POLYPHASIC CHARACTERIZATION OF THREE STRAINS OF ANABAENA RENIFORMIS AND APHANIZOMENON APHANIZOMENOIDES (CYANOBACTERIA) AND THEIR RECLASSIFICATION TO SPHAEROSPERMUM GEN. NOV. (INCL. ANABAENA KISSELEVIANA)1

Occurrences of rare cyanobacteria Anabaena reniformis Lemmerm. and Aphanizomenon aphanizomenoides (Forti) Horecká et Komárek were recently detected at several localities in the Czech Republic. Two monoclonal strains of An. reniformis and one strain of Aph. aphanizomenoides were isolated from distant localities and different sampling years. They were characterized by a combination of morphological, genetic, and biochemical approaches. For the first time, partial 16S rRNA gene sequences were obtained for these morphospecies. Based on this gene, all of these strains clustered separately from other planktonic Anabaena and Aphanizomenon strains. They appeared in a cluster with Cylindrospermopsis Seenaya et Subba Raju and Raphidiopsis F. E. Fritsch et M. F. Rich, clustered closely together with two An. kisseleviana Elenkin strains available from GenBank. A new generic entity was defined (Sphaerospermum gen. nov., with the type species S. reniforme, based on the traditional species An. reniformis). These results contribute significantly to the knowledge base about genetic heterogeneity among planktonic Anabaena–like and Aphanizomenon–like morphospecies. Accordingly, the subgenus Dolichospermum, previously proposed for the group of planktonic Anabaena, should be revaluated. Secondary metabolite profiles of the An. reniformis and Aph. aphanizomenoides strains differed considerably from 17 other planktonic Anabaena strains of eight morphospecies isolated from Czech water bodies. Production of puwainaphycin A was found in both of the An. reniformis strains. Despite the relatively short phylogenetic distance from Cylidrospermopsis, the production of cylindrospermopsin was not detected in any of our strains.     

Circular dichroism and guanine quadruplexes

Circular dichroism (CD) is remarkably sensitive to the conformational states of nucleic acids; therefore, CD spectroscopy has been used to study most features of DNA and RNA structures. Quadruplexes are among the significant noncanonical nucleic acids architectures that have received special attentions recently. This article presents examples on the contribution of CD spectroscopy to our knowledge of quadruplex structures and their polymorphism. The examples were selected to demonstrate the potential of this simple method in the quadruplex field. As CD spectroscopy detects only the global feature of a macromolecule, it should preferably be used in combination with other techniques. On the other hand, CD spectroscopy, often as a pioneering approach, can reveal the formation of particular structural arrangements, to search for the conditions stabilizing the structures, to follow the transitions between various structural states, to explore kinetics of their appearance, to determine thermodynamic parameters and also detect formation of higher order structures. This article aims to show that CD spectroscopy is an important complementary technique to NMR spectroscopy and X-ray diffraction in quadruplex studies.     

Poly AB for an Immobilized Trypsin

Pig polyclonal antibodies against the biospecific complex of trypsin with its inhibitor “antilysine” were prepared by affinity chromatography on trypsin-bound beaded cellulose. The antibodies were characterised by ion exchange FPLC and SDS PAGE as pure IgG. The catalytic activity of trypsin was not affected by interaction with these antibodies, even in the presence of excess of antibody. Trypsin, biospecifically bound to CNBr-activated Sepharose 4B, displayed full catalytic activity.     

