Trisubstituted flavonol glycosides in Coronilla emerus flowers

A novel trisubstituted kaempferol glycoside has been isolated from leaves and flowers of Coronilla emerus and identified as the 3-glucoside-7,4′-dirhamnoside. It co-occurs in the flowers with the 3-glucosides and 3-glucoside-7-rhamnosides of kaempferol and quercetin. A second kaempferol triglycoside based on glucose and xylose is also present. All six glycosides contribute to the UV patterning present in the wings of the flowers. This is the first report of kaempferol triglycosides with monosaccharide units substituting hydroxyl groups at the 3-, 7- and 4′-positions.     

Fat patterning in Papua New Guineans: effects of age, sex and acculturation

For the renewed interest in fat patterning to be fruitful, more information on the biology of fat patterning, particularly normative data are required. Nine skinfold thicknesses were measured in 2312 coastal and highland Papua New Guineans of all ages undergoing acculturation. Principal component analysis of the skinfold data revealed components of fatness and fat patterning. First component scores were typical of the changes in fatness with age and the sex differences expected in these populations. Second component scores of limb-trunk fat patterning showed marked changes with age in both sexes but they stabilised in adulthood. This pattern seemed more affected by maturity although it is usually thought to be a sex-specific pattern. The third component, upper-lower body patterning, was more sexually dimorphic in adults. Marked differences were not observed between the two communities but over a 15-year period fatness levels increased slightly and subcutaneous fat tended to be deposited more centrally.     

Arabidopsis Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development

SYP2 proteins are a sub-family of Qa-SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) that may be responsible for protein trafficking between pre-vacuolar compartments (PVC) and vacuoles. Arabidopsis thaliana SYP22/VAM3/SGR3 and SYP21/PEP12 proteins function independently, but are both reported to be essential for male gametophytic viability. Here, we systematically examined the redundancy of three SYP2 paralogs (i.e. SYP21, 22 and 23) using a Col-0 ecotype harboring a SYP2 paralog (SYP23/PLP) that lacked a transmembrane domain. Surprisingly, no visible phenotypes were observed, even in the double knockout syp21/pep12 syp23/plp. Deficiency of either SYP21/PEP12 or SYP23/PLP in the syp22 background resulted in a defect in vacuolar protein sorting, characterized by abnormal accumulation of protein precursors in seeds. SYP21/PEP12 knockdown enhanced the syp22 phenotype (i.e. semi-dwarfism, poor leaf vein development and abnormal development of myrosin cells), and additional knockout of SYP23/PLP further aggravated the phenotype. A GFP-SYP23/PLP fusion localized to the cytosol, but not to the PVC or vacuolar membrane, where SYP21/PEP12 or SYP22/VAM3, respectively, were localized. Immunoprecipitation analysis showed that SYP23/PLP interacted with the vacuolar Qb- and Qc-SNAREs, VTI11 and SYP5, respectively, suggesting that SYP23/PLP is able to form a SNARE complex anchoring the membrane. Unexpectedly, we found that expression of multiple copies of a genomic fragment of SYP23/PLP suppressed the abnormal syp22-3 phenotype. Thus, SYP2 proteins, including cytosolic SYP23/PLP, appear to function redundantly in vacuolar trafficking and plant development.     

HyBMP5-8b, a BMP5-8 orthologue, acts during axial patterning and tentacle formation in hydra

Developmental gradients play a central role in axial patterning in hydra. As part of the effort towards elucidating the molecular basis of these gradients as well as investigating the evolution of the mechanisms underlying axial patterning, genes encoding signaling molecules are under investigation. We report the isolation and characterization of HyBMP5-8b, a BMP5-8 orthologue, from hydra. Processes governing axial patterning are continuously active in adult hydra. Expression patterns of HyBMP5-8b in normal animals and during bud formation, hydra's asexual form of reproduction, were examined. These patterns, coupled with changes in patterns of expression in manipulated tissues during head regeneration, foot regeneration as well as under conditions that alter the positional value gradient indicate that the gene is active in two different processes. The gene plays a role in tentacle formation and in patterning the lower end of the body axis.     

Local variation in conspecific plant density influences plant–soil feedback in a natural grassland

Several studies have argued that under field conditions plant–soil feedback may be related to the local density of a plant species, but plant–soil feedback is often studied by comparing conspecific and heterospecific soils or by using mixed soil samples collected from different locations and plant densities. We examined whether the growth of the early successional species Jacobaea vulgaris in soil collected from the field is related to the local variation in plant density of this species. In a grassland restoration site, we selected eight 8 m × 8 m plots, four with high and four with low densities of J. vulgaris plants. In 16 subplots in each plot we recorded the density and size of J. vulgaris, and characteristics of the vegetation and the soil chemistry. Soil collected from each subplot was used in a greenhouse pot-experiment to study the growth of J. vulgaris, both in pure field soil and in sterile soil inoculated with a small part of field soil.In the field, flowering J. vulgaris plants were taller, the percentage of rosette plants was higher and seed density was larger in High- than in Low-density plots. In the pot experiment, J. vulgaris had a negative plant–soil feedback, but biomass was also lower in soil collected from High- than from Low-density plots, although only when growing in inoculated soil. Regression analyses showed that J. vulgaris biomass of plants growing in pure soil was related to soil nutrients, but also to J. vulgaris density in the field.We conclude that in the field there is local variation in the negative plant–soil feedback of J. vulgaris and that this variation can be explained by the local density of J. vulgaris, but also by other factors such as nutrient availability.     

