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BMC Immunology
Anti-inflammatory protein TNFα-stimulated gene-6 (TSG-6) reduces inflammatory response after brain injury in mice
Kazadi Nadine Mutoji1,
Mingxia Sun1,
Amanda Nash1 nAff3,
Sudan Puri1,
Vincent Hascall2 &
Vivien J. Coulson-Thomas ORCID: orcid.org/0000-0002-5848-02251
BMC Immunology volume 22, Article number: 52 (2021) Cite this article
Current research suggests that the glial scar surrounding penetrating brain injuries is instrumental in preserving the surrounding uninjured tissue by limiting the inflammatory response to the injury site. We recently showed that tumor necrosis factor (TNF)-stimulated gene-6 (TSG-6), a well-established anti-inflammatory molecule, is present within the glial scar. In the present study we investigated the role of TSG-6 within the glial scar using TSG-6 null and littermate control mice subjected to penetrating brain injuries.
Our findings show that mice lacking TSG-6 present a more severe inflammatory response after injury, which was correlated with an enlarged area of astrogliosis beyond the injury site.
Our data provides evidence that TSG-6 has an anti-inflammatory role within the glial scar.
Traumatic brain injury (TBI) is a major medical concern that affects over 10 million people in the world each year [1, 2]. A variety of injuries can cause TBI leading to a range of injury severities [3,4,5,6,7]. With improved medical interventions over the years, the mortality rate due to TBI has decreased, resulting in a significant number of people living with the long-term effects of TBI. It is well accepted that in addition to the immediate effects of TBI there are also multiple potential long-term gradually evolving sequelae that are influenced by the type of injury, severity of the injury and medical interventions at the time of injury [8, 9]. Additionally, a link between mild traumatic brain injuries and Alzheimer's disease or chronic traumatic encephalopathy has long been suspected [10]. At present, long-term effects of repeated TBI have been seen in multiple sports-related injuries, including post-traumatic parkinsonism, post-traumatic dementia and chronic post-concussion syndrome [11,12,13,14]. Thus, studying the short- and long-term consequences of TBI at a cellular and molecular level may lead to a new understanding and perhaps better long-term management of such injuries via new and/or refined treatment strategies.
Astrogliosis is a hallmark of TBI, which commences hours after injury and leads to an abnormal increase in the number of activated astrocytes in and around the injury site [15, 16]. Immediately after injury (acute phase), astrocytes are activated, becoming highly proliferative and up-regulating the production of extracellular proteins [17,18,19]. These astrocytes and their deposited extracellular matrix in and around the injury site form a glial scar. Over the years, a significant body of evidence has demonstrated that the glial scar contains molecules, such as chondroitin sulfate proteoglycans (CSPGs), that impede axonal growth, thus inhibiting neuronal regeneration [15, 20,21,22,23]. The intensity of the acute and chronic reactive astrogliosis, including the quantity and composition of the glial scar, affects immediate and long-term effects of TBI [6, 16, 24, 25]. Penetrating brain injuries (PBIs) cause direct parenchymal laceration, neuronal cell loss and hemorrhage, which lead to focal tissue damage at the injury site. Astrogliosis is triggered after TBIs forming a glial scar in and around the injury site [26,27,28,29]. Importantly, uninjured tissue bordering the injury site is also subject to astrogliosis, and the process of glial scarring therefore extends beyond the injury site [30]. Given the fact that glial scarring limits regeneration after injury, many studies have investigated whether limiting astrogliosis after injury, with particular focus on limiting deposition, could potentially promote regeneration [23, 27, 31,32,33,34]. Although many studies were able to demonstrate beneficial effects of limiting glial scarring on neuronal regeneration, many others were inconclusive or actually found there was an increased inflammatory response culminating in tissue damage beyond the injury site and an increase in neuronal loss. Thus, mounting evidence indicates that reactive astrocytes surrounding the injury site are instrumental in preserving the surrounding uninjured tissue by forming scar borders, which separate damaged and inflamed tissue from adjacent viable neural tissue [15, 16, 24, 35,36,37,38,39,40]. Sofroniew and colleagues elegantly demonstrated that targeting astrocytes after brain and spinal cord injury leads to increased inflammation, delayed recovery and increased neuronal loss [39, 41,42,43,44]. Moreover, the inhibition of astrocyte proliferation prolongs the healing period following central nervous system (CNS) injury [45]. Data from Hermann et al. show that GFAP-driven ablation of STAT3 in astrocytes leads to the loss of lesion demarcation and subsequent glial scar formation, and, in turn, results in increased invasion of inflammatory cells into adjacent viable tissue and further spread of inflammation [46]. This suggests that early glial scar formation by astrocytes restricts movement of inflammatory cells located within the lesion site into adjacent healthy tissue, thereby restricting tissue damage to the injury site. Thus, a recent body of evidence suggests scar tissue bordering the injury site is necessary for limiting inflammation and tissue damage to the injury site [37, 41]. There is currently a significant number of studies investigating how reactive astrocytes regulate and limit inflammation to the injury site, and which cellular components and major pathways could play a role in this process [20, 35, 41, 45, 47].