Stem cells: cross-talk and developmental programs

The thesis advanced in this essay is that stem cells-particularly those in the nervous system-are components in a series of inborn 'programs' that not only ensure normal development, but persist throughout life so as to maintain homeostasis in the face of perturbations-both small and great. These programs encode what has come to be called 'plasticity'. The stem cell is one of the repositories of this plasticity. This review examines the evidence that interaction between the neural stem cell (as a prototypical somatic stem cell) and the developing or injured brain is a dynamic, complex, ongoing reciprocal set of interactions where both entities are constantly in flux. We suggest that this interaction can be viewed almost from a 'systems biology' vantage point. We further advance the notion that clones of exogenous stem cells in transplantation paradigms may not only be viewed for their therapeutic potential, but also as biological tools for 'interrogating' the normal or abnormal central nervous system environment, indicating what salient cues (among the many present) are actually guiding the expression of these 'programs'; in other words, using the stem cell as a 'reporter cell'. Based on this type of analysis, we suggest some of the relevant molecular pathways responsible for this 'cross-talk' which, in turn, lead to proliferation, migration, cell genesis, trophic support, protection, guidance, detoxification, rescue, etc. This type of developmental insight, we propose, is required for the development of therapeutic strategies for neurodegenerative disease and other nervous system afflictions in humans. Understanding the relevant molecular pathways of stem cell repair phenotype should be a priority, in our view, for the entire stem cell field.     

The effect of reversion and duplication of leaf disks on the accuracy of photosynthesis determination by the dry weight method

Experiments with sunflower and fodder cabbage showed that reversion of the leaf disks lower side upwards (towards the radiation source) decreased the photosynthetic rate by as much as 26% both at full (2. 10⁵ erg. cm^?2. sec.^?1) and half density of irradiation. On placing two disks one on another the photosynthetic rate was decreased in the upper disk in a similar way as it was on reversion even if it were placed in the normal position (upper side upwards). But assimilation in the lower disk was only a fraction (0–40%) of that of the upper disk and in thick leaves was within the limits of the compensation point. The sum of the increase in dry weight in both disks due to photosynthesis exceeds the increment in one normally placed disk only in thin leaves and at high density of radiation. On the basis of the differences found it is advised to discard experimental samples where a quarter of the disks were reversed during exposure. On evaluating samples in which some of the disks were duplicated in error, it is most accurate to deduct the weight of one control disk from the weight obtained. The decrease in photosynthetic rate found in reversed and duplicated disks is evidently due to the simultaneous effect of differences in the supply of CO₂ and radiation energy to the assimilating tissue and apparatus and also to differences in the photosynthetic capacity of different leaf tissues.     

Virucidal Nanofiber Textiles Based on Photosensitized Production of Singlet Oxygen

Novel biomaterials based on hydrophilic polycaprolactone and polyurethane (Tecophilic®) nanofibers with an encapsulated 5,10,5,20-tetraphenylporphyrin photosensitizer were prepared by electrospinning. The doped nanofiber textiles efficiently photo-generate O₂(¹Δ_g), which oxidize external chemical and biological substrates/targets. Strong photo-virucidal effects toward non-enveloped polyomaviruses and enveloped baculoviruses were observed on the surface of these textiles. The photo-virucidal effect was confirmed by a decrease in virus infectivity. In contrast, no virucidal effect was detected in the absence of light and/or the encapsulated photosensitizer.     

The passive yet successful way of planktonic life: genomic and experimental analysis of the ecology of a free-living polynucleobacter population

Background

The bacterial taxon Polynucleobacter necessarius subspecies asymbioticus represents a group of planktonic freshwater bacteria with cosmopolitan and ubiquitous distribution in standing freshwater habitats. These bacteria comprise <1% to 70% (on average about 20%) of total bacterioplankton cells in various freshwater habitats. The ubiquity of this taxon was recently explained by intra-taxon ecological diversification, i.e. specialization of lineages to specific environmental conditions; however, details on specific adaptations are not known. Here we investigated by means of genomic and experimental analyses the ecological adaptation of a persistent population dwelling in a small acidic pond.

Findings

The investigated population (F10 lineage) contributed on average 11% to total bacterioplankton in the pond during the vegetation periods (ice-free period, usually May to November). Only a low degree of genetic diversification of the population could be revealed. These bacteria are characterized by a small genome size (2.1 Mb), a relatively small number of genes involved in transduction of environmental signals, and the lack of motility and quorum sensing. Experiments indicated that these bacteria live as chemoorganotrophs by mainly utilizing low-molecular-weight substrates derived from photooxidation of humic substances.

