Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease

Background: Sensitive and specific noninvasive biomarkers for tubulointerstitial injury are lacking, and proteomic techniques provide a powerful tool for biomarker discovery. Objective: The aim of this study was to identify novel urinary biomarkers of early tubulointerstitial injury in canine progressive renal disease using both 2‐dimensional differential in‐gel electrophoresis (2‐D DIGE), which identifies individual proteins, and surface‐enhanced laser desorption ionization time‐of‐flight mass spectrometry (SELDI‐TOF), which generates protein peak profiles. Methods: Urine was collected from 6 male dogs with X‐linked hereditary nephropathy (XLHN) at 2 time points (TP): 1) the onset of overt proteinuria (urine protein:creatinine ratio>2) and 2) the onset of azotemia (creatinine ≥1.2 mg/dL); corresponding renal biopsies were analyzed from 3 of the dogs. Urine samples from the 6 dogs were subjected to analysis by 2‐D DIGE and SELDI‐TOF. Urinary retinol‐binding protein (RBP) was evaluated in 25 male dogs with XLHN and normal control dogs by Western blot analysis. Results: Clinical data and histologic evaluation revealed reduced renal function and increased tubulointerstitial fibrosis at TP 2. A number of urine proteins and protein peaks were differentially present at the 2 time points, with several known biomarkers of renal disease identified in addition to several promising new biomarkers. RBP was first detected in urine approximately 2 months before onset of azotemia (TP 2), but after onset of overt proteinuria, and amounts increased with progression of disease. Conclusions: Proteomic techniques were successfully used to identify urinary biomarkers of renal disease in dogs with XLHN. Urinary RBP is a promising biomarker for early detection of tubulointerstitial damage and progression to end‐stage renal disease.     

Atrial Natriuretic Peptide as an Indicator of the Severity of Valvular Regurgitation and Heart Failure in Horses

Natriuretic peptides are cardiac biomarkers, routinely used for diagnosis, prognosis, and guidance for treatment in human and small animal cardiology. However, their diagnostic and prognostic value in horses has received little study. The aim of this study was to investigate the plasma atrial natriuretic concentration (ANP_Pl) in a large group of horses with various degrees of valvular regurgitation (VR) and congestive heart failure (CHF). Clinical examination and two-dimensional time-motion mode and Doppler echocardiography were performed on 91 horses admitted to the Equine Teaching Hospital of Liege University, with either no, mild, moderate, or severe VR and presenting various stages of CHF. Plasma atrial natriuretic concentration was measured using a commercially available human radioimmunoassay test. Mean values of body weight, age, ANP_Pl, and echocardiographic parameters were compared among horses with no, mild, moderate, and severe VR and among horses in different CHF stages. Correlation and linear regression of ANP_Pl with each echocardiographic parameter and with the percentage of dilation of each of cardiac chambers were assessed. Horses with severe or moderate VR had significantly higher ANP_Pl than those with mild or no VR. Plasma atrial natriuretic concentration was significantly higher in horses presenting CHF than those without CHF. Plasma atrial natriuretic concentration was significantly correlated with the left atrial diameter and its percentage of dilation. These results suggest a diagnostic value of ANP_Pl in horses with VR, especially with tricuspid or mitral insufficiency with dilation of the atria.     

稀有鮈鲫热休克因子1的分子克隆与表达分析

热休克因子1(Heat shock factor 1,HSF1)作为转录调控元件,能够调控热休克蛋白(heat shock proteins,HSPs)的表达,保护机体免受压力损伤。为评估稀有鮈鲫(Gobiocypris rarus)HSF1和环境污染物五氯酚(pentachlorophenol,PCP)的关系,本研究采用cDNA末端快速扩增(RACE)技术从稀有鮈鲫肝脏中克隆HSF1全长序列,并将其命名为GrHSF1;利用Real-time PCR检测稀有鮈鲫GrHSF1的组织分布;温度以及PCP暴露下稀有鮈鲫肝脏中的GrHSF1的表达情况。RACE结果显示:GrHSF1的开放阅读框(open reading frame,ORF)长度为1581 bp,共编码527个氨基酸。系统发育树分析表明,预测得到的GrHSF1氨基酸序列与其他鱼类HSF1具有广泛的序列相似性。组织分布显示GrHSF1在稀有鮈鲫的12种组织中均表达,在肝脏中表达量最高。在肝脏中的暴露实验表明,当80μg/L≥PCP≥8μg/L时,GrHSF1的mRNA与PCP之间具有明显的剂量依赖效应,当160μg/L≥PCP≥8μg/L时,GrHSF1的mRNA与PCP之间具有明显的时间依赖效应;在过高或过低温度下,GrHSF1的mRNA均显著下降。结果表明GrHSF1是已知鱼类HSF1家族的同源基因,并可作为评估环境污染物PCP的生物标志物。     

Candidate serum metabolite biomarkers of residual feed intake and carcass merit in sheep

