Pathological Urine Research Articles

Trends of Benzotriazoles and Benzothiazoles in Australian Pooled Urine Samples from 2012 to 2023.

Benzotriazoles (BZTs) and benzothiazoles (BTs) are high-production-volume chemicals utilized in many different commercial products and industrial processes, such as metal corrosion inhibitors, vulcanization accelerators, plastic-associated UV stabilizers, and pharmaceutical precursors. This study assessed age, gender, and temporal trends of BZTs and BTs in deidentified surplus pathology urine samples, pooled and stratified by age, gender, and sample collection year from a general Australian population (168 pools representing 16,800 individuals). Tolyltriazole (TTri), 5,6-dimethyl-1H-benzotriazole (DMBZT), 1,3-benzothiazole (BTH), 2-hydroxybenzothiazole (2-OH-BTH), and 2-aminobenzothiazole (2-amino-BTH) were detected in >50% of the pools. TTri was frequently detected in pooled samples representing ≤45-year-olds (both genders). Concentrations of DMBZT, BTH (females), 2-OH-BTH, and 2-amino-BTH (females) increased with age significantly, with adults (>15 years old) showing higher levels than children (≤15 years old). Gender differences in DMBZT concentrations (females > males) were observed across all sampling years and only in some for TTri (males > females: >45 years old), BTH (females > males), and 2-amino-BTH (males > females). A temporal increase in BTH, 2-OH-BTH, and 2-amino-BTH levels within the studied period (2012-2023) has been observed. Our findings suggest ongoing exposure of the Australian general population to BZTs and BTs, highlighting age, gender, and temporal trends of these compounds as measured via their urinary concentrations.

Abstract 1073: AI-assisted high-dimensional cytological single cell analysis distinguishes normal and malignant urine samples using cell morphology images with high accuracy at the single cell level

Abstract Background: Urothelial cancer (UCC) is the 4th most common malignancy in men. The current clinical problem in UCC management is the high interobserver variability of tissue UCC grading and the low sensitivity of urine cytology for early detection of recurrences and progression. We hypothesize that assessing urine samples and UCC tissues with detailed quantitative image analysis based on artificial intelligence has strong potential in both urine-based early detection of tumor recurrence and progression as well as tissue-based pathological grading. Methods: Here, we utilized the Deepcell® Human Foundation Model (HFM), a self-supervised deep learning model, to assess a cohort of urine samples from 54 normal healthy cases, 19 atypical cases, and 4 UCC cases. High resolution images of label-free single cells were captured and analyzed using the HFM to identify distinct morphological characteristics of cells from different urine origins. The urine samples were loaded onto a single-use microfluidic cartridge as part of the Deepcell® REM-I platform and brightfield images of single cells were captured in real-time. For data analysis, images of all cells run through the REM-I instrument are automatically sent to the Deepcell® Axon Suite, which provides an analysis platform to visualize cell images from specific runs, generate UMAPs of the embedding space, and run additional deep neural network models on samples in silico. Results: High-dimensional analysis of 115 deep learning and morphometric features reveals 12 morphotypes among the pooled urine samples, as represented by morphology UMAP plots. Morphology analysis reveals distinct morphotypes in normal compared to pathological urine samples. Confusion matrix showed deep learning model performance in classifying atypical versus UCC with 89.1% (Atypical (n = 8642), UCC (n = 12637)). This indicates high model performance accuracy for AI prediction of atypical versus UCC samples at the single cell level. Conclusions: The Deepcell® REM-I platform performs well with urine samples and identified numerous cell clusters among normal and pathological urines. AI-assisted high-dimensional morphology analysis distinguishes atypical urine samples from UCC cases with 89.1% accuracy at the single cell level. Future applications of this study include early detection or monitoring of UCC cases using non-invasive patient samples such as urine. Citation Format: Thierry P. van den Bosch, Geert J.L.H. van Leenders, Siamala Sinnadurai, Zhouyang Lian, Ryan Carelli, Matthew Nakaki, Vivian Lu, Manisha Ray, Janifer Cruz, Kelly R. Miller, Mahyar Salek, Maddison Masaeli, Peter J. van der Spek. AI-assisted high-dimensional cytological single cell analysis distinguishes normal and malignant urine samples using cell morphology images with high accuracy at the single cell level [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1073.

