Probing the S100 protein family through genomic and functional analysis

The EF-hand superfamily of calcium binding proteins includes the S100, calcium binding protein, and troponin subfamilies. This study represents a genome, structure, and expression analysis of the S100 protein family, in mouse, human, and rat. We confirm the high level of conservation between mammalian sequences but show that four members, including S100A12, are present only in the human genome. We describe three new members of the S100 family in the three species and their locations within the S100 genomic clusters and propose a revised nomenclature and phylogenetic relationship between members of the EF-hand superfamily. Two of the three new genes were induced in bone-marrow-derived macrophages activated with bacterial lipopolysaccharide, suggesting a role in inflammation. Normal human and murine tissue distribution profiles indicate that some members of the family are expressed in a specific manner, whereas others are more ubiquitous. Structure-function analysis of the chemotactic properties of murine S100A8 and human S100A12, particularly within the active hinge domain, suggests that the human protein is the functional homolog of the murine protein. Strong similarities between the promoter regions of human S100A12 and murine S100A8 support this possibility. This study provides insights into the possible processes of evolution of the EF-hand protein superfamily. Evolution of the S100 proteins appears to have occurred in a modular fashion, also seen in other protein families such as the C2H2-type zinc-finger family. (C) 2004 Elsevier Inc. All rights reserved.

Joining S100 proteins and migration:for better or for worse, in sickness and in health

The vast diversity of S100 proteins has demonstrated a multitude of biological correlations with cell growth, cell differentiation and cell survival in numerous physiological and pathological conditions in all cells of the body. This review summarises some of the reported regulatory functions of S100 proteins (namely S100A1, S100A2, S100A4, S100A6, S100A7, S100A8/S100A9, S100A10, S100A11, S100A12, S100B and S100P) on cellular migration and invasion, established in both culture and animal model systems and the possible mechanisms that have been proposed to be responsible. These mechanisms involve intracellular events and components of the cytoskeletal organisation (actin/myosin filaments, intermediate filaments and microtubules) as well as extracellular signalling at different cell surface receptors (RAGE and integrins). Finally, we shall attempt to demonstrate how aberrant expression of the S100 proteins may lead to pathological events and human disorders and furthermore provide a rationale to possibly explain why the expression of some of the S100 proteins (mainly S100A4 and S100P) has led to conflicting results on motility, depending on the cells used. © 2013 Springer Basel.

Transcriptome analysis of neutrophils after endurance exercise reveals novel signaling mechanisms in the immune response to physiological stress

Neutrophils serve as an intriguing model for the study of innate immune cellular activity induced by physiological stress. We measured changes in the transcriptome of circulating neutrophils following an experimental exercise trial (EXTRI) consisting of 1 h of intense cycling immediately followed by 1 h of intense running. Blood samples were taken at baseline, 3 h, 48 h, and 96 h post-EXTRI from eight healthy, endurance-trained, male subjects. RNA was extracted from isolated neutrophils. Differential gene expression was evaluated using Illumina microarrays and validated with quantitative PCR. Gene set enrichment analysis identified enriched molecular signatures chosen from the Molecular Signatures Database. Blood concentrations of muscle damage indexes, neutrophils, interleukin (IL)-6 and IL-10 were increased (P < 0.05) 3 h post-EXTRI. Upregulated groups of functionally related genes 3 h post-EXTRI included gene sets associated with the recognition of tissue damage, the IL-1 receptor, and Toll-like receptor (TLR) pathways (familywise error rate, P value < 0.05). The core enrichment for these pathways included TLRs, low-affinity immunoglobulin receptors, S100 calcium binding protein A12, and negative regulators of innate immunity, e.g., IL-1 receptor antagonist, and IL-1 receptor associated kinase-3. Plasma myoglobin changes correlated with neutrophil TLR4 gene expression (r = 0.74; P < 0.05). Neutrophils had returned to their nonactivated state 48 h post-EXTRI, indicating that their initial proinflammatory response was transient and rapidly counterregulated. This study provides novel insight into the signaling mechanisms underlying the neutrophil responses to endurance exercise, suggesting that their transcriptional activity was particularly induced by damage-associated molecule patterns, hypothetically originating from the leakage of muscle components into the circulation.

