The role of heterotrophic carbon acquisition by the hemiparasitic plant Rhinanthus alectorolophus in seedling establishment in natural communities: a physiological perspective

? Heterotrophic acquisition of substantial amounts of organic carbon by hemiparasitic plants was clearly demonstrated by numerous studies. Many hemiparasites are, however, also limited by competition for light preventing the establishment of their populations on highly productive sites. ? In a growth-chamber experiment, we investigated the effects of competition for light, simulated by shading, on growth and heterotrophic carbon acquisition by the hemiparasite Rhinanthus alectorolophus attached to C(3) and C(4) hosts using analyses of biomass production and stable isotopes of carbon. ? Shading had a detrimental effect on biomass production and vertical growth of the hemiparasites shaded from when they were seedlings, while shading imposed later caused only a moderate decrease of biomass production and had no effect on the height. Moreover, shading increased the proportion of host-derived carbon in hemiparasite biomass (up to 50% in shaded seedlings). ? These results demonstrate that host-derived carbon can play a crucial role in carbon budget of hemiparasites, especially if they grow in a productive environment with intense competition for light. The heterotrophic carbon acquisition can allow hemiparasite establishment in communities of moderate productivity, helping well-attached hemiparasites to escape from the critical seedling stage.     

Comparison of gemma production among three Lophozia species during the growing season

The extent and seasonal pattern of asexual reproduction and ability to germinate in the rare liverwort Lophozia ascendens and the common liverworts L. ventricosa and L. longiflora were studied in the Boubínský prales National Nature Reserve in Šumava Mts. (Bohemian Forest), South Bohemia, Czech Republic. Asexual reproduction was quantified as the number of gemmae produced per individual shoot. Numbers of gemmae per shoot among sampling months differed significantly; increase of gemma production was delayed in L. ascendens in comparison with gemma production of L. ventricosa and L. longiflora. We suggest that gemma production is influenced by environmental factors, mainly air humidity. Germinability of gemmae was low in early spring, highest in August and September and slightly depressed in October. This pattern suggests that rather mild winters in the Czech Republic cause the lower mortality of shoots during winter and the environmental pressure towards the production of dormant gemmae is not a prominent factor affecting the population dynamics of the species under study.     

Evening exposure to a light-emitting diodes (LED)-backlit computer screen affects circadian physiology and cognitive performance

Many people spend an increasing amount of time in front of computer screens equipped with light-emitting diodes (LED) with a short wavelength (blue range). Thus we investigated the repercussions on melatonin (a marker of the circadian clock), alertness, and cognitive performance levels in 13 young male volunteers under controlled laboratory conditions in a balanced crossover design. A 5-h evening exposure to a white LED-backlit screen with more than twice as much 464 nm light emission {irradiance of 0,241 Watt/(steradian × m(2)) [W/(sr × m(2))], 2.1 × 10(13) photons/(cm(2) × s), in the wavelength range of 454 and 474 nm} than a white non-LED-backlit screen [irradiance of 0,099 W/(sr × m(2)), 0.7 × 10(13) photons/(cm(2) × s), in the wavelength range of 454 and 474 nm] elicited a significant suppression of the evening rise in endogenous melatonin and subjective as well as objective sleepiness, as indexed by a reduced incidence of slow eye movements and EEG low-frequency activity (1-7 Hz) in frontal brain regions. Concomitantly, sustained attention, as determined by the GO/NOGO task; working memory/attention, as assessed by "explicit timing"; and declarative memory performance in a word-learning paradigm were significantly enhanced in the LED-backlit screen compared with the non-LED condition. Screen quality and visual comfort were rated the same in both screen conditions, whereas the non-LED screen tended to be considered brighter. Our data indicate that the spectral profile of light emitted by computer screens impacts on circadian physiology, alertness, and cognitive performance levels. The challenge will be to design a computer screen with a spectral profile that can be individually programmed to add timed, essential light information to the circadian system in humans.     

Factors Affecting Sleep/vigilance Behaviour in Incubating Mallards

Vigilance is a behavioural tactic that allows individuals to control their surroundings and to assess predation risk. In contrast, sleep is unique behavioural state with widely hypothesized restorative and energy‐saving functions, but reducing attentiveness and increasing susceptibility to predation. Sleeping birds resolve this conflict by interrupting sleep with short periods of eye opening (termed ‘scans’) during vigilant sleep. Miscellaneous environmental factors and sleeping postures may affect the perception of risk and corresponding vigilance level. Here, we investigated the influence of nest vegetation concealment, time of day and sleeping postures on the sleep/vigilance trade‐off in incubating Mallards (Anas platyrhynchos). We found that incubating females increased their vigilance with increasing nest vegetation cover facing the vigilant eye during both the day and the night periods; however, mean nest vegetation concealment did not affect female vigilance. Females also reduced their total vigilance along with scan frequency during the night period, while displaying the opposite pattern during the daylight. The rest‐sleeping position was preferred more during the night compared with the daylight period, and females were more vigilant in this position at night. Our data show that the nest vegetation concealment regardless of visual abilities during different light conditions, time of day and sleeping posture play an underlying role in antipredator vigilance during sleep in this cryptic ground‐nesting bird.     

hZwint-1 bridges the inner and outer kinetochore: identification of the kinetochore localization domain and the hZw10-interaction domain

