Secretion of CTLA4Ig by an SV40 T antigen-transformed islet cell line inhibits graft rejection against the neoantigen

We found that cytochrome c (Cyt c) could oxidize cardiolipin (CL), and detected monoepoxides of linoleic acid (LA) in the fatty acids constituting the oxidized CL. We also found that in the presence of CL and Cyt c, free LA was oxidized and LA monoepoxides were produced. The aim of this study was to elucidate the mechanism of this lipid peroxidation. We concluded that ferric Cyt c produced some radical species from water-soluble oxygen in the presence of CL (CL-Cyt c system) and that radicals oxidized free LA or CL. The CL-Cyt c system may be another LA monoepoxide producing system in the neutrophil and may account for the lipid peroxidation observed in the ischemia-reperfusion-induced cardiac injury.     

An ABC transporter plays a developmental aggregation role in Myxococcus xanthus

Myxococcus xanthus is a gram-negative bacterium which has a complex life cycle. Autochemotaxis, a process whereby cells release a self-generated signaling molecule, may be the principal mechanism facilitating directed motility in both the vegetative swarming and developmental aggregation stages of this life cycle. The process requires the Frz signal transduction system, including FrzZ, a protein which is composed of two domains, both showing homology to the enteric chemotaxis response regulator CheY. The first domain of FrzZ (FrzZ1), when expressed as bait in the yeast two-hybrid system and screened against a library, was shown to potentially interact with the C-terminal portion of a protein encoding an ATP-binding cassette (AbcA). The activation domain-AbcA fusion protein did not interact with the second domain of FrzZ (FrzZ2) or with two other M. xanthus response regulator-containing proteins presented as bait, suggesting that the FrzZ1-AbcA interaction may be specific. Cloning and sequencing of the upstream region of the abcA gene showed the ATP-binding cassette to be linked to a large hydrophobic, potentially membrane-spanning domain. This domain organization is characteristic of a subgroup of ABC transporters which perform export functions. Cloning and sequencing downstream of abcA indicated that the ABC transporter is at the start of an operon containing three open reading frames. An insertion mutation in the abcA gene resulted in cells displaying the frizzy aggregation phenotype, providing additional evidence that FrzZ and AbcA may be part of the same signal transduction pathway. Cells with mutations in genes downstream of abcA showed no developmental defects. Analysis of the proposed exporter role of AbcA in cell mixing experiments showed that the ABC transporter mutant could be rescued by extracellular complementation. We speculate that the AbcA protein may be involved in the export of a molecule required for the autochemotactic process.     

Display duration and stereoscopic depth discrimination.

Investigated the role of display duration in stereoscopic depth perception. The display consisted of a dynamic random-dot stereogram, with 2 disparity defined squares, one on the left and one on the right of a central (Nonius) fixation stimulus. The sign of the disparity (crossed or uncrossed) was always the same for both squares, and the magnitude of disparity was 0.25° for one square and either 0.125° or 0.375° for the other square. 100 participants (aged 18–43 yrs) indicated which square appeared closer. The display duration was varied adaptively between 20 and 1000 ms until participants performed at 75% accuracy. Results confirmed large individual differences in the display duration required for stereoscopic depth perception. Approximately half of the participants were able to perform the task at 20 ms, while the remaining participants required up to 1000 ms to perform at criterion. The present study shows that display duration is a critical variable in explaining wide differences in reported abilities of individuals to process stereoscopic depth information. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Interface structure in arsenide/phosphide heterostructun grown by gas-source MBE and low-pressure MOVPE

We have used cross-sectional scanning tunneling microscopy (STM) to study interface structure in arsenide/phosphide heterostructures grown by gas-source molecular beam epitaxy (GSMBE) and by low-pressure metalorganic vapor phase epitaxy (LP-MOVPE). High-resolution images of GSMBE samples consisting of GaAs interrupted at 200Å intervals with a 40 s P₂ flux reveal substantial, growth-temperature-dependent incorporation of phosphorus with nanometer-scale lateral variations in interface structure. STM images of InGaAs/InP multiple quantum well structures grown by LP-MOVPE show evidence of interface asymmetry and extensive atomic cross-incorporation at the interfaces. Data obtained by STM have been corroborated by high-resolution x-ray diffraction and reflection high-energy electron diffraction. Together, these studies provide direct information about nanometer-scale grading and lateral nonuniformity of arsenide/phosphide interfaces that can occur under these growth conditions.     

