Quantitative Analysis of a Promising Cancer Biomarker,Calretinin, by a Biosensing System Based on Simple and Effective Immobilization Process

Calretinin (CAL) is calcium binding protein, and its levels in blood and cerebrospinal fluids are increased, since its expression is increased various cancer types. A novel biosensor system fabricated by immobilization of a specific antibody to CAL, anti‐Calretinin (anti‐CAL), onto a gold electrode surface via an effective covalent binding method using mercaptohexanol, epichlorohydrin, and ethanolamine was reported for the sensitive, selective, and accurate analysis of CAL. The proposed biosensor showed a linear calibration range between 1 ng/mL and 5 ng/mL. LOD and LOQ values were determined as 0.11 ng/mL and 0.38 ng/mL, respectively. The standard deviation related to the reproducibility of the new biosensor system was calculated as 3.95 %. Lastly, in order to state the applicability of the biosensor to early diagnosis of CAL in practice, artificial serum samples spiked with CAL have been analyzed by the proposed biosensor.     

Quantitative Mass Spectrometry-Based Proteomics for Biomarker Development in Ovarian Cancer

Ovarian cancer is the most lethal gynecologic malignancy among women. Approximately 70–80% of patients with advanced ovarian cancer experience relapse within five years and develop platinum-resistance. The short life expectancy of patients with platinum-resistant or platinum-refractory disease underscores the need to develop new and more effective treatment strategies. Early detection is a critical step in mitigating the risk of disease progression from early to an advanced stage disease, and protein biomarkers have an integral role in this process. The best biological diagnostic tool for ovarian cancer will likely be a combination of biomarkers. Targeted proteomics methods, including mass spectrometry-based approaches, have emerged as robust methods that can address the chasm between initial biomarker discovery and the successful verification and validation of these biomarkers enabling their clinical translation due to the robust sensitivity, specificity, and reproducibility of these versatile methods. In this review, we provide background information on the fundamental principles of biomarkers and the need for improved treatment strategies in ovarian cancer. We also provide insight into the ways in which mass spectrometry-based targeted proteomics approaches can provide greatly needed solutions to many of the challenges related to ovarian cancer biomarker development.     

Clinical perspectives of high-resolution mass spectrometry-based proteomics in neuroscience: exemplified in amyotrophic lateral sclerosis biomarker discovery research

Biomarker discovery is a central application in today's proteomic research. There is an urgent need for valid biomarkers to improve diagnostic tools and treatment in many disorders, such as the rapidly progressing neurodegenerative disorder amyotrophic lateral sclerosis (ALS) that has a fatal outcome in about 3 years and yet no curative treatment. Screening for clinically relevant biomarkers puts high demands on high-throughput, rapid and precise proteomic techniques. There is a large variety in the methods of choice involving mainly gel-based approaches as well as chromatographic techniques for multi-dimensional protein and peptide separations followed by mass spectrometry (MS) analysis. This special feature article will discuss some important aspects of MS-based clinical proteomics and biomarker discovery in the field of neurodegenerative diseases and ALS research respectively, with the aim to provide a prospective view on current and future research aspects in the field. Furthermore, examples for application of high-resolution MS-based proteomic strategies for ALS biomarker discovery will be demonstrated with two studies previously reported by our group. These studies include among others, utilization of capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS) for advanced protein pattern classification in cerebrospinal fluid (CSF) samples of ALS patients as well as highly sensitive protein identification in minimal amounts of postmortem spinal cord tissue and laser micro-dissected motor neurons using FT-ICR-MS in conjunction with nanoflow LC coupled to matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (LC-MALDI-TOF-TOF-MS).     

Dye adsorbed few-layer thick Ti3C2Tx-MXenes for direct electrochemical detection of CD44 proteins

The direct electrochemical detection of cancer biomarkers using single single-component platforms is challenging. Herein, we propose constructing an efficient screen-printed electrode (SPE) based platform for selective detection of CD44 proteins, a non-kinase transmembrane glycoprotein. A sensing platform, MB-MX/HA/SPE, was developed by incorporating few-layered Ti₃C₂T_x nanosheets pre-loaded with methylene blue (MB) dye. The nanosheets were subsequently immobilized with hyaluronic acid (HA), which served as a ligand for the specific recognition of CD44. The simple electrode configuration and the highly conductive Ti₃C₂T_x facilitated the electrochemical oxidation of MB, generating a reference SWV signal that declined proportionally with the increasing concentration of CD44 owing to ligand (HA)-protein interaction. The sensor could register a sensitive inhibition response in the concentration range of 0.1 to 7.25 ng.mL⁻¹ with a detection limit of 1.2×10⁻² ng.mL⁻¹ for CD44 proteins. Moreover, the synergistic combination of the highly conductive/adsorptive Ti₃C₂T_x nanosheets and hyaluronic acid (HA) led to strong antifouling characteristics even in the presence of other common proteins, such as bovine serum albumin (BSA), haemoglobin (Ig), immunoglobulin G (IgG), prostate-specific antigen (PSA), and neuron-specific enolase (NSE). The proposed strategy eliminates the need for additional components in the electrode modification procedure. In addition, incorporating MXenes as electrode material paves the way for developing sensitive biosensors with prospective applications in cancer diagnosis.     

