Multiple Lytic Origins of Replication Are Required for Optimal Gammaherpesvirus Fitness In Vitro and In Vivo.

An unresolved question in herpesvirus biology is why some herpesviruses contain more than one lytic origin of replication (oriLyt). Using murine gammaherpesvirus 68 (MHV-68) as model virus containing two oriLyts, we demonstrate that loss of either of the two oriLyts was well tolerated in some situations but not in others both in vitro and in vivo. This was related to the cell type, the organ or the route of inoculation. Depending on the cell type, different cellular proteins, for example Hexim1 and Rbbp4, were found to be associated with oriLyt DNA. Overexpression or downregulation of these proteins differentially affected the growth of mutants lacking either the left or the right oriLyt. Thus, multiple oriLyts are required to ensure optimal fitness in different cell types and tissues.

Nanoparticle exposure reactivates latent herpesvirus and restores a signature of acute infection.

BACKGROUND: Inhalation of environmental (nano) particles (NP) as well as persistent herpesvirus-infection are potentially associated with chronic lung disease and as both are omnipresent in human society a coincidence of these two factors is highly likely. We hypothesized that NP-exposure of persistently herpesvirus-infected cells as a second hit might disrupt immune control of viral latency, provoke reactivation of latent virus and eventually lead to an inflammatory response and tissue damage. RESULTS: To test this hypothesis, we applied different NP to cells or mice latently infected with murine gammaherpesvirus 68 (MHV-68) which provides a small animal model for the study of gammaherpesvirus-pathogenesis in vitro and in vivo. In vitro, NP-exposure induced expression of the typically lytic viral gene ORF50 and production of lytic virus. In vivo, lytic viral proteins in the lung increased after intratracheal instillation with NP and elevated expression of the viral gene ORF50 could be detected in cells from bronchoalveolar lavage. Gene expression and metabolome analysis of whole lung tissue revealed patterns with striking similarities to acute infection. Likewise, NP-exposure of human cells latently infected with Epstein-Barr-Virus also induced virus production. CONCLUSIONS: Our results indicate that NP-exposure of persistently herpesvirus-infected cells - murine or human - restores molecular signatures found in acute virus infection, boosts production of lytic viral proteins, and induces an inflammatory response in the lung - a combination which might finally result in tissue damage and pathological alterations.

The COMTp.Val158Met Polymorphism and Cognitive Performance in Adult Development, Healthy Aging and Mild Cognitive Impairment.

BACKGROUND: The impact of genetic polymorphisms on cognition is assumed to increase with age as losses of brain resources have to be compensated for. We investigate the relation of catechol-O-methyltransferase (COMT)p.Val158Met polymorphism and cognitive capacity in the course of adult development, healthy aging and the development of mild cognitive impairment (MCI) in two birth cohorts of subjects born between 1930 and 1932 or between 1950 and 1952. METHODS: Thorough neuropsychological assessment was conducted in a total of 587 participants across three examination waves between 1993 and 2008. The COMT genotype was determined as a restriction fragment length polymorphism after PCR amplification and digestion with NlaIII. RESULTS: Significant effects of the COMTp.Val158Met polymorphism were identified for attention and cognitive flexibility in the younger but not the older cohort. CONCLUSION: These results confirm the importance of the COMTp.Val158Met genotype on tasks assessing attention and cognitive flexibility in midlife but not in healthy aging and the development of MCI. Our findings suggest that the influence of COMT changes as a function of age, decreasing from midlife to aging.

What to do and how to do it: action representations in tool use.

