Colony Interior Research Articles

Improved uniform ductility in a high-carbon pearlitic steel via microstructural homogeneity architecture

In this work, the nucleation and growth kinetics of pearlitic transformation are quantitatively investigated to unveil the distinctive microstructural evolution behavior under a high undercooling degree. Both intra-granular and inter-granular nucleation increasingly occur with 1-2 orders of magnitude increase in nucleation rate N˙. However, a moderate increase in the growth rate GV, namely a larger N˙/GV ratio, leads to a 30%–40% size reduction in pearlitic colonies. The avalanche-type increase in nucleation events induces a more random distribution of ferrite orientation among colonies that possess a slight increase in relative misorientation but a pronounced increase in accumulative misorientation, and an arbitrary alignment of Fe3C lamellas in colony interior. Such improved microstructural homogeneity ensures more uniform and localized deformability, which allows compatible deformation between colonies or within colonies and prevents the expansion of newly-formed cracks. These results provide guidance for industrial manufacture of high-carbon pearlitic products with high safety and reliability, resulting from the improvement in localized deformability, and for low-cost wires via reducing the drawing passes, stemming from the significant increase in uniform deformability.

Intra-colony spatial variance of oxyregulation and hypoxic thresholds for key Acropora coral species.

Oxygen (O2) availability is essential for healthy coral reef functioning, yet how continued loss of dissolved O2 via ocean deoxygenation impacts performance of reef building corals remains unclear. Here, we examine how intra-colony spatial geometry of important Great Barrier Reef (GBR) coral species Acropora may influence variation in hypoxic thresholds for upregulation, to better understand capacity to tolerate future reductions in O2 availability. We first evaluate the application of more streamlined models used to parameterise Hypoxia Response Curve data, models that have been used historically to identify variable oxyregulatory capacity. Using closed-system respirometry to analyse O2 drawdown rate, we show that a two-parameter model returns similar outputs as previous 12th-order models for descriptive statistics such as the average oxyregulation capacity (Tpos) and the ambient O2 level at which the coral exerts maximum regulation effort (Pcmax), for diverse Acropora species. Following an experiment to evaluate whether stress induced by coral fragmentation for respirometry affected O2 drawdown rate, we subsequently identify differences in hypoxic response for the interior and exterior colony locations for the species Acropora abrotanoides, Acropora cf. microphthalma and Acropora elseyi. Average regulation capacity across species was greater (0.78-1.03 ± SE 0.08) at the colony interior compared with exterior (0.60-0.85 ± SE 0.08). Moreover, Pcmax occurred at relatively low pO2 of <30% (±1.24; SE) air saturation for all species, across the colony. When compared against ambient O2 availability, these factors corresponded to differences in mean intra-colony oxyregulation, suggesting that lower variation in dissolved O2 corresponds with higher capacity for oxyregulation. Collectively, our data show that intra-colony spatial variation affects coral oxyregulation hypoxic thresholds, potentially driving differences in Acropora oxyregulatory capacity.

Force localization modes in dynamic epithelial colonies.

Collective cell behaviors, including tissue remodeling, morphogenesis, and cancer metastasis, rely on dynamics among cells, their neighbors, and the extracellular matrix. The lack of quantitative models precludes understanding of how cell–cell and cell–matrix interactions regulate tissue-scale force transmission to guide morphogenic processes. We integrate biophysical measurements on model epithelial tissues and computational modeling to explore how cell-level dynamics alter mechanical stress organization at multicellular scales. We show that traction stress distribution in epithelial colonies can vary widely for identical geometries. For colonies with peripheral localization of traction stresses, we recapitulate previously described mechanical behavior of cohesive tissues with a continuum model. By contrast, highly motile cells within colonies produce traction stresses that fluctuate in space and time. To predict the traction force dynamics, we introduce an active adherent vertex model (AAVM) for epithelial monolayers. AAVM predicts that increased cellular motility and reduced intercellular mechanical coupling localize traction stresses in the colony interior, in agreement with our experimental data. Furthermore, the model captures a wide spectrum of localized stress production modes that arise from individual cell activities including cell division, rotation, and polarized migration. This approach provides a robust quantitative framework to study how cell-scale dynamics influence force transmission in epithelial tissues.

A semi-interpenetrating network of polyacrylamide and recombinant basement membrane allows pluripotent cell culture in a soft, ligand-rich microenvironment.