We recently found that tumor necrosis factor (TNF)-stimulated gene-6 (TSG-6) is secreted by astrocytes after injury and is a major constituent of the glial scar, but the role it plays within the glial scar remains to be established [48]. TSG-6 is a 35-kDa protein that is secreted by a wide range of cell types in response to inflammatory mediators and growth factors [49], and was originally identified as a gene product induced in fibroblasts by TNF [12]. TSG-6 contains a link module domain that mediates its interaction with the glycosaminoglycans (GAGs) hyaluronan (HA) and CS [49,50,51]. Our recent study identified that TSG-6 is expressed in the CNS, where it catalyzes the transfer of heavy chains (HCs) from Inter-a-Inhibitor (IαI, also known as ITI) onto HA, forming a specialized HA/HC/TSG-6 matrix within the glial scar, but the role of this specialized matrix within the glial scar remains to be established [48, 52,53,54,55,56]. This specific HA/HC/TSG-6 matrix has previously been shown to be monocyte-adhesive in other tissues and is believed to be present in most, if not all, inflammatory processes [57, 58]. These TSG-6 modified HA matrices bind inflammatory cells, and the interaction of these cells with this matrix modulates their responses, which are central to pathological inflammation [59,60,61,62,63,64,65]. The main objective of this study was to investigate the role TSG-6, a constituent of the glial scar, has in astrogliosis after a PBI. Given the well-characterized anti-inflammatory role of TSG-6 in other sites, the premise of this study was that TSG-6 could participate in the formation of an immunosuppressive environment within the glial scar. Our findings show that TSG-6 null mice present a more severe inflammatory response and increased glial scar deposition after injury when compared to littermate control mice. This increased inflammatory response in TSG-6 null mice was correlated with an enlarged area of astrogliosis beyond the injury site.
TSG-6 expression after PBI
In this study, transgenic Tsg-6 null mice (Tnfip6Δ/Δ), hereafter referred to as Tsg-6−/− mice, and heterozygous mice, hereafter referred to as Tsg-6+/− mice, were used to investigate the role of TSG-6 in the glial scar. In order to investigate whether TSG-6 is present in the glial scar after brain injury, we analyzed the expression profile of Tsg6 in the injury site and injured hemisphere before and after a PBI in Tsg-6+/− mice (Fig. 1A). There was a twofold increase in Tsg-6 expression 5 days after injury when compared to uninjured mice. There was a further increase in Tsg-6 expression over time after injury, with expression increasing two fold from 5 to 10 days after injury (Fig. 1A). Interestingly, we did not find a difference in the expression levels of Tsg-6 between the injury site and the remaining hemisphere, indicating that Tsg-6 expression is not contained solely to the injury site (Fig. 1A). Therefore, there is also an increase in Tsg-6 expression in the surrounding tissue after injury. No Tsg-6 expression was identified in any of the samples from Tsg-6−/− mice confirming that these mice are indeed null for Tsg-6.
TSG-6 and GFAP expression after PBI. TSG-6 and GFAP mRNA expressions were quantified in the injury site and the injured hemisphere after PBI. A TSG+/− mice were subjected to PBI, and the injury site and remaining injured hemisphere were collected 5 and 10 days after injury for analysis of TSG-6 expression. B, C TSG+/− and TSG-6−/− mice were subjected to PBI, and the injury site and remaining injured hemisphere were collected 1 day (B) and 5 days (C) after injury for analysis of GFAP expression. * = p ≤ 0.05 comparing TSG-6−/+ and TSG-6−/− mice
Analysis of astrocyte recruitment after PBI
We assessed the level of astrogliosis in the injury site and in the remaining injured hemisphere by quantifying the levels of GFAP+ astrocytes using real-time PCR (Fig. 1B, C). For such, we isolated mRNA from the injury site and remaining injured hemisphere 1 and 5 days after injury of Tsg-6−/− and Tsg-6+/− mice. Both Tsg-6−/− and Tsg-6+/− mice presented an increase in the levels of GFAP expression in the injury site when compared to the remaining injured hemisphere, which corroborates literature [36, 63, 64]. Tsg-6−/− mice showed a significant increase in GFAP levels within the injury site at both 1 and 5 days post-injury when compared to Tsg-6+/− mice (Fig. 1B, C). This data indicates that Tsg-6−/− mice have more astrocytes in the injury site when compared to Tsg-6+/− mice. At 5 days after injury, there was a significant increase in GFAP expression in the injured hemisphere of Tsg-6−/− mice compared to Tsg-6+/− mice, indicating that Tsg-6−/− mice present astrogliosis beyond the injury site at 5 days post-injury.
The effect of TSG-6 on the secretion of inflammatory markers after PBI
The inflammatory response was also assessed in Tsg-6−/− and Tsg-6+/− mice 1, 5 and 10 days post-injury by quantifying the expression levels of NFκB, Rantes and IL1β (Fig. 2). Higher expression levels of NFκB, Rantes and IL1β were detected in Tsg-6−/− mice when compared to Tsg-6+/− mice during the acute phase after injury. Specifically, a ~ 2.5-fold and threefold increase in NfκB expression was found in the injury site and remaining injured hemisphere, respectively, in Tsg-6−/− mice compared to Tsg-6+/− mice 5 days after injury (Fig. 2B). 10 days after injury there was still a significant increase in NfκB expression in the surrounding hemisphere of Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 2C). No significant differences were found in the expression of NfκB between Tsg-6−/− and Tsg-6+/− mice 1 day after injury (Fig. 1A). The levels of Ccl5 (Rantes) were also assessed 1, 5 and 10 days post-injury. There was a significant increase in the expression of Rantes in the injured hemisphere of Tsg-6−/− mice when compared to Tsg-6+/− mice (a four fold increase) 5 days after injury; however, no difference was found between Tsg-6−/− and Tsg-6+/− mice 1 and 10 days post-injury (Fig. 2D–F). IL1β levels were increased in the injury site of TSG-6−/− mice when compared to Tsg-6+/− mice at 1 day post-injury (Fig. 2G). At 5 days post-injury a threefold and fourfold increase in the expression of IL1β were noted in the injury site and remaining injured hemisphere, respectively, of Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 2H). At 10 days post-injury, a 2.5-fold increase in the expression of IL1β was noted in the injury site of Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 2I).