Research on bimanual coordination has shown that the efficiency of programming an action is determined by the way the action is cognitively represented. In tool use, actions can be represented with respect to the spatial goal of the action (e.g., the nail that is to be hit by a hammer) or with respect to the tool and its transformation (i.e., the function that maps external target locations onto corresponding bodily movements). We investigated whether the way of cuing bimanual actions with tools affects their cognitive representation and the efficiency with which they are programmed. In one group of participants, tool transformations were specified by symbolic cues, whereas the targets were indicated by direct spatial cues. In another group of participants, symbolic cues specified the targets of the tool-use actions, whereas tool transformations were indicated by direct spatial cues. In a third group, both targets and tool transformations were cued directly by spatial cues. It was hypothesized that different cognitive representations would result in more or less efficient programming of the action. Results indicated longer reaction times and a higher error rate in the group with symbolic cuing of the targets as compared to the group with symbolic cuing of the transformations. The latter did not differ much from the direct cuing group. These results suggest that it is more efficient to represent bimanual tool-use actions in terms of the tool transformations involved than in terms of the targets at which they are directed.

Cognitive activity, education and socioeconomic status as preventive factors for mild cognitive impairment and Alzheimer's disease.

Growing epidemiological evidence suggests that premorbid participation in cognitive leisure activities (CLA) reduces the risk of dementia by increasing cognitive reserve. We investigated the differential effect of CLA, education, and socioeconomic status (SES) on the development of mild cognitive impairment (MCI) and Alzheimer's disease (AD). Participants in the prospective population-based ILSE study (*1930-1932; 12-year follow-up) were examined in three examination waves (t1:1993/94; t2:1997/98; t3:2005/07). In total, 381 subjects of the original cohort (n=500) were re-examined at t3. Of these subjects 29% received the diagnosis of MCI and 7% of AD. Subjects participated in a thorough psychogeriatric examination and neuropsychological testing. Moreover, they took part in a detailed autobiographical interview and completed questionnaires including socio-demographic data and current frequency of participation in CLA. Subjects who were highly cognitively active at t1 had a significantly reduced risk of developing MCI/AD at t3 (scores adjusted for education, SES, gender, and depressive symptoms). Additionally, high education and high SES separately reduced the risk of MCI and AD. Our results confirm the hypothesis that a high level of CLA acts as a protective factor against the development of MCI and AD by increasing cognitive reserve. This effect is not accounted for by important potential confounders.

Coordinative constraints in bimanual tool use.

This study investigates coordinative constraints when participants execute discrete bimanual tool use actions. Participants moved two levers to targets that were either presented near the proximal parts of the levers or near the distal tips of the levers. In the first case, the tool transformation (i.e. the relationship between hand movement direction and target direction) was compatible, whereas in the second case, it was incompatible. We hypothesized that an egocentric constraint (i.e. a preference for moving the hands and tools in a mirror-symmetrical fashion) would be dominant when targets are presented near the proximal parts of the levers because in this situation, movements can be coded in terms of body-related coordinates. Furthermore, an allocentric constraint (i.e. a preference to move the hands in the same (parallel) direction in extrinsic space) was expected to be dominant when one of the targets or both are presented near the distal parts of the levers because in this condition, movements have to be coded in an external reference frame. The results show that when both targets are presented near the proximal parts of the levers, participants are faster and produce less errors with mirror-symmetrical when compared to parallel movements. Furthermore, the RT mirror-symmetry advantage is eliminated, when both targets are presented near the distal parts of the levers, and it is reversed, when the target for one lever is presented near its distal part and the target for the other lever is presented near its proximal part. These results show that the dominance of egocentric and allocentric coordinative constraints in bimanual tool use depends on whether movements are coded in terms of body-related coordinates or in an external reference frame.

Impairment of T-regulatory cells in cord blood of atopic mothers.

