Morphogenetic Factors Research Articles

Retinoic acid induces prostatic bud formation

Formation of prostatic buds from the urogenital sinus (UGS) to initiate prostate development requires localized action of several morphogenetic factors. This report reveals all-trans-retinoic acid (RA) to be a powerful inducer of mouse prostatic budding that is associated with reciprocal changes in expression of two regulators of budding: sonic hedgehog (Shh) and bone morphogenetic protein 4 (Bmp4). Localization of retinoid signaling and expression of RA synthesis, metabolism, and receptor genes in the UGS on embryonic days 14.5-17.5 implicate RA in the mechanism of bud initiation. In UGS organ culture, RA increased prostatic budding, increased Shh expression, and decreased Bmp4. Prostatic budding was stimulated in the absence of RA by recombinant SHH, by blocking BMP4 signaling with NOGGIN, or by combined treatment with SHH and NOGGIN in UGS organ culture media. These observations suggest that reciprocal changes in hedgehog and BMP signaling by RA may regulate bud initiation.

Inventaire des stratégies cellulaires en ingénierie tissulaire de reconstruction osseuse

The objective of bone tissue engineering is to reconstruct bone stock using matrix structures, osteoinductor factors and osteogenic cells. Different types of natural or synthetic biomaterials are available or under development. The objective of recent work is to optimize matrix materials, particularly with better cell adhesion to the surface and better osteoconduction. For osteoinductors, most research is currently focused on bone morphogenetic protein (BMP) and angiogenic factors such as vascular endothelial growth factor (VEGF). Concerning the nature of the cells to be implanted, there is a clear dissociation between fundamental and clinical studies. Many clinical studies have demonstrated the strong osteogenic potential of fresh harvested total bone marrow. There has been nevertheless little fundamental work on the use of total bone marrow as a source of cells for bone tissue engineering. Most of the fundamental work has been focused on the use of mesenchymatous stromal cells selected from bone marrow and cultivated ex vivo. This approach which was first developed more than fifteen years ago has shown that the adjunction of these cells can improve the osteoformative capacity of bone substitutes. This strategy has, however, had almost no clinical impact to date since only two studies involving four patients have been reported. The purpose of this article is to review current research concerning bone tissue engineering using total bone marrow and mesenchymatous stromal cells.

Morphological and molecular characterization of a spontaneously tuberizing potato mutant: an insight into the regulatory mechanisms of tuber induction

BackgroundTuberization in potato (Solanum tuberosum L.) represents a morphogenetic transition of stolon growth to tuber formation, which is under complex environmental and endogenous regulation. In the present work, we studied the regulatory mechanisms and the role of different morphogenetic factors in a newly isolated potato mutant, which exhibited spontaneous tuberization (ST). The ST mutant was characterized in detail at morphological, physiological and biochemical levels.ResultsTuberization of the ST mutant grown in the soil was photoperiod-insensitive; predominantly sessile tubers formed directly from axillary buds even under continuous light. Single-node cuttings of the ST mutant cultured in vitro frequently formed tubers or basal tuber-like swellings instead of normal shoots under conditions routinely used for shoot propagation. The tuberization response of ST cuttings under light was dependent on sucrose, the concentration of which had to exceed certain threshold that inversely correlated with irradiance. Gibberellic acid prevented tuberization of ST cuttings, but failed to restore normal shoot phenotype and caused severe malformations. Carbohydrate analysis showed increased levels of both soluble sugars and starch in ST plants, with altered carbohydrate partitioning and metabolism. Comparative proteomic analysis revealed only a few differences between ST- and wild-type plants, primary amongst which seemed to be the absence of an isoform of manganese-stabilizing protein, a key subunit of photosystem II.ConclusionST mutant exhibits complex developmental and phenotypic modifications, with features that are typical for plants strongly induced to tuberize. These changes are likely to be related to altered regulation of photosynthesis and carbohydrate metabolism rather than impaired transduction of inhibitory gibberellin or photoperiod-based signals. The effect of gibberellins on tuberization of ST mutant suggests that gibberellins inhibit tuberization downstream of the inductive effects of sucrose and other positive factors.

