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Background Recombinant carbohydrases genes are used to produce transgenic woody plants with improved phenotypic traits. However, cultivation of such plants in open field is challenged by a number of problems. Therefore, additional research is needed to alleviate them. Results Results of successful cultivation of the transgenic aspens (Populus tremula) carrying the recombinant xyloglucanase gene (sp-Xeg) from Penicillium canescens in semi-natural conditions are reported in this paper for the first time. Change of carbohydrate composition of wood was observed in transgenic aspens carrying the sp-Xeg gene. The transformed transgenic line PtXVXeg1b demonstrated accelerated growth and increased content of cellulose in wood of trees growing in both greenhouse and outside in comparison with the control untransformed line Pt. The accelerated growth was observed also in the transgenic line PtXIVXeg1c. Thicker cell-wall and longer xylem fiber were also observed in both these transgenic lines. Undescribed earlier considerable reduction in the wood decomposition rate of the transgenic aspen stems was also revealed for the transformed transgenic lines. The decomposition rate was approximately twice as lower for the transgenic line PtXVXeg3b in comparison with the control untransformedline Pt. Conclusion Our results showed that the plants carrying the recombinant sp-Xeg gene do not demonstrate a decrease in growth parameters in semi-natural conditions. However, in some transgenic lines, a change in the carbohydrate composition of the wood, an increase in the cell wall thickness and a decrease in the rate of decomposition of wood are observed.

Using extraction with 0.7% aqueous ammonium oxalate, Sosnowskyi's hogweed pectins (yield 5–10%) with high molecular weight (200–700 kDa) and galacturonic acid content (70–85%), low degree of methylation (17–28%) and neutral sugar content (7–15%) were isolated from fresh, previously air-dried and frozen plant materials. All plant raw materials were pretreated with aqueous formaldehyde at 25 °C and dilute hydrochloric acid (pH 1.0 or 4.0, 50 °C). The different pretreatments of plant raw material slightly affected yield, chemical composition and molecular weight (average value and distribution) of hogweed pectin. The 13 C-NMR spectroscopy indicated the similar structure of the pectins isolated from fresh and air-dried hogweed raw materials. The pectins were differed slightly in thermal degradation depending on the type of hogweed raw material. The aqueous solutions of hogweed pectins had a pseudoplastic flow behavior. Dynamic oscillatory shear tests showed that the pectin hydrogels in the presence Ca 2+ (R∼0.6) represent true gels. The pectins from fresh plant material (pH 4.0) formed stronger gel networks (G' ∼2400 Pa) then compared to pectins from air-dried (G' ∼1400 Pa) and frozen (G' ∼900 Pa) plant raw materials. The pectin gelispheres of different size, particle shape, density and textural parameters were prepared by ionotropic gelation. The gelispheres derived from hogweed pectins (fresh and air-dried raw materials) were more stiffness compared to the gelispheres from the commercial low methoxyl apple pectin.

Background Recombinant carbohydrases genes are used to produce transgenic woody plants with improved phenotypic traits. However, cultivation of such plants in open field is challenging due to a number of problems. Therefore, additional research is needed to alleviate them. Results Results of successful cultivation of the transgenic aspens (Populus tremula) carrying the recombinant xyloglucanase gene (sp-Xeg) from Penicillium canescens in semi-natural conditions are reported in this paper for the first time. Change of carbohydrate composition of wood was observed in transgenic aspens carrying the sp-Xeg gene. The transformed transgenic line Xeg-2-1b demonstrated accelerated growth and increased content of cellulose in wood of trees growing in both greenhouse and outside in comparison with the control untransformed line Pt. The accelerated growth was observed also in the transgenic line Xeg-1-1c. Thicker cell-wall and longer xylem fiber were also observed in both these transgenic lines. Undescribed earlier considerable reduction in the wood decomposition rate of the transgenic aspen stems was also revealed for the transformed transgenic lines. The decomposition rate was approximately twice as lower for the transgenic line Xeg-2-3b in comparison with the control untransformed line Pt. Conclusion A direct dependence of the phenotypic and biochemical traits on the expression of the recombinant gene sp-Xeg was demonstrated. The higher was the level of the sp-Xeg gene expression, the more pronounced were changes in the phenotypic and biochemical traits. All lines showed phenotypic changes in the leave traits. Our results showed that the plants carrying the recombinant sp-Xeg gene do not demonstrate a decrease in growth parameters in semi-natural conditions. In some transgenic lines, a change in the carbohydrate composition of the wood, an increase in the cell wall thickness, and a decrease in the rate of decomposition of wood were observed.