Conclusions

Evolutionary genome streamlining resulted in a highly passive lifestyle so far only known among free-living bacteria from pelagic marine taxa dwelling in environmentally stable nutrient-poor off-shore systems. Surprisingly, such a lifestyle is also successful in a highly dynamic and nutrient-richer environment such as the water column of the investigated pond, which was undergoing complete mixis and pronounced stratification in diurnal cycles. Obviously, metabolic and ecological versatility is not a prerequisite for long-lasting establishment of abundant bacterial populations under highly dynamic environmental conditions. Caution should be exercised when generalizing the obtained insights into the ecology and adaptation of the investigated lineage to other Polynucleobacter lineages.     

Karyological analysis of an interspecific hybrid between the dioecious Silene latifolia and the hermaphroditic Silene viscosa.

The genus Silene is a good model for studying evolution of the sex chromosomes, since it includes species that are hermaphroditic and dioecious, while maintain a basic chromosome number of 2n = 24. For some combinations of Silene species it is possible to construct interspecific hybrids. Here, we present a detailed karyological analysis of a hybrid between the dioecious Silene latifolia as the maternal plant and a related species, hermaphroditic Silene viscosa, used as a pollen partner. Using genomic probes (the genomic in situ hybridization (GISH) technique), we were able to clearly discriminate parental genomes and to show that they are largely separated in distinct nuclear domains. Molecular GISH and fluorescence in situ hybridization (FISH) markers document that the hybrid genome of somatic cells was strictly additive and stable, and that it had 12 chromosomes originating from each parent, including the only X chromosome of S. latifolia. Meiotic analysis revealed that, although related, respective parental chromosomes did not pair or paired only partially, which resulted in frequent chromosome abnormalities such as bridges and irregular non-disjunctions. GISH and FISH markers clearly document that the larger genome of S. latifolia and its largest chromosome component, the X chromosome, were mostly employed in chromosome lagging and misdivision.     

Resprouting after disturbance: an experimental study with short-lived monocarpic herbs

We experimentally demonstrated the ability of three short-lived monocarpic species to vegetatively regenerate (resprout) from roots after severe disturbance. We assessed the relationship between resprouting ability and (1) timing of injury with respect to life-cycle stage (reproductive vs. vegetative plant), life-history mode (annual vs. winter annual) and phenological stage (flowering vs. fruiting plant), (2) nutrient availability, and (3) disturbance severity (removal of all axillary buds Yes/No). In a chamber experiment with the annual or potentially winter annualRorippa palustris, all injured plants resprouted in all nutrient levels and day-length regimes (day-length regimes simulated conditions of an annual and a winter annual cohort). The number of adventitious buds on roots was positively affected only by injury. The extent of regeneration and amount of regenerated biomass were higher at high nutrient level and long-day regime.     

Higher perceived dominance in Toxoplasma infected men--a new evidence for role of increased level of testosterone in toxoplasmosis-associated changes in human behavior

Toxoplasma is parasite of cats that uses any warm-blooded animals as intermediate hosts. It is known to induce shifts in behavior, physiology and even morphology of its intermediate hosts, including humans. The lower second to fourth digit ratio (2D:4D ratio) in infected man and women, and higher height in infected man suggest that sex steroid hormones like testosterone could play a role in these shifts. Here, we searched for another indirect indication for a higher postnatal testosterone level, i.e. increased perceived dominance and masculinity in infected men. We showed portrait pictures of 89 male students of which 18 were Toxoplasma-infected to 109 female students. When we statistically corrected for age, men with latent toxoplasmosis were perceived as more dominant (p=0.009) and masculine (p=0.052). These results support the idea that the higher level of testosterone could be responsible for at least some of the toxoplasmosis-associated shifts in human and animal behavior.