Structure, Assembly, and Function of the Spore Surface Layers

Endospores formed by Bacillus, Clostridia, and related genera are encased in a protein shell called the coat. In many species, including B. subtilis, the coat is the outermost spore structure, and in other species, such as the pathogenic organisms B. anthracis and B. cereus, the spore is encased in an additional layer called the exosporium. Both the coat and the exosporium have roles in protection of the spore and in its environmental interactions. Assembly of both structures is a function of the mother cell, one of two cellular compartments of the developing sporangium. Studies in B. subtilis have revealed that the timing of coat protein production, the guiding role of a small group of morphogenetic proteins, and several types of posttranslational modifications are essential for the fidelity of the assembly process. Assembly of the exosporium requires a set of novel proteins as well as homologues of proteins found in the outermost layers of the coat and of some of the coat morphogenetic factors, suggesting that the exosporium is a more specialized structure of a multifunctional coat. These and other insights into the molecular details of spore surface morphogenesis provide avenues for exploitation of the spore surface layers in applications for biotechnology and medicine.

Morphogenetic signals from chondrocytes promote chondrogenic and osteogenic differentiation of mesenchymal stem cells

Mesenchymal stem cells (MSCs) are potentially useful cells for musculoskeletal tissue engineering. However, controlling MSC differentiation and tissue formation in vivo remains a challenge. There is a significant need for well-defined and efficient protocols for directing MSC behaviors in vivo. We hypothesize that morphogenetic signals from chondrocytes may regulate MSC differentiation. In micromass culture of MSCs, incubation with chondrocyte-conditioned medium (CCM) significantly enhanced the production of cartilage specific matrix including type II collagen. In addition, incubation of MSCs with conditioned medium supplemented with osteogenic factors induced more osteogenesis and accumulation of calcium and increased ALP activity. These findings reveal that chondrocyte-secreted factors promote chondrogenesis as well as osteogenesis of MSCs during in vitro micromass culture. Moreover, when MSCs expanded with chondrocyte-conditioned medium were encapsulated in hydrogels and subsequently implanted into athymic mice, basophilic extracellular matrix deposition characteristic of neocartilage was evident. These results indicate that articular chondrocytes produce suitable morphogenetic factors that induce the differentiation program of MSCs in vitro and in vivo.

Hyaline Globules (Thanatosomes) in Gastrointestinal Epithelium

Hyaline globules (HGs; thanatosomes) are well-defined morphologic and functional entities representing a degenerative phenomenon common to all cell types. We present the first quantitative and qualitative study of HGs in normal and pathologic gastrointestinal (GI) epithelium from a series of 2,230 biopsies. HGs were very rarely found in normal epithelium (1.1%), but their number increased significantly in specimens with ischemic injury (47%) and benign regenerative proliferation (70%). Their incidence in adenomatous polyps and adenocarcinomas was about 11% to 27%. Of the HGs, 2.9% contained nuclear fragments. Our results entirely support the unifying morphogenetic concept for HGs. The role of 2 obligatory morphogenetic factors for the generation of thanatosomes (propensity to apoptosis and heterophagy/autophagy) is confirmed. The nature of the third factor, ischemic conditions, is specified. Although a nonspecific microscopic phenomenon, HGs in the GI tract represented a relatively constant and useful histologic marker of enhanced cell turnover and ischemic injury.

Bone morphogenetic proteins and growth factors: emerging role in regenerative orthopaedic surgery*

Bone morphogenetic proteins (BMPs) were discovered by Urist and colleagues in 1965 and later defined as multifunctional cytokines involved in osteoinduction. BMPs are members of the transforming growth factor-β superfamily, with the exception of the BMP-1. Presently, at least 20 BMPs have been identified and studied, but only BMP 2, 4 and 7 have been able in vitro to stimulate the entire process of stem cell differentiation into osteoblastic mature cells. In preclinical and clinical studies, BMPs have demonstrated potential in osteoinduction and have been approved for clinical use in treating open fractures of the long bones and nonunions and in vertebral arthrodesis. Additional clinical uses of these molecules are under investigation and the possibility of using gene therapy in selected pathologies seems the most appealing.