A novel method using intranasal delivery of EdU demonstrates that accessory olfactory ensheathing cells respond to injury by proliferation

Olfactory ensheathing cells (OECs) play an important role in the continuous regeneration of the primary olfactory nervous system throughout life and for regeneration of olfactory neurons after injury. While it is known that several individual OEC subpopulations with distinct properties exist in different anatomical locations, it remains unclear how these different subpopulations respond to a major injury. We have examined the proliferation of OECs from one distinct location, the peripheral accessory olfactory nervous system, following large-scale injury (bulbectomy) in mice. We used crosses of two transgenic reporter mouse lines, S100ß-DsRed and OMP-ZsGreen, to visualise OECs, and main/accessory olfactory neurons, respectively. We surgically removed one olfactory bulb including the accessory olfactory bulb to induce degeneration, and found that accessory OECs in the nerve bundles that terminate in the accessory olfactory bulb responded by increased proliferation with a peak occurring 2 days after the injury. To label proliferating cells we used the thymidine analogue ethynyl deoxyuridine (EdU) using intranasal delivery instead of intraperitoneal injection. We compared and quantified the number of proliferating cells at different regions at one and four days after EdU labelling by the two different methods and found that intranasal delivery method was as effective as intrapeitoneal injection. We demonstrated that accessory OECs actively respond to widespread degeneration of accessory olfactory axons by proliferating. These results have important implications for selecting the source of OECs for neural regeneration therapies and show that intranasal delivery of EdU is an efficient and reliable method for assessing proliferation of olfactory glia.

Web crippling tests of hollow flange channel beams : ETF AND ITF load cases

This paper presents the details of an experimental study of a cold-formed steel hollow flange channel beam known as LiteSteel Beam (LSB) subject to web crippling actions (ETF and ITF). Due to the geometry of the LSB, as well as its unique residual stress characteristics and initial geometric imperfections resultant of manufacturing processes, much of the existing research for common cold-formed steel sections is not directly applicable to LSB. Experimental and numerical studies have been carried out to evaluate the behaviour and design of LSBs subject to pure bending actions, predominant shear actions and combined actions. To date, however, no investigation has been conducted into the web crippling behaviour and strength of LSB sections under ETF and ITF load conditions. Hence experimental studies were conducted to assess the web crippling behaviour and strengths of LSBs. Twenty eight web crippling tests were conducted and the results were compared with the current AS/NZS 4600[1] and AISI S100 [2]design equations. Comparison of the ultimate web crippling capacities from tests showed that AS/NZS 4600[1] and AISI S100 [2] design equations are unconservative for LSB sections under ETF and ITF load cases. Hence new equations were proposed to determine the web crippling capacities of LSBs. Suitable design rules were also developed under the DSM format.

Flexural–torsional buckling behaviour and design of cold-formed steel compression members at elevated temperatures

Current design rules for the member capacities of cold-formed steel columns are based on the same non-dimensional strength curve for both fixed and pinned-ended columns at ambient temperature. This research has investigated the accuracy of using current ambient temperature design rules in Australia/New Zealand (AS/NZS 4600), American (AISI S100) and European (Eurocode 3 Part 1.3) standards in determining the flexural–torsional buckling capacities of cold-formed steel columns at uniform elevated temperatures using appropriately reduced mechanical properties. It was found that these design rules accurately predicted the member capacities of pin ended lipped channel columns undergoing flexural torsional buckling at elevated temperatures. However, for fixed ended columns with warping fixity undergoing flexural–torsional buckling, the current design rules significantly underestimated the column capacities as they disregard the beneficial effect of warping fixity. This paper has therefore recommended the use of improved design rules developed for ambient temperature conditions to predict the axial compression capacities of fixed ended columns subject to flexural–torsional buckling at elevated temperatures within AS/NZS 4600 and AISI S100 design provisions. The accuracy of the proposed fire design rules was verified using finite element analysis and test results of cold-formed lipped channel columns at elevated temperatures except for low strength steel columns with intermediate slenderness whose behaviour was influenced by the increased nonlinearity in the stress–strain curves at elevated temperatures. Further research is required to include these effects within AS/NZS 4600 and AISI S100 design rules. However, Eurocode 3 Part 1.3 design rules can be used for this purpose by using suitable buckling curves as recommended in this paper.