BACKGROUND: Maternal atopy is a strong predictor for the development of childhood allergic diseases. The underlying mechanisms are ill defined, yet regulatory T (Treg) and T(H)17 cells may play a key role potentially shaping the early immune system toward a proallergic or antiallergic immune regulation. OBJECTIVE: We examined T(H)1/T(H)2, Treg, and T(H)17 cell responses to innate (lipid A/peptidoglycan) and mitogen/adaptive (phytohemagglutinin/Dermatophagoides pteronyssinus 1) immune stimulation in cord blood from offspring of atopic/nonatopic mothers. METHODS: Cord blood mononuclear cells from 161 healthy neonates (59% nonatopic, 41% atopic mothers) were investigated regarding Treg and T(H)17 cells (mRNA/surface markers), suppressive function, and proliferation/cytokine secretion. RESULTS: Cord blood from offspring of atopic mothers showed fewer innate-induced Treg cells (CD4(+)CD25(+)high), lower mRNA expression of associated markers (glucocorticoid-induced tumor necrosis factor receptor-related protein/lymphocyte activation gene 3; P < .05), and a trend toward lower Forkhead box transcription factor 3 (Foxp3) expression. Treg cell function was impaired in mitogen-induced suppression of T effector cells in cord blood of offspring from atopic mothers (P = .03). Furthermore, IL-10 and IFN-gamma secretion were decreased in innate-stimulated cord blood of offspring from atopic mothers (P = .04/.05). Innate-induced IL-17 was independent of maternal atopy and highly correlated with IL-13 secretion. CONCLUSION: In offspring of atopic mothers, Treg cell numbers, expression, and function were impaired at birth. T(H)17 cells were correlated with T(H)2 cells, independently of maternal atopy.

Binding of multidimensional context information as a distinctive characteristic of remember judgments.

This research investigated the cognitive processes underlying remember-know judgments in terms of contextual binding in multidimensional source memory. Stochastic dependence between the retrieval of different context attributes, which formed the empirical criterion of binding, was observed for remembered items but not for known items. Experiment 1 showed that the qualitative difference in the stochastic relation holds even if quantitative source-memory performance is equated for items with remember and know judgments. Experiment 2 generalized the findings to context information from different modalities, and Experiment 3 ruled out a spurious stochastic dependence due to interindividual differences. Supporting recent dual-process models of remember-know judgments, the findings show that remember and know judgments differ with respect to binding processes that correspond to episodic recollection.

Herpesviruses and Their Host Cells: A Successful Liaison.

During a long history of coevolution, herpesviruses have reached a fine-tuned balance with their hosts, allowing them to successfully persist and spread to new hosts without causing too much damage. Only under certain circumstances, as in neonates or immunocompromised individuals, they may cause serious diseases. The delicate balance between herpesviruses and their hosts results from interactions of a great variety of viral and cellular factors which together shape the tropism for a particular host, tissue, or cell. Understanding these interactions will provide insight into the viral life cycle and cell biology in general. Moreover, it will also facilitate comprehension of herpesvirus pathogenesis, enabling the development of new strategies to combat herpesviruses in cases where they cause disease.

Diabetes mellitus Type II and cognitive capacity in healthy aging, mild cognitive impairment and Alzheimer's disease.

While diabetes mellitus (DM) Type II has repeatedly been linked to Alzheimer´s disease (AD) and mild cognitive impairment (MCI), longitudinal research is scarce and disease duration has not always been taken into account. In a birth cohort born between 1930 and 1932 we investigated the influence of DM Type II and disease duration on neuropsychological functioning (memory/learning, attention, verbal fluency, visuospatial thinking and abstract thinking) across 14 years. Subjects who developed MCI or AD performed significantly poorer on all neuropsychological tests applied. While significant main effects DM Type II did not arise, its presence led to a significant deterioration of performance in the digit symbol test and visuospatial thinking over time. Additionally, in visuospatial thinking this change was more pronounced for individuals suffering from MCI/AD. We found that, as a concomitant disease DM Type II does not affect memory functioning, which is typically compromised in MCI and early AD. Rather, it may lead to deficits in cognitive flexibility and visuospatial thinking. DM Type II can be considered a frequent comorbid condition which can aggravate the course of MCI and AD. In this respect it may serve as a model for other comorbid conditions in AD.

The small noncoding RNAs (sncRNAs) of murine gammaherpesvirus 68 (MHV-68) are involved in regulating the latent-to-lytic switch in vivo.