Regional and modular expression of morphogenetic factors in the demosponge Lubomirskia baicalensis

Some sponges [phylum Porifera], e.g. the demosponges Lubomirskia baicalensis or Axinella polypoides, show an arborescent growth form. In the freshwater sponge L. baicalensis this morphotype is seen mostly in depths below 4 m while in more shallow regions it grows as a crust. The different growth forms are determined in nature very likely by water current and/or light. The branches of this species are composed of modules, arranged along the apical–basal axis. The modules are delimited by a precise architecture of the spicule bundles; longitudinal bundles originate from the apex of the earlier module, while at the basis of each module these bundles are cross-linked by traverse bundles under formation of annuli. Genes encoding putative morphogenetic factors, myotrophin and epidermal growth factor (EGF)-like molecules, and one gene of an antagonist for the Wnt signaling pathway, the soluble frizzled molecule, have been identified and characterized. Their expression levels as well as those of silicatein, one major spicule-forming molecule, have been studied in the crusts and the modules. The data revealed that at the apices of each module higher level of expression of myotrophin and EGF can be detected, while the base of each module is characterized by a high steady-state expression level of soluble frizzled molecule. These results suggest that module formation in L. baicalensis is controlled by a tuned interaction of agonistic (e.g., myotrophin and EGF) as well as antagonistic morphogenetic factors (e.g., soluble frizzled molecule).

Efficient Generation of a Trisporoid Library by Combination of Synthesis and Biotransformation

Trisporic acids and their biosynthetic precursors represent a family of powerful fungal pheromones and morphogenetic factors. A highly flexible synthetic protocol is described that (i) provides rapid access to nonfunctionalized early trisporoids from beta-ionone, (ii) includes a regiospecific oxidative functionalization of beta-ionone leading to 1-acetoxy-beta-ionone giving access to functionalized trisporoids, and (iii) utilizes a biotransformation of early synthetic trisporoids by growing cells of Blakeslea trispora to prepare late trisporoids including trisporic acids. The same protocol also provides deuterium-labeled trisporoids such as trisporin B [2H3]-19. Administration of [2H3]-19 to growing cells of the (-)-mating type of B. trispora resulted in the formation of the labeled trisporols [2H3]-20 and [2H3]-21. Growing cultures containing both mating types can be used to prepare trisporic acids from early precursors.

The Timing ofcotEExpression AffectsBacillus subtilisSpore Coat Morphology but Not Lysozyme Resistance

The synthesis of structural components and morphogenetic factors required for the assembly of the Bacillus subtilis spore coat is governed by a mother cell-specific transcriptional cascade. The first two temporal classes of gene expression, which involve RNA polymerase sigma sigma(E) factor and the ancillary regulators GerR and SpoIIID, are deployed prior to engulfment of the prespore by the mother cell. The two last classes rely on sigma(K), whose activation follows engulfment completion, and GerE. The cotE gene codes for a morphogenetic protein essential for the assembly of the outer coat layer and spore resistance to lysozyme. cotE is expressed first from a sigma(E)-dependent promoter and, in a second stage, from a promoter that additionally requires SpoIIID and that remains active under sigma(K) control. CotE localizes prior to engulfment completion close to the surface of the developing spore, but formation of the outer coat is a late, sigma(K)-controlled event. We have transplanted cotE to progressively later classes of mother cell gene expression. This created an early class of mutants in which cotE is expressed prior to engulfment completion and a late class in which expression of cotE follows the complete engulfment of the prespore. Mutants of the early class assemble a nearly normal outer coat structure, whereas mutants of the late class do not. Hence, the early expression of CotE is essential for outer coat assembly. Surprisingly, however, all mutants were fully resistant to lysozyme. The results suggest that CotE has genetically separable functions in spore resistance to lysozyme and spore outer coat assembly.

