Identification of Novel Biomarker and Therapeutic Target Candidates for Diagnosis and Treatment of Follicular Adenoma

ollicular adenoma is a type of benign and encapsulated nodule in the thyroid gland, but some adenomas have the potential to progress to follicular carcinoma. Therefore, it is important to monitor the state and progress of follicular adenoma in the clinic and discover drug development targets for the treatment of follicular adenoma to prevent its worsening to follicular carcinoma. Currently, the study of biomarkers and therapeutic targets lacks applications of up-to-date technologies, including proteomics and bioinformatics. To discover novel protein biomarker and therapeutic target candidates, a liquid chromatography-tandem mass spectrometry approach was applied to directly compare follicular adenoma with normal thyroid tissue samples. The proteomics analysis revealed 114 protein biomarker candidates out of 1,780 identified and quantified proteins. A comprehensive approach to prioritize the biomarker candidates by category and rank revealed CD63, DDB1, TYMP, VDAC2, and DCXR as the top five biomarker candidates. Upstream regulator analysis using the Ingenuity Pathway Analysis (IPA) software discovered four therapeutic target candidates for follicular adenoma, including TGFB1, MYC, ANGPT2, and NFE2L2. This study provided biomarker and therapeutic target candidates for a follow-up study, which will facilitate monitoring and treatment of follicular adenoma

Wireless sensor integration for bridge model health monitoring

An integrated hardware and software system for a scalable wireless sensor network (WSN) is designed and developed for structural health monitoring. An extension sensor board is designed, developed, and calibrated to meet the requirements for structural vibration monitoring and modal identification. The extension sensor board has 3 axes of accelerometers in three directions and a temperature sensor. Software components have been implemented within the TinyOS operating system to provide a flexible software platform and scalable performance for structural health monitoring applications. The prototype WSN is deployed on a reduced-scale bridge model with 3 nodes in a single-hop network for performing dynamic monitoring civil engineering structures. Two output-only time-domain system identification methods are employed namely, the Frequency Domain Decomposition (FDD) method and the Natural Excitation Technique (NExT) combined with the Eigensystem Realization Algorithm (ERA). Testing results show that the WSN provides accurate vibration data for identifying vibration modes of a bridge

Identification of novel biomarker candidates for immunohistochemical diagnosis to distinguish low-grade chondrosarcoma from enchondroma

Chondrosarcoma is the third most common primary bone cancer, requiring surgical resection. However, differentiation of low-grade chondrosarcoma (grade 1) from enchondroma that is benign and only requires regular follow-up is one of the most frequent diagnostic dilemmas facing orthopedic oncologists in clinical management. Although multiple techniques are applied to make the distinction, immunohistochemistry is an important ancillary technique, especially when a histopathological stain of specimen must be obtained in order to guarantee an accurate confirmation. Currently, no adequate immunohistochemical diagnostic protein biomarkers are available to distinguish low-grade chondrosarcoma from enchondroma. To discover novel protein biomarker candidates, an LC-MS/MS approach was applied to directly compare formalin-fixed, paraffin-embedded low-grade chondrosarcoma with enchondroma tissue samples. The proteomics analysis revealed 17 protein biomarker candidates. A principle was developed to prioritize the candidates using category and ranking. An algorithm, prioritization index of biomarker candidates for immunohistochemistry on tissue specimens, was developed to rank the candidates inside each category. Using the proteomics data and bioinformatics results, the prioritization index of biomarker candidates for immunohistochemistry on tissue revealed periostin as a top candidate. Immunohistochemical staining of periostin in 23 low-grade chondrosarcoma and 31 enchondroma tissue specimens disclosed 87% specificity and 70% sensitivity

