated with CKD stages in patients with CKD stages 1–5. Multiple Cox regression analysis demonstrated that the higher concentration of serum total bilirubin was independently associated with better renal outcomes in CKD. ConclusionsOur results suggest that serum total bilirubin may have protective effects on kidneys.

Pathogen-specific leptospiral proteins in urine of patients with febrile illness aids in differential diagnosis of leptospirosis from dengue.

Leptospirosis and dengue are two commonly seen infectious diseases of the tropics. Differential diagnosis of leptospirosis from dengue fever is often difficult due to overlapping clinical symptoms and lack of economically viable and easy-to-perform laboratory tests. The gold standard for diagnosis is the microscopic agglutination test (MAT). In this study, the diagnostic potential of screening for pathogen-specific leptospiral antigens in urine samples is presented as a non-invasive method of disease diagnosis. In a study group of 40 patients, the serum was tested for anti-leptospiral antibodies by MAT and enzyme-linked immunosorbent assay (ELISA). Urine of these patients was screened for leptospiral antigens by ELISA using specific antibodies against LipL32, LipL41, Fla1, HbpA and sphingomyelinase. Group I patients (n=23) were classified as leptospirosis-positive based on MAT and high titres of circulating IgM-specific anti-leptospiral antibodies. All of these patients excreted all five leptospiral antigens in the urine. The 17 MAT-negative cases included six patients with pyrexia of unknown origin (PUO; Group II) and 11 confirmed dengue patients (Group III). The latter tested negative for both serum anti-leptospiral antibodies and urinary leptospiral antigens. A salient outcome of this study was highlighting the usefulness of screening for urinary leptospiral antigens in disease diagnosis, as their presence confirmed leptospiral aetiology in two PUO patients. Immunoblots of urinary antigens identified well-defined bands corresponding to LipL32, HbpA and sphingomyelinase; the significance of the 42- and 58-kDa sphingomyelinase bands is discussed.

Association of Interleukin-10 Polymorphisms (rs1800872, rs1800871, and rs1800896) with Predisposition to IgA Nephropathy in a Chinese Han Population: A Case-Control Study

Background/Aims: IgA nephropathy (IgAN) is a common form of primary glomerulonephritis worldwide. Previous studies indicated that IL-10 single nucleotide polymorphisms (SNP) play an important role in IgAN pathogenesis, but the results were controversy. This study aimed to investigate the association between IL-10 SNPs (rs1800872, rs1800871, and rs1800896) with IgAN in a Chinese Han population. Methods: We conducted a case–control study that included 351 patients with IgAN and 310 age-, gender- and ethnicity-matched healthy controls. Three promoter SNPs (rs1800872, rs1800871, and rs1800896) of IL-10 were genotyped by Sequenom MassARRAY. Odds ratios (ORs) with 95% confidence intervals (CI) were used to assess the relationship with IgAN. Results: We found that the rs1800896 did not correlate with IgAN risk, whereas rs1800872 and rs1800871 were significantly associated with increased IgAN risk in all genetic models. The haplotype analysis indicated that the CCA haplotype was associated with increased IgAN risk (OR = 1.36; 95% CI = 1.05–1.75). Moreover, there were no associations between these SNPs and blood pressure or gender, whereas the rs1800896 variant was correlated with higher 24-hour urine protein in patients with IgAN. Conclusion: Taken together, these results suggest that IL-10 is a susceptibility gene in patients with IgAN.

Quantification of angiotensin II-regulated proteins in urine of patients with polycystic and other chronic kidney diseases by selected reaction monitoring.