Experimental and numerical studies of fire exposed lipped channel columns subject to distortional buckling

Cold-formed steel sections are commonly used in low-rise commercial and residential buildings. During fire events, cold-formed steel structural elements in these buildings are exposed to elevated temperatures. Hence after such events there is a need to determine the residual strength of these structural elements. However, only limited information is available in relation to the residual strength of fire exposed cold-formed steel members. This research is aimed at investigating the residual distortional buckling capacities of fire exposed cold-formed steel lipped channel sections. A series of compression tests of fire exposed, short lipped channel columns made of varying steel grades and thicknesses was undertaken in this research. Test columns were exposed to different elevated temperatures up to 800 oC. They were then allowed to cool down at ambient temperature before they were tested to failure. Suitable finite element models of tested columns were also developed and validated using test results. The residual compression capacities of tested columns were predicted using the ambient temperature cold-formed steel design rules (AS/NZS 4600, AISI S100 and Direct Strength Method). Post-fire mechanical properties obtained from a previous study were used in this study. Comparison of results showed that ambient temperature design rules for compression members can be used to predict the residual compression capacities of fire exposed short or laterally restrained cold-formed steel columns provided the maximum temperature experienced by the columns can be estimated after a fire event. Such residual capacity assessments will allow structural and fire engineers to make an accurate prediction of the safety of buildings after fire events. This paper presents the details of these experimental and numerical studies and the results.

Web crippling studies of Duragal channel sections – ETF and ITF load cases

Cold-formed steel members have many advantages over hot-rolled steel members. However, they are susceptible to various buckling modes at stresses below the yield stress of the member because of their relatively high width-to-thickness ratio. Web crippling is one of the failure modes that can occur when the members are subjected to transverse high concentrated loadings and/or reactions. The four common loading conditions are the end-one-flange (EOF), interior-one-flange (IOF), end-two-flange (ETF) and interior-two-flange (ITF) loadings. Recently a new test method has been proposed by AISI to obtain the web crippling capacities under these four loading conditions. Using this test method 38 tests were conducted in this research to investigate the web crippling behaviour and strength of channel beams under ETF and ITF cases. Unlipped channel sections having a nominal yield stress of 450 MPa were tested with different web slenderness and bearing lengths. The flanges of these channel sections were not fastened to the supports. In this research the suitability of the current design rules in AS/NZS 4600 and the AISI S100 Specification for unlipped channels subject to web crippling was investigated, and suitable modifications were proposed where necessary. In addition to this, a new design rule was proposed based on the direct strength method to predict the web crippling capacities of tested beams. This paper presents the details of this experimental study and the results.

Experimental studies on web crippling behaviour of hollow flange channel beams under two flange load cases

This paper presents the details of an experimental study of a cold-formed steel hollow flange channel beam known as LiteSteel beam (LSB) subject to web crippling under End Two Flange (ETF) and Interior Two Flange (ITF) load cases. The LSB sections with two rectangular hollow flanges are made using a simultaneous cold-forming and electric resistance welding process. Due to the geometry of the LSB, and its unique residual stress characteristics and initial geometric imperfections, much of the existing research for common cold-formed steel sections is not directly applicable to LSB. Experimental and numerical studies have been carried out to evaluate the behaviour and design of LSBs subject to pure bending, predominant shear and combined actions. To date, however, no investigation has been conducted on the web crippling behaviour and strength of LSB sections. Hence an experimental study was conducted to investigate the web crippling behaviour and capacities of LSBs. Twenty-eight web crippling tests were conducted under ETF and ITF load cases, and the ultimate web crippling capacities were compared with the predictions from the design equations in AS/NZS 4600 and AISI S100. This comparison showed that AS/NZS 4600 and AISI S100 web crippling design equations are unconservative for LSB sections under ETF and ITF load cases. Hence new equations were proposed to determine the web crippling capacities of LSBs based on experimental results. Suitable design rules were also developed under the direct strength method (DSM) format.

Olfactory ensheathing cells are the main phagocytic cells that remove axon debris during early development of the olfactory system