An improved method for measuring catalase activity in biological samples.

Catalase (CAT) is an important enzyme that protects biomolecules against oxidative damage by breaking down hydrogen peroxide (H2O2) into water and oxygen. CAT is present in all aerobic microbes, animals, and plants. It is, however, absent from normal human urine but can be detected in pathological urine. CAT testing can thus help to detect such urine. This study presents a novel spectrophotometric method for determining CAT activity characterized by its simplicity, sensitivity, specificity, and rapidity. The method involves incubating enzyme-containing samples with a carefully chosen concentration of H2O2 for a specified incubation period. Subsequently, a solution containing ferrous ammonium sulfate (FAS) and sulfosalicylic acid (SSA) is added to terminate the enzyme activity. A distinctive maroon-colored ferrisulfosalicylate complex is formed. The formation of this complex is a direct result of the reaction between FAS and any residual peroxide present. This leads to the generation of ferric ions when coordinated with SSA. The complex has a maximum absorbance of 490 nm. This advanced method eliminates the need for concentrated acids to stop CAT activity, making it safer and easier to handle. A comparative analysis against the standard ferrithiocyanate method showed a correlation coefficient of 0.99, demonstrating the new method's comparable effectiveness and reliability. In conclusion, a simple and reliable protocol for assessing CAT activity, which utilizes a cuvette or microplate, has been demonstrated in this study. This interference-free protocol can easily be used in research and clinical analysis with considerable accuracy and precision.

Frequency of Urinary Tract Infection When Diagnosing Vesico Uretheral Reflux in Children in Tuzla Canton.

Vesicoureteral reflux (VUR) represents the return of urine from the bladder into the ureter and the renal canal system. Reflux can occur only on one or both kidneys. VUR most often occurs due to an incompetent ureterovesical junction, which consequently leads to hydronephrosis and dysfunction of the lower parts of the urinary system. The aim of the study was to determine the frequency of urinary infection when diagnosing vesicouretheral reflux in children in the Tuzla Canton, in the five-year period from 01.01.2016 to 01.01.2021. Through a retrospective study, we analyzed data from 256 children with vesiocouretheral reflux (VUR), examined in the Nephrology Outpatient Clinic, Clinic for Children's Diseases, University Clinical Center Tuzla, in the period from 01.01.2016 to 01.01.2021, from early neonatal to 15 years of age. The age and gender of children, the most common symptoms of urinary tract infections during the detection of VUR, and the degree of VUR were analyzed. From 256 children with VUR, 54% were male and 46% female. The highest prevalence of VUR was in the age group 0-2 years, and the lowest in the age of children > 15 years. There was no statistically significant difference between the groups of our respondents in relation to age groups, nor in relation to the gender of the children. Statistically significantly more children were without nonspecific symptoms and with asymptomatic bacteriuria in the group without UTI symptoms in children with VUR compared to the group with UTI symptoms in children with VUR. Pathological urine culture between the groups was without a statistically significant difference. Although urinary tract infection in children is common, the possibility of permanent consequences should always be kept in mind if VUR is not diagnosed and treated in time.

The Effect Of Centrifugation Speed On The Results Of Urine Sediment Examination

The process of centrifugation in urine examination is very important because it can be used in the manufacture of urine sediment. The purpose of this study was to determine the effect of centrifugation speed on the results of urine sediment examination. Centrifugation speeds used were 3000 rpm, 3500 rpm, 4000 rpm, and 4500 rpm using pathological urine samples. The samples used were 32. The research method used was an experimental laboratory at the Pusura Yos Sudarso clinical laboratory in Surabaya. The results of the study with a speed of 3000 rpm obtained an average of erythrocytes = 6.63/LPB, leukocytes = 13.63/LPB, and epithelial = 10.50/LPB. At a speed of 3500 rpm the average value of erythrocytes = 7.63/LPB, leukocytes = 15.13/LPB, and epithelium 11.63/PLB. At a speed of 4000 rpm the average value of erythrocytes = 8.63/LPB, leukocytes = 17.75/LPB and epithelial = 13.50/LPB. At a speed of 4500 rpm the average value of erythrocytes = 9.63/LPB, leukocytes = 20.50/LPB, and epithelial = 15.38/LPB. The results of the statistical test One Way Anova obtained p-value results of 0.001 < (0.05) in erythrocyte cells, p-value results for leukocytes 0.025 < (0.05), and p-value results for epithelial cells 0.002 < (0.05) means that there is an effect on the centrifugation speed on the results of urine sediment examination.