The physical properties of the extracellular matrix play an essential role in guiding stem cell differentiation and tissue morphogenesis both invivo and invitro. Existing work to investigate the role of matrix mechanics in directing stem cell proliferation, self-renewal, and differentiation has been limited by the poor attachment and survival of human pluripotent cells cultured on soft matrices (Young's modulus E ≲ 1000Pa). To address this limitation we developed a protocol for generating semi-interpenetrating networks of polyacrylamide and recombinant basement membrane. Using these materials, we found that human embryonic stem cells (hESCs) remained proliferative and pluripotent even when grown in small colonies and on surfaces ranging in stiffness from 150 to 12000Pa, spanning the range of tissue stiffnesses likely to be encountered in the embryo. Considerable recent attention has focused on the role of the transcriptional coactivator and Hippo effector YAP in regulating differentiation and cell proliferation both in the early embryo and invitro. We found that while YAP localized to the nucleus on substrates of E ≳ 1000Pa, its localization was heterogeneous on substrates of moduli ≲ 450Pa, with predominantly nuclear localization at the colony periphery and mixed cytoplasmic and nuclear localization for cells in the colony interior, a pattern reminiscent of YAP subcellular localization in the inner cell mass (ICM) of the early embryo. In addition, hESC colony dynamics were highly responsive to substrate stiffness, with cells assembling into monolayers, multilayer structures, and transient, hollow rosettes in response to decreasing substrate stiffnesses in the range of 12000 to 150Pa. We suggest that soft, ligand-rich substrates such as are described here provide a promising means of recapitulating aspects of early mammalian development that are otherwise inaccessible, and more broadly may be useful in the derivation of complex tissues from pluripotent cells in an invitro setting.

The surface structure of Botryococcus braunii colony prevents the entry of extraction solvents into the colony interior

The green colonial microalga Botryococcus braunii (race B) has attracted considerable attention because it is known to produce many hydrocarbons. The hydrocarbons are present in the extracellular matrix that connects the cells located in the interior of a B. braunii colony; however, the surface of each colony is covered with a retaining wall and a fibrillar colony sheath. This unique colony structure is thought to be associated with hydrocarbon storage and known to affect the extractability of hydrocarbons by using an organic solvent. In this study, we investigated the relation between hydrocarbon extractability and changes in the colony surface structure after two distinct treatments: culture of the algae in brackish medium or thermal pretreatment of the algae before solvent extraction. Both the treatments improved hydrocarbon extractability when non-polar solvents such as n-hexane were used. Thermal pretreatment inhibited the formation of fibrils on the surface of algal colonies, as revealed by rapid-freezing and freeze-substitution electron microscopy. However, when the alga was cultured in brackish medium, the fibrils on the surface shortened and became less dense. Thus, the fibrillar colony sheath that mainly consisted of saccharic components prevented the entry of organic solvents into the colony interior, and thus hydrocarbon extraction.

Molecular phylogenetic evidence supports a new family of octocorals and a new genus of Alcyoniidae (Octocorallia, Alcyonacea)

Molecular phylogenetic evidence indicates that the octocoral family Alcyoniidae is highly polyphyletic, with genera distributed across Octocorallia in more than 10 separate clades. Most alcyoniid taxa belong to the large and poorly resolved Holaxonia–Alcyoniina clade of octocorals, but members of at least four genera of Alcyoniidae fall outside of that group. As a first step towards revision of the family, we describe a new genus, Parasphaerasclera gen. n., and family, Parasphaerascleridae fam. n., of Alcyonacea to accommodate species of Eleutherobia Pütter, 1900 and Alcyonium Linnaeus, 1758 that have digitiform to digitate or lobate growth forms, completely lack sclerites in the polyps, and have radiates or spheroidal sclerites in the colony surface and interior. Parasphaerascleridae fam. n. constitutes a well-supported clade that is phylogenetically distinct from all other octocoral taxa. We also describe a new genus of Alcyoniidae, Sphaerasclera gen. n., for a species of Eleutherobia with a unique capitate growth form. Sphaerasclera gen. n. is a member of the Anthomastus–Corallium clade of octocorals, but is morphologically and genetically distinct from Anthomastus Verrill, 1878 and Paraminabea Williams & Alderslade, 1999, two similar but dimorphic genera of Alcyoniidae that are its sister taxa. In addition, we have re-assigned two species of Eleutherobia that have clavate to capitate growth forms, polyp sclerites arranged to form a collaret and points, and spindles in the colony interior to Alcyonium, a move that is supported by both morphological and molecular phylogenetic evidence.

