Expression of a functional C5a receptor in regenerating hepatocytes and its involvement in a proliferative signaling pathway in rat

Activation of the complement system generates the anaphylatoxin C5a whose activities are mediated through its binding to the widely expressed C5aR. C5aR mRNA and protein expressions are known to be induced in rat hepatocytes under inflammatory conditions. However, little is known about the role of the C5a/C5aR complex in liver and its involvement during a proliferative process. We have evaluated the expression of C5aR in regenerating rat hepatocytes following a partial hepatectomy and in hepatocyte cultures. C5aR induction was observed in hepatocytes from regenerating liver, as well as in normal hepatocytes under a culture-induced stress. The effect of a stimulation by a C5a agonist upon the synthesis of a growth factor/receptor pair (hepatocyte growth factor/c-Met) was also evaluated. Our data demonstrated an up-regulated expression of hepatocyte growth factor and c-Met mRNAs, but we failed to observe a direct mitogenic effect of C5a in culture. However, a significantly increased expression of cyclin E and D1mRNA levels, as well as an increased BrdU incorporation, were observed in rats given an i.v. C5a agonist injection following an 80% partial hepatectomy. These studies demonstrate for the first time that: 1) C5aR is up-regulated during liver regeneration, 2) the binding of C5a to C5aR promotes a growth response, and 3) C5aR is involved in a cell cycle signaling pathway. Taken together, these findings point to a novel role for the hepatic C5aR implicating this complement system in the context of normal or abnormal proliferative pathways.     

Identification and characterization of a eukaryotically encoded rubredoxin in a cryptomonad alga

We have identified an open reading frame with homology to prokaryotic rubredoxins (rds) on a nucleomorph chromosome of the cryptomonad alga Guillardia theta. cDNA analysis let us propose that the rd preprotein has an NH(2)-terminal extension that functions as a transit peptide for import into the plastid. Compared to rds found in non-photosynthetic prokaryotes or found in bacteria that exhibit an anoxigenic photosynthesis apparatus, nucleomorph rd has a COOH-terminal extension, which shows high homology exclusively to the COOH-termini of cyanobacterial rds as well as to a hypothetical rd in the Arabidopsis genome. This extension can be divided into a putative membrane anchor and a stretch of about 20 amino acids with unknown function linking the common rd fold to this anchor. Overexpression of nucleomorph rd in Escherichia coli using a T7 RNA polymerase/promotor system resulted in a mixture of iron-containing holorubredoxin and zinc-substituted protein. Preliminary spectroscopic studies of the iron form of nucleomorph rd suggest the existence of a native rd-type iron site. One-dimensional nuclear magnetic resonance spectroscopy of recombinant Zn-rd suggests the presence of a stable tertiary fold similar to that of other rd structures determined previously.     

Demography, life history and migrations in a Mexican mantled howler group in a rainforest fragment

This paper represents the results of a long-term study (1996-2003) on the demographic changes over time of a Mexican mantled howler (Alouatta palliata mexicana) group in a rainforest fragment (40 ha) in Los Tuxtlas, Mexico, with a follow-up census 3 years later (2006). In addition to demographic and life history parameters, we describe six dispersal events. Our results suggest that this group has been expanding during the study period, growing from six to 12 individuals, with an annual average intrinsic growth rate of 0.07, an infant survivorship of 67%, and an average immature to female ratio of 0.90. This increase in size is probably related to the high food availability in their home range. However, fragment isolation may be negatively affecting the dispersal patterns typical of the species, which could result in a loss of genetic variability over time.     

Synthesis and characterization of a biotin-alginate conjugate and its application in a biosensor construction

Biotin was covalently coupled with alginate in an aqueous-phase reaction by means of carbodiimide-mediated activation chemistry to provide a biotin-alginate conjugate for subsequent use in biosensor applications. The synthetic procedure was optimized with respect to pH of the reaction medium (pH 6.0), the degree of uronic acid activation (20%), and the order of addition of the reagents. The biotin-alginate conjugate was characterized by titration with 2-anilinonaphthalene-6-sulfonic acid (2,6-ANS), 4-hydroxyazobene-2'-carboxylic acid (HABA) and by an HPSEC-MALLS analytical method as well as by FTIR and 13C NMR spectroscopy. As a compromise between the need for a high percent of molar modification of the alginate, on one hand, and sufficient gelling capability, on the other hand, an optimal modification of 10-13% of biotin-alginate was used. The new biotin-alginate conjugate was used for the encapsulation of bioluminescent reporter cells into microspheres. A biosensor was prepared by conjugating these biotinylated alginate microspheres to the surface of a streptavidin-coated optical fiber, and the performance of the biosensor was demonstrated in the determination of the antibiotic, mitomycin C as a model toxin.     

