INTESTINAL TRANSPORT OF ANTIBODIES IN THE NEWBORN RAT

Evidence has been reported that the proximal small intestine of the neonatal rat selectively transports antibodies into the circulation. This study describes the morphology of the absorptive epithelial cells in this region of the intestine and their transport of several immunoglobulin tracers: ferritin-conjugated immunoglobulins (IgG-Ft) and antiperoxidase antibodies. Cells exposed to rat IgG-Ft bound the tracer on the membrane of tubular invaginations of the apical cell surface. Tubular and coated vesicles within the cell also contained the tracer, as did the intercellular spaces. Uptake of tracer was highly selective and occurred only with rat or cow IgG-Ft; when cells were exposed to chicken IgG-Ft, ferritin-conjugated bovine serum albumin, or free ferritin, tracer did not enter the cell or appear in the intercellular spaces. Experiments with rat and chicken antiperoxidase showed a similar selective uptake and transport of only the homologous antibody. When cells from the distal small intestine were exposed to the tracers, all tracers were absorbed nonselectively but none were released from the cells. Cells from the proximal small intestine of the 22-day-old rat failed to absorb even rat IgG-Ft. A model is presented for selective antibody transport in proximal cells of the neonatal rat in which antibodies are selectively absorbed at the apical cell surface by pinocytosis within tubular vesicles. The antibodies are then transferred to the intercellular space within coated vesicles. Distal cells function only to digest proteins nonselectively.     

Species-area curves and estimates of total species richness in an old-field chronosequence

Average species-area curves were generated for vascular plants in 20 old-fields that were sampled in 1983, 1989, and 1994. These curves were fit with a saturating function to estimate total species richness for each old-field. Additional estimates of total species richness were generated by fitting the same saturating function to subsets of the species area curves and with a first-order jackknife procedure. Estimates of total species richness were strongly correlated with observed species richness. There was limited evidence suggesting that greater sampling was necessary to identify the same proportion of species in older, more species-rich old-fields.     

Mlc1p Is a Light Chain for the Unconventional Myosin Myo2p in Saccharomyces cerevisiae

In Saccharomyces cerevisiae, the unconventional myosin Myo2p is of fundamental importance in polarized growth. We explore the role of the neck region and its associated light chains in regulating Myo2p function. Surprisingly, we find that precise deletion of the six IQ sites in the neck region results in a myosin, Myo2-Δ6IQp, that can support the growth of a yeast strain at 90% the rate of a wild-type isogenic strain. We exploit this mutant in a characterization of the light chains of Myo2p. First, we demonstrate that the localization of calmodulin to sites of polarized growth largely depends on the IQ sites in the neck of Myo2p. Second, we demonstrate that a previously uncharacterized protein, Mlc1p, is a myosin light chain of Myo2p. MLC1 (YGL106w) is an essential gene that exhibits haploinsufficiency. Reduced levels of MYO2 overcome the haploinsufficiency of MLC1. The mutant MYO2-Δ6IQ is able to suppress haploinsufficiency but not deletion of MLC1. We used a modified gel overlay assay to demonstrate a direct interaction between Mlc1p and the neck of Myo2p. Overexpression of MYO2 is toxic, causing a severe decrease in growth rate. When MYO2 is overexpressed, Myo2p is fourfold less stable than in a wild-type strain. High copies of MLC1 completely overcome the growth defects and increase the stability of Myo2p. Our results suggest that Mlc1p is responsible for stabilizing this myosin by binding to the neck region.     

Targeting Homologous Recombination in Notch-Driven C. elegans Stem Cell and Human Tumors

Mammalian NOTCH1-4 receptors are all associated with human malignancy, although exact roles remain enigmatic. Here we employ glp-1(ar202), a temperature-sensitive gain-of-function C. elegans NOTCH mutant, to delineate NOTCH-driven tumor responses to radiotherapy. At ≤20°C, glp-1(ar202) is wild-type, whereas at 25°C it forms a germline stem cell⁄progenitor cell tumor reminiscent of human cancer. We identify a NOTCH tumor phenotype in which all tumor cells traffic rapidly to G2⁄M post-irradiation, attempt to repair DNA strand breaks exclusively via homology-driven repair, and when this fails die by mitotic death. Homology-driven repair inactivation is dramatically radiosensitizing. We show that these concepts translate directly to human cancer models.     

A protocol for temporal calibration of general area cladograms

Aim To describe a protocol for incorporating a temporal dimension into historical biogeographical analysis, while maintaining the essential independence of all datasets, involving the generation of general area cladograms. Location Global. Methods General area cladograms (GACs) are a reconstruction of the evolutionary history of a set of areas and unrelated clades within those areas. Nodes on a GAC correspond to speciation events in a group of taxa; general nodes are those at which multiple unrelated clades speciate. We undertake temporal calibration of GACs using molecular clock estimates of splitting events between extant taxa as well as first appearance data from the fossil record. We present two examples based on re‐analysis of previously published data: first, a temporally calibrated GAC generated from secondary Brooks parsimony analysis (BPA) of six extant bird clades from the south‐west of North America using molecular clock estimates of divergence times; and second, an analysis of African Neogene mammals based on a phylogenetic analysis for comparing trees (PACT) analysis. Results A hypothetical example demonstrates how temporal calibration reveals potentially critical information about the timing of both unique and general events, while also illustrating instances of incongruence between dates generated from molecular clock estimates and fossils. For the African Neogene mammal dataset, our analysis reveals that most mammal clades underwent geodispersal associated with the Neogene climatic optimum (c. 16 Ma) and vicariant speciation in central Africa correlated with increased aridity and cooler temperatures around 2.5 Ma. Main conclusions Temporally calibrated GACs are valuable tools for assessing whether coordinated patterns of speciation are associated with large‐scale climatic or tectonic phenomena.     

The immunological network at the site of tumor rejection

The tumor mass irrespective of its type or location in the body has long been shrouded in mystery and even today we still have only a tentative handle on its secrets. Attempts to manipulate either the tumor cells per se or host-derived leukocytes have, on the whole, not been successful or at best questionable. The ability of the host to respond immunologically to TSTA is well documented, yet again attempts to manipulate this response have been disappointing. One of the problems has been a lack of knowledge concerning the tumor mass and its constituents, such as the intratumor leukocytes, and the significance of their presence to the biological properties of the neoplasm [8,9,80]. The purpose in studying the immunological network is, in part, to try to assign a function to these cells on the premise that lymphoid elements and macrophages have a potential role to play in recognition of TSTA. The advantage of adoptive immunotherapy model systems is that tumor rejection can be achieved under controlled conditions and this allows an analysis of the immunological network and its individual circuits. At the same time, valuable information on the mechanisms of action during adoptive immunotherapy and how best to improve therapeutic protocols is acquired.