Accurate chromosome segregation in mitosis is required to maintain genetic stability. hZwint-1 [human Zw10 (Zeste white 10)-interacting protein 1] is a kinetochore protein known to interact with the kinetochore checkpoint protein hZw10. hZw10, along with its partners Rod (Roughdeal) and hZwilch, form a complex which recruits dynein-dynactin and Mad1-Mad2 complexes to the kinetochore and are essential components of the mitotic checkpoint. hZwint-1 localizes to the kinetochore in prophase, before hZw10 localization, and remains at the kinetochore until anaphase, after hZw10 has dissociated. This difference in localization timing may reflect a role for hZwint-1 as a structural kinetochore protein. In addition to hZw10, we have found that hZwint-1 interacts with components of the conserved Ndc80 and Mis12 complexes in yeast two-hybrid and GST (glutathione transferase) pull-down assays. Furthermore, hZwint-1 was found to have stable FRAP (fluorescence recovery after photobleaching) dynamics similar to hHec1, hSpc24 and hMis12. As such, we proposed that hZwint-1 is a structural protein, part of the inner kinetochore scaffold and recruits hZw10 to the kinetochore. To test this, we performed mutagenesis-based domain mapping to determine which regions of hZwint-1 are necessary for kinetochore localization and which are required for interaction with hZw10. hZwint-1 localizes to the kinetochore through the N-terminal region and interacts with hZw10 through the C-terminal coiled-coil domain. The two domains are at opposite ends of the protein as expected for a protein that bridges the inner and outer kinetochore.     

Protein kinase A activity at the endoplasmic reticulum surface is responsible for augmentation of human ether-a-go-go-related gene product (HERG)

Human ether-a-go-go-related gene product (HERG) is a cardiac potassium channel commonly implicated in the pathogenesis of the long QT syndrome, type 2 (LQT2). LQT2 mutations typically have incomplete penetrance and affect individuals at various stages of their lives; this may mirror variations in intracellular signaling and HERG regulation. Previous work showed that sustained protein kinase A (PKA) activity augments HERG protein abundance by a mechanism that includes enhanced protein translation. To investigate the subcellular site of this regulation, we generated site-specific probes to the cytoplasmic surface of the endoplasmic reticulum (ER), the presumed locale of channel synthesis. Real-time FRET-based indicators demonstrated both cAMP and PKA activity at the ER. A PKA inhibitor targeted to the ER surface (termed p4PKIg) completely abolished PKA-mediated augmentation of HERG in HEK293 cells as well as rat neonatal cardiomyocytes. Immunofluorescence co-localization, targeted FRET-based PKA biosensors, phospho-specific antibodies, and in vivo phosphorylation experiments confirmed that p4PKIg is preferentially active at the ER surface rather than the plasma membrane. Rerouting this inhibitor to the outer mitochondrial membrane diminishes its ability to block cAMP-dependent HERG induction. Our results support a model where PKA-dependent regulation of HERG synthesis occurs at the ER surface. Furthermore, reagents generated for this study provide novel experimental tools to probe compartmentalized cAMP/PKA signaling within cells.     

Structure analysis of the Staphylococcus aureus UDP-N-acetyl-mannosamine dehydrogenase Cap5O involved in capsular polysaccharide biosynthesis

Bacterial UDP-sugar dehydrogenases are part of the biosynthesis pathway of extracellular polysaccharides. These compounds act as important virulence factors by protecting the cell from opsonophagocytosis and complement-mediated killing. In Staphylococcus aureus, the protein Cap5O catalyzes the oxidation of UDP-N-acetyl-mannosamine to UDP-N-acetyl-mannosaminuronic acid. Cap5O is crucial for the production of serotype 5 capsular polysaccharide that prevents the interaction of bacteria with both phagocytic and nonphagocytic eukaryotic cells. However, details of its catalytic mechanism remain unknown. We thus crystallized Cap5O and solved the first structure of an UDP-N-acetyl-mannosamine dehydrogenase. This study revealed that the catalytic cysteine makes a disulfide bond that has never been observed in other structurally characterized members of the NDP-sugar dehydrogenase family. Biochemical and mutagenesis experiments demonstrated that the formation of this disulfide bridge regulates the activity of Cap5O. We also identified two arginine residues essential for Cap5O activity. Previous data suggested that Cap5O is activated by tyrosine phosphorylation, so we characterized the phosphorylation site and examined the underlying regulatory mechanism.     

Primary hepatocytes from mice lacking cysteine dioxygenase show increased cysteine concentrations and higher rates of metabolism of cysteine to hydrogen sulfide and thiosulfate

The oxidation of cysteine in mammalian cells occurs by two routes: a highly regulated direct oxidation pathway in which the first step is catalyzed by cysteine dioxygenase (CDO) and by desulfhydration-oxidation pathways in which the sulfur is released in a reduced oxidation state. To assess the effect of a lack of CDO on production of hydrogen sulfide (H₂S) and thiosulfate (an intermediate in the oxidation of H₂S to sulfate) and to explore the roles of both cystathionine γ-lyase (CTH) and cystathionine β-synthase (CBS) in cysteine desulfhydration by liver, we investigated the metabolism of cysteine in hepatocytes isolated from Cdo1-null and wild-type mice. Hepatocytes from Cdo1-null mice produced more H₂S and thiosulfate than did hepatocytes from wild-type mice. The greater flux of cysteine through the cysteine desulfhydration reactions catalyzed by CTH and CBS in hepatocytes from Cdo1-null mice appeared to be the consequence of their higher cysteine levels, which were due to the lack of CDO and hence lack of catabolism of cysteine by the cysteinesulfinate-dependent pathways. Both CBS and CTH appeared to contribute substantially to cysteine desulfhydration, with estimates of 56 % by CBS and 44 % by CTH in hepatocytes from wild-type mice, and 63 % by CBS and 37 % by CTH in hepatocytes from Cdo1-null mice.