Mutton and lamb sales continue to grow globally at a rate of 5% per year. However, sheep farming struggles with low profit margins due to high feed costs and modest carcass yields. Selecting those sheep expected to convert feed efficiently and have high carcass merit, as early as possible in their life cycle, could significantly improve the profitability of sheep farming. Unfortunately, direct measurement of feed conversion efficiency (via residual feed intake [RFI]) and carcass merit is a labor-intensive and expensive procedure. Thus, indirect, marker-assisted evaluation of these traits has been explored as a means of reducing the cost of its direct measurement. One promising and potentially inexpensive route to discover biomarkers of RFI and/or carcass merit is metabolomics. Using quantitative metabolomics, we profiled the blood serum metabolome (i.e., the sum of all measurable metabolites) associated with sheep RFI and carcass merit and identified candidate biomarkers of these traits. The study included 165 crossbred ram-lambs that underwent direct measurement of feed consumption to determine their RFI classification (i.e., low vs. high) using the GrowSafe System over a period 40 d. Carcass merit was evaluated after slaughter using standardized methods. Prior to being sent to slaughter, one blood sample was drawn from each animal, and serum prepared and frozen at −80 °C to limit metabolite degradation. A subset of the serum samples was selected based on divergent RFI and carcass quality for further metabolomic analyses. The analyses were conducted using three analytical methods (nuclear magnetic resonance spectroscopy, liquid chromatography mass spectrometry, and inductively coupled mass spectrometry), which permitted the identification and quantification of 161 unique metabolites. Biomarker analyses identified three significant (P < 0.05) candidate biomarkers of sheep RFI (AUC = 0.80), seven candidate biomarkers of carcass yield grade (AUC = 0.77), and one candidate biomarker of carcass muscle-to-bone ratio (AUC = 0.74). The identified biomarkers appear to have roles in regulating energy metabolism and protein synthesis. These results suggest that serum metabolites could be used to categorize and predict sheep for their RFI and carcass merit. Further validation using a larger (3×) and more diverse cohort of sheep is required to confirm these findings.     

A Feedback Mechanism to Control Apoptosis Occurs in the Digestive Gland of the Oyster Crassostrea gigas Exposed to the Paralytic Shellfish Toxins Producer Alexandrium catenella

To better understand the effect of Paralytic Shellfish Toxins (PSTs) accumulation in the digestive gland of the Pacific oyster, Crassostrea gigas, we experimentally exposed individual oysters for 48 h to a PSTs producer, the dinoflagellate Alexandrium catenella. In comparison to the effect of the non-toxic Alexandrium tamarense, on the eight apoptotic related genes tested, Bax and BI.1 were significantly upregulated in oysters exposed 48 h to A. catenella. Among the five detoxification related genes tested, the expression of cytochrome P450 (CYP1A) was shown to be correlated with toxin concentration in the digestive gland of oysters exposed to the toxic dinoflagellate. Beside this, we observed a significant increase in ROS production, a decrease in caspase-3/7 activity and normal percentage of apoptotic cells in this tissue. Taken together, these results suggest a feedback mechanism, which may occur in the digestive gland where BI.1 could play a key role in preventing the induction of apoptosis by PSTs. Moreover, the expression of CYP1A, Bax and BI.1 were found to be significantly correlated to the occurrence of natural toxic events, suggesting that the expression of these genes together could be used as biomarker to assess the biological responses of oysters to stress caused by PSTs.     

Stability of needle‐ and root‐derived biomarkers during litter decomposition

Background and aims: Plant‐derived, ester‐bound substituted fatty acids have been used for decades as biomarkers to identify input of plant materials in sediments and soils. However, the long‐term decomposition patterns of these biomarker compounds under natural conditions are not well understood, although this is a basic prerequisite for quantitative biomarker applications. Methods: For this study, we analyzed the decomposition patterns of root‐ and needle‐specific compounds of Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) in a litterbag study conducted over 3 years. Samples were analyzed by methanolic KOH extraction with previous removal of free extractable lipids. Results: The concentrations of most detectable compounds had decreased after three years of incubation. The observed changes of concentrations followed a non‐linear path and cannot be explained by microbial uptake and metabolism alone. Other factors controlling the breakdown of ester‐bound lipids, like lipid oxidation must play a role. Between similar plant parts and different plant parts of the same species, the observed degradation patterns were heterogeneous. The estimated ratio of remaining root and needle biomass that may arise with the choice of a particular biomarker varies in this study between 0.6 and 40 times after three years. Conclusion: This range of variation does not allow reliable conclusions about the contribution of roots and needles to decomposed organic matter based on biomarkers ratios.     

Studies on the Structure Model of Coal and Its Polarization Characteristics

The macromolecular structures of coal samples collected from Yutianbao, Nantong and Qingnian coal mines in Sichuan were studied and a model of structure for these coals was set up. On the basis of above mentioned work,experimental studies and theoretical analyses were made to investigate the characteristics of coal polarization in applied electric field. The results indicate that the rotation polarzation and the space polarization are the essential reason for the attenuation of electric current through coal samples,and that the space polarization is the main form of coal polarization.     

先进功能高分子材料在设施农业中的应用与前景展望

发展设施农业既是我国农业农村经济发展新阶段的客观需求,也是加快农业结构调整、转型升级和发展低碳农业的必然选择。综述了光致变色高分子薄膜材料、可降解高分子膜材料、高透光薄膜、高分子半透膜材料、太阳能薄膜电池材料5种先进功能高分子材料的优越性能,并对其在现代设施农业中的应用前景进行了展望。