Low methyl-esterified pectin (AU701) was found to form gel beads with glycerol. Wet AU701-glycerol gel beads exhibited similar diameter and hardness compared to the AU701-Ca gel beads, prepared by ionotropic gelation with Ca2+ and used for comparison. The morphology of dry pectin gel beads determined by scanning electron microscopy revealed that the beads exhibited rough and grooved surface. The AU701-glycerol gel beads absorbed more grams of water than AU701-Ca gel beads (12.2 g vs 3.9 g per 1 g of the beads). Rheological properties and hardness of the AU701-glycerol gel beads improved with the increase of the pectin/glycerol ratio. Swelling behavior of the AU701-glycerol gel beads was determined after sequential incubation in simulated gastric (SGF) and intestinal (SIF) fluids. The AU701-glycerol gel beads swelled in SGF to a greater extent and revealed higher stability in SIF than the gel beads cross-linked by Ca2+.

Pectin hydrogel particles (PHPs) were prepared by ionotropic gelation of low methylesterified pectin of Tanacetum vulgare L. with calcium ions. Wet PHPs prepared from TVF exhibited a smaller diameter and the lower weight as well as exhibited the best textural properties in terms of hardness and elasticity compared to the PHPs prepared from commercial low methylesterified pectin (CU701) used for comparison. Upon air drying, PHPs prepared from CU701 became small and dense microspheres whereas the dry PHPs prepared from TVF exhibited a drop-like shape. The morphology of dry PHPs determined by scanning electron microscopy revealed that the surface of the TVF beads exhibited fibred structures, whereas the PHPs prepared from CU701 exhibited a smooth surface. The characterization of surface roughness using atomic force microscopy indicated less roughness profile of the PHPs prepared from TVF than CU701. PHPs prepared from TVF were found to possess in vitro resistance to successive incubations in simulated gastric (SGF), intestinal (SIF), and colonic fluid (SCF) at 37 °C for 2, 4 and 18 h, respectively. The PHPs prepared from CU701 swelled in SGF and then lost their spherical shape and were fully disintegrated after 4 h of incubation in SIF. The PHPs from TVF, which were subjected to treatment with SGF, SIF and SCF, were found to adsorb microbial β-glucuronidase (βG) in vitro. The data obtained offered the prospect for the development of the PHPs from TVF as sorbents of colonic βG for the inhibition of re-absorption of estrogens.

MDM2 and MDM4 are proto-oncoproteins that bind to and inhibit members of the p53 protein family, p53, p73 and possibly p63. p53 is a mammalian tumor suppressor and p63 and p73 are critical for development. With the sequencing of genomes from multiple organisms there is mounting evidence for a consensus scenario of p53 gene family evolution. A single p53/p63/p73 gene is in invertebrates and required for maintenance of germline DNA. Gene duplication occurred in an ancestor in common with cartilaginous fishes, giving rise to a separate p53 gene and at least one ancestral p63/p73 gene. In bony vertebrates, all three p53 gene family paralogs, p53, p63, and p73 are distinct genes. This raises the question of how MDM2 and MDM4 genes evolved. We show evidence that MDM2 and MDM4 arose from a gene duplication event prior to the emergence of bony vertebrates more than 440 millionyears ago. Comparative genome studies indicate that invertebrate organisms have only one MDM homolog. In jawed vertebrates, the p53-binding domains of MDM2 and MDM4 proteins evolved at a high rate, approaching the evolution rate of the MDM2-binding domain of p53. However, the MDM2-binding domain of p73 exhibits markedly stronger conservation suggesting novel p53-independent functions. The most conserved domain within all MDM2 family members is the RING domain of the MDM2 ortholog which is responsible for ubiquitination of p53 and heterodimerization with MDM4. We suggest a model where oligomerization is an ancient function of MDM and ubiquitination activity was acquired later near the MDM gene duplication event coinciding with the time of the emergence of p53 as a distinct gene.

Human populations contain a functional coding polymorphism (codon 72) in the p53 gene. To explore whether this polymorphism alters the transcriptional pattern of p53-regulated genes, the human isogenic cell lines harboring p53 with either the proline or arginine at codon 72 were employed to activate p53-mediated transcription. Thirty-four p53-regulated genes were assayed for their increased levels of mRNA using quantitative real-time PCR. The largest difference between p53-arginine and p53-proline was found with the PERP gene involved in cell-cell adhesion and apoptosis. The most common set of genes that are transcribed better by the p53-arginine protein than the p53-proline protein was found in the apoptotic function (DR-4, NOXA, PUMA, and PIG-3). LIF, a cytokine that is required for optimal reproductive function, was produced at 2x higher levels by the p53-arginine than the p53-proline allele. The genes that induced their mRNAs at the highest levels compared to the baseline tended to be synthesized better by the p53-arginine protein than the p53-proline protein. These molecular studies may help to explain the complicated associations observed between this polymorphism and the incidence of some cancers, the longevity of some populations, and the fecundity of different groups.