Discovery of Elaphuri Davidiani Cornu–specific peptide biomarkers by peptidomics analysis–based method

Elaphuri Davidiani Cornu (EDC) is the antler of the male Père David's deer, which has been reported to have multiple biological activities, and its use as a traditional Chinese medicine (TCM) in China has been known for thousands of years. However, EDC is difficult to distinguish from other related species–derived antlers in powder or extract form in TCM clinic use, such as Cervus elaphus Cornu (CEC) and Cervus nippon Cornu (CNC), both derived from Cervidae and easily confused with EDC. In this study, a strategy using peptidomics combined with mathematics set analysis was used to identify EDC-specific peptide biomarkers, and four specific peptide biomarkers (Pep-E1–E4) were identified and validated. Pep-E1, Pep-E3, and Pep-E4 could be exclusively detected in EDC samples, with relative peak areas of 0.298 ± 0.060, 0.039 ± 0.015, and 0.037 ± 0.008, whereas Pep-E2 showed relative peak area of 0.516 ± 0.101 in EDC, 0.132 ± 0.026 in CEC, and 0.136 ± 0.047 in CNC samples, respectively. These four peptides are applicable to distinguish EDC from CEC and CNC, which is of great significance for the quality control of EDC.     

低剂量甲基毒死蜱对鲫鱼肝脏主要酶活性的影响

生物防御过氧化损害系统能反映有毒污染物对水环境的早期影响,可作为早期警报指标应用于环境监测中.本文以幼龄鲫鱼为受试生物,研究了较低剂量(1μg/L)的甲基毒死蜱对其肝脏的毒理学效应,分析了29d中鲫鱼的生长指数、肝脏指数以及肝脏抗氧化酶系(包括超氧歧化物歧化酶活性、还原谷胱甘肽含量)和转氨酶活性等随暴露时间的变化趋势,综合考察了各项指标参数.结果表明:该生命体内的保护机制呈现相互竞争和相互代偿合作的关系;谷胱肝肽含量随暴露时间的变化较为显著,可作为敏感的生物标志物.     

应用代谢轮廓区分不同致病性副溶血性弧菌

基于tdh, trh和tlh 3个基因区分了不同的致病性副溶血性弧菌. 采用液相色谱(LC)和气相色谱-质谱联用(GC-MS)技术获得不同的致病性副溶血性弧菌的代谢轮廓, 并将其用于区分不同的致病性副溶血性弧菌; 同时以肠杆菌基因间重复共有序列聚合酶链反应技术(ERIC-PCR)及DNA重复序列PCR技术(REP-PCR)为对照, 采用NTsys2.10e软件计算所得结果的相似系数, 并对气相色谱-质谱联用结果进行解析. 结果表明, 根据所得代谢轮廓可以很好地区分不同的致病性副溶血性弧菌; 对气相色谱-质谱联用分析结果解析发现了不同致病性菌株的潜在生物标志物: tdh⁺, trh^-, tlh⁺菌株3种, tdh^-, trh⁺, tlh⁺菌株2种, tdh^-, trh^-, tlh⁺菌株 3种.     

蛋白质组驱动的精准医学研究进展

蛋白质是细胞的重要组成成分,参与人体各种代谢及生理功能的调控.蛋白质组学可以分析蛋白质表达水平、修饰状态和细胞内蛋白质相互作用,为研究这些分子在疾病发生和发展的不同阶段的变化提供了全局视角.近年来,临床蛋白质组学已被证实在疾病的早期诊断、预后和病情发展监测中发挥重要作用,并由此提出了蛋白质组学驱动的精准医学的概念.该文...     

顺铂对卵巢癌COC1细胞总蛋白表达的影响研究

对卵巢癌COC1细胞进行体外培养, 当培养至10⁹/mL浓度时裂解细胞, 获得细胞总蛋白, 利用二维电泳对顺铂诱导前后的细胞内总蛋白质进行分离, 利用PDQuest 7.1.1专业分析软件对二维凝胶图谱进行差异比较, 利用基质辅助激光解析飞行时间质谱(MALDI-TOF-MS)进行质量分析, 最后进行生物信息学检索, 以寻找卵巢癌细胞发生、发展和凋亡的分子标记物. 经二维电泳图谱和质谱分析, 鉴定了11种蛋白, 4种蛋白表达量有差异, 7种蛋白在诱导前后表现为有和无的关系. 有5种是“分子伴侣”蛋白, 有4种蛋白与人体能量代谢有关. 蛋白功能分析提示卵巢癌细胞的蛋白质组表达存在差异, 这些差异蛋白对于寻找卵巢癌生物标志物, 开发新药提供很好的理论依据.     

Fabrication and characterization of polyurethane patch loaded with palmarosa and cobalt nitrate for cardiac tissue engineering

Cardiac patches are attractive option in overcoming the morbidities associated with cardiac disorders. Nanofibrous scaffolds were fabricated using polyurethane (PU) added with palmarosa (PR) and cobalt nitrate (CoNO₃) using an electrospinning technique. Several characterizations were employed namely field emission scanning electron microscopy, wettability measurement, attenuated total reflectance infrared spectroscopy, thermal analysis, surface roughness measurements, and tensile testing. Further, biological response of the electrospun nanofibers were tested through coagulation study and MTS assay. As-spun composite mats showed smaller fibers than pure PU as depicted in morphology analysis. The interaction of PU with PR and CoNO₃ was confirmed in infrared spectrum and thermal analysis. The incorporation of the PR decreased the wettability and while CoNO₃ addition resulted in the hydrophilic nature as depicted in the contact angle measurements. Mechanical properties testing showed that elongation at break for the pristine PU was increased with the addition of PR and CoNO_3. The surface measurements depicted that the incorporation of PR resulted in the improvement of the surface roughness while the addition of CoNO₃ reduced the surface roughness of the pristine PU. The electrospun nanocomposites showed delayed blood clotting time compared to the pristine PU as shown in coagulation study. Both composites supported the better proliferation of fibroblast cells than pure PU. Therefore, novel composites with smaller fiber diameter, hydrophilicity, better mechanical properties, improved blood compatibility parameters, and good cell viability rates would be a promising candidate for cardiac tissue engineering.