Interrupting timing in interval production and discrimination: similarities and differences

Interruptions in human timing have been studied in the last few years using temporal production and discrimination tasks. Expecting a break shortened perceived duration in both paradigms but manipulating break duration affected time production only, suggesting that preparatory processes might not take place in time discrimination. In time production, using cues revealed that providing information about the break may modulate the effect of break expectancy. For example, time was perceived as shorter when a break was expected in trials with no breaks, but forewarning participants of the break absence with a cue almost abolished the effect. In the present experiment, a tone was classified as "short" or "long" in a discrimination task. Location and duration of breaks were varied and cues were provided in some trials with no breaks. Results showed an effect of break expectancy: perceived duration shortened with increasing pre-break duration. Reducing expectancy with cues in uninterrupted tones decreased the proportion of "short" responses in long-tone trials, but not in short-tone trials. As in previous discrimination experiments, perceived duration was unaffected by varying break duration. Similarities and differences in results as well as in their interpretation when breaks are used in time production and time discrimination tasks are discussed.     

Gender‐dependent regulation of G‐protein‐gated inwardly rectifying potassium current in dorsal raphe neurons in knock‐out mice devoid of the 5‐hydroxytryptamine transporter

Agonists at G‐protein‐coupled receptors in neurons of the dorsal raphe nucleus (DRN) of knock‐out mice devoid of the serotonin transporter (5‐HTT^?/?) exhibit lower efficacy to inhibit cellular discharge than in wild‐type counterparts. Using patch‐clamp whole‐cell recordings, we found that a G‐protein‐gated inwardly rectifying potassium (GIRK) current is involved in the inhibition of spike discharge induced by 5‐HT_1A agonists (5‐carboxamidotryptamine (5‐CT) and (±)‐2‐dipropylamino‐8‐hydroxy‐1,2,3,4‐tetrahydronaphthalene hydrobromide (8‐OH‐DPAT); 50 nM–30 μM) in both wild‐type and 5‐HTT^?/? female and male mice. These effects were mimicked by 5′‐guanylyl‐imido‐diphosphate (Gpp(NH)p; 400 μM) dialysis into cells with differences between genders. The 5‐HTT^?/? knock‐out mutation reduced the current density induced by Gpp(NH)p in females but not in males. These data suggest that the decreased response of 5‐HT_1A receptors to agonists in 5‐HTT^?/? mutants reflects notably alteration in the coupling between G‐proteins and GIRK channels in females but not in males. Accordingly, gender differences in central 5‐HT neurotransmission appear to depend—at least in part—on sex‐related variations in corresponding receptor‐G protein signaling mechanisms. © 2006 Wiley Periodicals, Inc. J Neurobiol, 2006     

Interference between Streptococcus pneumoniae and Staphylococcus aureus: In vitro hydrogen peroxide-mediated killing by Streptococcus pneumoniae

The bactericidal activity of Streptococcus pneumoniae toward Staphylococcus aureus is mediated by hydrogen peroxide. Catalase eliminated this activity. Pneumococci grown anaerobically or genetically lacking pyruvate oxidase (SpxB) were not bactericidal, nor were nonpneumococcal streptococci. These results provide a possible mechanistic explanation for the interspecies interference observed in epidemiologic studies.     

Mitochondrial Superoxide Radicals Differentially Affect Muscle Activity and Neural Function

