Antibodies specific for VB8 receptor peptide suppress experimental autoimmune encephalomyelitis.

Recent studies from our laboratory have shown, for the first time, that a synthetic peptide from that TCR VB chain used preferentially by encephalitogenic T cells induced the formation of protective, MHC class I-restricted T cells and prevented the development of EAE in Lewis rats. In this report we 1) demonstrate that immunization with the TCR-VB8-39-59 peptide generated peptide-specific antibodies that protect against experimental autoimmune encephalomyelitis induced by either of the two distinct encephalitogenic epitopes of basic protein, and 2) characterize the production and biologic functions of rat and rabbit antibody responses to the TCR peptide. The antibodies in both species increased in titer over time, were highly specific for the immunogen by direct reaction and inhibition assays, stained only VB8+ T cells, and suppressed clinical signs and to lesser extent the number of histologic lesions of experimental autoimmune encephalomyelitis mediated by VB8+ T cells. Coupled with our previous work, these results indicate that both humoral and cellular responses to the TCR-VB8-39-59 peptide can contribute independent immunoregulatory effects on encephalitogenic T lymphocytes that use common V region genes in response to epitopes of myelin basic protein.     

Properties of a simian virus 40 mutant T antigen substituted in the hydrophobic region: defective ATPase and oligomerization activities and altered phosphorylation accompany an inability to complex with cellular p53.

We have analyzed the biochemical properties of a nonviable simian virus 40 (SV40) mutant encoding a large T antigen (T) bearing an amino acid substitution (Pro-584-Leu) in its hydrophobic region. Mutant 5080 has an altered cell type specificity for transformation (transforming mouse C3H10T1/2 but not rat REF52 cells), is defective for viral DNA replication, and encodes a T that is unable to form a complex with the cellular p53 protein (K. Peden, A. Srinivasan, J. Farber, and J. Pipas, Virology 168:13-21, 1989). In this article, we show that 5080-transformed C3H10T1/2 cell lines express an altered T that is synthesized at a significantly higher rate but with a shorter half-life than normal T from wild-type SV40-transformed cells. 5080 T did not oligomerize beyond 5 to 10S in size compared with normal T, which oligomerized predominantly to 14 to 20S species. In addition, the 5080 T complex had significantly decreased ATPase activity and had a 10-fold-lower level of in vivo phosphorylation compared with that of normal T. Two-dimensional phosphopeptide analysis indicated several changes in the specific 32P labeling pattern, with altered phosphorylation occurring at both termini of the mutant protein compared with the wild-type T. Loss of p53 binding is therefore concomitant with changes in ATPase activity, oligomerization, stability, and in vivo phosphorylation of T and can be correlated with defective replication and restricted transformation functions. That so many biochemical changes are associated with a single substitution in the hydrophobic region of T is consistent with its importance in regulating higher-order structural and functional relationships in SV40 T.     

Purification, crystallization, and X-ray diffraction studies of lactotransferrin from buffalo colostrum.

Lactotransferrin is an iron-binding protein. It has been purified from buffalo colostrum. The purified lactotransferrin has been crystallized in 10% ethanol solution. The crystals are orthorhombic and the space group is P2(1)2(1)2(1) with unit cell dimensions a = 161.70 A, b = 155.75 A, c = 113.48 A. The asymmetric unit contains three molecules of the protein with a solvent content of about 59%. The crystals were stable in the X-ray beam and diffract beyond 3.5 A resolution. The native data have been collected and the structure determination is in progress.     

cDNA sequence of the small subunit of the hamster ribonucleotide reductase.

Ribonucleotide reductase activity is markedly elevated in cell lines selected for resistance to hydroxyurea, a cytotoxic drug known specifically to inhibit ribonucleotide reductase. From a cDNA library constructed from a highly hydroxyurea-resistant hamster lung cell line, 600H in which the activity is elevated more than 80-fold, we have isolated a full length cDNA for the small subunit of the reductase. The cDNA is 3.48 kb long with an open reading frame of 1158 nucleotides and a long 3' flanking region of 2169 nucleotides from the termination codon. The derived polypeptide sequence is closely similar to the small subunit of the mouse, differing from it in 20 amino acid positions. Most of these replacements occur in the N-terminal segment of the protein. The hamster subunit does not contain 4 amino acid residues found in the mouse small subunit near the C-terminal end. RNA blots probed with the cDNA show two poly(A)+ RNA species which are elevated in hydroxyurea-resistant cells.     