The human gammaherpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV), which are associated with a variety of diseases including tumors, produce various small noncoding RNAs (sncRNAs) such as microRNAs (miRNAs). Like all herpesviruses, they show two stages in their life cycle: lytic replication and latency. During latency, hardly any viral proteins are expressed to avoid recognition by the immune system. Thus, sncRNAs might be exploited since they are less likely to be recognized. Specifically, it has been proposed that sncRNAs might contribute to the maintenance of latency. This has already been shown in vitro, but the respective evidence in vivo is very limited. A natural model system to explore this question in vivo is infection of mice with murine gammaherpesvirus 68 (MHV-68). We used this model to analyze a MHV-68 mutant lacking the expression of all miRNAs. In the absence of the miRNAs, we observed a higher viral genomic load during late latency in the spleens of mice. We propose that this is due to a disturbed regulation of the latent-to-lytic switch, altering the balance between latent and lytic infection. Hence, we provide for the first time evidence that gammaherpesvirus sncRNAs contribute to the maintenance of latency in vivo.

Bimanual interference associated with handling different tool transformations.

Research on bimanual coordination of hand movements has identified several loci of bimanual interference, including interference because of programming different movement parameters or selecting different targets for the two hands. This study investigates the extent and origin of interference when participants execute bimanual actions with tools. In the experiments, participants moved two tools, one with each hand, to two directly cued target locations. One type of tool transformed the body movement into a spatially compatible movement of the effective part of the tool, whereas the other transformed it into a spatially incompatible movement. Tool transformations for the two hands were either the same or different. Furthermore, target locations were either in the same or in different spatial directions. Results indicated significantly shorter reaction times (RTs) and less errors when tool transformations were the same for both hands. In addition, movements were initiated more quickly and less error-prone when targets were in the same direction, but this effect was modulated by the congruency of the two lever transformations. Investigations of the time course of the effects revealed that they were not because of early perceptual processing (Experiment 2). Furthermore, the general pattern of results occurred for different grip positions (Experiment 3) and different lever types (Experiment 4), suggesting that it reflects rather general constraints in bimanual coordination of tool-use actions.

Physical fitness as a protective factor for cognitive impairment in a prospective population-based study in Germany.

To evaluate the predictive effects of subjective measures of physical activity (PA) and objective measures of physical fitness (PF) on dementia risk, Participants of the prospective population-based ILSE-study (*1930-1932; 12-year follow-up) were examined at three examination waves (t1 : 1993/94; t2 : 1997/98; t3 : 2005/07). 381 subjects of the original cohort (n = 500) were re-examined at t3. 29% of the subjects who were cognitively healthy at baseline received the diagnosis of mild cognitive impairment (MCI) and 7% of Alzheimer's disease (AD). Subjects were screened for physical and mental health using medical interviews, physical, and neuropsychological examinations. Participants completed a questionnaire on their current and past PA at t1. Subjects were classified as physically active if they reported a regular sport activity for at least 2 hours per week in the past year. Muscular strength (handgrip) and motor coordination (balance) served as objective indicators of PF. Subjects who passed the balance-test at t1 had a reduced risk of developing MCI/AD at t3 (OR = 0.35, 95%CI 0.19-0.66, p < 0.01) and performed significantly better on various neuropsychological measures. Muscular strength or subjective reports of PA did not predict MCI/AD development. Our results confirm the hypothesis that PF acts as a protective factor for the development of cognitive disorders. In our study, context, motor coordination served as a better predictor than muscular strength or self-rated PA. Since subjects with cognitive disorders due to cerebral and/or systemic disorders were excluded from the analyses, our findings suggest that the effect of skill-related PF extends beyond the reduction of cardiovascular risk factors.

Inhibition of T-cell proliferation by murine multipotent mesenchymal stromal cells is mediated by CD39 expression and adenosine generation.

