尿样中~(131)I 量的测定

本文介绍了人尿样品中~(131)I 含量的测定方法。用离子交换法浓集、CCl_4萃取,碘化银沉淀制源,在低本底β和 FJ-603γ测量装置上测量。本方法灵敏度为0.78Bq/L;对0.1L 尿样,加入17.1Bq~(131)l,化学回收率为86.0±4.3%,放化回收率与化学回收率基本一致;对总裂片核素的去污系数达10~4。     

尿中~(32)P的测定

本文介绍了尿中~(32)P 的测定方法。尿样经预处理后,用丁醇萃取、NH_4Cl-NH_4OH 反萃,将反萃液制成乳状液用切仑科夫法测量。方法的回收率为74.5±7.3%,对主要干扰核素的去污系数在10~3以上,当取样100ml,仪器本底为30cpm。仪器探测效率约为0.3,按本底计数率标准差三倍计算的探测下限为1.5Bq/L。     

Cobalt speciation in urine of hard metal workers. A study carried out by nuclear and radioanalytical techniques

A method for the determination of inorganic and organically-bound cobalt in human urine has been developed and applied to the urine of hard metal workers. The development was based on the use of the radionuclides ⁵⁷Co, ⁵⁸Co and ⁶⁰Co-labelled Co compounds such as Co-Vitamin B12 and CO²⁺ ions which allowed the study of their biotransformations in human and rat urine. The proposed procedure is based on the use of Chelex 100 resin which retains quantitatively the inorganic Co from the urine while the organic complexed form of the element is eluted. Cobalt is detected in both column and eluate by neutron activation analysis (NAA). The method has been applied to speciate inorganic and organically-bound Co in the urine of hard metal workers. There is a significant increase (P < 0.02) of the ratio inorganic/organic Co (2.3) in the urine of workers compared with controls (1.01), showing an increase of the inorganic fraction of Co in the urine of workers. The ratio was constant for the wide range of urinary Co analyzed (from 180 μg to 1254 μg Co/l). Therefore, the discrimination between inorganic and organic Co in urine should not represent progress in the biological monitoring of Co compared with the determination of total urinary Co. However, a large amount of organically-complexed Co is formed in the body of hard metal workers and excreted in urine, thus, investigationsof the nature of the organocobalt compounds are of fundamental importance in establishing their possible clinical significance.     

Solid-phase extraction method for preconcentration of trace amounts of some metal ions in environmental samples using silica gel modified by 2,4,6-trimorpholino-1,3,5-triazin

A method was proposed for the preconcentration of some transition elements at trace levels using a column packed with silica gel modified by a synthetic ligand. Metal ions were adsorbed on 2,4,6-trimorpholino-1,3,5-triazin modified silica gel, then analytes retained on the adsorbent were eluted by 1 mol L⁻¹ hydrochloric acid and determined by flame atomic absorption spectrometry (FAAS). The influences of some experimental parameters including pH of the sample solution, weight of adsorbent, type, concentration and volume of eluent, flow rates of the sample solution and eluent, and sample volume on the preconcentration efficiency have been investigated. The influences of some matrix elements were also examined. The method also was used for simultaneous preconcentration of these elements and the method was successfully applied to the preconcentration and determination of them. The detection limits of the method for Ni²⁺, Co²⁺, Cd²⁺ and Zn²⁺were 0.29, 0.20, 0.23 and, 0.30 ng mL⁻¹, respectively. The application of this modified silica gel to preconcentration of investigated cation from tap water, lake water, urine and apple leaves gave high accuracy and precision (relative standard deviation (R.S.D.) <3%).     

