Analysis of the interaction of monoclonal antibodies with surface IgM on neoplastic B-cells

In vitro studies identified three Burkitts lymphoma cell lines, Ramos, MUTU-I and Daudi, that were growth inhibited by anti-IgM antibody. However, only Ramos and MUTU-I were sensitive to monoclonal antibodies (mAb) recognizing the Fc region of surface IgM (anti-Fcμ). Experiments using anti-Fcμ mAb (single or non-crossblocking pairs), polyclonal anti-μ Ab, and hyper-crosslinking with a secondary layer of Ab, showed that growth inhibition of B-cell lines was highly dependent on the extent of IgM crosslinking. This was confirmed by using Fab′, F(ab′)2and F(ab′)3derivatives from anti-Fcμ mAb, where increasing valency caused corresponding increases in growth arrest and apoptosis, presumably as a result of more efficient BCR-crosslinking on the cell surface. The ability of a single mAb to induce growth arrest was highly dependent on epitope specificity, with mAb specific for the Fc region (Cμ2–Cμ4 domains) being much more effective than those recognizing the Fab region (anti-L chain, anti-Id and anti-Fdμ, or Cμ1). Only when hyper-crosslinked with polyclonal anti-mouse IgG did the latter result in appreciable growth inhibition. Binding studies showed that these differences in function were not related to differences in the affinity, but probably related to intrinsic crosslinking capacity of mAb. © 1999 Cancer Research Campaig

Structure and Novel Functional Mechanism of Drosophila SNF in Sex-Lethal Splicing

Sans-fille (SNF) is the Drosophila homologue of mammalian general splicing factors U1A and U2B″, and it is essential in Drosophila sex determination. We found that, besides its ability to bind U1 snRNA, SNF can also bind polyuridine RNA tracts flanking the male-specific exon of the master switch gene Sex-lethal (Sxl) pre-mRNA specifically, similar to Sex-lethal protein (SXL). The polyuridine RNA binding enables SNF directly inhibit Sxl exon 3 splicing, as the dominant negative mutant SNF1621 binds U1 snRNA but not polyuridine RNA. Unlike U1A, both RNA recognition motifs (RRMs) of SNF can recognize polyuridine RNA tracts independently, even though SNF and U1A share very high sequence identity and overall structure similarity. As SNF RRM1 tends to self-associate on the opposite side of the RNA binding surface, it is possible for SNF to bridge the formation of super-complexes between two introns flanking Sxl exon 3 or between a intron and U1 snRNP, which serves the molecular basis for SNF to directly regulate Sxl splicing. Taken together, a new functional model for SNF in Drosophila sex determination is proposed. The key of the new model is that SXL and SNF function similarly in promoting Sxl male-specific exon skipping with SNF being an auxiliary or backup to SXL, and it is the combined dose of SXL and SNF governs Drosophila sex determination

Biology-driven cancer drug development: back to the future

Most of the significant recent advances in cancer treatment have been based on the great strides that have been made in our understanding of the underlying biology of the disease. Nevertheless, the exploitation of biological insight in the oncology clinic has been haphazard and we believe that this needs to be enhanced and optimized if patients are to receive maximum benefit. Here, we discuss how research has driven cancer drug development in the past and describe how recent advances in biology, technology, our conceptual understanding of cell networks and removal of some roadblocks may facilitate therapeutic advances in the (hopefully) near future

Increasing altruistic and cooperative behaviour with simple moral nudges

The conflict between pro-self and pro-social behaviour is at the core of many key problems of our time, as, for example, the reduction of air pollution and the redistribution of scarce resources. For the well-being of our societies, it is thus crucial to find mechanisms to promote pro-social choices over egoistic ones. Particularly important, because cheap and easy to implement, are those mechanisms that can change people's behaviour without forbidding any options or significantly changing their economic incentives, the so-called "nudges". Previous research has found that moral nudges (e.g., making norms salient) can promote pro-social behaviour. However, little is known about whether their effect persists over time and spills across context. This question is key in light of research showing that pro-social actions are often followed by selfish actions, thus suggesting that some moral manipulations may backfire. Here we present a class of simple moral nudges that have a great positive impact on pro-sociality. In Studies 1-4 (total N = 1,400), we use economic games to demonstrate that asking subjects to self-report "what they think is the morally right thing to do" does not only increase pro-sociality in the choice immediately after, but also in subsequent choices, and even when the social context changes. In Study 5, we explore whether moral nudges promote charity donations to humanitarian organisations in a large (N = 1,800) crowdfunding campaign. We find that, in this context, moral nudges increase donations by about 44 percent

Organizational Existence From Different Approaches: Adaptation and Surviving of Organizational Form

Differential labelling techniques like differential in-gel electrophoresis (DIGE) enable mixing a control with an experimental sample prior to protein separation, thereby reducing complexity and greatly improving the resolution and analysis of changes in protein expression. Although the shift caused by phosphorylation to a more acidic pI can, in principle, reveal phosphorylation events using DIGE, analysis and verification of the phosphorylation are fraught with problems. Here we describe a differential phospho-labelling technique that obtains the same advantages as DIGE, which we named DIPPL, for differential phosphoprotein labelling. The technique involves labelling two samples, one with 32Pi (orthophosphate) and the other with 33Pi (orthophosphate). The two samples are mixed and proteins are separated on a single gel. Dried gels are exposed twice: once so that total radiation from 32P and 33P is collected on a film or screen; then acetate sheets are interposed between the gel and the screen such that 33P radiation is filtered out leaving 32P radiation to filter through. We demonstrate the utility of this approach by studying the MEK/ERK-dependent changes in stathmin phosphorylation induced by NGF in primary sympathetic neurons