Analysis of inflammatory markers after PBI. NFκB, RANTES and IL1β mRNA expressions were quantified in the injury site and the injured hemisphere after PBI. TSG+/− mice and TSG-6−/− mice were subjected to PBI and the injury site and remaining injured hemisphere were collected 1, 5 and 10 days after injury. mRNA was extracted and subjected to real-time PCR analysis for NFκB (A–C), RANTES (D–F) and IL1β (G–I) mRNA expression. * = p ≤ 0.05 comparing TSG-6−/+ and TSG-6−/− mice
The effect of TSG-6 on the activation of microglia and infiltration of macrophages into the injury site after PBI
In order to assess the inflammatory response in Tsg-6−/− and Tsg-6+/− mice, we also evaluated the number of CD68+ cells present within the injury site at 3 days post-injury (Fig. 3A, C). CD68 is routinely used as a marker for macrophages and activated microglia. There was a significant increase in the number of CD68+ cells in and around the injury site of Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 3A panels i and ii). Importantly, even when analyzing deeper regions of the injury site of Tsg-6+/− mice, the level of CD68+ cell infiltration was not as intense as that observed in Tsg-6−/− mice (Fig. 3A panel iii). The combined number of CD68+ cells in the injury site and within a range of 100 μm from the wound edge was counted from images obtained from 2 different sections from at least 5 mice from each experimental point (Fig. 3C). A two fold increase in CD68+ cells was found in Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 3C).
Analysis of inflammatory cell infiltration and neuronal cell loss after PBI. The distribution of macrophages and activated microglia was evaluated within the injury site of TSG+/− and TSG-6−/− mice 3 days post-injury (dpi) by anti-CD68 immunostaining (red) (A). Neuronal cells were immunostained with anti-β III tubulin (green) in the equivalent area of uninjured TSG+/− (i) and TSG-6−/− (ii) mice and within the injury site of TSG-6−/+ (iii) and TSG-6−/− (iv) mice 14 days post-injury (dpi). The number of CD68+ cells was counted in the injury site and within 100 μm of the wound edge of TSG-6−/+ and TSG-6−/− mice 3 days post-injury (C). The relative fluorescent units (RFU) of anti-β III tubulin staining were quantified in and around the injury site of TSG-6−/+ and TSG-6−/− mice 14 days post-injury (D). Nuclei were counterstained with DAPI. Scale bar represents 100 μm. * = p ≤ 0.05 comparing TSG-6−/+ and TSG-6−/− mice
Correlation between increased inflammatory response and neuronal damage
In order to verify whether the increased inflammatory response observed in Tsg-6−/− mice correlates with neuronal loss, the distribution of neurons in and around the injury site was analyzed in Tsg-6−/− and Tsg-6+/− mice 14 days post-injury (Fig. 3B). For such, β III tubulin was used as a tissue-specific marker for identifying neurons within injured and non-injured brains. The distribution of β III tubulin can be seen in the equivalent region of uninjured Tsg-6−/− and Tsg-6+/− mice (Fig. 3 B panels i and ii). A significant increase in the area devoid of β III tubulin staining can be observed in and around the injury site of Tsg-6−/− mice when compared to Tsg-6+/− mice 14 days post-injury (Fig. 3B iii and iv). The relative fluorescence units (RLU) were quantified from an image of the injury site captured from at least 3 mice per experimental point. There was a fourfold decrease in β III tubulin staining in and around the injury site of Tsg-6−/− mice when compared to Tsg-6+/− mice 14 days post-injury (Fig. 3D).
The effect of TSG-6 on the secretion of glial scar components after PBI
We also evaluated glial scar secretion within the injury site and injured hemisphere by evaluating the expression levels of the biosynthetic enzymes responsible for HA and CS chain elongation, specifically hyaluronan synthase 2 (Has2), carbohydrate (chondroitin 4) sulfotransferase (chst 11) and carbohydrate (chondroitin 4) sulfotransferase 12 (chst 12) (Fig. 4). Has2 expression increased in the injury site when compared to the remaining injured hemisphere 5 days post injury in both Tsg-6+/− and Tsg-6−/− mice, confirming the numerous previously published reports showing that HA is an integral component of the glial scar [17, 66,67,68]. Interestingly, there was a twofold increase in Has2 expression in the injury site of Tsg-6−/− mice when compared to Tsg-6+/− mice 5 days after injury, indicating that there is a higher rate of glial scar production in Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 4A). At 10 days post-injury, Has2 expression was still increased by twofold in Tsg-6−/− mice when compared to Tsg-6+/− mice, but at this time point there was also an increase in Has2 expression in the remaining injured hemisphere of Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 4B). Thus, at 10 days after injury, in Tsg-6−/− mice, the expression of glial scar components was no longer limited to the injury site, but was also present within the remaining injured hemisphere. Interestingly, this was also true for the expression of Chst11 and Chst12, which showed a five fold and four fold increase, respectively, within the injured hemisphere of Tsg-6−/− mice at 5 days post injury when compared to Tsg-6+/− mice (Fig. 4C, E). The increase in Chst11 and Chst12, in both the injury site and injured hemisphere, was maintained through to 10 days post-injury (Fig. 4D, F).