Many ssRNA/ssDNA viruses bind their genome by highly basic semiflexible peptide arms of capsid proteins. Here, we show that nonspecific electrostatic interactions control both the length of the genome and genome conformations. Analysis of available experimental data shows that the genome length is linear in the net charge on the capsid peptide arms, irrespective of the actual amino acid sequence, with a proportionality coefficient of 1.61 ± 0.03. This ratio is conserved across all ssRNA/ssDNA viruses with highly basic peptide arms, and is different from the one-to-one charge balance expected of specific binding. Genomic nucleotides are predicted to occupy a radially symmetric spherical shell detached from the viral capsid, in agreement with experimental data.

Vertebrate genomic assemblies were analyzed for endogenous sequences related to any known viruses with single-stranded DNA genomes. Numerous high-confidence examples related to the Circoviridae and two genera in the family Parvoviridae , the parvoviruses and dependoviruses, were found and were broadly distributed among 31 of the 49 vertebrate species tested. Our analyses indicate that the ages of both virus families may exceed 40 to 50 million years. Shared features of the replication strategies of these viruses may explain the high incidence of the integrations.

A common ancestor to the three p53 family members of human genes p53, p63, and p73 is first detected in the evolution of modern‐day sea anemones, in which both structurally and functionally it acts to protect the germ line from genomic instabilities in response to stresses. This p63/p73 common ancestor gene is found in almost all invertebrates and first duplicates to produce a p53 gene and a p63/p73 ancestor in cartilaginous fish. Bony fish contain all three genes, p53, p63, and p73, and the functions of these three transcription factors diversify in the higher vertebrates. Thus, this gene family has preserved its structural features and functional activities for over one billion years of evolution.

We report hitherto unrecognized cooperative behavior in the stochastic collapse of certain compressed lipid monolayers implicated in pulmonary function. The cooperativity emerges from a statistical analysis of the collapse events captured using fluorescence microscopy and digital image analysis. The collapse events involve folding of the monolayer on a micron scale, yet each event produces a macroscopic jerk of the layer. The cooperative collapse is striking for its temporal sharpness and large spatial extent.

We investigate asymptotic properties of long polymers grafted to convex cylindrical and spherical surfaces, and, in particular, distribution of chain free ends. The parabolic potential profile, predicted for flat and concave brushes, fails in convex brushes, and chain free ends span only a finite fraction of the brush thickness. In this paper, we extend the self-consistent model developed by Ball, Marko, Milner and Witten to determine the size of the exclusion zone, i.e. size of the region of the brush free from chain ends. We show that in the limit of strong stretching, the brush can be described by an alternative system of integral equations. This system can be solved exactly in the limit of weakly curved brushes, and numerically for the intermediate to strong curvatures. We find that going from melt state to theta solvent and then to marginal solvent decreases relative size of the exclusion zone. These relative differences grow exponentially as the curvature decreases to zero., 9 pages, 4 figures, submitted to J. Chem. Phys

We investigate spreading of phase separated copolymer films, where domain walls and thickness steps influence polymer flow. We show that at early stages of spreading its rate is determined by slow activated flow at terrace steps (i.e. thickness steps). At late stages of spreading, on the other hand, the rate is determined by the flow along terraces, with diffusion-like time dependence $t^{-1/2}$. This dependence is similar to de Gennes and Cazabat's prediction for generic layered liquids, as opposed to the classical Tanner's law of drop spreading. We also argue that chain hopping at the spreading terrace steps should lead to the formation of aligned, defect-free domain patterns on the growing terraces., Comment: 11 pages, 7 figures, submitted to J. Chem. Phys

Oligomerization of members of the p53 family of transcription factors (p53, p63, and p73) is essential for their distinct functions in cell-cycle control and development. To elucidate the molecular basis for tetramer formation of the various family members, we solved the crystal structure of the human p73 tetramerization domain (residues 351–399). Similarly to the canonical p53 tetramer, p73 forms a tetramer with D2 symmetry that can be described as a dimer of dimers. The most striking difference between the p53 and p73 tetramerization domain is the presence of an additional C-terminal helix in p73. This helix, which is conserved in p63, is essential for stabilizing the overall architecture of the tetramer, as evidenced by the different oligomeric structures observed for a shortened variant lacking this helix. The helices act as clamps, wrapping around the neighboring dimer and holding it in place. In addition, we show by mass spectrometry that the tetramerization domains of p63 and p73, but not p53, fully exchange, with different mixed tetramers present at equilibrium, albeit at a relatively slow rate. Taken together, these data provide intriguing insights into the divergent evolution of the oligomerization domain within the p53 family, from the ancestral p63/p73-like protein toward smaller, less promiscuous monomeric building blocks in human p53, allowing functional separation of the p53 pathway from that of its family members.