Chaos and synchrony in a model of a hypercolumn in visual cortex

Neurons in cortical slices emit spikes or bursts of spikes regularly in response to a suprathreshold current injection. This behavior is in marked contrast to the behavior of cortical neurons in vivo, whose response to electrical or sensory input displays a strong degree of irregularity. Correlation measurements show a significant degree of synchrony in the temporal fluctuations of neuronal activities in cortex. We explore the hypothesis that these phenomena are the result of the synchronized chaos generated by the deterministic dynamics of local cortical networks. A model of a hypercolumn in the visual cortex is studied. It consists of two populations of neurons, one inhibitory and one excitatory. The dynamics of the neurons is based on a Hodgkin-Huxley type model of excitable voltage-clamped cells with several cellular and synaptic conductances. A slow potassium current is included in the dynamics of the excitatory population to reproduce the observed adaptation of the spike trains emitted by these neurons. The pattern of connectivity has a spatial structure which is correlated with the internal organization of hypercolumns in orientation columns. Numerical simulations of the model show that in an appropriate parameter range, the network settles in a synchronous chaotic state, characterized by a strong temporal variability of the neural activity which is correlated across the hypercolumn. Strong inhibitory feedback is essential for the stabilization of this state. These results show that the cooperative dynamics of large neuronal networks are capable of generating variability and synchrony similar to those observed in cortex. Auto-correlation and cross-correlation functions of neuronal spike trains are computed, and their temporal and spatial features are analyzed. In other parameter regimes, the network exhibits two additional states: synchronized oscillations and an asynchronous state. We use our model to study cortical mechanisms for orientation selectivity. It is shown that in a suitable parameter regime, when the input is not oriented, the network has a continuum of states, each representing an inhomogeneous population activity which is peaked at one of the orientation columns. As a result, when a weakly oriented input stimulates the network, it yields a sharp orientation tuning. The properties of the network in this regime, including the appearance of virtual rotations and broad stimulus-dependent cross-correlations, are investigated. The results agree with the predictions of the mean field theory which was previously derived for a simplified model of stochastic, two-state neurons. The relation between the results of the model and experiments in visual cortex are discussed.     

Development of a method to evaluate caspase-3 activity in a single cell using a nanoneedle and a fluorescent probe

A method to detect an enzymatic reaction in a single living cell using an atomic force microscope equipped with an ultra-thin needle (a nanoneedle) and a fluorescent probe molecule was developed. The nanoneedle enables the low-invasive delivery of molecules attached onto its surface directly into a single cell. We hypothesized that an enzymatic reaction in a cell could be profiled by monitoring a probe immobilized on a nanoneedle introduced into the cell. In this study, a new probe substrate (NHGcas546) for caspase-3 activity based on fluorescent resonance energy transfer (FRET) was constructed and fixed on a nanoneedle. The NHGcas546-modified nanoneedle was inserted into apoptotic cells, in which caspase-3 is activated after apoptosis induction, and a change in the emission spectrum of the immobilized probe could be observed on the surface of the nanoneedle. Thus, we have developed a successful practical method for detecting a biological phenomenon in a single cell. We call the method MOlecular MEter with Nanoneedle Technology (MOMENT).     

