Urine as a source for clinical proteome analysis: From discovery to clinical application

The success of clinical proteome analysis should be assessed based on the clinical impact following implementation of findings. Although there have been several technological advancements in mass spectrometry in the last years, these have not resulted in similar advancements in clinical proteomics. In addition, application of proteomic biomarkers in clinical diagnostics and practical improvement in the disease management is extremely rare. In this review, we discuss the relevant issues associated with identification of robust biomarkers of clinical value. Urine appears to be an ideal source of biomarkers, for theoretical, methodological, and practical reasons. Therefore, this review is focused on the search for biomarkers in urine within the last decade. Urine can be used for non-invasive assessment of a variety of diseases including those affecting the urogenital tract and also other pathologies such as cardiovascular disease or appendicitis. We also discuss the importance of data validation, an essential step in translating biomarkers into the clinical practice. Furthermore, we examine several examples of apparently successful proteomic biomarker discovery studies and their implications for disease diagnosis, prognosis, and therapy evaluation. We also discuss some current challenges in this field and reflect on future research prospects. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge.     

Effects of Short-Distance Recreational Mushing on Oxytocin,Gastrin, and Creatinine Kinase in Sled Dogs

A considerable increase in creatinine kinase (CK) activity and gastrin hormone due to exercise has been observed in sled dogs during endurance mushing races; however, there have been no studies on sled dogs during recreational mushing. Although oxytocin hormone is involved in social behaviors and empathy, it has not been studied in sled dogs. This study aimed to assess changes in plasma CK activity, and gastrin and oxytocin concentrations in adult sled dogs used in touristic mushing in North Patagonia, Argentina. Blood samples were collected before, during, and after the winter season of 2017. Creatinine kinase activity measurement was done using an enzymatic assay. Hormone analyses were performed using commercial Enzyme-Linked InmunoSorbent Assay kits. Results showed an expected two-fold increase in CK activity during the winter, with recovering basal values after winter (< 400 UI/L), low and stable levels of gastrin (9.4 ± 8.8 pg/mL), and a slight increase in oxytocin (23%) after mushing activities. No evidence indicated gastrin alterations or muscular damage from touristic mushing, but an oxytocin increase would indicate a stimulation of the brain reward system.     

Hematobiochemical and histopathological alterations of kidney and testis due to exposure of 4G cell phone radiation in mice

The radiofrequency electromagnetic radiation emitted by smart phones on biological systems has wide media coverage and public concern in recent years. The aim of this study was to explore the effects of fourth-generation cell phone radiation exposure on hematological (Total leukocyte count, Total erythrocyte count, and hemoglobin %), biochemical (Serum creatinine) parameters, and histopathological changes in the kidney and testis of Swiss albino mice. A total of 30 male Swiss albino mice weighing 45–65 g was randomly divided into three groups (n = 10). The first group A was the control group, the second group B, was exposed to 40 minutes of mobile phone radiation daily, the third group C was exposed to 60 minutes of radiation daily from two 2400 Megahertz fourth-generation connected mobile phones for 60 days, respectively. The electromagnetic radiation frequency radiometer measured the frequency of electromagnetic radiation emitted from cell phones. The specific absorption rate was calculated as 0.087 W/kg. The control group was kept under similar conditions, but the electromagnetic field was not given for the same period. All the mice were sacrificed at the end of the experiment. The blood samples were collected for hematobiochemical study, and then kidney and testis tissues were collected for histopathological study. Results of the study showed that the body weight and total erythrocyte count values were significantly (p < 0.05) decreased while total leukocyte count, hemoglobin %, and serum creatinine values were significantly (p < 0.05) increased in both the radiation exposure groups relative to the control group. Histopathological observation showed the kidney of 60 minutes exposed mice interstitial inflammation that causes marked mononuclear cellular infiltration compared to the 40 minutes and control mice. Compared to control mice, histopathological examinations of testicular tissue from the exposed mice, showed irregular in shapes and non-uniform sizes and fewer spermatogenic cells layer that leads to the larger lumen in the seminiferous tubules. It is concluded that fourth-generation cell phone radiation exposure may affect blood hemostasis and inflammation of mice's kidney and testis tissue. Based on these studies, it is important to increase public consciousness of potential adverse effects of mobile phone radiofrequency electromagnetic radiation exposure.     

