Anaplasma phagocytophilum, the aetiologic agent of human granulocytic anaplasmosis, is an obligate intracellular bacterium that colonizes neutrophils and neutrophil precursors. The granulocytotropic bacterium uses multiple adhesins that cooperatively bind to the N-terminal region of P-selectin glycoprotein ligand-1 (PSGL-1) and to sialyl Lewis x (sLex) expressed on myeloid cell surfaces. Recognition of sLex occurs through interactions with α2,3-sialic acid and α1,3-fucose. It is unknown whether other bacteria–host cell interactions are involved. In this study, we have enriched for A. phagocytophilum organisms that do not rely on sialic acid for cellular adhesion and entry by maintaining strain NCH-1 in HL-60 cells that are severely undersialylated. The selected bacteria, termed NCH-1A, also exhibit lessened dependencies on PSGL-1 and α1,3-fucose. Optimal adhesion and invasion by NCH-1A require interactions with the known determinants (sialic acid, PSGL-1 and α1,3-fucose), but none of them is absolutely necessary. NCH-1A binding to sLex-modified PSGL-1 requires recognition of the known determinants in the same manners as other A. phagocytophilum strains. These data suggest that A. phagocytophilum expresses a separate adhesin from those targeting sialic acid, α1,3-fucose and the N-terminal region of PSGL-1. We propose that NCH-1A upregulates expression of this adhesin.

Anaplasma phagocytophilum, the causative agent of human granulocytic anaplasmosis, is an obligate intracellular bacterium that infects neutrophils and neutrophil precursors. Bacterial recognition of P-selectin glycoprotein ligand-1 (PSGL-1) and the ␣2,3-sialylatedand ␣1,3-fucosylated-moiety sialyl-Lewis x (sLe x ), which modifies the PSGL-1 N terminus, is important for adhesion to and invasion of myeloid cells. We have previously demonstrated that A. phagocytophilum organisms of the NCH-1 strain that utilize an sLe x -modified PSGL-1-independent means of entry can be enriched for by cultivation in undersialylated HL-60 cells that are unable to construct sLe x . Because it was unknown whether other A. phagocytophilum isolates share this ability, we extended our studies to the geographically diverse strains HZ and HGE1. HL-60 A2 is a clonal cell line that is defective for sialylation and ␣1,3-fucosyltransferase. HL-60 A2 cell surfaces, therefore, not only lack sLe x but also are virtually devoid of any other sialic acid-and/or ␣1,3-fucose-modified glycan. By cultivating HZ and HGE1 in HL-60 A2 cells, we enriched for bacterial subpopulations (termed HZA2 and HGE1A2) that bind and/or infect myeloid cells in the absence of sialic acid and ␣1,3-fucose and in the presence of antibody that blocks the N terminus of PSGL-1. Thus, multiple A. phagocytophilum isolates share the ability to use sLe xmodified PSGL-1-dependent and -independent routes of entry into myeloid cells. HZA2 and HGE1A2 represent enriched bacterial populations that will aid dissection of the complexities of the interactions between A. phagocytophilum and host myeloid cells.

Anaplasma phagocytophilum is the etiologic agent of human granulocytic anaplasmosis. MSP2(P44), the bacterium's major surface protein, is encoded by a paralogous gene family and has been implicated in a variety of pathobiological processes, including antigenic variation, host adaptation, adhesion, porin activity, and structural integrity. The consensus among several studies performed at the DNA and RNA levels is that a heterogeneous mix of a limited number of msp2(p44) transcripts is expressed by A. phagocytophilum during in vitro cultivation. Such analyses have yet to be extended to the protein level. In this study, we used proteomic and molecular approaches to determine that MSP2(P44)-18 is the predominant if not the only paralog expressed and is modified into multiple 42-to 44-kDa isoforms by A. phagocytophilum strain HGE1 during infection of HL-60 cells. The msp2(p44) expression profile was homogeneous for msp2(p44)-18. Thus, MSP2 (P44)-18 may have a fitness advantage in HL-60 cell culture in the absence of selective immune pressure. Several novel 22-to 27-kDa MSP2 isoforms lacking most of the N-terminal conserved region were also identified. A. phagocytophilum MSP2(P44) orthologs expressed by other pathogens in the family Anaplasmataceae are glycosylated. Gas chromatography revealed that recombinant MSP2(P44)-18 is modified by glucose, galactose, xylose, mannose, and trace amounts of other glycosyl residues. These data are the first to confirm differential modification of any A. phagocytophilum MSP2(P44) paralog and the first to provide evidence for expression of truncated versions of such proteins.