Proteomic Techniques in the Physiological Proteomics Core Facility

poster abstractA new software package, IdentiQuantXL, has been developed in the Physiological Proteomics Core Facility to provide large-scale protein identification and label-free quantification using either low or high resolution LC-MS/MS data. Though many software packages have been developed to perform label-free quantification of proteins in complex biological samples using peptide intensities generated by liquid chromatography - tandem mass spectrometry (LC-MS/MS), two important issues hinder the use of peptide intensity measurements: (i) It is difficult to accurately determine the retention time of each peptide peak, especially for low resolution data, and (ii) many peptides cannot be used for protein quantification. To address these two key issues, we have developed a new method to enable accurate peptide peak retention time determination and multiple filters to eliminate unqualified peptides for protein quantification. Repeatability and linearity have been tested using ion trap-derived low resolution data from six very different samples, i.e., standard peptides, kidney tissue lysates, HT29-MTX cell lysates, depleted human serum, human serum albumin-bound proteins, and standard proteins spiked in kidney tissue lysates. In all these unique experiments, at least 90.8% of proteins (up to 1,390) had CVs ≤ 3 0% across 10 technique replicates, and at least 92.1% of proteins (up to 2,013) had R2 ≥ 0.9500 across 7 concentrations. The performance of our strategy was verified using identical amounts of standard protein (lysozyme) spiked in complex biological samples (cell culture media containing secreted proteins) with a CV of 8.6% across eight injections. The excellent performance was further confirmed by comparing label-free mass spectrometry to Western blot detection of prolactin, which was decreased 17.1fold in dwarfed mice compared to wild-type using the label-free quantification strategy and very low or undetectable using Western blot. The results indicate that our new platform, named IdentiQuantXL, accurately quantifies thousands of peptides and proteins in complex samples. It has been applied in the aqueous humor proteome in patients with Fuchs endothelial corneal dystrophy. While many software packages focus only on high resolution data, our strategy is designed for both high and low resolution data. Consequently, it is very useful for data generated by low resolution mass spectrometers such as the LTQ, especially when the dynamic exclusion of ions in data acquisition is enabled to obtain more MS/MS fragments of low-abundance peptides to maximally identify proteins in a complex biological sample. Supported by NIEHS RC2ES018810 and NIGMS R01GM08521

Los factores que influyen en la satisfacción en el trabajo y su relación con la intención de baja laboral: tomando a los empleados que inician su carrera como ejemplo.

Job satisfaction of early-career employees has a significant effect on employee work behavior and work attitude. This study investigated the influence factors of job satisfaction and its relationship with turnover intention. Early-career employees were selected as the research subject. Questionnaires were distributed and the stratified random sampling method was used to analyze the 418 samples collected. The dimensions of job satisfaction were analyzed by exploratory factor analysis and confirmatory factor analysis, and a structural equation model was constructed to discuss the relationship between job satisfaction and turnover intention. First, for early-career employees, job satisfaction as a higher order factor consists of 6 first-order factors; personal growth’s influence on job satisfaction is the greatest, with a path coefficient reaching .918. Next come salary and welfare, work itself, interpersonal relationships and leader behavior. Additionally, job competency has a minimal effect on job satisfaction; its path coefficient is only .214. Second, job satisfaction has a significantly negative effect on turnover intention, indicating that the job satisfaction structure of early-career employees has its unique characteristics because they focus more on personal growth and development space. Through improving job satisfaction, turnover behavior of early-career employees can be effectively controlled.La satisfacción en el trabajo en empleados que inician su carrera tiene un efecto significativo en la conducta laboral y en la actitud ante el trabajo en los empleados. Este estudio investiga los factores que influyen en la satisfacción en el trabajo y su relación con el bajas laborales. Se seleccionaron empleados en su fase inicial como sujetos de investigación. Se distribuyeron cuestionarios según un método de muestreo aleatorizado estratificado para analizar 418 muestras recogidas. Las dimensiones de la satisfacción en el trabajo fueron analizadas mediantes análisis factorial exploratorio y análisis factorial confirmatorio, y con ello se construyó un modelo de ecuaciones estructurales para discutir la relación entre satisfacción en el trabajo y los motivos de bajas laborales. Primero, para empleados iniciales, la satisfacción en el trabajo como factor de orden superior consistía en seis factores de primer orden; la influencia del desarrollo personal en la satisfacción en el trabajo fue la mayor, con un coeficiente path de .918. El siguiente fue el salario y el bienestar, el trabajo por sí mismo, las relaciones interpersonales y la conducta de liderazgo. Adicionalmente, la competencia laboral tuvo un efecto mínimo sobre la satisfacción en el trabajo; su coeficiente path es sólo .214. En segundo lugar, la satisfacción en el trabajo tuvo un efecto significativamente negativo sobre los motivos de baja laboral, indicando que la estructura de satisfacción en el trabajo de los empleados iniciales tiene características propias, puesto que ellos se centran más en el desarrollo personales y el espacio de desarrollo. Mejorando la satisfacción laboral, la conducta de baja laboral de los empleados iniciales puede ser controlada efectivamente