Rapidly Developing Yeast Microcolonies Differentiate in a Similar Way to Aging Giant Colonies

During their development and aging on solid substrates, yeast giant colonies produce ammonia, which acts as a quorum sensing molecule. Ammonia production is connected with alkalization of the surrounding medium and with extensive reprogramming of cell metabolism. In addition, ammonia signaling is important for both horizontal (colony centre versus colony margin) and vertical (upper versus lower cell layers) colony differentiations. The centre of an aging differentiated giant colony is thus composed of two major cell subpopulations, the subpopulation of long-living, metabolically active and stress-resistant cells that form the upper layers of the colony and the subpopulation of stress-sensitive starving cells in the colony interior. Here, we show that microcolonies originating from one cell pass through similar developmental phases as giant colonies. Microcolony differentiation is linked to ammonia signaling, and cells similar to the upper and lower cells of aged giant colonies are formed even in relatively young microcolonies. A comparison of the properties of these cells revealed a number of features that are similar in microcolonies and giant colonies as well as a few that are only typical of chronologically aged giant colonies. These findings show that colony age per se is not crucial for colony differentiation.

Macroautophagy-mediated degradation of whole nuclei in the filamentous fungus Aspergillus oryzae.

Filamentous fungi consist of continuum of multinucleate cells called hyphae, and proliferate by means of hyphal tip growth. Accordingly, research interest has been focusing on hyphal tip cells, but little is known about basal cells in colony interior that do not directly contribute to proliferation. Here, we show that autophagy mediates degradation of basal cell components in the filamentous fungus Aspergillus oryzae. In basal cells, enhanced green fluorescent protein (EGFP)-labeled peroxisomes, mitochondria, and even nuclei were taken up into vacuoles in an autophagy-dependent manner. During this process, crescents of autophagosome precursors matured into ring-like autophagosomes to encircle apparently whole nuclei. The ring-like autophagosomes then disappeared, followed by dispersal of the nuclear material throughout the vacuoles, suggesting the autophagy-mediated degradation of whole nuclei. We also demonstrated that colony growth in a nutrient-depleted medium was significantly inhibited in the absence of functional autophagy. This is a first report describing autophagy-mediated degradation of whole nuclei, as well as suggesting a novel strategy of filamentous fungi to degrade components of existing hyphae for use as nutrients to support mycelial growth in order to counteract starvation.

The Rim101p/PacC Pathway and Alkaline pH Regulate Pattern Formation in Yeast Colonies

Multicellular organisms utilize cell-to-cell signals to build patterns of cell types within embryos, but the ability of fungi to form organized communities has been largely unexplored. Here we report that colonies of the yeast Saccharomyces cerevisiae formed sharply divided layers of sporulating and nonsporulating cells. Sporulation initiated in the colony's interior, and this region expanded upward as the colony matured. Two key activators of sporulation, IME1 and IME2, were initially transcribed in overlapping regions of the colony, and this overlap corresponded to the initial sporulation region. The development of colony sporulation patterns depended on cell-to-cell signals, as demonstrated by chimeric colonies, which contain a mixture of two strains. One such signal is alkaline pH, mediated through the Rim101p/PacC pathway. Meiotic-arrest mutants that increased alkali production stimulated expression of an early meiotic gene in neighboring cells, whereas a mutant that decreased alkali production (cit1Delta) decreased this expression. Addition of alkali to colonies accelerated the expansion of the interior region of sporulation, whereas inactivation of the Rim101p pathway inhibited this expansion. Thus, the Rim101 pathway mediates colony patterning by responding to cell-to-cell pH signals. Cell-to-cell signals coupled with nutrient gradients may allow efficient spore formation and spore dispersal in natural environments.

