Multiple pathways contribute to the hyperproliferative responses from truncated granulocyte colony-stimulating factor receptors

Mutations in the granulocyte colony-stimulating factor receptor (G-CSF-R) gene leading to a truncated protein have been identified in a cohort of neutropenia patients highly predisposed to acute myeloid leukemia. Such mutations act in a dominant manner resulting in hyperproliferation but impaired differentiation in response to G-CSF. This is due, at least in part, to defective internalization and loss of binding sites for several negative regulators, leading to sustained receptor activation. However, those signaling pathways responsible for mediating the hyperproliferative function have remained unclear. In this study, analysis of an additional G-CSF-R mutant confirmed the importance of residues downstream of Box 2 as important contributors to the sustained proliferation. However, maximal proliferation correlated with the ability to robustly activate signal transducer and activator of transcription (STAT) 5 in a sustained manner, whereas co-expression of dominant-negative STAT5, but not dominant-negative STAT3, was able to inhibit G-CSF-stimulated proliferation from a truncated receptor. Furthermore, a Janus kinase (JAK) inhibitor also strongly reduced the proliferative response, whereas inhibitors of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) or phosphatidylinositol (PI) 3-kinase reduced proliferation to a lesser degree. These data suggest that sustained JAK2/STAT5 activation is a major contributor to the hyperproliferative function of truncated G-CSF receptors, with pathways involving MEK and PI 3-kinase playing a reduced role.

Molecular characterization and analysis of a truncated serotonin receptor gene expressed in neural and reproductive tissues of abalone

In molluscs, the neurotransmitter serotonin (5-HT) has been linked to a variety of biological roles including gamete maturation and spawning. The possible involvement of 5-HT in abalone gamete release was demonstrated by a dose-dependent increase in Haliotis rubra gonad contractile bioactivity following 5-HT stimulation. Physiological functions associated with 5-HT, are mediated through binding to 5-HT receptors. A cDNA encoding a putative 5-HT receptor consisting of 359 amino acids was isolated from the tropical abalone H. asinina, termed 5-HT_{1 ha}. The 5-HT_{1 ha} shares G-protein-coupled receptor motifs with metazoan 5-HT receptors, including predicted transmembrane domains, active sites for protein kinase action, and N-linked glycosylation sites. However, the third intracellular loop of 5-HT_{1 ha} is relatively short, and only six transmembrane domains are predicted, implying a truncated receptor. Phylogenetic analysis with known 5-HT receptor genes suggests that 5-HT_{1 ha} belongs to the type 1 5-HT receptor family. Expression analysis by RT-PCR showed that 5-HT_{1 ha}  mRNA was present in all tissues examined, including the neural ganglia and gonad tissues. Immunocytochemistry revealed the presence of 5-HT_{1 ha} specifically within the soma of neuronal cells located in the outer cortex of both cerebral and pleuropedal ganglia. In ovarian and testicular tissues, 5-HT_{1 ha} immunoreactivity was observed in epithelial cells of the outer capsule and connective tissue of the trabeculae to which the gamete follicles adhere. Whether this receptor transcript is translated to a functional protein needs to be verified, but if so, it could play a role in reproduction.

Gaussian vault geometry : digital design and fabrication of scaled prototypes

This paper reports on three approaches to the translation of Gaussian surface models into scaled physical prototype models. Using the geometry of Eladio Dieste's Gaussian Vaults, the paper reports on the aspects encountered in the process of digital to physical prototype fabrication. The primary focus of the paper is on exploring the design geometry, investigating methods for preparing the geometry for fabrication and constructing physical prototypes. Three different approaches in the translation from digital to physical models are investigated: rapid prototyping, two dimensional surface models in paper and structural component models using Computer Numerical Controlled (CNC) fabrication. The three approaches identify a body of knowledge in the design and prototyping of Gaussian vaults. Finally the paper discusses the digital to. fabrication translation processes with regards to the characteristics, benefits and limitations of the three approaches of prototyping the ruled surface geometry of Gaussian Vaults.

Reliability and variability of day-to-day vault training measures in artistic gymnastics

Inter-day training reliability and variability in artistic gymnastics vaulting was determined using a customised infra-red timing gate and contact mat timing system. Thirteen Australian high performance gymnasts (eight males and five females) aged 11-23 years were assessed during two consecutive days of normal training. Each gymnast completed a number of vault repetitions per daily session. Inter-day variability of vault run-up velocities (at -18 to -12 m, -12 to -6 m, -6 to -2 m, and -2 to 0 m from the nearest edge of the beat board), and board contact, pre-flight, and table contact times were determined using mixed modelling statistics to account for random (within-subject variability) and fixed effects (gender, number of subjects, number of trials). The difference in the mean (Mdiff) and Cohen's effect sizes for reliability assessment and intra-class correlation coefficients, and the coefficient of variation percentage (CV%) were calculated for variability assessment. Approach velocity (-18 to -2 m, CV = 2.4-7.8%) and board contact time (CV = 3.5%) were less variable measures when accounting for day-to-day performance differences, than pre-flight time (CV = 17.7%) and table contact time (CV = 20.5%). While pre-flight and table contact times are relevant training measures, approach velocity and board contact time are more reliable when quantifying vaulting performance.

