Biosystems

Human Bocavirus (HBoV) is a novel virus which can cause respiratory tract disease in infants or children. In this study, the codon usage bias and the base composition variations in the available 11 complete HBoV genome sequences have been investigated. Although, there is a significant variation in codon usage bias among different HBoV genes, codon usage bias in HBoV is a little slight, which is mainly determined by the base compositions on the third codon position and the effective number of codons (ENC) value. The results of correspondence analysis (COA) and Spearman's rank correlation analysis reveals that the G+C compositional constraint is the main factor that determines the codon usage bias in HBoV and the gene's function also contributes to the codon usage in this virus. Moreover, it was found that the hydrophobicity of each protein and the gene length are also critical in affecting these viruses' codon usage, although they were less important than that of the mutational bias and the genes' function. At last, the relative synonymous codon usage (RSCU) of 44 genes from these 11 HBoV isolates is analyzed using a hierarchical cluster method. The result suggests that genes with same function yet from different isolates are classified into the same lineage and it does not depend on geographical location. These conclusions not only can offer an insight into the codon usage patterns and gene classification of HBoV, but also may help in increasing the efficiency of gene delivery/expression systems.

Spectral and kinetic transformations were studied in gelatin films made with 14-F wild type (WT) bacteriorhodopsin (BR) and 14-F D96N mutant BR. Unlike the recent study of water suspensions of the same pigments, where a red shifted species at 660 nm was shown to form under the light in 14-F WT only, there are no drastic differences in photoinduced behavior between gelatin films based on 14-F WT and 14-F D96N. It is not observed any photoinduced formation of red shifted species at 660 nm for both types of films as it is observed for corresponding pigments in water suspension. The observed results are explained in a terms of relationship between the rates of two photoinduced processes that occur in suspensions and films of corresponding pigments. Kinetic characteristics of the photoinduced processes for the films with chemical additives suggest that there are no advantages in using 14-F D96N films when compared to films based on 14-F WT.

Photosynthetic assimilation of CO(2) in plants results in the balance between the photochemical energy developed by light in chloroplasts, and the consumption of that energy by the oxygenation processes, mainly the photorespiration in C(3) plants. The analysis of classical biological models shows the difficulties to bring to fore the oxygenation rate due to the photorespiration pathway. As for other parameters, the most important key point is the estimation of the electron transport rate (ETR or J), i.e. the flux of biochemical energy, which is shared between the reductive and oxidative cycles of carbon. The only reliable method to quantify the linear electron flux responsible for the production of reductive energy is to directly measure the O(2) evolution by (18)O(2) labelling and mass spectrometry. The hypothesis that the respective rates of reductive and oxidative cycles of carbon are only determined by the kinetic parameters of Rubisco, the respective concentrations of CO(2) and O(2) at the Rubisco site and the available electron transport rate, ultimately leads to propose new expressions of biochemical model equations. The modelling of (18)O(2) and (16)O(2) unidirectional fluxes in plants shows that a simple model can fit the photosynthetic and photorespiration exchanges for a wide range of environmental conditions. Its originality is to express the carboxylation and the oxygenation as a function of external gas concentrations, by the definition of a plant specificity factor Sp that mimics the internal reactions of Rubisco in plants. The difference between the specificity factors of plant (Sp) and of Rubisco (Sr) is directly related to the conductance values to CO(2) transfer between the atmosphere and the Rubisco site. This clearly illustrates that the values and the variation of conductance are much more important, in higher C(3) plants, than the small variations of the Rubisco specificity factor. The simple model systematically expresses the reciprocal variations of carboxylation and oxygenation exchanges illustrated by a "mirror effect". It explains the protective sink effect of photorespiration, e.g. during water stress. The importance of the CO(2) compensation point, in classical models, is reduced at the benefit of the crossing points Cx and Ox, concentration values where carboxylation and oxygenation are equal or where the gross O(2) uptake is half of the gross O(2) evolution. This concept is useful to illustrate the feedback effects of photorespiration in the atmosphere regulation. The constancy of Sp and of Cx for a great variation of P under several irradiance levels shows that the regulation of the conductance maintains constant the internal CO(2) and the ratio of photorespiration to photosynthesis (PR/P). The maintenance of the ratio PR/P, in conditions of which PR could be reduced and the carboxylation increased, reinforces the hypothesis of a positive role of photorespiration and its involvement in the plant-atmosphere co-evolution.