A novel method for exploring elemental composition of microbial communities: Laser ablation-inductively coupled plasma-mass spectrometry of intact bacterial colonies

Bacterial colonies are spatially complex structures whose physiology is profoundly dependent on interactions between cells and with the underlying semi-solid substratum. Here, we use bacterial colonies as a model of a microbial community to evaluate the potential of laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) to delineate elemental distributions within colonies with minimal pre-treatment. To reduce water content of the colony and limit undesirable absorption of laser energy, we compared methods of preparing 24 h-old colonies of Escherichia coli TG1 on agar for laser ablation. Colonies on excised agar segments dried on chromatography paper were superior to colonies dried in a dessicator or by prolonged incubation, with respect to signal magnitude, signal:noise ratio and background signal. Having optimised laser scan speed (10 μm s − 1 ) and laser beam diameter (100 μm), further improvements were achieved by growing colonies on nylon membranes over agar, which were then transferred to the ablation chamber without further treatment. Repeated line rasters across individual membrane-supported colonies yielded three-dimensional elemental maps of colonies, revealing a convex morphology consistent with visual inspection. By normalising isotope counts for P, Mn, Zn, Fe and Ca against Mg, the most abundant cellular divalent cation, we sought elemental heterogeneity within the colony. The normalised concentration of Mn in the perimeter was higher than in the colony interior, whereas the converse was true for Ca. LA-ICP-MS is a novel and powerful method for probing elemental composition and organisation within microbial communities and should find numerous applications in, for example, biofilm studies.

Adaptive variation in coral geometry and the optimization of internal colony light climates

Summary The ability of photosynthetic organisms to adjust their light climate has high adaptive significance because irradiance can vary spatially by orders of magnitude. Using a plating (foliaceous) coral species (Turbinaria mesenterina), we tested the hypothesis that plasticity of colony geometry optimizes internal irradiance distributions. We developed a two‐dimensional model to predict the internal irradiance distribution of a foliaceous colony as a function of its geometry. Field tests showed that the model explained 85% of the variation in irradiance within colonies of T. mesenterina with minimal bias. Colony plate angle, plate spacing and range of tissue distributions into the colony were exponential functions of water depth. In shallow water plates tended to be nearly vertical, narrowly spaced, and had living tissue only near the growing edge of the plate. In deep water plates grew more horizontally, were more widely spaced, and had living tissue extending well into the colony interior. This pattern of phenotypic plasticity effectively evens out differences in within‐colony irradiances. We compared within‐colony irradiance distributions across light habitats (depth), based on the observed variation in colony geometry with water depth. Despite fourfold differences in environmental irradiance, within‐colony irradiances had a common mode of 100–200 µmol m−2 s−1. This is near the hypothesized photosynthetic optimum defined by the upper limit of the subsaturation parameter (Ek) of the photosynthesis–irradiance curve. Our study demonstrates that phenotypic plasticity of colony geometry is an important mechanism for regulating light capture during growth in T. mesenterina, and facilitates near‐optimal internal irradiances across a wide range of environmental light regimes.

The growth and form of bacterial colonies.

A simple method is described for measuring the profile of bacterial colonies. Profiles were determined for colonies of Bacillus cereus, Escherichia coli and Staphylococcus albus of different ages. In spite of differences in cell morphology, the colony profiles had a common basic structure consisting of steeply rising leading edge connected by a ridge to an interior region where height also rose, though less steeply, to a flat or domed centre. The colony mass increased exponentially through part of the growth phase. It is suggested that net colony growth consists of a combination of leading edge growth, which is unrestricted and approaches the maximum specific growth rate of the organism, and diffusion-limited growth in the colony interior. Common elements of profiles from each species may be a consequence of such differences in growth rate.

Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40.

Human epidermal keratinocytes were infected by simian virus 40 in vitro. The structure of the developing keratinocyte colony reflects the spatial separation of cell division and keratinization in intact skin; thymidine-incorporating cells were primarily localized at the colony periphery whereas nondividing, histologically differentiated cells accumulated in the interior. Viral infection produced a dramatic increase in the size of the proliferative population as, simultaneously, differentiation was reduced in the colony interior. These changes were manifest when simian virus 40 T-antigen synthesis was detectable in only a small percentage of the cells; differentiation became increasingly density dependent as the percentage of T-antigen-positive cells rose over serial passage. The disruption of the normal pattern of growth/differentiation localization coincided with a loss of dependence on serum for growth, but preceded the appearance of other virus-induced properties associated with transformation; i.e., the ability to form colonies in soft agar and independence of growth from fibroblasts.