BackgroundAngiotensin-II (Ang II) mediates progression of autosomal-dominant polycystic kidney disease (ADPKD) and other chronic kidney diseases (CKD). However, markers of kidney Ang II activity are lacking. We previously defined 83 Ang II-regulated proteins in vitro, which reflected kidney Ang II activity in vivo.MethodsIn this study, we developed selected reaction monitoring (SRM) assays for quantification of Ang II-regulated proteins in urine of ADPKD and CKD patients. We demonstrated that 47 of 83 Ang II-regulated transcripts were differentially expressed in cystic compared to normal kidney tissue. We then developed SRM assays for 18 Ang II-regulated proteins overexpressed in cysts and/or secreted in urine. Methods that yielded CV ≤ 6 % for control proteins, and recovery ~100 % were selected. Heavy-labeled peptides corresponding to 13 identified Ang II-regulated peptides were spiked into urine samples of 17 ADPKD patients, 9 patients with CKD predicted to have high kidney Ang II activity and 11 healthy subjects. Samples were then digested and analyzed on triple-quadrupole mass spectrometer in duplicates.ReslutsCalibration curves demonstrated linearity (R2 > 0.99) and within-run CVs < 9 % in the concentration range of 7/13 peptides. Peptide concentrations were normalized by urine creatinine. Deamidated peptide forms were monitored, and accounted for <15 % of the final concentrations. Urine excretion rates of proteins BST1, LAMB2, LYPA1, RHOB and TSP1 were significantly different (p < 0.05, one-way ANOVA) between patients with CKD, those with ADPKD and healthy controls. Urine protein excretion rates were highest in CKD patients and lowest in ADPKD patients. Univariate analysis demonstrated significant association between urine protein excretion rates of most proteins and disease group (p < 0.05, ANOVA) as well as sex (p < 0.05, unpaired t test). Multivariate analysis across protein concentration, age and sex demonstrated good separation between ADPKD and CKD patients.ConclusionsWe have optimized methods for quantification of Ang II-regulated proteins, and we demonstrated that they reflected differences in underlying kidney disease in this pilot study. High urine excretion of Ang II-regulated proteins in CKD patients likely reflects high kidney Ang II activity. Low excretion in ADPKD appears related to lack of communication between cysts and tubules. Future studies will determine whether urine excretion rate of Ang II-regulated proteins correlates with kidney Ang II activity in larger cohorts of chronic kidney disease patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-016-9117-x) contains supplementary material, which is available to authorized users.

Evaluation of random urine sample protein: creatinine ratio as an index of 24 hour urine protein in patients with various renal disorders in tertiary care center

Background: Commonly used methods to measure protein are 24 hours urine collection, which is time consuming cumbersome and often in accurate, the other method, infrequently used, is estimation of proteinuria from protein-creatinine ratio. The objective of the study was to compare spot urine protein-creatinine ratio with 24 hours urine protein as an index of quantitative proteinuria. Methods: 110 patients with persistent dipstick positive proteinuria with varying degrees of renal dysfunction were included in this study. First morning spot urine sample were used to estimate protein creatinine ratio and then 24 hours urine protein estimation was done and compared. Results: There was significant correlation between 24 hours urine protein and protein creatinine ratio (r = 0.70) (P<0.01) However maximum correlation was in patients with normal or mild renal dysfunction and non nephrotic range proteinuria (r = 0.92) (p<0.01). Conclusions: Protein creatinine ratio in a spot morning urine sample is a precise indicator of proteinuria and represents a simple and inexpensive procedure in establishing severity of proteinuria.

Evaluation of Rapid Diagnostic Methods of Urinary Protein Estimation in Patients of Preeclampsia of Advanced Gestational Age

24-h urine protein is traditionally used as a gold standard method for protein estimation. Because of the operational difficulty, there is the necessity to use rapid, convenient, and reliable method of proteinuria estimation. We carried out this study to compare the two rapid methods of protein estimation: dipstick method and spot urine protein creatinine ratio (UPCR) with that of 24-h urine protein in patients of preeclampsia with advanced gestational period. The values of proteinuria estimated by dipstick method and spot UPCR were compared with that of 24-h urine protein. The strength of correlation was measured by Pearson's correlation coefficient (r). A p value of <0.05 is considered to be statistically significant. The most discriminant spot UPCR value for detecting significant proteinuria (≥300mg/day) was determined by plotting receiver-operator curve (ROC). The value of spot UPCR strongly correlated with 24-h urine protein (r=0.88 with p value <0.001). The most discriminant spot UPCR value for detecting significant proteinuria (≥300mg/day) was 0.3. The estimation of proteinuria by dipstick method was poorly correlated with 24-h urine protein with r=-0.09. Spot UPCR can be used as a rapid and reliable alternative method in preference to 24-h proteinuria in patients of preeclampsia of advanced gestational age.