成都地区中老年居民血清尿酸相关危险因素分析

目的：分析成都地区中老年居民血清尿酸水平及其影响因素,为防治心脑血管事件提供策略支持。方法：利用2007代谢综合征研究调查资料（共1061人）,把人群依据血尿酸水平分为增高组（男性〉420umol/L,女性〉357umol/L）及正常组（男性≤420umol/L,女性≤357umol/L）,分析两组人群的多个代谢性指标的分布特征,并采用logistic回归分析寻找与尿酸相关的危险因素。结果：①两个分组间年龄、收缩压、男性比重、高血压家族史、人体质量指数、腰围、臀围、空腹血糖、肾功能等指标尿酸增高组明显高于尿酸正常组,舒张压则是尿酸增高组明显低于尿酸正常组。②血脂各成分中,总胆固醇、低密度脂蛋白水平尿酸增高组高于正常组,其余甘油三酯、高密度脂蛋白分组间无统计学差异。③尿酸增高组其代谢综合征、高血压、糖尿病、肥胖、腹型肥胖、血脂异常等患病率皆高于尿酸正常组,差异有统计学意义。④Logistic回归分析提示尿酸与你男性性别、年龄、收缩压、BMI、腰围、臀围、空腹血糖、肌酐、总胆固醇、低密度脂蛋白水平呈正相关,与女性性别及舒张压呈负相关。结论：成都地区尿酸与代谢性指标及肾功能相关指标关系密切,可能可以通过减少尿酸来减少心脑血管疾病、代谢性疾病及肾脏疾病的发生。     

Molecular genetic assay of mucopolysaccharidosis IVA in South China

Mucopolysaccharidosis IVA (MPS IVA) is an autosomal recessive lysosomal storage disorder caused by the deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Molecular mutational analysis was performed by PCR product sequencing for fourteen exons and exon–intron boundaries of GALNS gene in 21 patients from 19 unrelated families with severe MPS IVA in South China. We identified fifteen different mutations, including 10 reported mutations (p.P125L, p.G290S, p.M318R, p.G340D, p.L366P, p.R386C, p.A392V, c.1243-1G>C, p.L440RfsX54 and p.X523E) and five novel mutations (p.N177S, p.G290R, p.F306S, p.W403_T404delinsCS, p.W520X). All five novel mutations were inherited from parents of the patients and not found in 100 normal control alleles. Three mutations, p.M318R, p.L366P and p.R386C were common, accounting for 36.8% of mutant alleles investigated. One patient homozygous of p.A392V and the other two unrelated patients homozygous of p.L366P presented classical disease course. The results show that the GALNS gene has a different mutational spectrum in South China as compared to other regions. The p.A392V and p.L366P mutations were associated with severe phenotype of MPS IVA.     

Multiorgan autoimmunity in a Turner syndrome patient with partial monosomy 2q and trisomy 10p

Turner syndrome is a condition caused by numeric and structural abnormalities of the X chromosome, and is characterized by a series of clinical features, the most common being short stature and gonadal dysgenesis. An increased frequency of autoimmune diseases as well as an elevated incidence of autoantibodies has been observed in Turner patients.     

高原寒环境暴露对急进入高原健康人群肾功能的影响

目的:探讨高原低压低氧寒环境对急进高原的平原健康人群肾功能的影响。方法:选取30例长期居住兰州地区急进高原(海拔3300米、日平均气温-4度)健康人群,对比分析所有受试者进驻高原前和进驻高原后第24 h、72 h、7天、14天、28天尿IL-18(白介素-18)、尿NAGL(中性粒细胞明胶酶相关性脂质运载蛋白)、尿KIM-1(肾损伤因子-1)及血肌酐水平的变化。结果:共28人完成全部6次尿液及血标本采集并检测,其中2名女性因某次采集点处于月经期剔除,其中男性17人、女性11人,平均年龄26.8±3.2岁(男性28.3±4.2岁,女性24.8±2.6岁)。进入高原前和进驻高原后第24 h、72 h、7天、14天、28天尿IL-18检测值分别为3.62±0.32 ng/L、11.20±0.65 ng/L(P<0.001,较进入前组比较)、6.32±0.46 ng/L(P<0.001)、4.36±0.68 ng/L(P<0.05)、3.58±0.71 ng/L、3.32±0.46 ng/L;尿NAGL水平分别为0.126±0.20μg/L、0.513±0.003μg/L(P<0.001)、0.116±0.006μg/L、0.009±0.001μg/L、0.121±0.010μg/L、0.632±0.009μg/L;尿KIM-1水平分别为2.61±0.22 ng/L、18.20±0.69 ng/L(P<0.001)、6.32±0.46 ng/L(P<0.001)、6.36±0.68 ng/L(P<0.001)、2.58±0.31 ng/L、2.32±0.26 ng/L;血肌酐水平分别为61.0±9.16μmol/L、58.5±8.13μmol/L、80.3±10.38μmol/L(P<0.05)、76.5±12.04μmol/L(P<0.05)、62.6±10.14μmol/l、62.3±8.18μmol/L。结论:急进高原健康入群早期肾功能有改变,其中尿IL-18、尿KIM-1、尿NAGL水平较血肌酐水平变化更早、更敏感。     

Evaluation of urinary biomarkers of oxidative/nitrosative stress in adolescents and adults with Down syndrome