Anaplasma phagocytophilum is an obligate intracellular bacterium that infects neutrophils to cause granulocytic anaplasmosis in humans and mammals. P-selectin glycoprotein ligand-1 (PSGL-1) and the tetrasaccharide sialyl Lewis x (sLex), which caps the PSGL-1 N-terminus, are confirmed A. phagocytophilum receptors. A. phagocytophilum is capable of sLex-modified PSGL-1-dependent and -independent infection. PSGL-1 N-terminus-mediated entry is dependent on spleen tyrosine kinase (Syk). Here, we determined that PSGL-1-independent entry does not alter bacterial replication and investigated whether it involves Syk using NCH-1A2, an enriched subpopulation of A. phagocytophilum NCH-1 obtained through cultivation in a sLex-deficient HL-60 cell line, HL-60 A2. Pharmacological inhibition of Syk nearly abolishes NCH-1 infection, but does not alter NCH-1A2 invasion and only marginally reduces NCH-1A2 propagation. This phenomenon was confirmed by a competitive infection assay using PSGL-1-dependent and -independent A. phagocytophilum organisms transformed to express mCherry or green fluorescent protein respectively. We also assayed for delivery and tyrosine phosphorylation of the A. phagocytophilum effector, AnkA, following NCH-1or NCH-1A2 incubation with HL-60 or HL-60 A2 cells in the presence of PSGL-1 blocking antibody. PSGL-1 N-terminus recognition promotes optimal AnkA delivery while binding to sLex or the unknown receptor is comparably less important for this process.

Blast-induced traumatic brain injury (bTBI) is the "signature wound" of the current wars in Iraq and Afghanistan. However, with no objective information of relative blast exposure, warfighters with bTBI may not receive appropriate medical care and are at risk of being returned to the battlefield. Accordingly, we have created a colorimetric blast injury dosimeter (BID) that exploits material failure of photonic crystals to detect blast exposure. Appearing like a colored sticker, the BID is fabricated in photosensitive polymers via multi-beam interference lithography. Although very stable in the presence of heat, cold or physical impact, sculpted micro-and nano-structures of the BID are physically altered in a precise manner by blast exposure, resulting in color changes that correspond with blast intensity. This approach offers a lightweight, power-free sensor that can be readily interpreted by the naked eye. Importantly, with future refinement this technology may be deployed to identify soldiers exposed to blast at levels suggested to be supra-threshold for nonimpact blast-induced mild TBI.

Blast-induced traumatic brain injury (bTBI) has been described as the defining injury of Operations Enduring Freedom and Iraqi Freedom (OEF/OIF). Previously, most blast injury research has focused on the effects of blast on internal, gas filled organs due to their increased susceptibility. However, due to a change in enemy tactics combined with better armor and front-line medical care, bTBI has become one of the most common injuries due to blast. Though there has been a significant amount of research characterizing the brain injury produced by blast, a sound understanding of the contribution of each component of the shockwave to the injury is needed. Large animal models of bTBI utilize chemical explosives as their shockwave source while small animal models predominantly utilize compressed air-driven membrane rupture as their shockwave source. We designed and built a multi-mode shock tube capable of utilizing compressed gas (air or helium)-driven membrane rupture or chemical explosives (oxyhydrogen-a 2:1 mixture of hydrogen and oxygen gasses, or RDX-high order explosive) to produce a shockwave. Analysis of the shockwaves produced by each mode of the McMillan Blast Device (MBD) revealed that compressed air-driven shockwaves exhibited longer duration positive phases than compressed helium-, oxyhydrogen-, or RDX-driven shockwaves of similar peak overpressure. The longer duration of compressed air-driven shockwaves results in greater energy being imparted on a test subject than would be imparted by shockwaves of identical peak overpressures from the other sources. Animals exposed to compressed air-driven shockwaves exhibited more extensive brain surface hematoma, more blood-brain barrier compromise, more extensive reactive astrocytosis, and greater numbers of activated microglia in their brains than did animals exposed to oxyhydrogen-driven shockwaves of even greater peak overpressure. Taken together, these data suggest that compressed air-driven shockwaves contain more energy than their chemical explosive-derived counterparts of equal peak overpressure and vi TABLE OF CONTENTS