Analysis of glial scar extracellular matrix components after PBI. HAS2, Chst 11 and Chst 12 mRNA expression levels were quantified in the injury site and the injured hemisphere after PBI. TSG+/− and TSG-6−/− mice were subjected to PBI, and the injury site and remaining injured hemisphere were collected 5 and 10 days after injury. mRNA was extracted and subjected to real-time PCR analysis for HAS2 (A, B), Chst11 (C, D) and Chst 12 (E, F) mRNA expression. * = p ≤ 0.05 comparing TSG-6−/+ and TSG-6−/− mice
The effect of TSG-6 on astrocyte activation and recruitment after PBI
In order to further investigate the process of astrogliosis in Tsg-6+/− and Tsg-6−/− mice, injured brains were harvested and processed for histology. Sections were stained for GFAP in order to assess the distribution of astrocytes in and around the injury site, and, also, throughout the remaining brain tissue. The number of astrocytes (GFAP+ cells) was counted within the injury site, throughout the injured hemisphere, and, also, throughout the contralateral hemisphere 3 and 14 days post-injury (Fig. 5A, B). At 3 and 14 days post-injury there was a significant increase in the number of astrocytes within the injury site when compared to the injured hemisphere and contralateral hemisphere in both Tsg-6+/− and Tsg-6−/− mice. At 3 days post-injury there was no significant difference between the number of astrocytes within the injury site between Tsg-6+/− and Tsg-6−/− mice; however, there was a significant increase in the number of astrocytes within the injured hemisphere in Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 5A). At 14 days post-injury there was a significant increase in the number of astrocytes within the injury site and injured hemisphere in Tsg-6−/− mice when compared to Tsg-6+/− mice (Fig. 5B, D). The increase in astrocytes can be seen beyond the injury site in Tsg-6−/− mice (Fig. 5C, D panel iv).
Analysis of astrocyte activation and recruitment after PBI. Brain sections from TSG-6+/− and TSG-6−/− mice were analyzed by immunofluorescence. Astrocytes were identified with anti-GFAP (green) and the glial scar with HABP (red). Nuclei were counterstained with DAPI (blue). Z-stacks were captured of the entire brain section using the tilling mode, and images were stitched together using Zen software. Thereafter, the number of astrocytes was counted within the injury site, within the injured hemisphere and in the contralateral hemisphere of brains 3 (A) and 14 dpi (B) in a double blinded manner. The distribution of astrocytes throughout the brain sections shows that in TSG-6−/− mice the increase in astrocytes is not restricted to the injury site (C). Magnified images of the areas demarcated in (C) can be seen in (D). At least 3 mice were analyzed per genotype for each time point. * = p ≤ 0.05 comparing TSG+/− and TSG-6−/− mice
Chondroitin sulfate proteoglycans (CSPGs) are well established as major extracellular matrix components in the central nervous system [69]. Over a decade ago, Silver et al. identified that CSPGs within the glial scar inhibit axonal growth, and this triggered a great deal of interest in targeting CS within the scar tissue as a means to promote axonal regeneration [32, 70,71,72]. Over the years, strategies utilizing the enzymes chondroitinase ABC (ChABC) and ChAC have been used to remove the CS component of the glial scar as a means to promote axonal growth and regeneration [50, 73,74,75,76,77]. Many studies have shown that specifically removing CS within the glial scar is enough for axons to grow across the injury site [32, 70, 78, 79]. However, significant regeneration was never observed in these studies, and many groups found limited or no improvement after targeting CS within the glial scar [70]. One unique characteristic of TSG-6 is its known ability to bind to a number of ligands including HA, CS and core proteins of proteoglycans (i.e. versican and aggrecan), forming specific HA/HC/TSG-6 and/or CS/HA/HC/TSG-6 matrices with immunosuppressive characteristics [61, 80,81,82,83,84]. Our previous study suggests these HA/HC/TSG-6 matrices are also present within the glial scar [48]. Therefore, given that TSG-6 directly binds to both HA and CS to form specific anti-inflammatory matrices, the ChABC and ChAC treatments used over the years to target the glial scar as a means to promote regeneration would also have removed TSG-6, a known anti-inflammatory molecule that is also a component of the glial scar [82]. The loss of TSG-6 by these treatments could, in part, explain why significant functional recovery was never obtained after ChABC and/or ChAC treatments.
