Characterisation of the anti-bladder-cancer monoclonal antibody BLCA-8: Identification of its antigen as a neutral glycolipid

A monoclonal antibody, BLCA-8, was raised against the human bladder cancer cell line, UCRU-BL-17CL. By flow cytometry and immunoperoxidase staining, this antibody was found to possess high specificity for bladder tumours, some reactivity with fetal tissues, and no reactivity with normal bladder, or any normal or malignant tissue. This high specificity and the stability of the antigen to the urinary environment suggest that BLCA-8 may have potential for use as an anti-bladder-cancer therapeutic agent. By thin-layer chromatography and autoradiography, BLCA-8 was found to bind four components within the neutral lipid fraction of a bladder cancer cell line, UCRU-BL-17/23. These components hadR _F values of 0.22, 0.16/0.15 (doublet), 0.12 and 0.08, and migrated below globoside, indicating the presence of more than four sugars. By enzyme-linked immunosorbant assay and thin-layer chromatography it was found that the binding of BLCA-8 to the lipid extract was increased by both mild alkaline hydrolysis and enzymatic treatments, indicating that adjacent phospholipids and glycolipids interfere with the accessibility of the antibody-binding site. Full biochemical characterisation of the BLCA-8 antigen is currently underway.     

Measures of geographic range size: the effects of sample size

A number of methods have been used for quantifying the sizes of the geographic ranges of species. The consequences of different levels of sampling (the proportion of actual spatial occurrences) are explored for eight of these, using data on the occurrences of butterfly species on a 10 × 10 km grid across Britain. For all methods, the percentage error of estimation (PEE) decreases with the number of 10 × 10 km squares which a species occupies, most rapidly for extent measures, and more rapidly for area measures than for measures of numbers of units occupied. The rate of decline in PEE itself falls as sampling effort increases. At a given sampling level, rank correlations between range sizes measured by different methods are generally high, but there is no consistent change in the magnitude of these correlations as the level of sampling increases. The composition of the set of species with the smallest range sizes changes with the level of sampling.     

The effects of glycophorin A on the expression of the human red cell anion transporter (band 3) in Xenopus oocytes

The effects of human red cell glycophorin A (GPA) on the translocation to the plasma membrane and anion transport activity of the human erythrocyte anion transporter (band 3; AE1) have been examined using the Xenopus oocyte expression system. We show that band 3 accumulates steadily at the oocyte surface with time in the presence or absence of GPA, but this occurs more quickly when GPA is coexpressed. The amount of band 3 at the surface is determined by the concentrations of band 3 and GPA cRNA that are injected, with a higher proportion of total band 3 being translocated to the surface in the presence of GPA cRNA. The increased expression of DNDS-sensitive chloride transport is highly specific to GPA, and is not observed when the cRNA to the putative glycophorin E or a very high concentration of the cRNA to glycophorin C are coexpressed with band 3 in oocytes.We thank Dr. Kay Ridgwell and Charlotte Ratcliffe for supplying plasmids and Dr. David Anstee for antibodies. This work was supported by grants from the Medical Research Council.     

ExbB acts as a chaperone-like protein to stabilize TonB in the cytoplasm

The TonB protein is required to transduce energy from the cytoplasmic membrane to outer membrane transport proteins of Gram-negative bacteria. Two accessory proteins, ExbB and ExbD, are required for TonB function and it has been suggested that TonB and ExbBD form a complex in the membrane. In this paper we demonstrate that there are two spatially distinct, functional interactions between ExbBD and TonB. First, there is an interaction between ExbBD and the N-terminal signal-like peptide of TonB, probabiy the formation of a stable complex in the membrane. Second, ExbB interacts with TonB in the cytoplasm. This interaction involves the domain of TonB that is normally periplasmic. Thus, this is a transient interaction which occurs during the synthesis and/or localization of TonB, implying a chaperone-like role for ExbB. The transmembrane topology of ExbB was shown to be consistent with this role.     

Parthenogenesis in an elasmobranch in the presence of conspecific males

Parthenogenesis has been observed in several elasmobranch species, primarily in public aquaria. The majority of cases of parthenogenesis have occurred either when females were held without males or once a male was removed from a female's habitat. Here we report a second instance of parthenogenesis in a zebra shark female that was housed with conspecific mature males. This study calls into question the conditions under which elasmobranch females undergo parthenogenesis.     

Enzymology in vivo using NMR and molecular genetics

Models of metabolic flux regulation are frequently based on an extrapolation of the kinetic properties of enzymes measured in vitro to the intact cell. Such an extrapolation assumes a detailed knowledge of the intracellular environment of these enzymes in terms of their free substates and effectors concentrations and possible interaction with other cellular macromolecules, which may modify their kinetic properties. These is a considerable incentive, therefore, to study the properties of enzymes directly in vivo. We have been using non-invasive NMR techniques, in conjunction with molecular genetic manipulation of enzyme levels, to study the kinetic properties of individual enzymes in vivo. We have also developed a novel strategy which has allowed us to monitor, by NMR, the ligand binding properties and mobilities of enzymes in the intact cell. This technique may also allow us to measured the diffusion coefficients of these proteins in the cell. These studies should give new insight into the properties of enzymes in vivo     

Mechancial properties of bat wing membrane skin

The skin of the bat wing in functionally unique among mammals: it serves as a major locomotor organ in addition to its protective and regulatory functions. We used tensile testing to investigate the mechanical capabilities of wing membrane skin, and compared stiffness, strength, load at failure, and energy absorption among specific wing regions and among a variety of bat taxa. We related these characteristics to the highly architectural fibrous supporting network of the wing membrane. We found that all material properties showed a strong anisotropy. In particular, wing membrane skin shows maximum stiffness and stregth parallel to the wing skeleton, and greatest extensibility parallel to the wing's trailing edge. We also found significant variation among wing regions. The uropatagium (tail membrane) supported the greatest load at failure, and the plagiopatagium (proximal wing membrane between laterl body wall and hand skeleton) is the weakest and most extensible part of the wing. We believe that the increased load bearing ability of the uropatagium relats to its key role in capture of insect prey, and that the great extensibility of the plagiopatagium promotes development of camber near the wing's centre of lift. In interspecific comparisons, energy absorpion and load to failure were greatest in Artibeus jamaicensis , the largest bat in our sample and the species with the highest wing loading, suggesting that wing loading may play a role in dictating the fuctional design of wing membranes.     

Electric field-induced concentration gradients in planar supported bilayers.

A simple method of generating electric field-induced concentration gradients in planar supported bilayers has been developed. Gradients of charged, fluorescently labeled probes were visualized by epifluorescence microscopy and could be observed at field strengths as low as 1 V/cm. Steady-state concentration gradients can be described by a simple competition between random diffusion and electric field-induced drift. A model based on this principle has been used to determine the diffusion coefficient of the fluorescent probes. This technique achieves a degree of electrical manipulation of supported bilayers that offers a variety of possibilities for the development of new molecular architectures and the study of biological membranes.