Safety and efficacy of use of demineralised bone matrix in orthopaedic and trauma surgery

Demineralised bone matrix (DBM) acts as an osteoconductive, and possibly as an osteoinductive, material. It is widely used in orthopaedic, neurosurgical, plastic and dental areas. More than 500,000 bone grafting procedures with DBM are performed annually in the US. It does not offer structural support, but it is well suited for filling bone defects and cavities. The osteoinductive nature of DBM is presumably attributed to the presence of matrix-associated bone morphogenetic proteins (BMPs) and growth factors, which are made available to the host environment by the demineralisation process. Clinical results have not been uniformly favourable; however, a variable clinical response is attributed partly to nonuniform processing methods found among numerous bone banks and commercial suppliers. DBMs remain reasonably safe and effective products. The ultimate safe bone-graft substitute, one that is osteoconductive, osteoinductive, osteogenic and mechanically strong, remains elusive.

Growth and Developmental Control in the Model and Pathogenic Aspergilli

The filamentous fungi comprise a ubiquitous group of heterotrophic organisms living as saprophytes, parasites, or symbionts. The basis for fungal vegetative growth is the continued and coordinated expansion of a series of fungal cell tips into a linear or complex structure. Fungi differentiate into a variety of structures including spores, which are the effective means of genome protection, survival, and propagation. Spores are also the primary agent for infecting host organisms for many human- and plant-pathogenic fungi. Asexual sporulation is a prevalent mode of reproduction for a diverse group of fungi, which results in the production of vast numbers of mitotically derived spores (reviewed in reference 2). The genus Aspergillus represents the most widespread fungi in our environment, which all reproduce asexually by forming long chains of conidiospores (or conidia) radiating from a central structure known as a conidiophore (Fig. 1) (reviewed in reference 3). The impact of various Aspergillus species on humans runs the range from “good” to “bad.” For instance, several species such as Aspergillus oryzae and Aspergillus niger are used in industry for enzyme production and food processing. In contrast, Aspergillus flavus and Aspergillus parasiticus can produce the most potent naturally present carcinogen, aflatoxin, which can contaminate various foods and feeds (reviewed in reference 43). Moreover, the opportunistic human pathogen Aspergillus fumigatus has become the most prevalent airborne fungal pathogen, causing severe and usually fatal invasive aspergillosis in immunocompromised patients (reviewed in reference 22). Aspergillus nidulans has served as an excellent model system for studying various biological questions, primarily due to the ease of genetic analysis through meiotic (sexual) recombination and the development of sophisticated molecular tools (32). These properties have provided a better understanding of the mechanisms controlling growth, development, secondary metabolism, and other aspects of cell biology in filamentous fungi (3, 35, 50, 57). The availability of the genome sequences of several aspergilli facilitates comparative genomic, genetic, and functional studies. In particular, knowledge and information obtained from a model fungus can be effectively tested in less genetically tractable aspergilli. Recent studies of a number of Aspergillus species have proven that a model fungus can provide a useful framework for understanding the biology of agriculturally and/or medically relevant aspergilli (6, 7, 38, 41, 49, 57). For instance, the novel nuclear protein LaeA has been shown to function as a global regulator of secondary metabolism as well as a regulator of morphogenetic virulence factors in the genus Aspergillus (6, 7). This review summarizes our current understanding of the genetic mechanisms controlling asexual development (conidiation) and vegetative growth in the model (A. nidulans) and pathogenic (A. fumigatus) aspergilli.

Influence of Physico-Chemical Properties, Macro- and Microstructure on Osteoinductive Potential of Calcium-Phosphate Ceramics

Osteoinduction by biomaterials that initially do not contain bone morphogenetic proteins and other growth factors has been shown to be a real phenomenon by many investigators in the past two decades. Although it is well-known that a material needs to meet very specific requirements in terms of physico-chemical and structural properties in order to be osteoinductive, the underlying mechanism of osteoinduction is not fully unraveled yet. In the present study we investigated parameters which are of importance for the osteoinductive potential of biomaterials by comparing four biphasic calcium-phosphate and a carbonated apatite ceramic. The results showed that the presence of micropores, by which the specific surface area of a material is increased, is essential for the material’s osteoinductivity. However, if the surface area is too high, or material is too resorbable because of its chemical composition, the implant might degrade and lose its shape. In that case, ectopic bone formation does not occur, as a relatively stable surface is needed to facilitate new bone growth.

