No abstract

Species identification of cell lines and detection of cross-contamination are crucial for scientific research accuracy and reproducibility. Whereas short tandem repeat profiling offers a solution for a limited number of species, primarily human and mouse, the standard method for species identification of cell lines is enzyme polymorphism. Isoezymology, however, has its own drawbacks; it is cumbersome and the data interpretation is often difficult. Furthermore, the detection sensitivity for cross-contamination is low; it requires large amounts of the contaminant present and cross-contamination within closely related species may go undetected. In this paper, we describe a two-pronged molecular approach that addresses these issues by targeting the mitochondrial genome. First, we developed a multiplex PCR-based assay to rapidly identify the most common cell culture species and quickly detect cross-contaminations among these species. Second, for speciation and identification of a wider variety of cell lines, we amplified and sequenced a 648-bp region, often described as the "barcode region" by using a universal primer mix targeted at conserved sequences of the cytochrome C oxidase I gene (COI). This method was challenged with a panel of 67 cell lines from 45 diverse species. Implementation of these assays will accurately determine the species of cell lines and will reduce the problems of misidentification and cross-contamination that plague research efforts.

Differential gene expression in the adipose tissue of morbidly obese patients includes genes related to lipid and glucose metabolism, membrane transport, and genes promoting the cell cycle. These findings are a first step toward clarifying the molecular pathogenesis of obesity and identifying potential targets for therapeutic intervention.

To eliminate the human anti-mouse antibody (HAMA) response seen in patients treated with murine and chimeric antibodies, fully human monoclonal antibodies (MAbs) are now being developed. Tumor Necrosis Therapy (TNT) is an approach to tumor targeting that utilizes MAbs directed against common intracellular antigens such as nucleic acids, accessible only in necrotic areas of solid tumors. By binding to the necrotic core of tumors, these TNT MAbs can circumvent many of the limitations of MAbs directed against tumor cell surface antigens. Chimeric TNT-1 (chTNT-1) was first developed from the parent murine antibody by genetically engineering the murine variable regions to the human IgG(1) and kappa constant regions. Although the chimeric antibody's behavior was similar to that of the murine version, the 35% murine homology it shares allows for the potential of a HAMA response. A human antibody derived from a phage display library, designated NHS76, has been developed with similar binding characteristics to the TNT-1. To demonstrate that this genetically engineered human counterpart to chTNT-1 has similar pharmacokinetic characteristics, in vivo behavior, and targeting abilities, both antibodies were rigorously tested in parallel. For these studies, biodistribution analysis in LS174T human colon tumor-bearing nude mice was performed to compare the uptake levels in tumor and normal organs. In addition, mouse imaging and autoradiographic studies were conducted to demonstrate positive uptake in necrotic regions of tumor and negative uptake in viable tissues and organs. The results of these studies confirm the comparable nature of both antibodies and provide the necessary preclinical data to show the suitability of NHS76 as an improved product for the therapy of solid tumors in man.

Human services professionals will undoubtedly work with the dying and bereaved populations at one time or other. Yet, they are poorly prepared to do so since death education, that is, lessons about the human and emotional aspects of death, its implications, and subsequent bereavement issues, is often not part of their curriculum. This nonequivalent comparison group study (N = 86) examined death fear and death anxiety among human services students before and after receiving death education using the Multidimensional Fear of Death Scale. The results showed a statistically significant decrease in death anxiety among the group of students who participated in death education compared to those who did not.