Pattern recognition in honeybees: chromatic properties of orientation analysis

The spectral properties of the discrimination of pattern orientation in freely flying honeybees (Apis mellifera) were examined. Bees were trained to discriminate between two random black/white gratings oriented perpendicularly to each other, one of which was associated with a reward. Subsequently the bees were tested on two-colour gratings or gratings consisting of grey and coloured stripes, providing a range of different chromatic contrasts, luminance contrasts and specific channel contrasts. The results of these experiments indicate that orientation analysis in the honeybee is mediated almost exclusively by the green receptor channel, although the bee's visual system as a whole is endowed with excellent trichromatic colour vision.     

Computer simulation of the effect of dDAVP with saline loading on fluid balance after 24-hour head-down tilt

Fluid loading (FL) before Shuttle reentry is a countermeasure currently in use by NASA to improve the orthostatic tolerance of astronauts during reentry and postflight. The fluid load consists of water and salt tablets equivalent to 32 oz (946 ml) of isotonic saline. However, the effectiveness of this countermeasure has been observed to decrease with the duration of spaceflight. The countermeasure's effectiveness may be improved by enhancing fluid retention using analogs of vasopressin such as lypressin (LVP) and desmopressin (dDAVP). In a computer simulation study reported previously, we attempted to assess the improvement in fluid retention obtained by the use of LVP administered before FL. The present study is concerned with the use of dDAVP. In a recent 24-hour, 6 degree head-down tilt (HDT) study involving seven men, dDAVP was found to improve orthostatic tolerance as assessed by both lower body negative pressure (LBNP) and stand tests. The treatment restored Luft's cumulative stress index (cumulative product of magnitude and duration of LBNP) to nearly pre-bedrest level. The heart rate was lower and stroke volume was marginally higher at the same LBNP levels with administration of dDAVP compared to placebo. Lower heart rates were also observed with dDAVP during stand test, despite the lower level of cardiovascular stress. These improvements were seen with only a small but significant increase in plasma volume of approximately 3 percent. This paper presents a computer simulation analysis of some of the results of this HDT study.     

Object detection by honeybees: Why do they land on edges?

Behavioural experiments using a variety of experimental situations (Figs. 1, 3, 6, 7, 9) were conducted to investigate the visual cues which bees use in the task of object-ground discrimination. The bees' flight and landing behaviour was video-filmed throughout the experiments. The evaluation of the video data shows that bees trained to find a randomly textured figure raised above a similarly textured ground land mainly on the boundaries of the figure, facing its inner surface (Fig. 2a, b). Bees can also be trained to find a hole, i.e. a low texture viewed through a window cut in a raised texture, but these bees are not attracted to the edges of the hole (Fig. 5a, b). Bees trained to a single edge between a low and a raised random texture land at the edge mainly facing the raised side (Table 1). Bees approaching the edge from the high side cross the edge in most cases without landing on it (Table 1). Bees trained to an edge between 2 striped patterns, one raised above the other, again land on the edge facing the raised pattern, regardless of whether the stripes on the 2 patterns run parallel or perpendicular to each other or to the edge (Fig. 8). In this case, the bees acquire range information by flying in oblique directions with respect to the orientation of the stripes (Fig. 10). All of the results suggest that the edge elicits landings when the bee perceives a local increase in the speed of image motion, signalling an abrupt decrease in range. This is corroborated by the results of further experiments in which artificial motion was used to simulate range differences between the two sides of an edge (Table 2). We conclude that image speed is a powerful cue in range discrimination as well as object detection. Dedicated to G. Adrian Horridge on the occasion of his retirement     

Perception of heterochromatic flicker by honeybees: a behavioural study

Freely flying honeybees were trained to discriminate a stimulus consisting of two alternating chromatic lights (heterochromatic flicker) from a steady mixture of the same two lights, using 3 different pairs of lights: blue-UV, UV-green, and green-UV. With each light pair, training to the heterochromatic flicker was conducted at several flicker frequencies, using experimentally naive bees in each training. In subsequent tests, the trained bees were given a choice between the two lights that constituted the flicker, presented steady, as well as between either of them and the steady mixture. We find that bees trained to particular frequencies of heterochromatic flicker prefer one of the component lights over the other as well as over the steady mixture, suggesting that the colour they perceive in the heterochromatic flicker to which they have been trained is shifted in the direction of one of the lights contained in the flicker. The colour shift occurs at flicker frequencies that depend on the pair of lights used. We propose that the shift is generated by an effect similar to the Brücke-Bartley phenomenon known from human vision. This effect is based on the enhancement of the photoreceptors' response upon onset of stimulation, causing an intermittent light to appear brighter than a steady light of identical physical intensity. We propose that the degree of enhancement might differ among the 3 spectral classes of photoreceptor, causing the colour perceived in a heterochromatic flicker to differ from that perceived in a steady mixture of its two light components.