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

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.

Exploratory study of proteins in urine of patients with histoplasma antigenuria

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.

The Relationship Between Serum Free Light Chain Levels and Urine Protein in Patients with IgG or IgA Myeloma.

Abstract 1810Poster Board I-836The serum free light chain (SFLC) assay is often though to be a replacement for the cumbersome process of 24-hour urine collection for monoclonal protein estimation (http://www.freelite.co.uk/initialinvestigationpro-24.asp; accessed 16 July 2009) despite evidence suggesting that SFLC estimation cannot replace urine protein quantification (Singhal et al. Blood 2007; 109:3611-2). We studied results from patients with IgG or IgA myeloma if all the following criteria were satisfied: concomitant SFLC and 24-hour urine specimens analyzed, no oligoclonal proteins, no diminished free light chain levels (involved or uninvolved). The aim was to study the correlation between SFLC and 24-hour urine total protein (24UTP), 24-hour urine monoclonal protein (24UMP) and urine immunofixation electrophoresis (UIFE). Only concordant abnormal SFLC ratios were considered abnormal (Singhal et al. Blood 2009; 114:38-9). 558 samples in 114 patients were identified. SFLC ratio was abnormal in 242 and normal in 316 (including 2 discordant abnormal). UIFE (available in 540) was negative in 290. The proportion of samples with normal SFLC ratio decreased as 24UTP increased (P<0.001): ≤200 mg - 75% of 243, 200-500 mg 33% of 129, 501-1000 mg - 24% of 54, and >1000 mg - 6% of 32. Similarly, the proportion of samples with normal SFLC ratio declined as 24UMP increased (P=0.001): ≤200 mg (including negative) - 41% of 145, 200-500 mg 4% of 28, 501-1000 mg - 0% of 14, and >1000 mg - 0% of 15. SFLC ratio was normal in 84% of samples with negative UIFE and in 26% of samples with positive UIFE (P<0.0001). Among the 47 samples with abnormal SFLC ratio and negative UIFE, the 24UTP was 47-288 mg (median 112). 24UMP was 35 mg in 1, “faint” in 1, and no restricted bands were seen in ther remaining 45. The table shows the relationship between UPEP, UIFE and SFLC amongst the 540 samples where all 3 tests were available. The presence of any restricted band on UPEP, even if too small to quantify, was considered positive.n=540Positive UPEPNegative UPEPPositive UIFENegative UIFEPositive UIFE22228Negative UIFE17273Abnormal SFLC ratio1725918447Normal SFLC ratio6724266243If a positive UPEP is considered the “gold standard” evidence of the presence monoclonal light chains in the urine, the sensitivity of UIFE in detecting urine monoclonal protein is 93% and that of an abnormal SFLC ratio is 72%. On the other hand, if a positive UIFE is considered the “gold standard” evidence of monoclonal light chains in the urine, the sensitivity of an abnormal SFLC ratio in detecting urine monoclonal protein is 74%. These data, obtained from a much large number of homogeneous clinical samples, confirm our previous observations (Singhal et al. Blood 2007; 109:3611-2) that an abnormal SFLC ratio cannot consistently predict the presence of either non-specific or monoclonal proteinuria. Fortunately, the proportion of urine samples with large aounts of non-specific or monoclonal proteinuria that is associated with normal SFLC ratios is small. The sensitivity of the SFLC ratio in detecting urine monoclonal protein is less than that of UIFE. The SFLC assay cannot replace 24-hour urine collection in clinical practice without compromising patient care. Whether a less cumbersome technique such as a spot (random) urine protein-creatinine ratio - used in conjunction with the SFLC assay - can avert the need for 24-hour urine collections in myeloma patients remains to be investigated. DisclosuresNo relevant conflicts of interest to declare.