Vegan diet to slow the progression of diabetic nephropathy: A review

Diabetic nephropathy (DN) is the major cause of renal disease that affects 40% of patients with diabetes. DN is characterized by the pathological urine albumin excretion rate (UAER), diabetic glomerular lesions, and loss of glomerular filtration rate (GFR). Several studies have found strong association of animal protein with the glucagon activation, insulin resistance, proteinuria, microalbuminuria, and progression of renal disease in diabetic patients. Vegan diet that excludes all animal foods such as meat (including fish, shellfish, and insects), dairy, eggs, and honey, as well as products such as leather has been found significantly beneficial on enhancing insulin sensitivity, reducing glucagon activation and incidence of chronic kidney disease (CKD), and slow the progression of DN. Other studies have suggested that avoiding animal food and shifting the source of dietary protein from animal-derived to plant-based products may be a better dietary approach than dietary protein restriction to reduce kidney disease and cardiovascular disease (CVD), particularly in CKD and diabetes patients with profound insulin resistance. Based on the literature search conducted in this study we concluded that the vegan diet is beneficial in patients with diabetes and CKD to slow the progression of the DN and renal failure. However, additional interventional studies in humans are needed to understand the mechanisms underlying the increased insulin sensitivity by vegan diets and further studies of efficacy and safety of vegan diets in patients with diabetes and DN are recommended.

Accuracy of an urine-based liquid biopsy genetic test in detecting renal pelvic cancer.

e16500 Background: Renal pelvis cancer is accounting for about two-thirds of upper tract urinary carcinoma. Lesions in renal pelvis often lead to a differential diagnosis that requires ureteroscopic biopsy. Besides causing discomfort and pain, ureteroscopy prior to surgery increases risk of tumor implantation and recurrence. Although urine cytology and FISH were applied, more effective non-invasive diagnostic method is imperative for diagnosis of renal pelvic cancer. Methods: A total of 141 patients with one or more lesions in renal pelvis detected by ultrasound were enrolled. 50-ml first-void urine sample was collection before ureteroscopic biopsy and was analysis by Genetron uro-18 genes panel, by which mutations in 17 genes and methylation status of ONECUT2 were analysis using high-throughput sequencing. If mutations and/or ONECUT2 methylation were detected in urine sediment DNA, the patients were considered as high risk of renal pelvis cancer. The results of the urine-based liquid biopsy genetic test were compared with pathology report and urine FISH results of urinary cells. Results: According to pathological analysis with ureteroscopic biopsy, 42 out of 141 patients (29.7%) were diagnosis with renal pelvic cancer: 13 low-grade, 24 high-grade, 5 equivocal. For 33 renal pelvic cancer patients with FISH testing result of urinary cells, only 54.5% (18/33) samples were positive. By Genetron uro-18 genes test, 95 patients (67.4%) were classified as high risk and 46 (32.6%) patients were low risk. The overall specificity was 88.0% (88/95) and overall sensitivity was 92.9% (39/42), which is significant higher than FISH test of urinary cells. Conclusions: With high specificity and sensitivity, Genetron uro-18 genes test could be incorporated in renal pelvic cancer diagnosis, especially for patients with a ultrasound detectable lesions. Ureteroscopy prior to surgery could be reduced, which may lead to decrease risk of tumor implantation and recurrence.