Computation and construction of vault geometry prototypes

Physical models and scaled prototypes of architecture play an important role in design. They enable architects and designers to investigate the formal, functional, and material attributes of the design. Understanding digital processes of realizing scaled prototypes is a significant problem confronting design practice. This paper reports on three approaches to the translation of Gaussian surface models into scaled physical prototype models. Based on the geometry of Eladio Dieste’s Gaussian Vaults, the paper reports on the aspects encountered in the process of digital to physical construction using scaled prototypes. The primary focus of the paper is on computing the design geometry, investigating methods for preparing the geometry for fabrication and physical construction. Three different approaches in the translation from digital to physical models are investigated: rapid prototyping, two-dimensional surface models in paper and structural component models using CNC fabrication. The three approaches identify a body of knowledge in the design and prototyping of Gaussian vaults. Finally the paper discusses the digital to fabrication translation processes with regards to the characteristics, benefits and limitations of the three approaches of prototyping the ruled surface geometry of Gaussian Vaults. The results of each of three fabrication processes allowed for a better understanding of the digital to physical translation process. The use of rapid prototyping permits the production of form models that provide a representation of the physical characteristics such as size, shape and proportion of the Gaussian Vault.

Gaussian vault geometry : integrated approach in design and fabrication of physical prototypes

This paper reports the second part of a study on the digital design and fabrication of scaled architectural prototypes. The first paper reported techniques in the realization of a double curved vault surface, the Gaussian Vault. The aims of the research here further extend this body of knowledge to a better understanding of constructible components. It addresses the problem of fabricating complex curved forms through the integration of the basic building elements, skin and structure, to achieve a scaled physical prototype. The focus of the experimentation is to investigate the process from which a digital surface form is conceived, to its preparation for fabrication and eventual construction in the fashion of a scaled model or workable prototype.

Synthesis and characterization of hexagonal and truncated hexagonal shaped MoO3 nanoplates

Hexagonal and truncated hexagonal shaped MoO₃ nanoplates (MoO₃ HNP) were synthesized through a simple vapor-deposition method in Ar atmosphere under ambient pressure without the assistant of any catalysts. The structure and morphology of MoO₃ HNP were investigated by XRD, EDX, SEM, TEM, and HRTEM. The results reveal that the HNP are α-MoO₃ and have a large area surface. The Raman spectrum shows a significant size effect on the vibrational property of MoO₃ HNP. The photoluminescence (PL) spectrum was carried out, and two peaks at 351 and 410 nm were observed in the spectrum. In addition, a possible growth mechanism proposed as VS is discussed in detail.

A truncated fragment of Src protein kinase generated by calpain-mediated cleavage is a mediator of neuronal death in excitotoxicity

Excitotoxicity resulting from overstimulation of glutamate receptors is a major cause of neuronal death in cerebral ischemic stroke. The overstimulated ionotropic glutamate receptors exert their neurotoxic effects in part by overactivation of calpains, which induce neuronal death by catalyzing limited proteolysis of specific cellular proteins. Here, we report that in cultured cortical neurons and in vivo in a rat model of focal ischemic stroke, the tyrosine kinase Src is cleaved by calpains at a site in the N-terminal unique domain. This generates a truncated Src fragment of ?52 kDa, which we localized predominantly to the cytosol. A cell membrane-permeable fusion peptide derived from the unique domain of Src prevents calpain from cleaving Src in neurons and protects against excitotoxic neuronal death. To explore the role of the truncated Src fragment in neuronal death, we expressed a recombinant truncated Src fragment in cultured neurons and examined how it affects neuronal survival. Expression of this fragment, which lacks the myristoylation motif and unique domain, was sufficient to induce neuronal death. Furthermore, inactivation of the prosurvival kinase Akt is a key step in its neurotoxic signaling pathway. Because Src maintains neuronal survival, our results implicate calpain cleavage as a molecular switch converting Src from a promoter of cell survival to a mediator of neuronal death in excitotoxicity. Besides unveiling a new pathological action of Src, our discovery of the neurotoxic action of the truncated Src fragment suggests new therapeutic strategies with the potential to minimize brain damage in ischemic stroke.