Development, validation, and comparison of four methods to simultaneously quantify L-arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometry

To understand the role of l-arginine depletion in impaired nitric oxide synthesis in disease, it is important to simultaneously quantify arginine, citrulline, and ornithine in the plasma. Because the three amino acids are endogenous analytes, true blank matrix for them is not available. It is necessary and valuable to compare the performance of different approaches due to lack of regulatory clarity for validation. A two-step sample preparation method using methanol as protein precipitation reagent was developed in this study is used for sample preparation. Because true blank matrix for endogenous analytes is not available, water as blank matrix, 1% BSA in PBS as blank matrix, surrogate analyte, and background subtraction were designed to establish successful quantification methods. Four methods to simultaneously quantify arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometry were developed, validated, and compared. The developed two-step sample preparation method using methanol as protein precipitation reagent in this study needs less time and provides higher recovery comparing with other approaches. Three of the four methods, water as blank matrix, 1% BSA in PBS as blank matrix, and surrogate analyte, have been successful in fulfilling all the criteria, while background subtraction has failed. Results of the measured concentrations in 97 human plasma samples using the three methods show that the difference between any two methods or among the three methods presents 100% of samples with less than 20% for all the three amino acids and majority of them are under 10%. The developed two-step sample preparation method using methanol as protein precipitation reagent is simple and convenient. Three of the four methods are fully validated and the validation is successful. The BSA functioned effectively as a blank matrix for these three amino acids, considering cost, data quality, matrix similarity, and practicality

Teacher burnout and turnover intention in higher education: The mediating role of job satisfaction and the moderating role of proactive personality

IntroductionTeacher burnout and frequent turnover negatively affect stability and productivity in the context of higher education. Despite the fact that the relationship between burnout and turnover intention has been thoroughly studied, the role of other factors in this relationship should be evaluated in order to better clarify underlying mechanisms, particularly in the context of higher education.MethodsIn this study, we first aim to bridge a research gap by utilizing job satisfaction as a mediating variable for the relationship between burnout and turnover intention. Moreover, we uniquely evaluate the role of proactive personality as a moderating variable, first in terms of the relationship between burnout and job satisfaction, and then for the relationship between job satisfaction and turnover intention. Based on 296 valid questionnaires collected from university faculty members in China, proposed hypotheses were evaluated empirically.ResultsThe results demonstrate that, as expected, burnout has a significant and positive impact on turnover intention, and job satisfaction has significantly negative impact on turnover intention, with job satisfaction partially mediating the relationship between burnout and turnover intention. Moreover, proactive personality moderated the relationship between job satisfaction and turnover intention, with this relationship being stronger for individuals with high proactive personality as compared to low proactive personality.DiscussionThese findings provide a better understanding of the relationship between burnout and turnover intention of university instructors. Theoretical and practical implications, limitations, and recommendations for further research are provided

Metabolic and Molecular Regulation of Dietary Polyunsaturated Fatty Acids on Prostate Cancer

Purpose The aim of this study is to investigate the role of n-3 and n-9 fatty acids in crucial processes involved in prostate cancer cell growth through a large-scale proteomic analysis. Experimental design We used a label-free protein quantification method to profile global protein expression of fish oil and oleic acid treated PCa cells and validated a panel of differentially expressed proteins by either Western blot or multiple reaction monitoring. Bioinformatic analysis was also performed to uncover the pathways involved in fatty acid metabolism. Results Fish oil, not oleic acid, suppresses prostate cancer cell viability. Assessment of fatty acid synthesis pathway activity also shows that oleic acid is a more potent inhibitor than fish oil on de novo fatty acid synthesis. Although fatty acid synthase activity decreases with fish oil treatment, the inhibition of its activity occurs over time while reduction in viability occurs within 24 h. Bioinformatic analysis revealed the pathways altered by these fatty acid treatments. Conclusions and clinical relevance This study suggests that suppression of cell viability by fish oil is independent of fatty acid synthase and fish oil regulates prostate cancer cells through activation of other pathways depending upon length of exposure to fish oil