2nd Combined Working Group and Management Committee Meeting of Urine and Kidney Proteomics COST Action 29-30 March 2009, Nafplio, Greece

EuroKUP (Urine and Kidney Proteomics; www.eurokup.org) is a COST (European Cooperation in the field of Scientific and Technical research: www.cost.esf.org Action fostering a multi-disciplinary network of investigators from 25 countries and focusing on facilitating translational proteomic research in kidney diseases. Four Working Groups focusing respectively on defining clinically important research questions in kidney diseases, kidney tissue proteomics, urine proteomics and bioinformatics have been generated. The EuroKUP members had their second combined Working Group and Management Committee (MC) meeting in Nafplio, Greece from March 29 to 30, 2009. This report summarizes the main presentations, discussions and agreed action points during this meeting. These refer to the design of collaborative projects and clinical center networks for specific kidney diseases; establishment of guidelines for kidney tissue proteomics analysis by laser-based imaging- and laser capture microdissection-MS; development and characterization of a "standard" urine specimen to be used for assessment of platform capability and data comparability in clinical proteomics applications; definition of statistical requirements in biomarker discovery studies; and development of a specialized kidney and urine ontology. Various training activities are planned involving training schools on laser capture microdissection- and imaging-MS, workshops on ontologies as well as short-term travel grants for junior investigators.     

Urinary extracellular vesicles for biomarker source to monitor polycystic kidney disease

Extracellular vesicles (EVs) are bilayered lipid vesicles, 50–1000 nm in diameter and secreted by most types of cells. They contain many proteins, mRNAs, miRNAs, and lipids that reflect the pathophysiological state of the cells they originate from, and are therefore considered to be a rich source of potential biomarkers. In this issue (Pocsfalvi, G. et al., Proteomics Clin. Appl. 2015, 9, 552–567), Pocsfalvi et al. conducted pioneering investigations to determine whether changes in the protein content of EVs occur during progression of autosomal dominant polycystic kidney disease (ADPKD), a common genetic disorder that predominantly affects the kidneys. Most significantly, iTRAQ-based quantitative proteomics showed that cytoskeleton-regulating and Ca²⁺-binding proteins are differentially expressed in urinary EVs of ADPKD patients. Impressively, these proteins are involved in biological processes that are closely related to the pathogenic state of tubular epithelial cells in ADPKD, demonstrating the possibility to monitor the status of patients using urinary EVs.     

Sample handling for mass spectrometric proteomic investigations of human urine

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.     

Discovery and validation of urinary biomarkers for prostate cancer

Only 30% of patients with elevated serum prostate specific antigen (PSA) levels who undergo prostate biopsy are diagnosed with prostate cancer (PCa). Novel methods are needed to reduce the number of unnecessary biopsies. We report on the identification and validation of a panel of 12 novel biomarkers for prostate cancer (PCaP), using CE coupled MS. The biomarkers could be defined by comparing first void urine of 51 men with PCa and 35 with negative prostate biopsy. In contrast, midstream urine samples did not allow the identification of discriminatory molecules, suggesting that prostatic fluids may be the source of the defined biomarkers. Consequently, first void urine samples were tested for sufficient amounts of prostatic fluid, using a prostatic fluid indicative panel (“informative” polypeptide panel; IPP). A combination of IPP and PCaP to predict positive prostate biopsy was evaluated in a blinded prospective study. Two hundred thirteen of 264 samples matched the IPP criterion. PCa was detected with 89% sensitivity, 51% specificity. Including age and percent free PSA to the proteomic signatures resulted in 91% sensitivity, 69% specificity.     

The variability in tissue proteomics

Variability is one of the most critical issues of concern in clinical proteomics. In this issue of Proteomics Clinical Applications, Yoshida et al. [Proteomics Clin. Appl. 2012, 6, 412-417] describe the effects of blood and number of washes on the human glomerular proteome isolated from the kidney by laser microdissection. The blood-derived proteins occupied almost 50% of all the identified proteins in the unwashed samples, whereas varying the number of washes (from 1-5) with PBS yielded only 43-80% of the proteins identified in each sample that were common in all samples. This urges caution to all proteomists to carefully consider sample preservation and preparation for tissue proteome analysis.