Identification of Candidate Serum Proteins for Classifying Well-Differentiated Small Intestinal Neuroendocrine Tumors

Background

Patients with well-differentiated small intestine neuroendocrine tumors (WD-SI-NETs) are most often diagnosed at a metastatic stage of disease, which reduces possibilities for a curative treatment. Thus new approaches for earlier detection and improved monitoring of the disease are required.

Materials and Methods

Suspension bead arrays targeting 124 unique proteins with antibodies from the Human Protein Atlas were used to profile biotinylated serum samples. Discoveries from a cohort of 77 individuals were followed up in a cohort of 132 individuals both including healthy controls as well as patients with untreated primary WD-SI-NETs, lymph node metastases and liver metastases.

Results

A set of 20 antibodies suggested promising proteins for further verification based on technically verified statistical significance. Proceeding, we assessed the classification performance in an independent cohort of patient serum, achieving, classification accuracy of up to 85% with different subsets of antibodies in respective pairwise group comparisons. The protein profiles of nine targets, namely IGFBP2, IGF1, SHKBP1, ETS1, IL1α, STX2, MAML3, EGR3 and XIAP were verified as significant contributors to tumor classification.

Conclusions

We propose new potential protein biomarker candidates for classifying WD-SI-NETs at different stage of disease. Further evaluation of these proteins in larger sample sets and with alternative approaches is needed in order to further improve our understanding of their functional relation to WD-SI-NETs and their eventual use in diagnostics.     

Genomic Analysis of Sleeping Beauty Transposon Integration in Human Somatic Cells

The Sleeping Beauty (SB) transposon is a non-viral integrating vector system with proven efficacy for gene transfer and functional genomics. However, integration efficiency is negatively affected by the length of the transposon. To optimize the SB transposon machinery, the inverted repeats and the transposase gene underwent several modifications, resulting in the generation of the hyperactive SB100X transposase and of the high-capacity “sandwich” (SA) transposon. In this study, we report a side-by-side comparison of the SA and the widely used T2 arrangement of transposon vectors carrying increasing DNA cargoes, up to 18 kb. Clonal analysis of SA integrants in human epithelial cells and in immortalized keratinocytes demonstrates stability and integrity of the transposon independently from the cargo size and copy number-dependent expression of the cargo cassette. A genome-wide analysis of unambiguously mapped SA integrations in keratinocytes showed an almost random distribution, with an overrepresentation in repetitive elements (satellite, LINE and small RNAs) compared to a library representing insertions of the first-generation transposon vector and to gammaretroviral and lentiviral libraries. The SA transposon/SB100X integrating system therefore shows important features as a system for delivering large gene constructs for gene therapy applications.     

Molecular studies in three patients with isodicentric Y chromosome

Three patients carrying an isodicentric (idic) Y chromosome associated with a mosaic 45,X cell line were studied using molecular techniques. Genotype-phenotype correlations suggested an effect of the 45,X cell line on sexual differentiation. A relationship was established between instability of the idic(Y) chromosome and localization of the breakpoint on Yq, and between azoospermia and deletion of interval 6 on Yq. Received: 31 May 1996 / Revised: 12 July 1996     

Quantitative evaluation of chromosomal rearrangements in gene-edited human stem cells by CAST-Seq

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Résultats hémodynamiques et cliniques de la dissection, assistée par lunettes chirurgicales, des veines spermatiques au niveau de l’orifice inguinal interne

Inversion of venous flow is well visualized by colour duplex ultrasound and is the basic haemodynamic feature of varicocele. We tried to assess the value of dissection of branches of the spermatic vein in the internal inguinal ring using an operating microscope with a magnification of 4.5x. 16 patients were operated according to the ultrasound classification proposed by Hirsh et al. [8]. Varicocele was grade III in 2 cases, grade IV in 11 cases and grade V in 3 cases and was situated on the left in 13 patients and on the right in 3 patients. The diagnosis was based on clinical examination and colour duplex ultrasound. The operating time was 25–35 min. The incision was 1.5 to 2 cm long. No short-term complications or long-term hydrocele were observed. No reno-spermatic flow was observed on colour duplex ultrasound after six months. The mean testicular volume was 22 ml before surgery and did not change after the operation. This surgical technique is less invasive than the conventional retroperitoneal or laparoscopic procedures, and the incision is smaller than with the percutaneous technique. It is less radical than conventional microsurgery, but the operating time is shorter, and the surgical wound is smaller, sparing the artery and lymphatic vessels. In conclusion, this operation can be considered to be a good option in the surgical treatment of varicocele in view of its low complication rate, the absence of hydrocele, a minimally invasive procedure and the good haemodynamic results.     

Alien flora in Calabria (Southern Italy): an updated checklist

Biological Invasions - An updated checklist of the Calabrian alien vascular flora is presented. By way of field, bibliographic, and herbarium research, we recorded 382 alien taxa (representing...     

Phylogeography of the common pandora Pagellus erythrinus in the central Mediterranean Sea: sympatric mitochondrial lineages and genetic homogeneity

The distribution of the genetic diversity and the population structure of Pagellus erythrinus were analysed using mitochondrial control region sequences and cytochrome b restriction profiles in a total of 128 and 508 individuals, respectively, that were collected from 15 sampling sites in the central Mediterranean Sea and from one site in the Atlantic Ocean. No population genetic structure was detected within the central Mediterranean and thus, the commonly recognized transition zones in the area do not seem to affect population connectivity. The comparison between the Mediterranean samples and the single Atlantic sample suggests weak differentiation between the two basins. Three mitochondrial lineages were identified, each including individuals from almost every sampling site. The haplotype and nucleotide diversity values, mismatch distribution and demographic parameters indicate that the sympatry of these lineages can be ascribed to a period of isolation followed by genetic divergence, population expansion and secondary contact, all of which are likely to be associated with climatic oscillations that occurred during the middle and late Pleistocene.     

Effect of oxygen on radiation-induced DNA damage in isolated nuclei.

In intact mammalian cells, ionizing radiation causes substantially less damage to DNA in the absence of oxygen than in the presence of oxygen. In contrast, when DNA is isolated (usually from viruses) and irradiated in solution, the absence of oxygen does not lead to a decrease in damage unless low-molecular-weight thiols are also present. We investigated an intermediate condition: that of DNA irradiated in isolated nuclei. Using an HPLC-based assay of thiols with electrochemical detection, we have determined that the nuclear isolation procedure leads to the elimination of virtually all low-molecular-weight thiols (predominantly glutathione and cysteine). Thus it was our expectation that the thiol-depleted state would concurrently eliminate the OER, and thereby mimic the isolated DNA system, while retaining structural characteristics of chromosomal DNA. We evaluated radiation-induced DNA damage in isolated nuclei by measuring single-strand breaks using alkaline elution and by measuring double-strand breaks using neutral elution and pulsed-field gel electrophoresis. Despite the removal of low-molecular-weight thiol compounds, the oxygen dependence of radiation-induced damage more closely paralleled that of whole cells than that of DNA in solution. Thus damage of DNA irradiated in isolated nuclei is dependent on oxygen.