Osteogenic Protein-1 Injection Into a Degenerated Disc Induces the Restoration of Disc Height and Structural Changes in the Rabbit Anular Puncture Model

In vivo study of the effect of injection of osteogenic protein-1 (OP-1) on a rabbit anular needle puncture model of intervertebral disc (IVD) degeneration. To study radiographic, magnetic resonance imaging (MRI), biochemical, and histologic changes in the rabbit IVD after injection of OP-1 into the nucleus pulposus in a needle puncture disc degeneration model. Growth factors, such as OP-1, have the ability to stimulate synthesis of proteoglycans and collagen in vitro. The in vivo injection of OP-1 into the normal rabbit IVD has increased disc height and proteoglycan content in the anulus fibrosus and nucleus pulposus. However, to our knowledge, no attempts have yet been made to determine the effects of these growth factors in an in vivo model of disc degeneration. New Zealand adolescent white rabbits (n = 90, 8 for baseline evaluation, 82 at 8 times) received an anular puncture in 2 noncontiguous discs with an 18-gauge needle to induce disc degeneration. Four weeks later, either 5% lactose (10 microL) or OP-1 (100 microg in 10 microL 5% lactose) was injected into the center of the nucleus pulposus. The disc height was followed radiographically for up to 24 weeks after the injections. At the 2, 4, 8, 12, and 24-week times after the injection, rabbits were euthanized, and MRI of the harvested spinal columns was obtained to grade the degeneration. The discs injected with OP-1 or lactose and noninjected discs were subjected to biochemical and histologic analysis. The specimens at the 24-week time were limited to histologic evaluation. The anular puncture with a needle induced a consistent disc narrowing within 4 weeks. The injection of OP-1 induced a restoration of disc height at 6 weeks, which was sustained for the entire experimental period, up to 24 weeks after the injection. The injection of lactose alone did not change the course of disc narrowing over the same time. MRI grading score showed significant differences between the OP-1 and lactose groups at the 8, 12, and 24-week times, suggesting an increase in water content in the nucleus pulposus of the OP-1 group. The proteoglycan content of the nucleus pulposus and anulus fibrosus was significantly higher in the OP-1 group than in the control group. The degeneration grades of the punctured discs in the OP-1 group were significantly lower than those in the lactose group. The results of this study show the feasibility of restoring degenerative rabbit discs by a single injection of OP-1 into the nucleus pulposus. Importantly, the effects of the OP-1 injection on disc height were sustained for up to 24 weeks. The metabolic changes in the cells, following a single injection, might be sustained and, thus, induce long-term changes in disc structure. An efficacy study in large animals is required to show further that the intradiscal injection of OP-1, or bone morphogenetic proteins or growth factors with similar properties would be useful for the structural restoration of the IVD in humans.

Quantitative trait loci for adventitious and lateral roots in rice

AbstractThe mass of a root is determined by two underlying morphogenetic factors: (1) the length of the main root axis (root axis length: RAL), and (2) the amount of lateral root development generated by a unit length of the axis (branching index: BI). The mass of a rice adventitious root was partitioned into these two factors at the quantitative trait locus (QTL) level, using ‘Akihikari’ (a lowland cultivar as a recurrent parent) × ‘IRAT 109’ (an upland cultivar) backcross inbred lines. Four QTLs were detected for the dry weight of individual adventitious roots (root dry weight: RDW), which were associated with a QTL for RAL (chromosomes 1 and 9) or a QTL for BI (chromosomes 6 and 11). For BI, an additional QTL was detected on chromosome 1. The absence of co‐located QTLs suggested that RAL and BI are under separate genetic control. Despite its small population size and few replications, this study suggests that the partitioning of root mass into these underlying components could be useful for understanding the complexity involved in the genetic control of root growth.