Numerous studies focus on the measurement of conductances for CO2 transfer in plants and especially on their regulatory effects on photosynthesis. Measurement accuracy is strongly dependent on the model used and on the knowledge of the flow of photochemical energy generated by light in chloroplasts. The only accurate and precise method to quantify the linear electron flux (responsible for the production of reductive energy) is the direct measurement of O2 evolution, by 18O2 labelling and mass spectrometry. The sharing of this energy between the carboxylation (P) and the oxygenation of photorespiration (PR) depends on the plant specificity factor (Sp) and on the corresponding atmospheric concentrations of CO2 and O2 (André, 2013). The concept of plant specificity factor simplifies the equations of the model. It gives a new expression of the effect of the conductance (g) between atmosphere and chloroplasts. Its quantitative effect on photosynthesis is easy to understand because it intervenes in the ratio of the plant specificity factor (Sp) to the specificity of Rubisco (Sr). Using this ‘simple’ model with the data of 18O2 experiments, the calculation of conductance variations in response to CO2 and light was carried out.

The search for genes in a newly sequenced DNA is a well known problem. Among other factors, the gene-searching process is hampered by a number of ambiguities which may remain unresolved experimentally for a long time. A computer method that is able to predict genes in a DNA sequence containing ambiguities has been developed, based on the non-homogeneous Markov chain technique. The reliability of the method has been tested using a set of sequences generated by a Monte-Carlo procedure and a set of 425 E. coli sequences with ambiguities introduced artificially.

The studies of Rubisco characteristics observed during plant evolution show that the variation of the Rubisco specificity factor only improved by two times from cyanobacteria to modern C3 plants. However we note important variations of the ratio between the maximum rates of oxygenation and carboxylation (V(O)/V(C)). Modelling in vivo ¹⁸O₂ data in plant gas exchange shows that the oxygenation reaction of Rubisco plays a regulating role when the photochemical energy exceeds the carboxylation capacity. A protective index 'oxygenation capacity' is postulated, related to the ratio V(O)/V(C) of Rubisco, and hence to the sink energy effect of photorespiration. Analysing the trends of Rubisco parameters along the evolutionary scale, we show: (1) the increase of both V(C) and V(O); (2) the enhancement of CO₂ affinity; and (3) the rise in oxygenation capacity at the expense of the CO₂ specificity. Hence, the factors of evolutionary pressure have not only directed the enzyme towards a more efficient utilisation of CO₂, but mainly to positively use the unavoidable great loss of energy and assimilated carbon in the process of photorespiration. These observations reinforce the hypothesis of plant-atmosphere co-evolution and of the complex role of Rubisco, which seems to be selected to develop both better CO₂ affinity and oxygenation capacity. The latter increases the capacity of sink of photorespiration, in particular, during water stress or under high irradiance, the two conditions experienced by plants in terrestrial environments. These observations help to explain some handicaps of C4 plants, and the supremacy of CAM and C3 perennial higher plants in arid environments.

In closed systems, the O₂) compensation point (Γ₀) was previously defined as the upper limit of O₂ level, at a given CO₂ level, above which plants cannot have positive carbon balance and survive. Studies with ¹⁸O₂ measure the actual O₂ uptake by photorespiration due to the dual function of Rubisco, the enzyme that fixes CO₂ and takes O₂ as an alternative substrate. One-step modelling of CO₂ and O₂ uptakes allows calculating a plant specificity factor (Sp) as the sum of the biochemical specificity of Rubisco and a biophysical specificity, function of the resistance to CO₂ transfer from the atmosphere to Rubisco. The crossing points (Cx, Ox) are defined as CO₂ and O₂ concentrations for which O₂ and CO₂ uptakes are equal. It is observed that: (1) under the preindustrial atmosphere, photorespiration of C3 plants uses as much photochemical energy as photosynthesis, i.e. the Cx and Ox are equal or near the CO₂ and O₂ concentrations of that epoch; (2) contrarily to Γ(C), a Γ₀ does not practically limit the plant growth, i.e. the plant net CO₂ balance is positive up to very high O₂ levels; (3) however, in a closed biosystem, Γ₀ exists; it is not the limit of plant growth, but the equilibrium point between photosynthesis and the opposite respiratory processes; (4) a reciprocal relationship exists between Γ₀ and Γ(C), as unique functions of the respective CO₂ and O₂ concentrations and of Sp, this invalidates some results showing two different functions for Γ₀ and Γ(C), and, consequently, the associated analyses related to greenhouse effects in the past; (5) the pre-industrial atmosphere levels of O₂ and CO₂ are the Γ₀ and Γ(C) of the global bio-system. They are equal to or near the values of Cx and Ox of C3 plants, the majority of land plants in preindustrial period. We assume that the crossing points represent favourable feedback conditions for the biosphere-atmosphere equilibrium and could result from co-evolution of plants-atmosphere-climate. We suggest that the evolution of Rubisco and associated pathways is directed by an optimisation between photosynthesis and photorespiration.

