Analysis of IGF and IGFBP as prognostic serum biomarkers for adrenocortical carcinoma.

BACKGROUND: Adrenocortical carcinoma (ACC) lacks diagnostic and prognostic biomarkers to guide treatment. A consistently dysregulated pathway in ACC is the IGF signaling pathway, specifically overexpression of IGF2, IGF-I-receptor, and IGFBP2. The objective of this study was to perform a comprehensive analysis of serum IGF and IGFBP levels and to determine their utility as diagnostic and prognostic biomarkers in ACC. METHODS: Preoperative serum samples from 53 patients who underwent surgery for adrenocortical adenomas, 3 patients who underwent initial surgery for ACC, 16 patients who underwent reoperative surgery for ACC, and 5 healthy volunteer controls were analyzed. The serum concentration of IGF1, IGF2, IGFBP1, IGFBP2, and IGFBP3 was determined by enzyme-linked immunosorbent assay. RESULTS: No difference in the levels of IGF2 (p = .231) and IGFBP2 (p = .511) was observed between patients with ACC, benign adrenocortical tumors, and healthy volunteers. IGF1, IGFBP1, and IGFBP3 levels were not detected. High IGFBP2 levels were associated with better overall survival (OS) (p = .001) and showed a trend toward better abdominal progression-free (APFS) survival (p = .070) in patients with ACC. A subanalysis of patients undergoing reoperation for recurrent ACC showed better OS with high levels of IGFBP2 (p = .003) and a trend toward better APFS (p = .107). There was no significant difference in IGF2 and IGFBP2 levels by extent of disease. CONCLUSIONS: IGF2 and IGFBP2 are not elevated in the serum of patients with ACC compared with patients with benign neoplasms and healthy volunteers. Elevated serum IGFBP2 is associated with better survival in patients with ACC and those undergoing reoperative surgery for recurrent ACC.

A method for determining the age of a bloodstain.

DNA allows for the unambiguous identification of the person from whom a biological sample was derived but provides little information about when the sample was deposited. This information only indicated that the biological material was deposited at the crime scene prior to the collection of evidence. The ability to determine the age of a biological sample would greatly benefit the forensic science community. If there were independent evidence that the biological sample was deposited at the time of the crime, then its age would reveal when the crime occurred. If the time of the crime were known through another means, then the age of the biological sample could potentially exclude the human source as a suspect. We have used real-time reverse transcriptase PCR to show that the ratio between different types of RNA (mRNA versus rRNA) changes over time in a linear fashion when dried human blood from eight individuals was examined over the course of 150 days. Although other approaches have been used in the past to estimate the age of a biological sample, our approach offers the following advantages: enhanced detectability of small samples, simultaneous isolation of DNA and RNA from the same sample, species-specific probes, and an increased window of usefulness.

Integrated analysis of genome-wide methylation and gene expression shows epigenetic regulation of CYP11B2 in aldosteronomas.

CONTEXT: Differential methylation of CpG regions is the best-defined mechanism of epigenetic regulation of gene expression. OBJECTIVE: Our objective was to determine whether any changes in methylation are associated with aldosteronomas. METHODS: We performed integrated genome-wide methylation and gene expression profiling in aldosteronomas (n = 25) as compared with normal adrenal cortical tissue (n = 10) and nonfunctioning adrenocortical tumors (n = 13). To determine the effect of demethylation on gene expression of CYP11B2, the H295R cell line was used. RESULTS: The methylome of aldosteronomas, normal adrenal cortex, and nonfunctioning adrenocortical tumors was distinct, with hypomethylation of aldosteronomas. Integrated analysis of gene expression and methylation status showed that 53 of 60 genes were hypermethylated and downregulated, or hypomethylated and upregulated, in aldosteronomas. Of these, 3 genes that regulate steroidogenic signals and synthesis in adrenocortical cells were differentially methylated: AVPR1α and PRKCA were downregulated and hypermethylated, and CYP11B2 was upregulated and hypomethylated. Demethylation treatment resulted in upregulation of these genes, with direct hypomethylation of CpG sites associated with the genes. The CpG island in the promoter region of CYP11B2 was hypomethylated in aldosteronomas but not in blood DNA from the same patients (P = .0004). CONCLUSIONS: Altered methylation in aldosteronomas is associated with dysregulated expression of genes involved in steroid biosynthesis. Aldosteronomas are hypomethylated, and CYP11B2 is overexpressed and hypomethylated in these tumors.

Local anesthetic syringe ergonomics and student preferences.

With a diverse population of dentists, dental hygienists, and dental assistants, there is a wide range of sizes and shapes of individuals. Ergonomic considerations have resulted in the design of many adjustable delivery systems, chairs, and pieces of equipment. Companies have marketed instrumentation specifically for people with small hands, yet little research has been done regarding the advertising claims. The objective of this study was to compare the perceptions of dental and dental hygiene students after they used a conventional syringe and a petite syringe that has been marketed as advantageous for individuals with smaller hands. The two syringes were compared for aspiration, injection, and sense of control. A total of 181 students were invited to serve as subjects in the study, and ninety participated. The study involved two phases. During phase I, students used conventional and petite syringes to give a simulated inferior alveolar nerve block injection. In phase II, students gave a simulated palatal injection using both sizes of syringes. After performing the simulations, the students responded to a questionnaire. The students reported that the petite syringe gave them control, and 62.2 percent of them preferred it to the conventional syringe. There were differences in preferences between female and male students and also differences in preferences based on students' glove sizes. The female students preferred the petite syringe when aspiration was required. Overall, the students who wore small and extra small gloves had a preference for the petite syringe and felt it provided a sense of control, a key component when learning to provide injections.