Multipotent mesenchymal stromal cells (MSCs) are bone marrow-derived cells of nonhematopoietic origin with immunoregulatory properties. Although some functions of MSCs have been identified, there are still features that are not explained thus far. The aim of the present study was to identify novel factors involved in MSC-mediated inhibition of T-cell proliferation. We here demonstrate that the surface molecule CD39 is coexpressed in concert with CD73 on murine MSCs catalyzing the generation of adenosine, which can directly act on activated T cells via the adenosine A2A receptor. Blocking of the adenosine pathway either by the A2A receptor antagonist SCH58261 or the specific CD39 inhibitor polyoxotungstate 1 (POM-1) blocked MSC-mediated suppression of T-cell proliferation almost completely. We conclude that CD39/CD73 coexpression is a novel important component of the immunoregulatory functions of murine MSCs. Our findings may both be important to improve our understanding of MSC function and for the development of immunomodulatory cellular therapies.

Bimanual interference with compatible and incompatible tool transformations.

The present study investigates bimanual interference in a tool-use task, in which two target locations had to be touched concurrently with two tools, one for each hand. Target locations were either in the same, or in different directions for the two hands. Furthermore, the tools implemented either a compatible or an incompatible relationship between the direction of target locations and the direction of associated bodily movements. Results indicated bimanual interference when the tools had to be moved to targets in different directions. Furthermore, this interference was much more pronounced when the tools required body movements that were spatially incompatible to the cued target locations as compared to when they were compatible. These results show that incompatible relationships between target directions and bodily movement directions can aggravate bimanual interference in tool use.

Macrophages driven to a novel state of activation have anti-inflammatory properties in mice.

Recurrent episodes of inflammation underlie numerous pathologies, notably those of inflammatory bowel diseases. In this study, we describe a population of macrophages in a novel state of activation that mitigates colitis in mice. The cells responsible for this effect, called IFN-gamma-stimulated monocyte-derived cells (IFNgamma-MdC), derive from mouse spleen, blood, and bone marrow monocytes and are distinguished from known macrophage populations by mode of generation, cell surface phenotype, and function. IFNgamma-MdC only arise when macrophages are cultivated in the presence of CD40L-expressing CD4+ T cells, M-CSF, and IFN-gamma. IFNgamma-MdC express markers including F4/80, CD11b/c, CD86, and CD274; they are negative for CD4, CD8, Gr1, CD19, CD80, and CD207. Functionally, IFNgamma-MdC are defined by their capacity to enrich cocultured T cell populations for CD4+CD25+Foxp3+ regulatory cells; this enrichment, constituting up to 60% or more of residual lymphocytes, is attributed to an expansion, but also to a cell contact and caspase-dependent depletion of activated T cells. In mice, IFNgamma-MdC delivered i.v. traffic to gut-associated peripheral lymphoid tissues, including the mesenteric lymph nodes, Peyer's patches, and colonic mucosa, and promote the clinical and histological resolution of chronic colitis. We conclude that IFNgamma-MdC represent macrophages in a novel state of activation, possessing multiple T cell-suppressive effects with therapeutic potential for the treatment of autoimmune inflammation.

Boundaries of the relation between conscious recollection and source memory for perceptual details.

The relation between conscious recollection and source memory for perceptual details was investigated in three experiments that combined the remember-know paradigm with a multidimensional source monitoring test. Experiment 1 replicated that source memory for perceptual details is better in the case of "remember" than "know" judgments. Experiment 2 showed that the relation between "remember" judgments and source memory for perceptual details is diminished by a semantic orienting task during encoding. Experiment 3 demonstrated that "remember" judgments are related to enhanced source memory for specific and unique kinds of perceptual source information, whereas memory for incomplete and global perceptual source information does not differentiate between "remember" and "know" judgments. The results show that the attentional focus during encoding and the specificity of retrieved source information form boundary conditions for the use of source memory for perceptual details as a basis of "remember" judgments.

Metacognitive inferences in source memory judgements: the role of perceived differences in item recognition.