OF DISSERTATION BLAST-INDUCED TRAUMATIC BRAIN INJURY: INFLUENCE OF SHOCKWAVE COMPONENTS Blast-induced traumatic brain injury (bTBI) has been described as the defining injury of Operations Enduring Freedom and Iraqi Freedom (OEF/OIF). Previously, most blast injury research has focused on the effects of blast on internal, gas filled organs due to their increased susceptibility. However, due to a change in enemy tactics combined with better armor and front-line medical care, bTBI has become one of the most common injuries due to blast. Though there has been a significant amount of research characterizing the brain injury produced by blast, a sound understanding of the contribution of each component of the shockwave to the injury is needed. Large animal models of bTBI utilize chemical explosives as their shockwave source while small animal models predominantly utilize compressed air-driven membrane rupture as their shockwave source. We designed and built a multi-mode shock tube capable ...

Individuals with semantic dementia (SD) were differentiated neuropsychologically from individuals with dementia of the Alzheimer type (DAT) at very mild-to-mild stages (clinical dementia rating 0.5 or 1). A picture naming and recognition memory experiment provided a particularly useful probe for early identification, with SD individuals showing preserved picture recognition memory and impaired naming, and DAT individuals tending to show the reverse dissociation. The identification of an early SD group provided the opportunity to inform models of reading by exploring the influence of isolated lexical semantic impairment on reading regular words. Results demonstrated prolonged latency in both SD and DAT group reading compared to a control group but exaggerated influence of frequency and length only for the SD group. The SD reading pattern was associated with focal atrophy of the left temporal pole. These cognitive-neuroanatomical findings suggest a role for the left temporal pole in lexical/semantic components of reading and demonstrate that cortical thickness differences in the left temporal pole correlate with prolonged latency associated with increased reliance on sublexical components of reading.

This study was designed to identify the neural network supporting the semantic processing of visual words in a patient with large-scale damage to left-hemisphere (LH) language structures. Patient GP, and a control subject, RT, performed semantic and orthographic tasks while brain-activation patterns were recorded using functional magnetic resonance imaging. In RT, the semantic-orthographic comparison activated LH perisylvian and extrasylvian temporal regions comparable to the network of areas activated by non-brain-damaged subjects in other neuroimaging studies of semantic discrimination. In GP, the same comparison activated homologous right-hemisphere regions, demonstrating the ability of the right hemisphere to subserve visual lexicosemantic processes. The results are discussed within the context of the normal right hemisphere's capacity for semantic processing of visual words. Examining results from functional neuroimaging studies on recovery in the context of innate hemispheric abilities may enable reconciliation of disparate claims about mechanisms supporting recovery from aphasia.

By employing visual lexical decision and functional MRI, we studied the neural correlates of morphological decomposition in a highly inXected language (Finnish) where most inXected noun forms elicit a consistent processing cost during word recognition. This behavioral eVect could reXect suYx stripping at the visual word form level and/or subsequent meaning integration at the semantic-syntactic level. The Wrst alternative predicts increased activation for inXected vs. monomorphemic words in the left occipitotemporal cortex while the second alternative predicts left inferior frontal gyrus and/or left posterior temporal activation increases. The results show signiWcant activation eVects in the latter areas. This provides support for the second alternative, i.e., that the morphological processing cost stems from the semantic-syntactic level.