Cellular superoxide radicals (O₂⁻) are mostly generated during mitochondrial oxygen metabolism. O₂⁻ serves as the raw material for many reactive oxygen species (ROS) members like H₂O₂ and OH^.− radicals following its catalysis by superoxide dismutase (SOD) enzymes and also by autocatalysis (autodismutation) reactions. Mitochondrial ROS generation could have serious implications on degenerative diseases. In model systems overproduction of mitochondrial O₂⁻ resulting from the loss of SOD2 function leads to movement disorders and drastic reduction in life span in vertebrates and invertebrates alike. With the help of a mitochondrial SOD2 loss-of-function mutant, Sod2ⁿ²⁸³, we measured the sensitivity of muscles and neurons to ROS attack. Neural outputs from flight motor neurons and sensory neurons were unchanged in Sod2ⁿ²⁸³ and the entire neural circuitry between the giant fiber (GF) and the dorsal longitudinal muscles (DLM) showed no overt defect due to elevated ROS. Such insensitivity of neurons to mitochondrial superoxides was further established through neuronal expression of SOD2, which failed to improve survival or locomotive ability of Sod2ⁿ²⁸³. On the other hand, ultrastructural analysis of Sod2ⁿ²⁸³ muscles revealed fewer mitochondria and reduced muscle ATP production. By targeting the SOD2 expression to the muscle we demonstrate that the early mortality phenotype of Sod2ⁿ²⁸³ can be ameliorated along with signs of improved mobility. In summary, muscles appear to be more sensitive to superoxide attack relative to the neurons and such overt phenotypes observed in SOD2-deficient animals can be directly attributed to the muscle.BETWEEN Drosophila, mouse, and human, the enzymatic antioxidant defense system shares similar organization both structurally (Landis and Tower 2005) and functionally. Besides having a good degree of homology (Duttaroy et al. 1994; Landis and Tower 2005), other significant similarities include the presence of a single copy of Sod1 and Sod2 genes in each with no degree of functional complementation between these enzymes (Copin et al. 2000). While vertebrates have developed additional antioxidant defense enzymes such as glutathione peroxidase (Gpx) and extracellular superoxide dismutase (EcSOD or Sod3), neither Gpx nor an active SOD3 has been demonstrated in Drosophila, although a Sod3-like sequence has been identified (Landis and Tower 2005). Complete loss of SOD2 function is fatally injurious for both mice and Drosophila (Li et al. 1995; Lebovitz et al. 1996; Kirby et al. 2002; Duttaroy et al. 2003). The severe phenotypic effects of SOD2 loss of function have been attributed to elevated DNA damage and protein carbonylation (Golden and Melov 2001). SOD2 loss of function has also been attributed to “free radical attack” or “oxidative insult” on mitochondria where obvious mitochondrial damage was apparent from the inactivation of mitochondrial Fe-S cluster enzymes aconitase and succinate dehydrogenase (Melov et al. 1999; Kirby et al. 2002; Paul et al. 2007). Furthermore, impairment of cellular signaling, specifically those induced by reactive oxygen species (ROS) (Klotz 2005), might also play a very significant role in the early mortality effects of SOD2-deficient flies as indicated recently (Wicks et al. 2009).Sod2 null mice with damaged mitochondria display a number of pathologies including cardiomyopathy (Li et al. 1995), neurodegeneration, and seizures (Melov et al. 1998). Drosophila mutants of mitochondrial dysfunction are also claimed to be associated with neurodegeneration (Kretzschmar et al. 1997; Min and Benzer 1997, 1999; Rogina et al. 1997; Palladino et al. 2002, 2003; Celotto et al. 2006). In addition to the neurons, muscles are important targets for oxidative modification (Choksi and Papaconstantinou 2008; Choksi et al. 2008). Aerobic muscles with high mitochondrial content and high myoglobin levels, for example, show a significant increase in oxidative modification of all electron transport chain proteins compared to muscles with fewer mitochondria and less myoglobin (anaerobic muscle) (Choksi and Papaconstantinou 2008; Choksi et al. 2008). Mice lacking the Cu-ZnSOD enzyme suffer from a rapid loss of skeletal muscle mass, resembling an accelerated sarcopenia (Jackson 2006; Muller et al. 2006). We therefore set out to measure the impact of heightened superoxide concentration on neurons and muscles of Sod2ⁿ²⁸³ flies that are devoid of SOD2, the principal scavenger of superoxide radicals in mitochondria (Duttaroy et al. 2003; Belton et al. 2006).     

Conjugated Linoleic Acid Is a Preferential Substrate for Fatty Acid Nitration

The oxidation and nitration of unsaturated fatty acids by oxides of nitrogen yield electrophilic derivatives that can modulate protein function via post-translational protein modifications. The biological mechanisms accounting for fatty acid nitration and the specific structural characteristics of products remain to be defined. Herein, conjugated linoleic acid (CLA) is identified as the primary endogenous substrate for fatty acid nitration in vitro and in vivo, yielding up to 10⁵ greater extent of nitration products as compared with bis-allylic linoleic acid. Multiple enzymatic and cellular mechanisms account for CLA nitration, including reactions catalyzed by mitochondria, activated macrophages, and gastric acidification. Nitroalkene derivatives of CLA and their metabolites are detected in the plasma of healthy humans and are increased in tissues undergoing episodes of ischemia reperfusion. Dietary CLA and nitrite supplementation in rodents elevates NO₂-CLA levels in plasma, urine, and tissues, which in turn induces heme oxygenase-1 (HO-1) expression in the colonic epithelium. These results affirm that metabolic and inflammatory reactions yield electrophilic products that can modulate adaptive cell signaling mechanisms.     