Sleep in amphibians and reptiles: a review and a preliminary analysis of evolutionary patterns

Despite the ubiquitous nature of sleep, its functions remain a mystery. In an attempt to address this, many researchers have studied behavioural and electrophysiological phenomena associated with sleep in a diversity of animals. The great majority of vertebrates and invertebrates display a phase of immobility that could be considered as a sort of sleep. Terrestrial mammals and birds, both homeotherms, show two sleep states with distinct behavioural and electrophysiological features. However, whether these features have evolved independently in each clade or were inherited from a common ancestor remains unknown. Unfortunately, amphibians and reptiles, key taxa in understanding the evolution of sleep given their position at the base of the tetrapod and amniote tree, respectively, remain poorly studied in the context of sleep. This review presents an overview of what is known about sleep in amphibians and reptiles and uses the existing data to provide a preliminary analysis of the evolution of behavioural and electrophysiological features of sleep in amphibians and reptiles. We also discuss the problems associated with analysing existing data, as well as the difficulty in inferring homologies of sleep stages based on limited data in the context of an essentially mammalian‐centric definition of sleep. Finally, we highlight the importance of developing comparative approaches to sleep research that may benefit from the great diversity of species with different ecologies and morphologies in order to understand the evolution and functions of sleep.     

Synthesis and characterization of a pyrrole-alginate conjugate and its application in a biosensor construction

N-(3-Aminopropyl)pyrrole was covalently coupled with alginate in an aqueous-phase reaction by means of carbodiimide-mediated activation chemistry to provide a pyrrole-alginate conjugate for subsequent use in biosensor applications. The pyrrole-alginate conjugate was quantified by UV spectroscopy at 230 nm, by an HPSEC-MALLS analytical method, as well as by FTIR and 13C NMR spectroscopies. The new pyrrole-alginate conjugate was used for the immobilization of polyphenol oxidase (PPO) onto an electrode surface by physical entrapment resulting from the gellification process and electrochemical polymerization of the pyrrole groups. The efficiency of this cross-linking approach (chemical and electrochemical) was investigated by comparing the amount of enzyme released from polypyrrole-alginate and regular alginate. In addition, biosensors were prepared by entrapment of the PPO in polypyrrole-alginate and regular alginate matrixes and their performance for the amperometric determination of catechol chosen as a model analyte was examined, yielding a sensitivity of 350 and 80 microA M(-1) cm(-2), respectively, for polypyrrole-alginate and alginate biosensors.     

Hunting and predation in a fiddler crab

Fiddler crabs are known primarily to be deposit feeders. They eat detritus, bacteria, and other small particles of organic material found in the sandy or muddy substrate on which they live. They have highly specialized mouthparts used to separate edible matter from nondigestable material. Here we provide evidence of cannibalism and predation in a fiddler crab, Uca annulipes. We additionally provide the first evidence of a fiddler crab hunting shrimp and insects. This study is an exemplary reminder that, even though an animal may have evolved highly specialized feeding traits, this need not preclude it from opportunistically acting as a generalist feeder.     

Incidence and remediation of fungi in a sick building in Malaysia: a case study

Problems caused by indoor microbial, especially fungal growth, have been further exaggerated by the increased incidence of water intrusions, condensation from air-conditioning system, and other factors. While fungal contamination in a building can be one of the indicators of indoor air quality (IAQ), air quality assessment and remediation should be considered as important and should be carried out systematically. This study reports the incidence and remediation of an excessive fungal growth in a building in Universiti Sains Malaysia (USM), Pulau Pinang, Malaysia. Acting on an official report by the building owner and occupants, an immediate walk-through investigation was carried out between June and December 2009. A thorough sampling comprising swab and spore impactor techniques showed that the colony-forming unit per cubic meter of air (CFU/m³) levels were far above those in the guidelines of most developed countries. Thus, the building was declared a ‘sick building’ and closed to the publics. However, through immediate action with recommended methods to overcome the problem, the premise has been pronounced safe. This is a good example of successful teamwork that involved a continuous investigation by a team of experts along with comprehensive action taken by the occupants and other authorities responsible for the building.     

Development of tool use in a macaque and a gorilla

The development of the capacity to use a stick as a tool was tested in a macaque (Macaca fuscata) and a gorilla (Gorilla gorilla gorilla) infants that had previously shown to be able to use strings and supports as dragging tools. Subjects were tested between 15 and 38 months of age. Different levels of competence between the subjects emerged over testing. The macaque developed a stereotyped strategy to cope with the problem, only getting random successes, whilst the gorilla developed a flexible strategy and revealed to be able to mentally represent the solution of the problem. In fact, when not successful using the stick, the gorilla thought out an alternative strategy, choosing and adapting a new object to use it as a tool.     