Adhesion properties of mutants of Staphylococcus aureus defective in fibronectin-binding proteins and studies on the expression of fnb genes

Staphylococcus aureus 8325-4 has the potential to express two distinct cell wall-associated fibronectin-binding proteins called FnBPA and FnBPB. In order to test if both proteins are expressed in S. aureus and if both are required for promoting bacterial adhesion to fibronectin-coated surfaces, insertion mutations were isolated in each gene. A DNA fragment encoding tetracycline resistance was inserted into fnbA and a fragment encoding erythromycin resistance was inserted into fnbB. A double fnbAfnbB mutant was also constructed. The fnbA and fnbB single mutants showed no significant reduction in their adhesion to polymethylmethacrylate coverslips that had been coated in vitro with fibronectin. However, the double mutant was completely defective in adhesion. Monospecific antibodies directed against the non-conserved N-terminal regions of both proteins confirmed the lack of expression of FnBPs in the mutant strains. Wild-type fnbA and fnbB genes cloned seperately on a multicopy plasmid were each able to restore fully the adhesion-defective phenotype of the 8325-4 fnbAfnbB mutant. This demonstrates that both fnb genes are expressed in S. aureus and that both contribute to the ability of strain 8325-4 to adhere to fibronectin-coated surfaces. The double mutant was also defective in adhesion to coverslips that had been removed from tissue cages implanted subcutaneously in guinea-pigs, which suggests that fibronectin is important in promoting attachment of S. aureus to biomaterial in vivo.     

Attentional modulation of visual processes in motion perception.

Examined the effects of spatially directed attention on the temporal characteristics of information transmission in the visual system. A 2-stimulus, 2-flash, long-range motion display was used. The results showed that attending to 1 of the 2 stimuli altered the perceived direction of motion, the pattern of motion and no-motion responses, and ratings of motion quality. The results were compared with predictions derived from 2 conceptual frameworks of attention: a temporal-profile model and an additive account. The results support the temporal-profile model. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Synthetic inhibitors of endopeptidase EC 3.4.24.15: potency and stability in vitro and in vivo

1. The role of the metalloendopeptidase EC 3.4.24.15 (EP 24.15) in peptide metabolism in vivo is unknown, in part reflecting the lack of a stable enzyme inhibitor. The most commonly used inhibitor, N-[1-(R,S)-carboxy-3-phenylpropyl]-Ala-Ala-Tyr-p-aminobenzoate (cFP-AAY-pAB, Ki = 16 nM), although selective in vitro, is rapidly degraded in the circulation to cFP-Ala-Ala, an angiotensin converting enzyme (ACE) inhibitor. This metabolite is thought to be generated by neutral endopeptidase (NEP; EC 3.4.24.11), as the Ala-Tyr bond of cFP-AAY-pAB is cleaved by NEP in vitro. In the present study, we have examined the role of NEP in the metabolism of cFP-AAY-pAB in vivo, and have tested a series of inhibitor analogues, substituted at the second alanine, for both potency and stability relative to the parent compound. 2. Analogues were screened for inhibition of fluorescent substrate cleavage by recombinant rat testes EP 24.15. D-Ala or Asp substitution abolished inhibitory activity, while Val-, Ser- and Leu-substituted analogues retained activity, albeit at a reduced potency. A relative potency order of Ala (1) > Val (0.3) > Ser (0.16) > Leu (0.06) was observed. Resistance to cleavage by NEP was assessed by incubation of the analogues with rabbit kidney membranes. The parent compound was readily degraded, but the analogues were twice (Ser) and greater than 10 fold (Leu and Val) more resistant to cleavage. 3. Metabolism of cFP-AAY-pAB and the Val-substituted analogue was also examined in conscious rabbits. A bolus injection of cFP-AAY-pAB (5 mg kg-1, i.v.) significantly reduced the blood pressure response to angiotensin I, indicating ACE inhibition. Pretreatment with NEP inhibitors, SCH 39370 or phosphoramidon, slowed the loss of cFP-AAY-pAB from the plasma, but did not prevent inhibition of ACE. Injection of 1 mg kg-1 inhibitor resulted in plasma concentrations at 10 s of 23.5 microM (cFP-AAY-pAB) and 18.0 microM (cFP-AVY-pAB), which fell 100 fold over 5 min. Co-injection of 125I-labelled inhibitor revealed that 80-85% of the radioactivity had disappeared from the circulation within 5 min, and h.p.l.c. analysis demonstrated that only 25-30% of the radiolabel remained as intact inhibitor at this time. Both analogues were cleared from the circulation at the same rate, and both inhibitors blunted the pressor response to angiotensin I, indicative of ACE inhibition. 4. These results suggest that both NEP and other clearance/degradation mechanisms severely limit the usefulness of peptide-based inhibitors such as cFP-AAY-pAB. To examine further EP 24.15 function in vivo, more stable inhibitors, preferably non-peptide, must be developed, for which these peptide-based inhibitors may serve as useful molecular templates.     

Attentional and ocular movements.

The relationship between eye movements and movements of attention was examined in a series of 6 experiments. A temporal order judgment technique was used to index attentional allocation. The results showed that endogenous movements of attention were slower than movements of the eyes and that the participants were able to hold attention at one location while executing an eye movement to another location. Under conditions of exogenous cueing, attention moved rapidly to the cued location, in advance of the eyes. These findings challenge the prevailing view that ocular movements must necessarily be preceded by a movement of attention. (PsycINFO Database Record (c) 2010 APA, all rights reserved)