Engineering cell alignment in vitro

Cell alignment plays a critical role in various cell behaviors including cytoskeleton reorganization, membrane protein relocation, nucleus gene expression, and ECM remodeling. Cell alignment is also known to exert significant effects on tissue regeneration (e.g., neuron) and modulate mechanical properties of tissues including skeleton, cardiac muscle and tendon. Therefore, it is essential to engineer cell alignment in vitro for biomechanics, cell biology, tissue engineering and regenerative medicine applications. With advances in nano- and micro-scale technologies, a variety of approaches have been developed to engineer cell alignment in vitro, including mechanical loading, topographical patterning, and surface chemical treatment. In this review, we first present alignments of various cell types and their functionality in different tissues in vivo including muscle and nerve tissues. Then, we provide an overview of recent approaches for engineering cell alignment in vitro. Finally, concluding remarks and perspectives are addressed for future improvement of engineering cell alignment.     

Functional characterization and genetic mapping of alk8

The novel type I TGFβ family member receptor alk8 is expressed both maternally and zygotically. Functional characterization of alk8 was performed using microinjection studies of constitutively active (CA), kinase modified/dominant negative (DN), and truncated alk8 mRNAs. CA Alk8 expression produces ventralized embryos while DN Alk8 expression results in dorsalized phenotypes. Truncated alk8 expressing embryos display a subtle dorsalized phenotype closely resembling that of the identified zebrafish dorsalized mutant, lost-a-fin (laf). Single-strand conformation polymorphism (SSCP) analysis was used to map alk8 to zebrafish LG02 in a region demonstrating significant conserved synteny to Hsa2, and which contains the human alk2 gene, ACVRI. Altogether, these functional, gene mapping and phylogenetic analyses suggest that alk8 may be the zebrafish orthologue to human ACVRI (alk2), and therefore extend previous studies of Alk2 conducted in Xenopus.     

Over a Century of Neuron Culture: From the Hanging Drop to Microfluidic Devices

The brain is the most intricate, energetically active, and plastic organ in the body. These features extend to its cellular elements, the neurons and glia. Understanding neurons, or nerve cells, at the cellular and molecular levels is the cornerstone of modern neuroscience. The complexities of neuron structure and function require unusual methods of culture to determine how aberrations in or between cells give rise to brain dysfunction and disease. Here we review the methods that have emerged over the past century for culturing neurons in vitro, from the landmark finding by Harrison (1910) — that neurons can be cultured outside the body — to studies utilizing culture vessels, micro-islands, Campenot and brain slice chambers, and microfluidic technologies. We conclude with future prospects for neuronal culture and considerations for advancement. We anticipate that continued innovation in culture methods will enhance design capabilities for temporal control of media and reagents (chemotemporal control) within sub-cellular environments of three-dimensional fluidic spaces (microfluidic devices) and materials (e.g., hydrogels). They will enable new insights into the complexities of neuronal development and pathology.     

Cells on the move: Modulation of guidance cues during germ cell migration

In Drosophila melanogaster the progenitors of the germ-line stem cells, the primordial germ cells (PGCs) are formed on the outside surface of the early embryo, while the somatic gonadal precursor cells (SGPs) are specified during mid-embryogenesis. To form the primitive embryonic gonad, the PGCs travel from outside of the embryo, across the mid-gut and then migrate through the mesoderm to the SGPs. The migratory path of PGCs is dictated by a series of attractive and repulsive cues. Studies in our laboratory have shown that one of the key chemoattractants is the Hedgehog (Hh) ligand. Although, Hh is expressed in other cell types, the long-distance transmission of this ligand is specifically potentiated in the SGPs by the hmgcr isoprenoid biosynthetic pathway. The distant transmission of the Hh ligand is gated by restricting expression of hmgcr to the SGPs. This is particularly relevant in light of the recent findings that an ABC transporter, mdr49 also acts in a mesoderm specific manner to release the germ cell attractant. Our studies have demonstrated that mdr49 functions in hh signaling likely via its role in the transport of cholesterol. Given the importance of cholesterol in the processing and long distance transmission of the Hh ligand, this observation has opened up an exciting avenue concerning the possible role of components of the sterol transport machinery in PGC migration.