A model for the process of translation in gene expression is proposed. The model is based on the assumption that the ribosome decodes the mRNA sequences using consecutive subsequences of the 3(')-end of its 16S rRNA subunit. The biological consistency of the model is validated by successful detection of the Shine-Dalgarno signal and the start codon. Furthermore, implications on the role of the 3(')-end in the complete process of prokaryotic translation are presented and discussed. Interestingly, the results obtained support the possibility of an involvement of this part of the ribosome in the process of translation termination. Subsequently, results obtained via the proposed model are compared with published experimental results for different mutations of the last 13 bases of the 16S rRNA molecule. Agreement between predictions and experimental results validate the biological relevance of the proposed model. By means of simulated nucleotide mutations, a global analysis of this part of the ribosome in the process of translation is conducted.

I propose two postulates that allow precise matching of an indefinite number of neurites when repairing a transected nerve. The first one, based on the principle of de Saint-Venant, indicates how to keep nerve fibers in proper longitudinal alignment, by diverting stress to an appropriate distance from the tip of the stumps. The second postulate, based on the symmetrical mechanical resistance of the stumps, indicates how to restore correct transversal alignment of the fibers, by circularizing the severed nerve extremities.

The decline in stocks of commercial fish species has been documented in several regions of the world. This decline is due partially to the effect of evolutionary pressure caused by the management of fishing activity, which reduces the size of fish after a few generations. In this paper, the population dynamics of the Pintado Pseudoplatystoma corruscans, one of the main commercial species of freshwater fish in Brazil, were simulated considering different scenarios of fishing mortality and different minimum and maximum lengths of capture. The results show that selective fishing based on the different proposed selectivity curves can result in an evolution-mediated increase in the growth rate of the fish, the biomass and the catch. This suggests that appropriate changes in Brazilian legislation can contribute to the sustainability of fisheries and to conservation of the fish stocks exploited by man.

For the true fungi, phylogenetic relationships inferred from 18S ribosomal DNA sequence data agree with morphology when (1) the fungi exhibit diagnostic morphological characters, (2) the sequence-based phylogenetic groups are statistically supported, and (3) the ribosomal DNA evolves at roughly the same rate in the lineages being compared. 18S ribosomal RNA gene sequence data and biochemical data provide a congruent definition of true fungi. Sequence data support the traditional fungal subdivisions Ascomycotina and Basidiomycotina. In conflict with morphology, some zygomycetes group with chytrid water molds rather than with other terrestrial fungi, possibly owing to unequal rates of nucleotide substitutions among zygomycete lineages. Within the ascomycetes, the taxonomic consequence of simple or reduced morphology has been a proliferation of mutually incongruent classification systems. Sequence data provide plausible resolution of relationships for some cases where reduced morphology has created confusion. For example, phylogenetic trees from rDNA indicate that those morphologically simple ascomycetes classified as yeasts are polyphyletic and that forcible spore discharge was lost convergently from three lineages of ascomycetes producing flask-like fruiting bodies.

A study of phylogenetic relationships of the colonial green algal flagellates based on nuclear 18S and 26S rRNA sequence data suggests that the colonial habit has had at least two independent origins. All colonial taxa included in the analysis, except Stephanosphaera, are allied in a clade with Chlamydomonas reinhardtii and other Chlamydomonas taxa ascribed to the Euchlamydomonas group by Ettl. In contrast, Stephanosphaera is allied with other unicellular flagellates including Haematococcus. Comparison of the 18S and 26S data shows that the two sets of data yield different results following cladistic analysis. The 18S data provide the principal signal that supports the more basal divergences, but the data do not unambiguously address relationships among taxa in the clade that includes most colonial flagellates and Chlamydomonas taxa representative of the Euchlamydomonas group (sensu Ettl). In contrast, the 26S data have fewer informative sites that support basal divergences than the 18S data, but provide much of the signal that supports resolution of taxa in the colonial flagellate clade in an analysis of the combined 18S and 26S rRNA sequence data. Additional sequence data from the 26S molecule and additional taxa may reduce the topological ambiguity inferred from the sequence data for the colonial flagellates. Alternatively, an ancient and rapid radiation of taxa in the colonial lineage could account for the topological ambiguity. Despite some unresolved questions of relationships, cladistic analysis of the combined data sets provides some robustly supported concepts of evolution in these flagellates.

