Nt. The lipid contents were analyzed using the Clarus 500 gas chromatograph with a FTY720 built-in Autosampler (PerkinElmer), and the total runtime for each sample was 34 min. 1uL FA methyl esters (FAME) was injected in split injection mode with a 100:1 split ratio, and the resultant peaks were identified and quantified by comparison with the standard Supelco 37-component FAME Mix (Sigma-Aldrich, USA).Fluid Percussion Injury (FPI)FPI was performed as previously described [10]. Under deep anesthesia, A 3.0 mm diameter craniotomy was made 3.0 mm posterior to the bregma and 6.0 mm lateral (left) to the midline, and a cap was glued over the craniotomy. The cap was filled with 0.9 saline solution and a mild fluid percussion pulse (1.5 atm) was administered. The severity of injury was confirmed based on the unconscious time (less than 120 seconds) of the animal following injury before the first response to a paw inch. Sham animals underwent an identical preparation with the exception of the FPI.Statistical AnalysisProtein data were expressed as a mean percentage of the control (sham/n-3 adq) group DHA and AA levels were expressed as a ratio of fatty acid (mg)/tissue (g). Significance among groups was determined using SPSS software. One-way analysis of variance (ANOVA) was performed followed by Fisher’s Least Significance Difference test. Results were expressed as mean 6 standard error of the mean (SEM), n = 5?/group. Significant difference was considered at P,0.05.Tissue CollectionOne week following FPI, rats were sacrificed by decapitation, and the cervical enlargement (C3 6) and lumbar enlargement (L2 6) regions were collected and stored in 270uC for lipids and protein measurements.Protein AnalysisBDNF, phosphorylated tyrosine kinase (pTrkB), phosphorylated cAMP response element-binding (pCREB), syntaxin-3, calciumindependent phospholipase A2 (iPLA-2), and 4-Hydroxynonenal (4-HNE) proteins of the right lumbar SC were analyzed usingAuthor ContributionsConceived and designed the experiments: ZY RA FGP. Performed the experiments: CF RA YZ. Analyzed the data: CF YZ. Wrote the paper: ZY CF FGP.
Colon cancer ranks among the most common cancers in terms of both cancer incidence and cancer-related deaths in Western countries [1]. Early-stage colon cancer can be managed successfully by surgical resection; however, metastatic disease is often refractory to treatment and responsible for the majority of morbidity and mortality. Clinical decision-making is guided by the American Joint Committee on Cancer TNM (tumor-nodemetastasis) staging that is imperfect for Fexaramine manufacturer prognosis and does not predict response to therapy. A critical need exists to identify objective markers of malignancy that could be used for early detection, prognostication, intervention, and/or targeting of cancerous cells. As an example, tumor-associated antigens (TAAs) have been utilized for the detection of circulating tumor cells (CTCs) in the bloodstream as well as disseminated tumor cells (DTCs) in the bone marrow with the ability to monitor tumor burden, predict risk of progression, and measure chemotherapy response. These technologies include clinically-approved products such as CellSearchTM (Veridex) that utilize immunodetection of CTCs on the basis of Epithelial Cell Adhesion Molecule (EpCAM)membranous expression [2]. Cell-surface TAAs in particular are also valuable because they are accessible to systemically-delivered targeting molecules (e.g. antibodies, aptamers, etc.) that could be used.Nt. The lipid contents were analyzed using the Clarus 500 gas chromatograph with a built-in Autosampler (PerkinElmer), and the total runtime for each sample was 34 min. 1uL FA methyl esters (FAME) was injected in split injection mode with a 100:1 split ratio, and the resultant peaks were identified and quantified by comparison with the standard Supelco 37-component FAME Mix (Sigma-Aldrich, USA).Fluid Percussion Injury (FPI)FPI was performed as previously described [10]. Under deep anesthesia, A 3.0 mm diameter craniotomy was made 3.0 mm posterior to the bregma and 6.0 mm lateral (left) to the midline, and a cap was glued over the craniotomy. The cap was filled with 0.9 saline solution and a mild fluid percussion pulse (1.5 atm) was administered. The severity of injury was confirmed based on the unconscious time (less than 120 seconds) of the animal following injury before the first response to a paw inch. Sham animals underwent an identical preparation with the exception of the FPI.Statistical AnalysisProtein data were expressed as a mean percentage of the control (sham/n-3 adq) group DHA and AA levels were expressed as a ratio of fatty acid (mg)/tissue (g). Significance among groups was determined using SPSS software. One-way analysis of variance (ANOVA) was performed followed by Fisher’s Least Significance Difference test. Results were expressed as mean 6 standard error of the mean (SEM), n = 5?/group. Significant difference was considered at P,0.05.Tissue CollectionOne week following FPI, rats were sacrificed by decapitation, and the cervical enlargement (C3 6) and lumbar enlargement (L2 6) regions were collected and stored in 270uC for lipids and protein measurements.Protein AnalysisBDNF, phosphorylated tyrosine kinase (pTrkB), phosphorylated cAMP response element-binding (pCREB), syntaxin-3, calciumindependent phospholipase A2 (iPLA-2), and 4-Hydroxynonenal (4-HNE) proteins of the right lumbar SC were analyzed usingAuthor ContributionsConceived and designed the experiments: ZY RA FGP. Performed the experiments: CF RA YZ. Analyzed the data: CF YZ. Wrote the paper: ZY CF FGP.
Colon cancer ranks among the most common cancers in terms of both cancer incidence and cancer-related deaths in Western countries [1]. Early-stage colon cancer can be managed successfully by surgical resection; however, metastatic disease is often refractory to treatment and responsible for the majority of morbidity and mortality. Clinical decision-making is guided by the American Joint Committee on Cancer TNM (tumor-nodemetastasis) staging that is imperfect for prognosis and does not predict response to therapy. A critical need exists to identify objective markers of malignancy that could be used for early detection, prognostication, intervention, and/or targeting of cancerous cells. As an example, tumor-associated antigens (TAAs) have been utilized for the detection of circulating tumor cells (CTCs) in the bloodstream as well as disseminated tumor cells (DTCs) in the bone marrow with the ability to monitor tumor burden, predict risk of progression, and measure chemotherapy response. These technologies include clinically-approved products such as CellSearchTM (Veridex) that utilize immunodetection of CTCs on the basis of Epithelial Cell Adhesion Molecule (EpCAM)membranous expression [2]. Cell-surface TAAs in particular are also valuable because they are accessible to systemically-delivered targeting molecules (e.g. antibodies, aptamers, etc.) that could be used.