This research investigated the hypothesis that metacognitive inferences in source memory judgements are based on the recognition or nonrecognition of an event together with perceived or expected differences in the recognizability of events from different sources. The hypothesis was tested with a multinomial source-monitoring model that allowed separation of source-guessing tendencies for recognized and unrecognized items. Experiments 1A and 1B manipulated the number of item presentations as relevant source information and revealed differential guessing tendencies for recognized and unrecognized items, with a bias to attribute unrecognized items to the source associated with poor item recognition. Experiments 2A and 2B replicated the findings with a manipulation of presentation time and extended the analysis to subjective differences in item recognition. Experiments 3A and 3B used more natural source information by varying type of acoustic signal and demonstrated that subjective theories about differences in item recognition are sufficient to elicit differential source-guessing biases for recognized and unrecognized items. Together the findings provide new insights into the cognitive processes underlying source memory decisions, which involve episodic memory and reconstructive tendencies based on metacognitive beliefs and general world knowledge.

DX5+ NKT cells induce the death of colitis-associated cells: involvement of programmed death ligand-1.

NKT cells are activated by CD1d and show an immune regulating function. Here, we investigated whether DX5+ NKT cells could be used to reduce colitis in a chronic colitis mouse model and studied the potential immunological mechanisms involved. Chronic colitis was induced either by transfer of enriched CD62L+ CD4+ T cells to severe-combined-immunodeficient mice or by feeding dextran sodium sulfate to immune competent mice. DX5+ NKT cells were transferred to mice with chronic colitis. Co-transfer of DX5+ NKT cells, but not CD8+ control cells, prevented the onset of colitis, and the immune regulatory effect of DX5+ NKT cells was completely abrogated by injecting CD1d blocking antibody. Moreover, DX5+ NKT cells reduced established colitis in both chronic colitis models. In vitro, DX5+ NKT cells induced cell death of colon-infiltrating lymphocytes isolated from diseased mice. This effect was inhibited in the presence of either anti-CD1d or anti-programmed death ligand-1 (PD-L1) blocking antibodies. The specific potency of DX5+ NKT cells in regulating chronic colitis in two mouse models is demonstrated. In vitro testing suggests that DX5+ NKT cells activated by CD1d induce cell death of colitis-inducing lymphocytes, which is mediated through PD-L1. Therefore, DX5+ NKT cells could be important in the regulation of immune responses associated with chronic colitis.

Blocking MAdCAM-1 in vivo reduces leukocyte extravasation and reverses chronic inflammation in experimental colitis.

BACKGROUND: Leukocyte recruitment to sites of intestinal inflammation is a crucial multi-step process, leading ultimately to the accumulation of cells in the inflamed tissue. These interactions in the gut are critically dependent on the mucosal addressin cell adhesion molecule-1 (MAdCAM-1), which is expressed on endothelial cells within the mesenteric lymph nodes and the lamina propria of the intestine. Here, we investigate the pathophysiologic role of MAdCAM-1 in the intestinal microcirculation in vivo. METHODS: Using a standard mouse model, chronic colitis was established after four cycles of dextran sodium sulfate (DSS) application. MAdCAM-1 expression was investigated by immunohistochemistry and Western blotting, as well as real-time polymerase chain reaction (PCR). Intravital microscopy was used to study the role of MAdCAM-1 on leukocyte-endothelium interactions and leukocyte extravasation. RESULTS: Significant changes in MAdCAM-1 were observed in mice with chronic DSS-induced colitis. Upregulation of MAdCAM-1 expression in chronic colitis was demonstrated on a protein and messenger ribonucleic acid (mRNA) level. Anti-MAdCAM-1 treatment lead to a marked reduction (>60%) of leukocyte sticking and extravasation in vivo, compared to the controls. This was parallelled by a significant reduction (45%) of intestinal inflammation, as measured by the histologic grading score. CONCLUSION: These in vivo results demonstrate a distinct role of MAdCAM-1 in inflammatory intestinal diseases, and suggest that therapeutic strategies targeting this adhesion molecule could be useful in the treatment of chronic colitis.