Kleptoparasitism and complexity in a multi-trophic web

In this work the effects of kleptoparitism in a multi-trophic food-web, in which an omnivorous species scavenges the kills of a top predator, are investigated. Scavengers are assumed to be able to steal predators’ kills by direct interference and aggression. The amount of prey shared depends on the relative competitive abilities of omnivores and predators and on their abundances. To make the proposed model consistent, scavenging is accompanied by other features. First, the predator can prey only on the juvenile life stage of scavengers (i.e. not on adult individuals) as well as on a different species of herbivores. Hence, an age structure is enforced within scavengers, so that they belong to the guild of preys when they are young and vulnerable. Second, predators can switch between prey species selecting the most abundant one. This condition grants the competitive exclusion of the two vegetation eaters, allowing them to co-exist: the omnivore and the herbivore compete for the same vegetation pool but pure herbivores are indeed assumed to be more efficient in its exploitation. The omnivorous species is not able to kill the herbivores, but nonetheless it may exploit their carcasses after they have been killed by predators. Finally, the juvenile/adult ratio of omnivores varies depending on the available resources and predators’ amount. In such a scenario it is shown that kleptoparasitism can modify to a large extent the stability of the system, leading either to a regularization or to chaos, depending both on the scavenger's and the predator's functional response. Moreover, an excess of kleptoparasitism is shown to compromise the whole trophic web, making extinct both predators and scavengers. As far as the authors are aware of, this paper is the first one to explicitly introduce kleptoparasitism in trophic web models.     

Isomerization and oxidation of vitamin a in cone-dominant retinas: a novel pathway for visual-pigment regeneration in daylight

The first step toward light perception is 11-cis to all-trans photoisomerization of the retinaldehyde chromophore in a rod or cone opsin-pigment molecule. Light sensitivity of the opsin pigment is restored through a multistep pathway called the visual cycle, which effects all-trans to 11-cis re-isomerization of the retinoid chromophore. The maximum throughput of the known visual cycle, however, is too slow to explain sustained photosensitivity in bright light. Here, we demonstrate three novel enzymatic activities in cone-dominant ground-squirrel and chicken retinas: an all-trans-retinol isomerase, an 11-cis-retinyl-ester synthase, and an 11-cis-retinol dehydrogenase. Together these activities comprise a novel pathway that regenerates opsin photopigments at a rate 20-fold faster than the known visual cycle. We suggest that this pathway is responsible for sustained daylight vision in vertebrates.     

Human C3a and C3a desArg anaphylatoxins have conserved structures,in contrast to C5a and C5a desArg

Complement is a part of innate immunity that has a critical role in the protection against microbial infections, bridges the innate with the adaptive immunity and initiates inflammation. Activation of the complement, by specific recognition of molecular patterns presented by an activator, for example, a pathogen cell, in the classical and lectin pathways or spontaneously in the alternative pathway, leads to the opsonization of the activator and the production of pro‐inflammatory molecules such as the C3a anaphylatoxin. The biological function of this anaphylatoxin is regulated by carboxypeptidase B, a plasma protease that cleaves off the C‐terminal arginine yielding C3a desArg, an inactive form. While functional assays demonstrate strikingly different physiological effects between C3a and C3a desArg, no structural information is available on the possible conformational differences between the two proteins. Here, we report a novel and simple expression and purification protocol for recombinant human C3a and C3a desArg anaphylatoxins, as well as their crystal structures at 2.3 and 2.6 Å, respectively. Structural analysis revealed no significant conformational differences between the two anaphylatoxins in contrast to what has been reported for C5a and C5a desArg. We compare the structures of different anaphylatoxins and discuss the relevance of their observed conformations to complement activation and binding of the anaphylatoxins to their cognate receptors.     