To explore the role of TSG-6 in TBI, specifically in astrogliosis, we compared the differences in injury outcomes in Tsg-6−/− and Tsg-6+/− mice after PBIs. Our data show an increase in TSG-6 expression in the injured hemisphere of Tsg-6+/− mice after TBI. This increase in expression of TSG-6 after CNS insults supports our earlier findings in a rat model that astrocytes secrete high levels of TSG-6 upon injury, which aids in the formation of a specialized HA/HC/TSG-6 matrix as part of an inflammatory response [48]. Since TSG-6 is known for having anti-inflammatory properties, to further study whether high levels of TSG-6 serve a purpose of rapidly suppressing inflammation after injury, we performed similar penetrating injuries in Tsg-6−/− mice. We used immunofluorescence and RNA expression analyses of inflammatory and glial scar markers to elucidate the outcome during the acute phase and chronic phase of TBI. During the acute phase after injury, the observed increase in astrocyte activation, inflammatory cell infiltration and expression of inflammatory cytokines in Tsg-6−/− mice indicate that the loss of TSG-6 results in a greater inflammatory response. Moreover, during the chronic phase of injury, unrestricted inflammatory response was observed throughout the injured hemisphere and was not limited to the injury site, as is seen after normal glial scar formation. Thus, injured Tsg-6−/− mice appear to experience more severe tissue damage than their Tsg-6+/− counterpart, both within and around the injury site. Thus, the loss of TSG-6 allows the damage to spread from the injury site to neighboring healthy tissues. We postulate that the cause of such widespread damage is due to the lack of the specialized HA-TSG6 or HA/HC/TSG-6 matrix, which could possibly serve to stabilize the glial scar and form an immunosuppressive environment, thereby protecting adjacent tissue from further damage. This hypothesis is further supported by the increase in CSPG and HA biosynthesis, both glial scar components, in Tsg-6−/− mice. Specifically, these mice show increased Has2, Chst11 and Chst12 expression levels in tissues collected after the onset of glial scarring, and, also, during the chronic phase of astrogliosis, indicating an increase in scar tissue formation. This increase in expression was not only observed at the injury site, but also throughout the whole injured hemisphere, suggesting that the tissue damage spreads beyond the injury site in the absence of TSG-6. Collectively, these results demonstrate that the loss of TSG-6 leads to a more severe inflammatory response and, consequently, increased scarring after TBI. Thus, our results support the hypothesis put forward by many groups over the past decade that preventing the formation of the glial scar leads to inflammation and damage beyond the injury site. We also provide experimental evidence that shows that the glial scar functions to restrict the damage to the injury site. Importantly, these findings should be taken into account when attempts are made to disrupt the glial scar as a means to promote neuronal regeneration, since preventing formation of the glial scar may not have the beneficial outcomes as previously presumed.
Our results show that TSG-6 has an anti-inflammatory role in the glial scar. Our study further supports the hypothesis that the glial scar forms a protective border surrounding the injury site thereby preventing the spread of inflammation and damage beyond the injury site.
TSG6 null (TSG6−/−) or heterozygous (TSG6+/−) mice and animal maintenance
Transgenic Tsg-6 null mice (Tnfip6Δ/Δ), herein referred to as Tsg-6−/− mice, and heterozygous mice, herein referred to as Tsg-6+/− mice, were maintained as previously described [56]. Our previously published work demonstrated that Tsg-6+/− mice present a similar distribution of astrocytes throughout the brain to wild-type (wt) mice [48]. Moreover, Tsg-6+/− mice have previously been shown not to display a phenotype and present similar TSG-6 expression levels as wt mice, and were therefore used as littermate controls in our study [56]. Experimental procedures for handling the mice were approved by the Institutional Animal Care and Use Committee (IACUC), University of Houston under protocol 16-036.
Mice (7 to 8 weeks old) were anesthetized with ketamine (80–100 mg/kg—Vedco INC, Catalog# 07-890-8598) and xylazine (5–10 mg/kg, Akorn INC, Catalog# 07-808-1947) by IP injection and allowed to go into full anesthetic state. A sterile surgical drill (Precision Tools, Model Craft PPV2237) was used to make a hole of approximately 1.5 mm in diameter in the skull over the right frontal cortex at the stereotaxic coordinates AP: 1.0 mm, ML: 1.5 mm, and DV: 1.5 mm, according to Franklin and Paxinos (85). A 30-gauge needle (Exel, Catalog# 26437) was then used to make a puncture wound at a depth of 2 mm. After injury, the skin at the surgical site was closed with two sutures. The area was then cleaned with 70% ethanol, and mice were placed on a heating pad and monitored until they regained consciousness prior to being transferred to a clean cage. All surgeries were carried out at the same time of day to minimize bias. Mice were monitored daily and did not show any decrease in weight ≥ 15% when compared to their pre-surgical weight. Mice were euthanized, as outlined below, at 1, 3 and 5 days post injury to study the acute effects of brain injury, and at 10 and 14 days to study long-term/chronic effects. Five mice per experimental group were used for the real-time PCR analysis and at least seven mice per experimental group were used for immunofluorescence analysis.
Perfusion fixation and brain tissue processing
Brain samples were collected at 1, 3, 5, 10, and 14 days post injury for immunofluorescence analyses. Briefly, mice were initially injected with a lethal dose of combined anesthetics containing 200 mg of ketamine and 40 mg xylazine. Final dosage received was 3 mg of ketamine and 0.6 mg of xylazine per mouse. Once mice were under deep anesthesia, abdominal and thoracic excisions were performed to expose the heart, which was used to perfuse 2% formalin (Fisher Scientific, Catalog# SF100-4) throughout the whole body via a gravity-driven flow system for whole body fixation. Subsequently, the brain was isolated from the skull and further immerse fixed for 2 days in 2% paraformaldehyde (Electron Microscopy Sciences, Catalog# 15710). For cryosection processing, brains were immersed in 30% sucrose for 2 days, embedded in OCT embedding medium (Fisher Healthcare, Catalog# 4585) and frozen. Sections 10 μm thick were obtained, mounted on superfrost slides (VWR, Catalog# 48311-703) and stored at − 20 °C until use.