Differential epithelial expression of SHH and FOXF1 in usual and nonspecific interstitial pneumonia

Morphogenetic factors have been shown to play a role in embryogenesis and post-embryonic disease. Interstitial pulmonary fibrosis is a chronic and often progressive disorder that can lead to end-stage cystic lung. Its two major subtypes, usual interstitial pneumonitis (UIP) and nonspecific interstitial pneumonitis (NSIP) differ in their response to immunosuppressive regimens, with UIP having a significantly worse prognosis. The clinical and histologic overlap between these disorders is substantial, and there are no ancillary findings that can accurately distinguish them. We examined surgical and autopsy specimens of lung in 13 cases of patients with either UIP or the fibrotic variant of NSIP (NSIP-F) for their expression of Sonic hedgehog (Shh) and Foxf1 in situ. We identified a pattern of strong Shh expression with weak expression of Foxf1 in all cases of UIP and a complementary expression of Shh and Foxf1 in cases of NSIP-F. We conclude that morphogenetic genes may participate differentially in the pathogenesis of UIP and NSIP-F.

Osteogenic differentiation induced by bone morphogenetic proteins can be suppressed by platelet‐released supernatant in vitro

Both bone morphogenetic proteins (BMPs) and growth factors released from activated platelets both occur at sites of bone regeneration but their functional relationship to regulate the temporal and spatial sequence of cellular events is not well defined. Here we investigated whether supernatants derived from activated platelets can modulate the response of the osteogenic cell line MC3T3-E1 to BMPs, and whether BMPs have an effect on MC3T3-E1 cells stimulated with platelet-released supernatant. Platelet-released supernatant suppressed BMP-2-, BMP-6-, and BMP-7-induced osteogenic differentiation of MC3T3-E1 cells, as indicated by the significant decrease in alkaline phosphatase activity and lower levels of osteocalcin transcripts, whereas BMP-2, BMP-6, and BMP-7 did not modulate migration and proliferation of MC3T3-E1 cells, which were stimulated with platelet-released supernatant. Osteogenic differentiation in response to BMPs was not affected after precultivation of MC3T3-E1 cells with platelet-released supernatant. These data suggest that activated platelets can provide a microenvironment that temporarily suppresses the differentiation of osteogenic cells in the presence of BMPs. Future strategies to stimulate bone regeneration should take the suppression of BMP-induced osteogenic differentiation during the existence of the blood clot into account.

Zonal gene expression in murine liver: Are tumors helping us to solve the mystery?