P1838THE CONTRIBUTION OF MICROSCOPE IN THE KNOWLEDGE OF KIDNEY DISEASES AS DESCRIBED BY GIULIO BIZZOZERO

Abstract Background and Aims Giulio Bizzozero (20 March 1846 - 8 April 1901) was an eminent Italian scientist, the father of the modern microscopy. In 1879 he published his masterpiece “Manuale di Microscopia Clinica “(Handbook of Clinical Microscopy). Here he underlined how the microscopic examination of the urine gave to the physicians of the time indicative criteria of alteration of the kidney, often essential for diagnosis. Method Bizzozero operates in his book a detailed analysis of the methods of time, laying the foundations of microscopy and diagnosis of renal diseases. He proposed a classification of pathological urine and gave large space to the role of the presence of red blood as a great diagnostic sign in urinary sediment (Figure 1 and Figure 2). The author emphasized the use of microscopy against the obsoleted vision of old academics and promulgated the experimental methods in opposition to the vitalistic philosophy of the time. Bizzozero underlined the idea of “teamwork” and the strong connection with his close collaborators, he says that on a laboratory “a mind should give the idea and the direction of work while other minds join together dealing with particular executions”. Results Bizzozero's revolutionary vision of medicine had the purpose to allow every scientist to reach new discoveries in their field, previously being the privilege of an elite, making him a model both as a doctor as a humanist. In one of his memorable speeches he declared: “We need to undress the science of the cloak of mystery and authority: the professor in the school must not offer a series of dogmas supported by the prestige of his name, he must instead expose science in the real state in which it is found, with its doubts and its unresolved mysteries”. Conclusion This Bizzozero’s masterpiece definitely represents a milestone of scientific literature, a witness of the radical evolution of modern medicine and experimental research.

Chemotherapy readiness in pediatric oncology: Assessing an automated method to measure urine specific gravity

As part of the evaluation for chemotherapy readiness, urine specific gravity is measured to assess the patient’s overall hydration status. Depending on the accuracy of the methods used, patients may be adversely affected by having their chemotherapy delayed or prematurely started. To evaluate the diagnostic accuracy of a new automated urine dipstick readout device (Clinitek), we tested 196 consecutive urine samples for urine specific gravity and compared them with the practical gold standard, a urine refractometer. We found a high correlation between both tools among clean urine samples, but a poor correlation among the pathological urine samples.

Impaired urinary concentration ability is a sensitive predictor of renal disease progression in Joubert syndrome.

BackgroundJoubert syndrome (JS) is an inherited ciliopathy characterized by a complex midbrain–hindbrain malformation and multiorgan involvement. Renal disease, mainly juvenile nephronophthisis (NPH), was reported in 25–30% patients although only ∼18% had a confirmed diagnosis of chronic kidney disease (CKD). NPH often remains asymptomatic for many years, resulting in delayed diagnosis. The aim of the study was to identify a biomarker able to quantify the risk of progressive CKD in young children with JS.MethodsRenal features were investigated in 93 Italian patients, including biochemical tests, ultrasound and 1-deamino-8D-arginine vasopressin test in children with reduced basal urine osmolality. A subset of patients was followed-up over time.ResultsAt last examination, 27 of 93 subjects (29%) presented with CKD, ranging from isolated urinary concentration defect (UCD) to end-stage renal disease. Both normal and pathological urine osmolality levels remained stable over time, even when obtained at very early ages. Follow-up data showed that the probability of developing CKD can be modelled as a function of the urine osmolality value, exceeding 75% for levels <600 mOsm/kg H2O, and significantly increased in patients with an early diagnosis of isolated UCD.ConclusionsWe conclude that the frequency of CKD in JS increases with age and is higher than previously reported. Urine osmolality represents an early sensitive quantitative biomarker of the risk of CKD progression.