Upon use, the slides were heated at 65 °C for 30 min and, subsequently, sections were washed with PBS to remove tissue freezing medium. Sections were then treated with 0.1% glycine (Fisher Chemical, Catalog# G46-500), blocked with 5% FBS (Seradigm, Catalog# 3100-500) and permeabilized with 0.1% saponin prepared in PBS. Sections were then incubated with the primary antibodies anti-Tenascin (Abcam, Ab108930), anti-GFAP (Abcam, Ab4647), anti-CD68 (Abcam, Ab31630) and anti-β III tubulin (Covance, PRB-435P-100). Sections were washed and incubated with appropriate secondary donkey antibodies conjugated with Alexa Fluor® 488 (Life Technologies) or Alexa Fluor® 555 (Life Technologies) for one hour at 18 °C. For HA staining, tissues were incubated with biotinylated HA binding protein (385911, Millipore) followed by NeutrAvidin®Alexa 555 (Life Technologies). The tissues were then washed and nuclei stained with 4',6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich). Sections were mounted in Prolong®Gold (Molecular Probes) and imaged using a ZEISS LSM 800 Confocal microscope with Airyscan. Secondary controls were done with a goat IgG isotype control (ab37388; Abcam) in place of the primary antibody and did not yield any significant staining (results not shown). For imaging, multiple z-stack tiles were captured of entire brain sections and frames were processed together into a single image (using the stitching mode followed by full orthogonal projection) using Zen Software (Zeiss). The number of GFAP+ and CD68+ cells in and around the injury site were counted by two independent investigators in a blinded manner and the relative Fluorescent intensity was measured using the Zen Software (Zeiss). At least 2 sections were scanned and analyzed from each animal for each set of antibodies and representative images shown in the figures.
RNA extraction from brains and real-time PCR analysis
Brains collected from injured mice at 1, 5 and 10 days post injury were used for RNA extraction. At least 5 mice were used per experimental group and each animal was analyzed separately. Briefly, mice were euthanized and brain tissue was immediately isolated from each mouse. Injury sites (A samples) were dissected from the rest of the injured right hemisphere, transferred into a labeled Eppendorf tube and immediately immersed in liquid nitrogen. The remaining right hemisphere brain tissue (B samples) from each animal was transferred into a different tube and frozen as described. The samples were kept at − 80 °C until RNA extraction. Total RNA was isolated from these tissue samples using Trizol® Reagent (Invitrogen, Carlsbad, CA) and chloroform extraction (Sigma-Aldrich, Catalog# 650498). First strand cDNA was reverse transcribed using 1.5 to 2 μg of total RNA and the high capacity cDNA Reverse Transcription kit (Applied Biosystems, catalog# 4368814, lot 00593854), according to the manufacturer's instructions. Quantitative real-time PCR amplification was performed on 1 μg or 50 ng of the cDNA (1:5) using the PowerUp SYBR Green Master Mix kit (Applied Biosystems, Catalog# A25918) in a CXF Connect Real-time System from BIO-RAD, using an activation cycle of 95 °C for 10 min, 40 cycles of 95 °C for 15 s and 60 °C for 1 min. A complete list of primers used in this study is shown in Table 1. Gene expression levels were normalized against Actb and Gapdh using the 2−ΔCt and/or 2−ΔΔCt methods.
Table 1 Primers used for real time PCR analysis
All values are presented as the mean ± standard deviation of the mean. The difference between the two groups was compared by means of the Student's t-test. p ≤ 0.05 was considered to be statistically significant. Statistical analysis was performed using the GraphPad Prism version 7 software package (GraphPad Software, San Diego, CA, USA). * was used to indicate statistical differences of ≤ 0.05. Unless indicated otherwise, * indicates the statistical difference of Tsg-6−/− mice compared to Tsg-6+/− mice for each time point.
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
TBI:
TSG-6:
TNF-stimulated gene-6
GFAP:
Glial fibrillary acidic protein
Vimentin
CSPGs:
Chondroitin sulfate proteoglycans
STAT3:
Signal transducer and activator of Transcription 3
HA:
Hyaluronan
GAGs:
HC:
ITI or IαI:
Inter-alpha inhibitor
hUMSCs:
Human umbilical cord mesenchymal stem cells
PTX3:
Pentraxin-3
NFKB:
Nuclear factor kappa-light-chain-enhancer
IL1β:
Interleukin 1 beta
Ccl5:
Chemokine (C–C motif) ligand 1 or Rantes
HAS:
Hyaluronan synthase
Chst:
Carbohydrate (chondroitin 4) sulfotransferase
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The authors would like to thank Denise Lerma, Nicole Grimmes and Garrett Elliot who contributed in the early stages of the study. The authors are in debt to Tarsis F. Gesteira and Yvette May Coulson- Thomas for their invaluable discussions and suggestions throughout the study. Transgenic Tsg-6 null mice (Tnfip6Δ/Δ) were kindly provided by Dr. Mark Lauer.