Two brain regions with established roles in reading are the posterior middle temporal gyrus and the posterior fusiform gyrus. Lesion studies have also suggested that the region located between them, the posterior inferior temporal gyrus (pITG), plays a central role in word recognition. However, these lesion results could reflect disconnection effects since neuroimaging studies have not reported consistent lexicality effects in pITG. Here we tested whether these reported pITG lesion effects are due to disconnection effects or not using parallel ERP/fMRI studies. We predicted that the Recognition Potential (RP), a left-lateralized ERP negativity that peaks at about 200-250 ms, might be the electrophysiological correlate of pITG activity and that conditions that evoke the RP (perceptual degradation) might therefore also evoke pITG activity. In Experiment 1, twenty-three participants performed a lexical decision task (temporally flanked by supraliminal masks) while having high-density 129-channel ERP data collected. In Experiment 2, a separate group of fifteen participants underwent the same task while having fMRI data collected in a 3T scanner. Examination of the ERP data suggested that a canonical Recognition Potential effect was produced. The strongest corresponding effect in the fMRI data was in the vicinity of the pITG. In addition, results indicated stimulus-dependent functional connectivity between pITG and a region of the posterior fusiform gyrus near the visual word form area (VWFA) during word compared to nonword processing. These results provide convergent spatiotemporal evidence that the pITG contributes to early lexical access through interaction with the VWFA.

Volume losses in the medial temporal lobe, posterior cingulated, and orbitofrontal region have been observed in Alzheimer's disease (AD). Smaller reductions in similar regions have also been reported in amnestic mild cognitive impairment (aMCI), a canonical precursor to AD. We previously demonstrated that volume loss in bilateral anteromedial temporal lobe is present at baseline in longitudinally followed normal subjects who later developed MCI or AD. In this study we compared grey matter volumes within this predefined anteromedial temporal region (AMTR) at baseline between: 1) normal subjects enrolled in the Alzheimer's Disease Neuroimaging Initiative (ADNI) who subsequently developed cognitive complaints as reflected in a CDR memory box score of 0.5; and 2) normal subjects who remained normal over a median of 48 months of follow-up (CDR sum of boxes 0). We found significantly decreased volume within AMTR in the ADNI memory complainers. To relate AMTR results to those from conventional anatomy, we demonstrate that volumes extracted with the ICBM amygdala region had the best correspondence with AMTR volumes. In contrast, regions that have demonstrated volume loss in frank MCI and AD in ADNI, e.g., the posterior cingulate, did not show volume loss. These findings provide independent confirmation that volume changes preceding MCI occur in AMTR, a region of overlap between amygdala and anterior hippocampus.

Alzheimer's disease (AD) is a progressive brain disorder that initially affects medial temporal lobe circuitry and memory functions. Current drug treatments have only modest effects on the symptomatic course of the disease. In contrast, a growing body of evidence suggests that lifelong bilingualism may delay the onset of clinical AD symptoms by several years. The purpose of the present review is to summarize evidence for bilingualism as a reserve variable against AD and discuss potential underlying neurocognitive mechanisms. Evidence is reviewed suggesting that bilingualism may delay clinical AD symptoms by protecting frontostriatal and frontoparietal executive control circuitry rather than medial temporal lobe memory circuitry. Cellular and molecular mechanisms that may contribute to bilingual cognitive reserve effects are discussed, including those that may affect neuronal metabolic functions, dynamic neuronal-glial interactions, vascular factors, myelin structure and neuroche...

Recent evidence suggests specialization of anterior left inferior prefrontal cortex (aLIPC;~BA 45/47) for controlled semantics and of posterior LIPC (pLIPC;~BA 44/6) for controlled phonology. However, the more automated phonological tasks commonly used raise the possibility that some of the typically extensive aLIPC activation during semantic tasks may relate to controlled language processing beyond the semantic domain. In the present study, an event-related fMRI adaptation paradigm was employed that used a standard controlled semantic task and a phonological task that also emphasized controlled processing. When compared with letter (baseline) processing, significant fMRI task and adaptation effects in the aLIPC and pLIPC regions (~BA 45/47,~BA 44) were observed during both semantic and phonological processing, with aLIPC showing the strongest effects during semantic processing. A left frontal region (~BA 6) showed task and relative adaptation effects preferential for phonological processing, and a left temporal region (~BA 21) showed task and relative adaptation effects preferential for semantic processing. Our results demonstrate that aLIPC and pLIPC regions are involved in controlled processing across multiple language domains, arguing against a domain-specific LIPC model and for domain-preferentiality in left posterior frontal and temporal regions.