Ion chromatographic method for the determination of orotic acid in urine

Excess urinary orotic acid excretion occurs in patients with some inborn errors of metabolic pathways such as pyrimidine synthesis and urea cycle. Thus, rapid diagnosis of orotic aciduria has a vital importance for patients. In this paper, a novel method for determination of orotic acid in urine samples by ion chromatography with suppressed conductivity detection was investigated. The separation of orotic acid from urine matrix was accomplished by using an anion exchange column with optimized isocratic eluent program which utilized 50 mM NaOH. The other chromatographic conditions were as follows: the suppressor current was 31 mA; the flow rate of mobile phase was 0.25 mL min−1; the column temperature was 30 °C; sample loop volume was 10 μL. Under optimized conditions, the limit of detection was 0.2 μmol L‾1. Dramatically elevated orotic acid concentration was observed on pathological urine samples comparing to healthy urines, as expected. There are a good many advantages of the proposed method, but using an environmentally friendly reagent free system, no organic solvent employment and its quick nature, and being a sensitive and reliable method are the most obvious ones. The proposed method, therefore, may be utilized as an alternative technique for clinical laboratories.

DIFFERENCES IN LABORATORY FINDINGS OF HYPERTENSIVE PATIENTS DEPENDING ON RISK FACTORS, SETTLEMENT AND AGE

Objective: Hypertension is a chronic disease with growing incidence. Monitoring of laboratory parameters is important in global cardiovascular risk assessment of hypertensive patients and prevention of complications. Evaluation and comparison of laboratory results and risks in patients included in distinct age groups and those from urban and rural settlements. Design and method: The study was conducted on 127 hypertensive patients during the first three months of 2017 at an urban outpatient unit specialized on cardiovascular diseases. Serum samples of patients were processed with Konelab20XTi analyzer using photometric method. Glycemia and lipid profile, kidney and liver function were determined. Urine tests were performed using the HandUReader equipment. Statistical processing of data was made using the GraphPad InStat program. Threshold of significance was set at p < 0.05. Results: Mean age of the patients was 63 years, 56.7% were male subjects. Significant difference (p = 0.0475) occured between serum HDL-cholesterol concentration of rural (average 75.6 mg/dl ± 12.5 SD) and urban patients (average 49.1 mg/dl ± 13.0 SD). Significantly higher serum urea levels were found in the rural subgroup (mean: 44.3 mg/dl ± 13.0 SD) compared to those from urban settlements (mean: 36.4 mg/dl ± 15.0 SD). Pathological urine compounds were found in 43.3% of rural patients and in 29.9% of urban subjects. Significant difference could be observed between the comorbidities present in these subgroups: diabetes was twice as frequent in the rural subgroup compared to urban subjects, prevalence of dislipidemia was three times higher in the urban group compared to rural patients. Significant difference could be observed between creatinine-based glomerular filtration rate (average 57.7 mL/min) of elderly subjects (>70 years) compared to higher values of younger patients, especially those under 60 years of age. No significant difference could be noticed between glycemia, serum uric acid, triglyceride, total cholesterol concentration and transaminase activity of different subgroups based on age or settlement. Conclusions: We can conclude that rural patients present higher protective HDL-cholesterol level compared to urban subjects and lower prevalence of dislipidemia, probably related to their different diet, this might also be the background of their higher serum urea concentration.

Urine cytology screening of French workers exposed to occupational urinary tract carcinogens: a prospective cohort study over a 20-year period

ObjectivesTo demonstrate that urine cytology screening can provide relevant epidemiological data for earlier detection of urothelial cancer caused by occupational exposure.DesignProspective cohort study.SettingIndustries using urothelial carcinogens in France. Urine samples...

Sarcoidosis : Renal manifestations

Renal involvement in sarcoidosis is much more common than generally assumed from old epidemiological studies and is often only detected when actively searched for. Many patients with renal sarcoidosis present with no or only few symptoms. The diagnostic work-up of sarcoidosis should always include apossible renal involvement. In cases of impaired renal function, proteinuria or a pathological urine sediment, arenal biopsy specimen should be obtained to assess the type, severity and prognosis of the kidney disease. Treatment is primarily based on the use of corticosteroids. Steroid-sparing agents, such as disease-modifying antirheumatic drugs and infliximab can be applied; however, the evidence for efficacy of these therapies is mostly based on case series and expert opinions. Discontinuation of immunosuppression therapy bears ahigh risk of relapse.

Adrenocortical cancer: mortality, hormone secretion, proliferation and urine steroids - experience from a single centre spanning three decades.