Hailfinger and colleagues1 put forward an interesting hypothesis to explain the mystery of hepatocyte heterogeneity. Based on the observation that tumors carrying mutations of either the H-Ras or Catnb genes express sets of genes resembling those predominating in periportal and pericentral regions of liver lobuli, they propose that hepatocyte heterogeneity is determined by two opposing gradients. The first of these would involve soluble factors from the bloodstream inducing Ras activation prevailing in the periportal zone, while the second would consist of an inverse gradient of Wnt signals inducing activation of β-catenin prevailing in the pericentral zone. Although this hypothesis is attractive for a number of reasons, the experimental evidence provided by Hailfinger et al.1 does not in itself constitute substantial support. Recent observations have left no doubt that expression of glutamine synthetase (GS), the most pronounced marker enzyme of pericentral cells,2 is somehow dependent on β-catenin signaling.3-5 However, although activation of β-catenin seems necessary, it does not appear to be sufficient, since many liver tumors with activated β-catenin do not express GS. The experimental approach taken by Hailfinger et al.1 (to induce pericentral genes in cultured hepatocytes using Wnt factors or inhibitors of GSK3β) is not conclusive, since whole hepatocyte preparations contain both periportal and pericentral cells of which only the pericentral subpopulation might respond. Moreover, the mechanism by which β-catenin signaling might influence GS expression remains obscure, since indication of a permanent activation of β-catenin in the GS-positive zone of the lobuli in adult liver is missing. Rather, during regeneration following liver damage in zone 3 we recently observed the transient activation and cytoplasmic accumulation of β-catenin in the most distal pericentral hepatocytes immediately before GS expression is switched on.6 This suggests some kind of zonal reprogramming of otherwise similarly differentiated hepatocytes which we would like to call postdifferentiation tissue patterning consistent with the role of Wnts in many similar phenomena during embryogenesis and tissue formation.7 This is also consistent with the early observation that cultured GS-positive hepatocytes maintain their phenotype throughout their lifespan in vitro despite cultivation in media mimicking periportal conditions.8 The evidence provided by Hailfinger et al.1 concerning periportal signals is less clear. Firstly, the proposed influence of Ras activation was not demonstrated in vitro. Secondly, the finding that hepatocellular tumors develop rarely in response to β-catenin mutation alone, but frequently in the presence of an additional mutation in the H-Ras gene,9 renders it doubtful that all of the tumors exhibiting β-catenin mutations in the study by Hailfinger et al.1 were really lacking Ras mutations. Thirdly, Hailfinger et al. maintain that periportal Ras activation results from blood-borne factors.1 This assumption neglects earlier evidence that liver zonation in ectopic explants develops correctly even without portal blood or the natural orientation of blood flow.10 Thus, the underlying mechanism of liver parenchymal heterogeneity remains an intrahepatic issue controlled by as yet unknown (morphogenetic) factors. Wnts and β-catenin-dependent signaling certainly have a role to play, but this seems unlikely to be in the context proposed by Hailfinger et al.1 Rolf Gebhardt*, Elke Ueberham*, * Institute of Biochemistry, University of Leipzig, Leipzig, Germany.

LaeA, a Regulator of Morphogenetic Fungal Virulence Factors

Opportunistic animal and plant pathogens, well represented by the genus Aspergillus, have evolved unique mechanisms to adapt to and avoid host defenses. Aspergillus fumigatus, an increasingly serious pathogen owing to expanding numbers of immunocompromised patients, causes the majority of human infections; however, an inability to identify bona fide virulence factors has impeded therapeutic advances. We show that an A. fumigatus mutation in a developmentally expressed transcriptional regulator (deltalaeA) coordinating morphological and chemical differentiation reduces virulence in a murine model; impaired virulence is associated with decreased levels of pulmonary gliotoxin and multiple changes in conidial and hyphal susceptibility to host phagocytes ex vivo. LaeA, a conserved protein in filamentous fungi, is a developmental regulator of virulence genes and, possibly, the first antimicrobial target specific to filamentous fungi that are pathogenic to plants and animals.

Automated 3-D Reconstruction of the Surface of Live Early-Stage Amphibian Embryos

Although three-dimensional (3-D) reconstructions of the surfaces of live embyos are vital to understanding embryo development, morphogenetic tissue movements and other factors have prevented the automation of this task. Here, we report an integrated set of software algorithms that overcome these challenges, making it possible to completely automate the reconstruction of embryo surfaces and other textured surfaces from multiview images. The process involves: 1) building accurate point correspondences using a robust deformable template block matching algorithm; 2) removing outliers using fundamental matrix calculations in conjunction with a RANSAC algorithm; 3) generating 3-D point clouds using a bundle adjustment algorithm that includes camera position and distortion corrections; 4) meshing the point clouds into triangulated surfaces using a Tight Cocone algorithm that produces water tight models; 5) refining surfaces using midpoint insertion and Laplacian smoothing algorithms; and 6) repeating these steps until a measure of convergence G, the rms difference between successive reconstructions, is below a specified threshold. Reconstructions were made of 2.2-mm diameter, neurulation-stage axolotl (amphibian) embryos using 44 multiview images collected with a robotic microscope. A typical final model (sixth iteration) contained 3787 points and 7562 triangles and had an error measure of G = 5.9 microm.