Identification of prognostic immune-related signature predicting the overall survival for colorectal cancer.

OBJECTIVE: The morbidity and mortality of patients with colorectal cancer, one of the most common malignant tumors worldwide, is steadily increasing. The aim of this study was to investigate the association between prognostic immune-related gene profile and the outcome of colorectal cancer in patients by analyzing datasets from The Cancer Genome Atlas (TCGA). MATERIALS AND METHODS: Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) further demonstrated that these genes were enriched in many immune-related biological processes. Univariate Cox regression analysis was applied to examine the association of immune-related genes with the prognosis in patients with colorectal cancer. The least absolute shrinkage and selection operation (LASSO) Cox regression model was then used to establish the immune-related signature for the prognostic evaluation of colorectal cancer in patients. Survival differences were assessed by the Kaplan-Meier method along with the log-rank test. RESULTS: A total of 133 prognostic immune-related signatures were identified by using the univariate Cox proportional hazards regression analysis. A 14-gene signature-based risk score was constructed using the LASSO Cox regression. According to the cut-off of the risk-score, patients were assigned to the low-risk and high-risk groups. The log-rank test suggested that the survival time of the low-risk group was significantly higher than that of the high-risk group. In the time-dependent ROC curve analysis, the AUC for 1-year, 3-year, and 5-year overall survival (OS) were 0.781, 0.742, and 0.791, respectively. GO and KEGG analysis further revealed that the gene sets were actively involved in immune and inflammatory response, as well as the cytokine-cytokine receptor interaction pathway. CONCLUSIONS: To summarize, we identified a novel 14-gene immune-related signature that may potentially serve as a prognostic predictor for colorectal cancer, thereby contributing to patient personalized treatment decisions. Further research needs to be conducted to validate the prognostic value of the selected genes.

Para-phenylenediamine induced DNA damage and apoptosis through oxidative stress and enhanced caspase-8 and -9 activities in Mardin-Darby canine kidney cells.

Para-phenylenediamine (p-PD), a suspected carcinogen, is a component of permanent hair dyes. In this study we examined the mechanism of cytotoxicity and genotoxicity in Mardin-Darby canine kidney cells (MDCK)-treated with p-PD. Our results showed that p-PD decreased cell viability in a dose- and time-dependent manner. In addition, p-PD induced DNA damage was confirmed by the comet and TUNEL assays. Pre-treatment of MDCK cells with antioxidants vitamin C or E significantly inhibited p-PD induced cytotoxicity and reactive oxygen species (ROS) generation. Furthermore, p-PD induced apoptosis through activated initiator caspase-8 and -9, and effector caspase-3/7. Based on these results, we suggested that p-PD induce apoptosis which was mediated with caspase-8, caspase-9 and caspase-3/7 activation via the involvement of ROS.

A 28-kilodalton pod storage protein of French bean plants. Purification, characterization, and primary structure.

When French bean (Phaseolus vulgaris) plants were depodded in the early stages of fruit development, relative levels of a specific protein with a relative molecular weight of 28,000 were enhanced in the young pods that formed later. The protein, designated pod storage protein (PSP), was purified from extracts of newly formed pods from plants that had been previously depodded four times at intervals of 2 weeks. Two-dimensional polyacrylamide gel electrophoresis showed the presence of three forms (designated A, B, and C) of PSP with identical electrophoretic mobilities but different charges. The molecular mass of native PSP was estimated by gel filtration to be 67 kD; therefore, the protein was most likely present as a dimer. The antisera raised against forms A and C were crossreactive with each other. Form B lacked the N-terminal alanine of forms A and C. An expression library from French bean pods was screened using the antiserum against form A, and a full-length cDNA clone was isolated. The cDNA insert included 765 bp potentially encoding a polypeptide with 255 amino acid residues (and a calculated molecular mass of 28,854 D). The amino acid sequence deduced from the PSP cDNA had 65 to 71% identity with soybean (Glycine max) vegetative storage protein sequences (P.E. Staswick [1988] Plant Physiol 87: 250-254; and Correction [1989] Plant Physiol 89: 717). Genomic Southern blot analysis suggested that PSP is derived from a single-copy gene.

Cofractionation of the 17-kD PK 14105 binding site protein with solubilized peripheral-type benzodiazepine binding sites.

To examine the relationship between PKBS, a 17-kD protein covalently photolabeled by [3H]PK 14105, and its association with peripheral-type benzodiazepine binding sites, rat adrenal mitochondrial fractions were photolabeled with [3H]PK 14105, solubilized in digitonin, and subjected to anion-exchange chromatography over Q-Sepharose. The chromatographic behavior of PKBS was evident as principally two major fractions, signified as Q-I and Q-II. Specific binding sites for [3H]Ro5-4864 and [3H]PK 11195 were also assayed and found to cofractionate with each other and in a manner which coincided with the photolabeled PKBS profile. The Q-I and Q-II fractions were further distinguished based on their different molecular sizes observed by gel filtration, yet both fractions were characterized as containing peripheral-type benzodiazepine recognition sites according to the following criteria. Scatchard analysis of both subpopulations revealed a single class of binding sites for [3H]Ro5-4864 with an apparent KD of 14 nM for Q-I and 22 nM for Q-II; these affinities were slightly lower than those found in mitochondrial membrane preparations used as the starting material for solubilization. The rank order of potency to inhibit [3H]Ro5-4864 binding in both subpopulations was PK 11195 greater than Ro5-4864 greater than diazepam greater than clonazepam, in connection with the pharmacological specificity of membrane-associated peripheral-type benzodiazepine binding sites. These studies provide direct biochemical evidence that the recognition sites for benzodiazepines and isoquinoline carboxamides cofractionate in unison with the 17-kD PKBS protein, demonstrating an intimate relationship between this protein and the binding domains for peripheral-type benzodiazepine ligands.

Ventrolateral medulla: an important site of action for the hypotensive effect of drugs that activate serotonin-1A receptors.

The purpose of our study was to test the hypothesis that drugs that activate the 5-HT1A receptor lower arterial pressure by interacting with 5-HT1A receptors located in the ventrolateral medulla. For this purpose, cats were anesthetized with alpha-chloralose while arterial pressure, heart rate, and several indices of respiratory activity were being monitored during either topical application or microinjection of 5-HT1A receptor agonist and antagonist drugs in the region of the intermediate area of the ventrolateral medulla. Topical application of drugs that activate 5-HT1A receptors (e.g., 8-OH-DPAT and urapidil) produced decreases in arterial pressure, heart rate, and tidal volume but an increase in respiratory rate. These effects were prevented by prior topical application of the 5-HT1A antagonist drug, spiperone. Intravenous administration of 5-HT1A receptor agonist drugs produced similar cardiorespiratory effects, and these effects were counteracted by topical application of spiperone to the intermediate area of the ventrolateral medulla. Microinjection of 8-OH-DPAT into two sites associated with the intermediate area, namely the subretrofacial nucleus and the bicuculline-sensitive hypotensive site, also resulted in decreases in arterial pressure (-40 +/- 10 mm Hg, n = 5; and -21 +/- 4 mm Hg, n = 3, respectively). These data indicate that hypotension produced by drugs that activate 5-HT1A receptors occurs, at least in part, from an action in the ventrolateral medulla, presumably in the subretrofacial nucleus and/or the bicuculline-sensitive hypotensive site.