Proteomics reveals potential biomarkers of seed vigor in sugarbeet

To unravel biomarkers of seed vigor, an important trait conditioning crop yield, a comparative proteomic study was conducted with sugarbeet seed samples of varying vigor as generated by an invigoration treatment called hydropriming and an aging treatment called controlled deterioration. Comparative proteomics revealed proteins exhibiting contrasting behavior between seed samples. Thus, 18 proteins were up-regulated during priming and down-regulated during aging and further displayed an up-regulation upon priming of the aged seeds, meaning that down-regulation of these spot volumes during aging was reversible upon subsequent priming. Also, 11 proteins exhibited the converse behavior characterized by a decrease and an increase of the spot volumes during priming and aging of the control seeds, respectively, and a decrease in the spot volumes upon priming of the aged seeds. The results underpinned the role in seed vigor of several metabolic pathways involved in lipid and starch mobilization, protein synthesis or the methyl cycle. They also corroborate previous studies suggesting that the glyoxylate enzyme isocitrate lyase, the capacity of protein synthesis and components of abscisic acid signaling pathways are likely contributors of seed vigor.     

High resolution imaging of vascular function in zebrafish

Rationale

The role of the endothelium in the pathogenesis of cardiovascular disease is an emerging field of study, necessitating the development of appropriate model systems and methodologies to investigate the multifaceted nature of endothelial dysfunction including disturbed barrier function and impaired vascular reactivity.

Objective

We aimed to develop and test an optimized high-speed imaging platform to obtain quantitative real-time measures of blood flow, vessel diameter and endothelial barrier function in order to assess vascular function in live vertebrate models.

Methods and Results

We used a combination of cutting-edge optical imaging techniques, including high-speed, camera-based imaging (up to 1000 frames/second), and 3D confocal methods to collect real time metrics of vascular performance and assess the dynamic response to the thromboxane A₂ (TXA₂) analogue, U-46619 (1 µM), in transgenic zebrafish larvae. Data obtained in 3 and 5 day post-fertilization larvae show that these methods are capable of imaging blood flow in a large (1 mm) segment of the vessel of interest over many cardiac cycles, with sufficient speed and sensitivity such that the trajectories of individual erythrocytes can be resolved in real time. Further, we are able to map changes in the three dimensional sizes of vessels and assess barrier function by visualizing the continuity of the endothelial layer combined with measurements of extravasation of fluorescent microspheres.

Conclusions

We propose that this system-based microscopic approach can be used to combine measures of physiologic function with molecular behavior in zebrafish models of human vascular disease.     

Distinct Effects on Diversifying Selection by Two Mechanisms of Immunity against Streptococcus pneumoniae

Antigenic variation to evade host immunity has long been assumed to be a driving force of diversifying selection in pathogens. Colonization by Streptococcus pneumoniae, which is central to the organism''s transmission and therefore evolution, is limited by two arms of the immune system: antibody- and T cell- mediated immunity. In particular, the effector activity of CD4⁺ T_H17 cell mediated immunity has been shown to act in trans, clearing co-colonizing pneumococci that do not bear the relevant antigen. It is thus unclear whether T_H17 cell immunity allows benefit of antigenic variation and contributes to diversifying selection. Here we show that antigen-specific CD4⁺ T_H17 cell immunity almost equally reduces colonization by both an antigen-positive strain and a co-colonized, antigen-negative strain in a mouse model of pneumococcal carriage, thus potentially minimizing the advantage of escape from this type of immunity. Using a proteomic screening approach, we identified a list of candidate human CD4⁺ T_H17 cell antigens. Using this list and a previously published list of pneumococcal Antibody antigens, we bioinformatically assessed the signals of diversifying selection among the identified antigens compared to non-antigens. We found that Antibody antigen genes were significantly more likely to be under diversifying selection than the T_H17 cell antigen genes, which were indistinguishable from non-antigens. Within the Antibody antigens, epitopes recognized by human antibodies showed stronger evidence of diversifying selection. Taken together, the data suggest that T_H17 cell-mediated immunity, one form of T cell immunity that is important to limit carriage of antigen-positive pneumococcus, favors little diversifying selection in the targeted antigen. The results could provide new insight into pneumococcal vaccine design.