Technical efficiency in Australian textile and clothing firms: evidence from the business longitudinal survey

This paper estimates technical efficiency of Australian textile and clothing firms based on the data obtained from the Business Longitudinal Survey (BLS) conducted by the Australian Bureau of Statistics (ABS). Using a Cobb Douglas stochastic production frontier the paper examines firm level technical efficiency in the time varying inefficiency effect model with technical inefficiency effects assumed as an independently distributed truncated normal variable. Estimates of the production frontier revealed significant but small elasticities of labour and capital for textile and clothing firms, respectively, and a negative (but insignificant) Hicks neutral technical change for both. Estimated coefficients of the explanatory variables for inefficiency effects indicated that technical efficiency varied significantly according to firms’ age, size, capital intensity, proportion of non-production to total workers and type of legal status. Predicted firm specific efficiency varied from 16 per cent to 95 per cent and mean efficiency ranged between 30 to 70 per cent. In view of these results policies have been suggested to improve technical efficiency of the firms as well as productivity growth of the sub sectors.

ORP3 splice variants and their expression in human tissues and hematopoietic cells

ORP3 is a member of the newly described family of oxysterol-binding protein (OSBP)-related proteins (ORPs). We previously demonstrated that this gene is highly expressed in CD34+ hematopoietic progenitor cells, and deduced that the "full-length" ORP3 gene comprises 23 exons and encodes a predicted protein of 887 amino acids with a C-terminal OSBP domain and an N-terminal pleckstrin homology domain. To further characterize the gene, we cloned ORP3 cDNA from PCR products and identified multiple splice variants. A total of eight isoforms were demonstrated with alternative splicing of exons 9, 12, and 15. Isoforms with an extension to exon 15 truncate the OSBP domain of the predicted protein sequence. In human tissues there was specific isoform distribution, with most tissues expressing varied levels of isoforms with the complete OSBP domain; while only whole brain, kidney, spleen, thymus, and thyroid expressed high levels of the isoforms associated with the truncated OSBP domain. Interestingly, the expression in cerebellum, heart, and liver of most isoforms was negligible. These data suggest that differential mRNA splicing may have resulted in functionally distinct forms of the ORP3 gene.

Growth and structure of prismatic boron nitride nanorods

Prismatic boron nitride nanorods have been grown on single crystal silicon substrates by mechanical ball-milling followed by annealing at 1300 °C. Growth takes place by rapid surface diffusion of BN molecules, and follows heterogeneous nucleation at catalytic particles of an Fe/Si alloy. Lattice imaging transmission electron microscopy studies reveal a central axial row of rather small truncated pyramidal nanovoids on each nanorod, surrounded by three basal planar BN domains which, with successive deposition of epitaxial layers adapt to the void geometry by crystallographic faceting. The bulk strain in the nanorods is taken up by the presence of what appear to be simple nanostacking faults in the external, near-surface domains which, like the nanovoids are regularly repetitive along the nanorod length. Growth terminates with a clear cuneiform tip for each nanorod. Lateral nanorod dimensions are essentially determined by the size of the catalytic particle, which remains as a foundation essentially responsible for base growth. Growth, structure, and dominating facets are shown to be consistent with a system which seeks lowest bulk and surface energies according to the well-known thermodynamics of the capillarity of solids.

Functional interaction between mutations in the granulocyte colony-stimulating factor receptor in severe congenital neutropenia

Most severe congenital neutropenia (SCN) cases possess constitutive neutrophil elastase mutations; a smaller cohort has acquired mutations truncating the granulocyte colony-stimulating factor receptor (G-CSF-R). We have described a case with constitutive extracellular G-CSF-R mutation hyporesponsive to ligand. Here we report two independent acquired G-CSF-R truncation mutations and a novel constitutive neutrophil elastase mutation in this patient. Co-expression of a truncated receptor chain restored STAT5 signalling responses of the extracellular G-CSF-R mutant, while constitutively-active STAT5 enhanced its proliferative capacity. These data add to our knowledge of SCN and further highlight the importance of STAT5 in mediating proliferative responses to G-CSF.

Truncation of Plasmodium berghei merozoite surface protein 8 does not affect in vivo blood-stage development

Merozoite surface protein 8 (MSP8) has shown promise as a vaccine candidate in the Plasmodium yoelii rodent malaria model and has a proposed role in merozoite invasion of erythrocytes. However, the temporal expression and localisation of MSP8 are unusual for a merozoite antigen. Moreover, in Plasmodium falciparum the MSP8 gene could be disrupted with no apparent effect on in vitro growth. To address the in vivo function of full-length MSP8, we truncated MSP8 in the rodent parasite Plasmodium berghei. PbΔMSP8 disruptant parasites displayed a normal blood-stage growth rate but no increase in reticulocyte preference, a phenomenon observed in P. yoelii MSP8 vaccinated mice. Expression levels of erythrocyte surface antigens were similar in P. berghei wild-type and PbΔMSP8-infected erythrocytes, suggesting that a parasitophorous vacuole function for MSP8 does not involve global trafficking of such antigens. These data demonstrate that a full-length membrane-associated form of PbMSP8 is not essential for blood-stage growth.