This study was supported by start-up funds from the University of Houston to VJCT, The Mizutani Foundation grant to VJCT and the National Institute of Health/National Eye Institute R01 EY029289 to VJCT and Core grant P30 EY07551.
Amanda Nash
Present address: Department of Bioengineering, Rice University, Houston, TX, 77030, USA
College of Optometry, University of Houston, 4901 Calhoun Road, Houston, TX, 77204-2020, USA
Kazadi Nadine Mutoji, Mingxia Sun, Amanda Nash, Sudan Puri & Vivien J. Coulson-Thomas
Cleveland Clinic, Cleveland, OH, USA
Vincent Hascall
Kazadi Nadine Mutoji
Mingxia Sun
Sudan Puri
Vivien J. Coulson-Thomas
VH and VJCT generated the hypothesis and experimental design. VJCT, KNM and MS contributed to the experimental design. KNM, MS, AN and VJCT conducted the experiments and helped with the data analysis. KNM and VJCT wrote the manuscript. KNM, MS, AN, SP, VH and VCT read and approved the final manuscript.
Correspondence to Vivien J. Coulson-Thomas.
Experimental procedures for handling the mice and animal care were in accordance to regulations of the National Institute of Health and were approved by the Institutional Animal Care and Use Committee, University of Houston under protocols 16-025, 16-035, 16-036, 16-044.
The authors grant their consent to publish the material presented herein.
Mutoji, K.N., Sun, M., Nash, A. et al. Anti-inflammatory protein TNFα-stimulated gene-6 (TSG-6) reduces inflammatory response after brain injury in mice. BMC Immunol 22, 52 (2021). https://doi.org/10.1186/s12865-021-00443-7
TSG-6
Glial scar
Glycosaminoglycans and inflammation
Submission enquiries: bmcimmunology@biomedcentral.com


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Welcome to the fourth post in my Hogwarts House series, if you missed the other posts about Hufflepuff – Ravenclaw – or Gryffindor be sure to click them to read it and let me know what you think!
The whole premise of this book revolves around the fact that one of the London's was too ambitious for its own good. They sought more power than they could handle and it ultimately led to their downfall. The characters in this series are ambitious, determined, cunning and sometimes ruthless – but they are also incredibly loyal. I think that Slytherins would be fascinated by the struggle each character faces as they walk the line between letting their traits be a positive or a negative force in their lives. If this series sounds intriguing, check out my review here!
Nemesis must become (or masquerade as) Sidonia, the most important person in the worlds to Nemesis and the person she was created to protect, essentially infiltrate the center of the galactic Empire and allow no one to discover the deception. I feel it's best to go into this book without a lot of preconcieved notions or information about the book beyond a quick summary, it's an interesting ride when you're not quite sure what's going to happen.
Psst, there may or may not be more posts coming in this series… any guesses?


Solid Sands and UPMEM SAS announced that UPMEM has acquired a SuperTest license to test and validate the compiler for its memory-embedded general-purpose processor. UPMEM's processing-in-memory technology is designed to speed up big-data analytics by a factor of 20 or more while also reducing power consumption, making it highly attractive for high-throughput data-intensive applications, such as human genome sequencing/mapping or string searching on compressed document databases. UPMEM will use SuperTest to test and validate the LLVM compiler it is developing for its unique embedded processor architecture.


I shared last week about our trip to the Museum of Fine Arts. After the museum we went to the Chalk Art walk which I saved to share today. The kids and I were amazed at the chalk creations and how they transformed the sidewalks. I have to admit I don't envy the chalk artists life. They are a tough breed. It was brutally hot outside and all their hard work could be ruined in minutes if it rained.(which it did that night) I would much rather create something that would last but some of the artists I talked to have been doing this all their life so I guess they must enjoy it.
We were given tickets to use for voting on which chalk art we like the best. We chose the blue heron. They had chalk for the kids to use and they were very inspired by the professionals. Thankfully Luke was inspired to take a nap!
On our walk back to our van we took a last look at our pier which is being torn down for a new one. It has been an iconic landmark here for years and was a point of controversy as to tearing it down or not and then what to put in it's place. I think the city has finally decided on what to replace it with. I am excited about it as I wasn't to crazy about the old one.
We truly live in such a beautiful place. Our whole county is a peninsula, so pretty much any way you go you will run into the water. We have bad areas like everywhere else but the downtown is really gorgeous. It's right on Tampa Bay and has the best view of the sun rising in the morning.
If you are ever in Florida and want to visit a great place you really need to check out St.Petersburg or as we call it St.Pete. Most of our weekends are filled with cleaning, preparing for the school week, birthday parties and family get togethers but I love it when we are able to leave the dirty dishes and laundry and get out and explore. I am trying to get used to taking them out all by myself when Josh is away and I have to say that sometimes it's way easier than staying inside with all their craziness.
Linking up at these great link parties!
Wow! That chalk art is amazing! The 3D quality of the faces is stunning, I didn't realise chalk could give so much detail. Your children's fish are very good too, were they drawn from memory? A gorgeous spot as well, right by the sea for fantastic views over the water. You certainly live in a fantastic area.