During development of the primary olfactory system, axon targeting is inaccurate and axons inappropriately project within the target layer or overproject into the deeper layers of the olfactory bulb. As a consequence there is considerable apoptosis of primary olfactory neurons during embryonic and postnatal development and axons of the degraded neurons need to be removed. Olfactory ensheathing cells (OECs) are the glia of the primary olfactory nerve and are known to phagocytose axon debris in the adult and postnatal animal. However, it is unclear when phagocytosis by OECs first commences. We investigated the onset of phagocytosis by OECs in the developing mouse olfactory system by utilizing two transgenic reporter lines: OMP-ZsGreen mice which express bright green fluorescent protein in primary olfactory neurons, and S100β-DsRed mice which express red fluorescent protein in OECs. In crosses of these mice, the fate of the degraded axon debris is easily visualized. We found evidence of axon degradation at embryonic day (E)13.5. Phagocytosis of the primary olfactory axon debris by OECs was first detected at E14.5. Phagocytosis of axon debris continued into the postnatal animal during the period when there was extensive mistargeting of olfactory axons. Macrophages were often present in close proximity to OECs but they contributed only a minor role to clearing the axon debris, even after widespread degeneration of olfactory neurons by unilateral bulbectomy and methimazole treatment. These results demonstrate that from early in embryonic development OECs are the primary phagocytic cells of the primary olfactory nerve.

Web crippling tests of rivet fastened rectangular hollow flange channel beams under two flange load cases

Recent research on hollow flange beams has led to the development of an innovative rectangular hollow flange channel beam (RHFCB) for use in floor systems. The new RHFCB is a mono-symmetric structural section made by intermittently rivet fastening two torsionally rigid closed rectangular hollow flanges to a web plate element, which allows section optimisation by selecting appropriate combinations of web and flange widths and thicknesses. However, the current design rules for cold-formed steel sections are not directly applicable to rivet fastened RHFCBs. To date, no investigation has been conducted on their web crippling behaviour and strengths. Hence an experimental study was conducted to investigate the web crippling behaviour and capacities of rivet fastened RHFCBs under End Two Flange (ETF) and Interior Two Flange (ITF) load cases. It showed that RHFCBs failed by web crippling, flange crushing and their combinations. Comparison of ultimate web crippling capacities with the predictions from the design equations in AS/NZS 4600 and AISI S100 showed that the current design equations are unconservative for rivet fastened RHFCB sections under ETF and ITF load cases. Hence new equations were proposed to determine the web crippling capacities of rivet fastened RHFCBs. These equations can also be used to predict the capacities of RHFCBs subject to combined web crippling and flange crushing conservatively. However, new capacity equations were proposed in the case of flange crushing failures that occurred in thinner flanges with smaller bearing lengths. This paper presents the details of this web crippling experimental study of RHFCB sections and the results.

Experimental studies of lipped channel beams subject to web crippling under ETF and ITF load cases

Lipped channel beams (LCBs) are commonly used as floor joists and bearers in buildings. However, they are subjected to specific failure modes such as web crippling. Despite considerable web crippling research, recent studies [1-6] have shown that the current web crippling design rules are unable to predict the test capacities under ETF and ITF load cases. In many instances, the predictions by the available design standards such as AISI S100, AS/NZS 4600 and Eurocode 3 Part 1-3 [7-9] are inconsistent. Hence thirty-six tests were conducted to assess the web crippling behaviour and strengths of LCBs under two flange load cases. Experimental web crippling capacities were then compared with the predictions from the current design rules. These comparisons showed that AS/NZS 4600 and AISI S100 design equations are very unconservative for LCB sections under ETF load case and are conservative for ITF load case. Hence improved equations were proposed to determine the web crippling capacities of LCBs. Suitable design rules were also developed using the direct strength method. This paper presents the details of this study and the results including improved design rules.

Bearing capacity of cold-formed unlipped channels with restrained flanges under EOF and IOF load cases

Bearing failure is a form of localized failure that occurs when thin-walled cold-formed steel sections are subjected to concentrated loads or support reactions. To determine the bearing capacity of cold-formed channel sections, a unified design equation with different bearing coefficients is given in the current North American specification AISI S100 and the Australian/New Zealand standard AS/NZS 4600. However, coefficients are not available for unlipped channel sections that are normally fastened to supports through their flanges. Eurocode 3 Part 1.3 includes bearing capacity equations for different load cases, but does not distinguish between fastened and unfastened support conditions. Therefore, an experimental study was conducted to determine the bearing capacities of these sections as used in floor systems. Twenty-eight web bearing tests on unlipped channel sections with restrained flanges were conducted under End One Flange (EOF) and Interior One Flange (IOF) load cases. Using the results from this study, a new equation was proposed within the AISI S100 and AS/NZS 4600 guidelines to determine the bearing capacities of cold-formed unlipped channels with flanges fastened to supports. A new design rule was also proposed based on the direct strength method.