Oxygen Isotope Fractionation during Photosynthesis in a Blue-Green and a Green Alga

Oxygen isotope fractionation (¹⁸O/¹⁶O) at the natural abundance level has been measured during photosynthesis of a blue-green and a green alga. When sufficient attention is paid to removal of contaminating air O₂ before and during the experiments, then the photosynthetic O₂ evolved, as compared to the water O₂, had an average difference of −0.36% for a blue-green alga and −0.80% for a green alga. These experiments suggest that there is no reason to invoke an inverse isotope effect in photosynthesis as part of the explanation for the ¹⁸O enrichment in atmospheric O₂ relative to O₂ in oceanic waters. In addition, in an indirect way, the experiments also support the argument that the bulk of O₂ evolved during photosynthesis comes from water. A 10% contribution of O₂ arising from CO₂ would have been detectable in the present work.     

Bifurcation and chaos for a mutating autocatalator in a CSTR

In this paper, we develop a conceptual model for autocatalytic reactions in which the autocatalyst undergoes a mutation process in a completely stirred tank reactor. Three cases with different mutation coefficient, !, and mutation efficiency, #, have been considered. The analysis of these cases shows the importance of the two parameters and the complexity of the resulting bifurcation diagrams. Co-existence of a mushroom and an isola have been determined for the first case. The isola portion is a low main product yield solution and is to be avoided. A sub-region of 7 co-existing steady states, with three stable static branches, has been determined for the second case. Chaos developing through period doubling and type 1 intermittency has been shown to occur for a fourth case.     

Nutrient retention and release in a floodplain's aquifer and in the hyporheic zone of a lowland river

Fertilizer applications and other non-point sources result in an increasing diffuse N and P pollution of receiving waters degrading water quality by eutrophication with several adverse impacts. Floodplains are regarded as reactive interfaces between uplands and receiving waters. In the present study groundwater quality on its subsurface flow from an upland area through a lowland floodplain towards the receiving water body of the Spree River was monitored biweekly over 2 years with two transects of 18 groundwater observation wells. Within the floodplain reaction rates of the nutrients are unevenly distributed. On a scale smaller than the floodplain, the hyporheic zone is regarded as reactive interface with unproportional high reaction rates. Therefore, phosphate and dissolved iron were measured with high spatial resolution in the pore water of the riverbed and the oxbow bed to investigate turnover processes and their small-scale spatial variability at the immediate surface–subsurface interface. The biogeochemical composition of subsurface water is characterized by little temporal variability while spatial heterogeneity is high on the hectametre scale of the study site as well as on the centimetre scale of the bed sediments. Nitrate is eliminated very efficiently by denitrification in the anoxic aquifer of the floodplain while ammonium and phosphate concentrations increase under anoxic conditions. Phosphate and ammonium originate from the mineralization of organic matter and phosphate is additionally released by reductive dissolution of iron-bound phosphorus and weathering of bedrock. Sorption–desorption processes equalize temporal fluctuations of phosphate concentrations. Phosphate uptake by plants is assumed as an important process at only one of the groundwater observation wells. Redox conditions required for a phosphate sink are opposite to those involved in nitrate removal by denitrification. Thus, redox patchiness of floodplain aquifers favours nitrate and phosphate removal, i.e. a temporal and spatial sequence of anoxic and oxic conditions eliminates nitrogen and causes phosphate storage. On the groundwater's path from the upland to the river further phosphate is released in the bed sediments. It originates from previously settled particulate compounds containing phosphorus. While the release of iron-bound phosphorus clearly predominates in the riverbed sediments the mineralization of organic matter is an important additional phosphorus release process in the oxbow bed sediments.     

Imitation of a bimanual task in preschool- and school-age children: a hierarchical construction

The present study examined the development of bimanual interaction during the imitation of a live demonstration. To this end, children of five different age groups observed an adult model performing in an object manipulation task consisting to open a box with one hand, taking out an object with the other hand, and closing the box again, before they were asked to imitate this motor task under different imitation conditions. The children's responses were videotaped, coded in dichotomous data, and then transformed in percentage scores. The main results showed that all children were able to imitate/attain the goal of the task. However, differences were observed for the different imitation conditions, which were also reflected in some age effects, while hand dominance was a strong constraint on imitation. Also, practice did not seem to increase the likelihood of model imitation. These findings confirm that imitation is a reconstruction mechanism hierarchically organized.