BackgroundAdrenocortical carcinoma (ACC) is a rare malignant disease with a poor prognosis. Our aims were to study survival and to explore prognostic markers.MethodsWe retrospectively investigated the medical records of all 50 ACC patients at a single centre diagnosed between 1985 and 2012 and followed them up until 31/12/2014.ResultsOf this cohort, twenty six (52 %) were females. Adrenalectomy was performed in 48 patients (96 %), and twenty seven (54 %) were treated with adjuvant cytotoxic agents. The tumor sizes ranged from 6 to 20 cm. Overall survival time was 5.5 years (0.3–19.8), the two and five-year survival was 64 and 40 %, respectively. In ENSAT stage II 25/48 patients had a median survival of 7.0 years (0.7–15.5), in stage III 8/48 this was 1.9 (0.4 – 19.8), and in stage IV 15/48 it was 1.2 (0.3–3.6) years. Seventeen patients (34 %) were still alive at the end of 2014. The total follow-up time was 8.4 (0.3–19.8) years. Cell proliferation measured with Ki-67 had a median value of 15 % (2–80) and the urinary steroid profile was clearly pathologic in 29 of 43 (67 %) tested patients. The proliferation index did not significantly predict mortality (Ki-67 ≤ 10 vs. >10 %, 9.0 vs. 3.2 years, P = 0.0833), but resection margins did (R1 vs. R2, P = 0.0066; R0 vs. R2, P < 0.0001). The urinary steroid profile did not predict mortality (normal vs. pathologic urine profile: median survival 6.6 vs. 3.3 years, P = 0.261).ConclusionsThe prognosis was generally poor and macroscopically positive resection margins resulted in a worse prognosis. However, some patients were still alive many years following primary surgery with no sign of residual disease.

The effectiveness of BD Vacutainer® Plus Urinalysis Preservative Tubes in preservation of urine for chemical strip analysis and particle counting.

IntroductionThe aim of this study was to evaluate the stability of urine collected in preservative tubes for chemistry strip analyses and particle counting to determine whether the transport of urine samples with all of their constituents is possible.Materials and methods275 pathologic urine specimens were included. Each urine sample was evaluated after 4, 8, 12, 24, and 48 hours of storage in BD Vacutainer® Plus Urinalysis Preservative (BD UAP) tubes and compared with refrigeration at 4 °C. All analyses were peformed on H-800 and FUS-200 automatic modular urine analyzers (Dirui Industry, Changchun, China). The kappa coefficients (κ), false positive (FP) and false negative (FN) rates were evaluated. κ > 0.8 was accepted as good agreement.ResultsHaemoglobin (Hb), leucocyte esterase (LE), and protein (Pro) analyses should be performed within 4 hours, whereas glucose (Glc) was stable until the end of 48 hours in both storage conditions. Nitrite (Nit) was well preserved in BD UAP tubes for 24 hours but was stable only up to 8 hours at 4 °C. Bilirubin (Bil) had very high FN rates even at 4 hours in both conditions. The particle counting showed high FN rates for white blood cells (WBC) and red blood cells (RBC), whereas squamous epithelial cells (EC) were stable up to 8 hours in both conditions.ConclusionsPreanalytical requirements for both urine chemical strip analyses and particle counting in a unique sample were not met in either condition. Thus, the transfer of urine samples for centralization of urinalysis is not yet feasible.

Pilot Study of Canine Urine Analysis Using Fluorescent Fingerprint

ABSTRACT Urine contains a variety of compounds including a number of natural fluorophores. Fluorescence spectroscopy has proven to be a useful tool in analytical science. Fluorescence detection has three major advantages over other light-based investigation methods: high sensitivity, speed, and safety. Our work presents the newest approach to analysis of dog urine. Fluorescent fingerprint can very quickly reveal differences between complicated mixtures without adding any reagents. We describe autofluorescence characteristics of the urine of healthy dogs and of those with various disorders using fluorescent fingerprint. Fluorescent analysis has given good results to distinguish between pathological urine and the healthy standard in dogs. Our results suggest that this method can be used to characterize fluorescent properties of canine urine and to reveal some pathological changes in dogs.