Recent classifications of the phylum Ciliophora are compared with the system proposed by Small and Lynn in 1981 and revised in 1985. The classes COLPODEA, PHYLLOPHARYNGEA, OLIGOHYMENOPHOREA and NASSOPHOREA sensu Small & Lynn are considered monophyletic by a number of researchers. The classes LITOSTOMATEA and PROSTOMATEA sensu Small & Lynn are not as well accepted. Some divergence of opinion arises in recognition of the categories included in the classes KARYORELICTEA and SPIROTRICHEA sensu Small & Lynn, in which the great diversity of cortical ultrastructure is still not recognized in the taxonomy. Numerous minor groups, for example the phacodiniids, plagiopylids and schizocaryids, present taxonomic problems that may not be solved by ultrastructural examination. This is also true of the subphyletic divisions where there are substantial differences of opinion about relationships among the classes.

A new macrosystem for the phylum Ciliophora is described. It is based primarily on the concept of the structural conservatism of the cortical fibrillar structures. Three subphyla, the Postciliodesmatophora, Rhabdophora, and Cyrtophora, are described. These three subphyla are divided respectively into the following classes: (1) the Postciliodesmatophora, into the Karyorelictea and Spirotrichea; (2) Rhabdophora, into the Prostomea and the Litostomea n, nov.; and (3) Cyrtophorea, into the Nassophorea n. nov., Phyllopharyngea, Coipodea, and Oligohymenophorea. The subclass divisions of these eight classes are briefly discussed. Problem areas are indicated where further research will test the relationship proposed by this new macrosystem.

The ultrastructure of Coleps bicuspis Noland, 1925 is described. The ciliate is a typical prostomate: the somatic kinetid is a monokinetid with a postciliary ribbon at triple 9, a kinetodesmal fibril originating near triplets 5, 6, 7 and an apparently radial transverse ribbon at triplet 4. The oral area is circular and has three brosse kineties associated with it. The brosse kineties are composed of dikinetids whose anterior kinetosome bears a tangential transverse ribbon and whose posterior kinetosome bears the fibrillar associates typical of a somatic monokinetid. The oral dikinetids are oriented parallel to the circumference of the oral cavity, which is surrounded by oral papillae and oral ridges. Pairs of nematodesmata, originating from oral dikinetid kinetosomes, are typically triangular in transection. A phylogeny of rhabdophoran ciliates is presented using the mixed parsimony algorithm and is discussed with reference to the systematic revisions of the phylum Ciliophora.

This paper summarizes Michael Conrad's academic and professional career from the time he began his Ph.D. studies in 1964 to his appointment at Wayne State University in 1979. It describes the origins of several of his major research interests and presents a personal evaluation of how this early work continues to be of fundamental importance.

The notion of hypercycles has been introduced (Eigen, 1971) in order to characterize a functional entity which integrates information stored in individual self-replicating elements. Commonly, these self-replicating elements compete, like polynucleotides do in test tube experiments (Spiegelman, 1971; Biebricher et al., 1981) or, like species do in nature. A single species is selected eventually together with its most frequent mutants and the less efficient competitors disappear. In order to suppress competition, specific coupling terms of second order - these are coupling terms which are proportional to the product of two population numbers or concentration - have to be introduced into the kinetic equations such a link has to extend to all members, requiring the formation of a closed catalytic cycle (Eigen and Schuster, 1979). The copying of polynucleotide sequences, like every transfer of information, can only occur with finite accuracy (Eigen et al., 1981). This physical restriction imposes a limit upon the content of information that can be transmitted. A higher content of information requires more precise replication. A more involved replication machinery, however, presupposes more information to build it. Thus, a single autocatalyst will not be able to increase its content of information over the given threshold. How does one escape from this vicious circle? Hypercycles present a solution to the problem: many information carriers coexist and may cooperate to build a more sophisticated and more precisely copying replication machinery. Optimization of growth rates has never been the goal for which hypercycles were conceived. Any first order process can proceed at a higher rate than a second order process if low enough concentrations are chosen. On the other hand, any realistic hypercycle involves both first and second order terms. At low concentrations first order terms provide for accumulation of material while at higher concentration second order terms stabilize cooperation. At which concentration level such a prevalence changes is a matter of quantitative values of rate parameters rather than of basic principle. An autocatalyst may outgrow a hypercycle under certain conditions but it can never solve the problem of integrating information.

Chronic hepatitis C patients display many genotype-specific clinical features of HCV infection. The core proteins encoded by different genotypes dysregulate numerous sets of distinct host genes. In this study we tested the hypothesis that HCV core proteins 1b and 3a would actually act on a limited number of independent cellular players, as well as on several functionally linked gene products. Structural and functional tests identified a core set of host genes dysregulated by HCV core genotypes 1b and 3a. The core proteins of HCV genotypes 1b and 3a target specifically limited sets of functionally related gene products, which may be responsible for the variations in the clinical spectra associated with HCV infection.