elements riust be two-directionaL The Problems of reversibility andaddressing can be overcome with the syninetrical switch. Figure 1 shows anoptical binary tree riade b the syrmietrical switch As can be seen, in thiscase the addressing is the tree-structure and the Paths are always reversible.An architecture for an integrated optical version of the tree structurewith holograi*iics intercounnection is show in Fig, 2. Light from each input

The paper presents a study of a single-mode Ti and Fe in-diffusion waveguide. We have reported in this letter the experimental observation of diffraction effects due to the formation of a transitory photorefractive grating. The interference of a guided-wave with light scattering inside the waveguide can form a spatial phase grating due to the photorefractive effect, analogous to the storage process of holograms in bulk crystals. The storage mechanism in these materials is based on light induced permanent changes of the refractive index--the photorefractive effect. In this paper some advantages and limitations of this method are discussed.

We have discovered spectacular patterns formed by drying aqueous drops of sodium poly(styrene sulfonate) containing fixed amounts of sodium chloride. Crystallization of salt molecules takes place at a critical concentration and several factors such as the polyelectrolyte concentration and humidity affect the final patterns formed. The observed patterns include concentric rings, needle-like structures, chains of triangular-shaped crystallites, etc. We describe the observed phenomenology accompanied by a theoretical model for the occurrence of concentric rings.

Vertebrate genomes contain numerous copies of retroviral sequences, acquired over the course of evolution. Until recently they were thought to be the only type of RNA viruses to be so represented, because integration of a DNA copy of their genome is required for their replication. In this study, an extensive sequence comparison was conducted in which 5,666 viral genes from all known non-retroviral families with single-stranded RNA genomes were matched against the germline genomes of 48 vertebrate species, to determine if such viruses could also contribute to the vertebrate genetic heritage. In 19 of the tested vertebrate species, we discovered as many as 80 high-confidence examples of genomic DNA sequences that appear to be derived, as long ago as 40 million years, from ancestral members of 4 currently circulating virus families with single strand RNA genomes. Surprisingly, almost all of the sequences are related to only two families in the Order Mononegavirales: the Bornaviruses and the Filoviruses, which cause lethal neurological disease and hemorrhagic fevers, respectively. Based on signature landmarks some, and perhaps all, of the endogenous virus-like DNA sequences appear to be LINE element-facilitated integrations derived from viral mRNAs. The integrations represent genes that encode viral nucleocapsid, RNA-dependent-RNA-polymerase, matrix and, possibly, glycoproteins. Integrations are generally limited to one or very few copies of a related viral gene per species, suggesting that once the initial germline integration was obtained (or selected), later integrations failed or provided little advantage to the host. The conservation of relatively long open reading frames for several of the endogenous sequences, the virus-like protein regions represented, and a potential correlation between their presence and a species' resistance to the diseases caused by these pathogens, are consistent with the notion that their products provide some important biological advantage to the species. In addition, the viruses could also benefit, as some resistant species (e.g. bats) may serve as natural reservoirs for their persistence and transmission. Given the stringent limitations imposed in this informatics search, the examples described here should be considered a low estimate of the number of such integration events that have persisted over evolutionary time scales. Clearly, the sources of genetic information in vertebrate genomes are much more diverse than previously suspected., Author Summary Vertebrate genomes contain numerous copies of retroviral sequences, acquired over the course of evolution. Until recently they were thought to be the only type of RNA viruses to be so represented. In this comprehensive study, we compared sequences representing all known non-retroviruses containing single stranded RNA genomes, with the genomes of 48 vertebrate species. We discovered that as long ago as 40 million years, almost half of these species acquired sequences related to the genes of certain of these RNA viruses. Surprisingly, almost all of the nearly 80 integrations identified are related to only two viral families, the Ebola/ Marburgviruses, and Bornaviruses, which are deadly pathogens that cause lethal hemorrhagic fevers and neurological disease, respectively. The conservation and expression of some of these endogenous sequences, and a potential correlation between their presence and a species' resistance to the diseases caused by the related viruses, suggest that they may afford an important selective advantage in these vertebrate populations. The related viruses could also benefit, as some resistant species may provide natural reservoirs for their persistence and transmission. This first comprehensive study of its kind demonstrates that the sources of genetic inheritance in vertebrate genomes are considerably more diverse than previously appreciated.