Correlation between Muscle Tone and Motor Delay and Biochemical Levels of Carbohydrates in Urine

Aims: To establish a possible correlation between children’s clinical status and carbohydrates in their urine samples as they could be a useful additional tool for objective assessment of muscle tone and motor development. Place and Duration of Study: The Department of Child, Adolescent and Developmental Neurology, University Children’s Hospital in Ljubljana between December 1, 2011 and August 31, 2013. Methodology: The retrospective part of the study comprised children aged 0-5 years (average age, 29.3 months) with motor developmental delay and/or abnormal muscle tone. A group of healthy individuals comprised the prospective part. Besides clinical assessment, electrochemical detection of ten sugars in the urine samples was performed using ion chromatography combined with electrochemical detection and tandem mass spectrometry. Results: 72.0% of 82 hospitalised children had a disorder of muscle tone and/or DQ < 70, 28.0% had a pathological urine sample and an abnormal clinical examination, while 44.0% of children had Original Research Article Anzic et al.; BJMMR, 10(10): 1-8, 2015; Article no.BJMMR.20650 2 a clinical abnormality and a normal urine sample. In the control group, 22.7% had pathological urine samples. There was a statistically significant difference between patients and control subjects in the value of fucose (p = 0.020), sucrose (p = 0.051) and ribose (p = 0.000). Conclusion: A significant correlation was found between abnormalities in clinical status and pathological urine samples (p = 0.002). Absolute and relative values of specific carbohydrates and the number of elevated sugars showed a strong correlation with disease severity.

Age as a determinant of phosphate flame retardant exposure of the Australian population and identification of novel urinary PFR metabolites

The demand for alternative flame retardant materials such as phosphate flame retardants and plasticizers (PFRs) is increasing, although little is known of their possible effects on human health and development. To date, no information on the exposure of children or general Australian population to PFRs is available. The objectives of this study were to characterize the average levels and age-related patterns of PFR metabolites in urine in the general Australian population and to identify novel hydroxylated PFR metabolites in urine. Surplus pathology urine samples from Queensland, Australia were stratified and pooled by age and sex (3224 individuals aged 0 to 75years into 95 pools) according to two different pooling strategies at two different time periods. Samples were analyzed by solid phase extraction and liquid chromatography–tandem mass spectrometry following enzymatic treatment. Nine PFR metabolites were measured in the Australian population, including the first report of a hydroxylated metabolite of TCIPP (BCIPHIPP). Diphenyl phosphate (DPHP), BCIPHIPP and bis(1,3-dichloro-2-propyl) phosphate (BDCIPP) were detected in >95% of samples. DPHP, a metabolite of aryl-PFRs, was found in several samples at levels which were one order of magnitude higher than previously reported (up to 730ng/mL). Weighted linear regression revealed a significant negative association between log-normalized BDCIPP and DPHP levels and age (p<0.001). Significantly greater levels of BDCIPP and DPHP were found in children's urine compared with adults, suggesting higher exposure to PFRs in young children. BCIPHIPP was identified for inclusion in future PFR biomonitoring studies.

Automated chip-nanoelectrospray mass spectrometry for glycourinomics in Schindler disease type I

In this study an integrative mass spectrometry (MS) approach based on fully automated chip-nanoelectrospray quadrupole time-of-flight was optimized and applied for the discovery and structural characterization of O-glycopeptides in a fraction from the urine of a patient diagnosed with Schindler disease type I. A mixture of O-glycopeptides extracted and purified from an age matched healthy subject served as the control. 49 glycoforms were discovered in the investigated urine fraction from Schindler disease versus only 14 in control urine. Structures with relevant biological significance, previously not described, such as O-fucosylated tetrasaccharides and chains up to pentadecamers O-linked to serine, threonine, or threonine-proline were identified in the pathological urine and characterized by tandem MS (MS/MS). A number of 29 species discovered here, most of which with long chain glycans, were not previously reported as associated to this condition. All glycopeptides were detected in only 1min analysis time, with a sample consumption situated in the femtomole range.