Urinary biomarkers of oxidative stress have been little studied in adults with Down syndrome (DS), usually no more than two biomarkers have been measured in the population studied and controversial results are reported in literature. Thus, we aimed to assess a set of oxidative and nitrosative stress biomarkers in urine samples of adolescents and adults with DS, with and without hypothyroidism, which comprise: 8-hydroxy-2′-deoxyguanosine (8-OHdG), isoprostane 15-F_2t-IsoP, thiobarbituric acid-reacting substances (TBARS), advanced glycation end products (AGEs), dityrosine (diTyr), hydrogen peroxide (H₂O₂) and nitrite/nitrate (NOx). Fluorimetric and spectrophotometric assays were performed in DS (n = 78), some of them taking levothyroxine for hypothyroidism (n = 24), and in their healthy age-matched controls (n = 65). We found that levels of AGEs, diTyr, H₂O₂ and NOx are increased in DS patients in any or in all age groups, whereas Cr levels were lower in DS than in controls in all age groups. Besides, correlations with age in DS were positive for diTyr and negative for Cr, TBARS, 15-F_2t-IsoP and NOx. We also found lower levels of Cr from 15 to 19 years, higher levels of TBARS and AGEs from 20 to 40 years and higher levels of diTyr from 15 to 40 years in DS patients receiving levothyroxine than in DS without hypothyroidism diagnosed. We conclude that AGEs, diTyr, H₂O₂ and NOx could be used as oxidative stress biomarkers in DS in contrast to 8-OHdG, 15-F_2t-IsoP and TBARS, at least with the methods used. However, renal impairment could occur in DS and Cr adjustment may bias the results, particularly in hypothyroid patients.     

Impaired DNA repair and genomic stability in hereditary tyrosinemia type 1

The autosomal recessive disorder, hereditary tyrosinemia type 1 (HT1), is caused by a defective fumarylacetoacetate hydrolase enzyme. Consequently intermediate metabolites such as fumarylacetoacetate, succinylacetone and p-hydroxyphenylpyruvic acid accumulate. Characteristic to HT1 is the development of hepatocellular carcinoma, irrespective of dietary intervention or pharmacological treatment. Carcinogenesis may occur through a chromosomal instability mutator phenotype or a microsatellite instability phenotype, and deficient DNA repair may be a contributing factor thereof. The purpose of this study was to investigate the expression of DNA repair proteins, and the possible occurrence of microsatellite instability in HT1. Gene expression analyses show low expression of hOGG1 and ERCC1 in HT1 patient lymphocytes. Results from microsatellite instability analyses show allelic imbalance on chromosome 7 of the fah^−/− mouse genome, and instability of the D2S123, D5S346 and (possibly) D17S250 microsatellite markers, in HT1 patient lymphocytes.     

Evaluation of the health status outcome among inpatients treated for Amphetamine Addiction

Amphetamine is one of the most abuser drugs in Saudi Arabia. The aim of this study was to evaluate health status outcome at baseline and after detoxification in amphetamine users through the evaluation of the body mass index, renal function tests, cardiac biomarkers, gonadal hormonal levels, and oxidative stress markers. A cross-sectional study was conducted on 90 participants. Sixty participants were hospitalized patients for treatment of addiction and 30 participants were healthy volunteers. This study was performed at a psychiatric and rehabilitation center, in Qassim region, in the Kingdom of Saudi Arabia. Participants were divided into: group I = control; group II = amphetamine users and group III = amphetamine plus cannabis users. Socio-demographic data was collected. The urinary amphetamine level, Severity Dependence Scale (SDS), body mass index (BMI), vital signs; serum levels of troponin T (TnT), immunoglobulin M (IgM), immunoglobulin G (IgG), luteinizing Hormone (LH), testosterone Hormone (TSTS), urea, creatinine, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were measured on admission and after detoxification. The results showed that the BMI was significantly decreased while, vital signs such as heart rate, blood pressure and respiratory rate were significantly increased in all abusers and returned to normal values after the detoxification period. The cardiac biomarker troponin T was significantly increased and reversed after detoxification. The immune system was evaluated through assessing serum levels of immunoglobulin (Ig) M and IgG. The immune system remained immunocompromised in drug users, and IgM and IgG levels did not reach the level of control group after treatment. Luteinizing and testosterone hormones were evaluated. Both hormones were increased on admission and improved after the detoxification period. Renal function showed no significant differences between drug users and the control group. In the evaluation of the antioxidant system, there was a significant increase in serum MDA, SOD, GPx, and CAT levels compared to healthy controls. After the detoxification phase, these oxidative stress biomarkers still remained elevated. The current results have shown the addiction of amphetamine and cannabis exert detrimental effects on different body organs and the exert major consequences on the health status of drug users. The present study showed that, there was no improvement in the levels of oxidative stress biomarkers, although an improvement was observed in the other parameters after the detoxification phase.