Functional-anatomic correlates of individual differences in memory

Memory abilities differ greatly across individuals. To explore a source of these differences, we characterized the varied strategies people adopt during unconstrained encoding. Participants intentionally encoded object pairs during functional MRI. Principal components analysis applied to a strategy questionnaire revealed that participants variably used four main strategies to aid learning. Individuals' use of verbal elaboration and visual inspection strategies independently correlated with their memory performance. Verbal elaboration correlated with activity in a network of regions that included prefrontal regions associated with controlled verbal processing, while visual inspection correlated with activity in a network of regions that included an extrastriate region associated with object processing. Activity in regions associated with use of these strategies was also correlated with memory performance. This study reveals functional-anatomic correlates of verbal and perceptual strategies that are variably used by individuals during encoding. These strategies engage distinct brain regions and may separately influence memory performance.     

Structure of a ribulose 5-phosphate 3-epimerase from Plasmodium falciparum

The crystal structure of Pfal009167AAA, a putative ribulose 5-phosphate 3-epimerase (PfalRPE) from Plasmodium falciparum, has been determined to 2 A resolution. RPE represents an exciting potential drug target for developing antimalarials because it is involved in the shikimate and the pentose phosphate pathways. The structure is a classic TIM-barrel fold. A coordinated Zn ion and a bound sulfate ion in the active site of the enzyme allow for a greater understanding of the mechanism of action of this enzyme. This structure is solved in the framework of the Structural Genomics of Pathogenic Protozoa (SGPP) consortium.     

Cuticular lipids and silverleaf whitefly stage affect conidial germination of Beauveria bassiana and Paecilomyces fumosoroseus

Beauveria bassiana and Paecilomyces fumosoroseus are generalist entomopathogenic fungi that infect the silverleaf whitefly (Bemisia argentifolii). We found second and third instar whiteflies to be the most susceptible larval stage to both fungi. Conidia of B. bassiana germinated most readily on the cuticle of second instars (54% germinated) and P. fumosoroseus germination was highest on third instar cuticle (45%). Fourth instars (the ultimate instar) had low susceptibility to these pathogens, and spore germination on the cuticle of fourth instars was very low for B. bassiana (7%) and intermediate for P. fumosoroseus (33%). Cuticular lipids were found to have toxic or inhibitory effects on conidia of B. bassiana and P. fumosoroseus when the spores were germinated on nutrient agar in the presence of the lipids. In the absence of added nutrients, P. fumosoroseus conidial germination increased in the presence of the lipids. To test if the inhibitory effects of the lipids were due solely to hydrophobicity (preventing water from coming into contact with the conidia) we tested the effects of synthetic long-chain wax esters. The synthetic wax esters inhibited germination of P. fumosoroseus to a degree that was similar to the effect of the cuticular lipid extracts, but the synthetic lipids did not have a significant effect on B. bassiana. Thus, the thick coating of long-chain wax esters produced by whitefly nymphs affect spore germination of fungal pathogens, but whether they play a significant role in defense against disease is not clear.     

Structure-activity relationship of a novel class of naphthyl amide KATP channel openers

We have discovered a novel series of N-[2-(2,2,2-trifluoro-1-hydroxy-1-trifluoromethyl-ethyl)-naphthalen-1-yl] amides that are potent openers of K(ATP) channels and investigated structure-activity relationships (SAR) around the 1,2-disubstituted naphthyl core. A-151892, a prototype compound of this series, was found to be a potent and efficacious potassium channel opener in vitro in transfected Kir6.2/SUR2B cells and pig bladder strips. Additionally, A-151892 was found to selectively inhibit unstable bladder contractions in vivo in an obstructed rat model of myogenic bladder function     

Differential antigen sensitivity and costimulatory requirements in human Th1 and Th2 antigen-specific CD4+ cells with similar TCR avidity

The differentiation of naive CD4(+) Th cells into Th1 and Th2 phenotypes is influenced by cytokines, concentration of Ag, accessory molecules, and the affinity of the MHC-TCR interaction. To study these factors in human memory T cells, T cell lines with Th1 or Th2 phenotypes specific for the peptide hemagglutinin (HA)(307-319) in the context of DRB1*0401 were established from the peripheral blood of an individual previously vaccinated for influenza virus. Flow cytometric analysis with fluorescent-labeled MHC class II tetramers was used to analyze TCR avidity: the Th2 line bound the HLA-DR*0401-HA(307-319) tetramers with higher mean avidity, although the range of binding avidity largely overlapped with the Th1 line. High-affinity Th1 and Th2 lines were established for further study by FACS sorting. When activated with plate-bound HLA-DR*0401-HA(307-319) monomers, the Th1 line proliferated and produced IFN-gamma without additional costimulation whereas the Th2 line required the addition of soluble anti-CD28 Ab to induce proliferation and IL-5 production, but this requirement could be overcome with high concentrations of plate-bound monomer alone. IL-2 production was dependent on costimulation in both cell lines. These findings demonstrate that upon antigenic rechallenge, Th1 and Th2 cells differ in their response to Ag-specific stimulation. Th2 cells were sensitive to the strength of signal to a greater degree than Th1 cells and required costimulation through CD28 for maximal proliferation. These distinctions between Th1 and Th2 activation are not consistent with a simple avidity model of Ag recognition and indicate both qualitative and quantitative differences in determining cell lineage commitment.     

Camouflage Patterning in Maize Leaves Results from a Defect in Porphobilinogen Deaminase

Maize leaves are produced from polarized cell divisions that result in clonal cell lineages arrayed along the long axis of the leaf. We utilized this stereotypical division pattern to identify a collection of mutants that form chloroplast pigmentation sectors that violate the clonal cell lineages. Here, we describe the camouflage1 （cfl） mutant, which develops nonclonal, yellow-green sectors in its leaves. We cloned the cfl gene by transposon tagging and determined that it encodes porphobilinogen deaminase （PBGD）, an enzyme that functions early in chlorophyll and heme biosynthesis. While PBGD has been characterized biochemically, no viable mutations in this gene have been reported in plants. To investigate the in vivo function of PBGD, we characterized the cfl mutant. Histological analyses revealed that cfl yellow sectors display the novel phenotype of bundle sheath cell-specific death. Light-shift experiments determined that constant light suppressed cfl sector formation, a dark/light transition is required to induce yellow sectors, and that sectors form only during a limited time of leaf development. Biochemical experiments determined that of 1 mutant leaves have decreased PBGD activity and increased levels of the enzyme substrate in both green and yellow regions. Furthermore, the cfl yellow regions displayed a reduction in catalase activity. A threshold model is hypothesized to explain the cfl variegation and incorporates photosynthetic cell differentiation, reactive oxygen species scavenging, and PBGD function.