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Tag: NAPABA
National Affinity Bar Associations Join AAPIs in Demanding Apology from Fox News for Watters' World Segment
Jesse Watters interviews people in NYC's Chinatown during a segment that aired on October 3, 2016. (Photo credit: Fox News)
The Hispanic National Bar Association (HNBA), the National LGBT Bar Association (National LGBT Bar), the National Native American Bar Association (NNABA), the National Asian Pacific American Bar Association (NAPABA) and the National Bar Association (NBA) issued a joint statement late last week expressing support for efforts by the AAPI community to hold Fox News accountable for a segment of Watters' World that aired in early October and which has been widely decried as racist against the Asian American community.
Combined, the five Affinity Bar Associations represent nearly two hundred thousand Black, Hispanic, AAPI, Native and LGBT lawyers and hundreds of local chapters across the country.
In the statement released publicly on Friday and sent to Fox News headquarters, the coalition of Affinity Bar Associations expressed "deep dissatisfaction" with Fox News' failure to adequately address charges of racism. The coalition added their voices to the growing chorus insisting that Fox News meet the demands outlined in a recent joint letter signed by 120+ AAPI organizations. That letter was delivered, along with a petition signed by over 20,000 community members, to Fox News representatives earlier this month.
Continue reading "National Affinity Bar Associations Join AAPIs in Demanding Apology from Fox News for Watters' World Segment"
Tags Tags Bill O'Reilly, Fox News, HNBA, NAPABA, National Bar Association, National LGBT Bar, NNABA, Watters World
Federal Court of Appeals Rules Denial of "Slants" Trademark is Unconstitutional
Asian American rock band, The Slants.
For the last several years, Asian American rock band, "The Slants", has been in a knockdown fight with the US Patent and Trademarks Office (USPTO) over their band's name.
The fight stems from a decision made by the USPTO to refuse registration of "The Slants" as a trademark on the grounds that the bands' name is racially offensive to Asian Americans. The USPTO was empowered to make their decision based on the 1946 Lanaham Act, which prohibits the registration of any mark that "consists of or comprises of immoral, deceptive, or scandalous matter; or matter which may disparage or falsely suggest a connection with persons, living or dead, institutions, beliefs or national symbols, or bring them into contempt, or disrepute."
In practice, the USPTO frequently makes such decisions unilaterally through the determination of a single case officer who by happenstance receives an application to register a questionable mark. Consequently, the USPTO has been notoriously inconsistent in their determination of a mark's "immoral", "scandalous" or "disparaging" content. "The Slants" is a group of Asian American musicians seeking to reclaim an offensive word they recognize as traditionally a slur; despite the USPTO's refusal of their application, the Office has permitted the registration of "Dykes on Bikes", which is a group named under similar reasoning.
Continue reading "Federal Court of Appeals Rules Denial of "Slants" Trademark is Unconstitutional"
Tags Tags ChangetheName, NAPABA, Simon Tam, The Slants, Trademarks, USPTO, Washington Redskins
Reappropriate Joins AANHPI Organizations in Filing as SCOTUS Amici for Affirmative Action | #Edu4All
Three national Asian American, Native Hawaiian & Pacific Islander civil rights organizations filed separate amicus briefs today in support of affirmative action; the briefs were filed in relation to the upcoming Fisher v. University of Texas case scheduled to be heard by the Supreme Court on December 9th.
In addition to briefs filed by the Asian American Legal Defense Fund (AALDEF) and the National Asian Pacific American Bar Association (NAPABA), the brief filed by Asian Americans Advancing Justice (AAAJ) was signed by over 160 national AANHPI groups and individuals, including by this blog. Other signatories hail from all parts of the AANHPI diaspora, in terms of ethnicity, gender and group focus.
Continue reading "Reappropriate Joins AANHPI Organizations in Filing as SCOTUS Amici for Affirmative Action | #Edu4All"
Categories Categories Affirmative Action, Asian Americans & Pacific Islanders
Tags Tags AAAJ, AALDEF, Abigail Fisher, Amicus Brief, Campus Diversity, Edu4All, Educational Access, Fisher v University of Texas, Holistic Review, IAmNotYourWedge, NAPABA, SCOTUS
AAPI Civil Rights Groups File Amicus Brief Opposing Asian American Band "The Slants"
It's Asian vs. Asian, once again.
Earlier this year, I profiled the Oregon-based Asian American rock band, "The Slants", whose fight to trademark their name had gone all the way to the Federal Circuit Court of Appeals. According to their website, The Slants has been making music together since their 2007 debut album "Slanted Eyes, Slanted Hearts".
Recently, however, The Slants have found themselves embroiled in a legal battle with the US Patent and Trademarks Office (USPTO), which denied them registration for the trademark of their band name on the grounds that it is derogatory towards Asian Americans. The 1946 Lanham Act prohibits registered trademarks from "consisting of or comprising of immoral, deceptive, or scandalous matter; or matter which may disparage or falsely suggest a connection with persons, living or dead, institutions, beliefs or national symbols, or bring them into contempt, or disrepute."
It is on this basis that opponents of the Washington Redskins and other racist sports mascots have demanded that the NFL #ChangeTheName. And indeed, when the federal government allows an entity to register a racist, sexist, or otherwise derogatory trademark, they in so doing condone the language by granting it institutional recognition and legal protection.
But, should all uses of all historically derogatory slurs simply become — in essence — blacklisted — prohibited regardless of context?
Continue reading "AAPI Civil Rights Groups File Amicus Brief Opposing Asian American Band "The Slants""
Categories Categories Act Now!, Asian Americans & Pacific Islanders
Tags Tags ChangetheName, Korematsu Institute, NAPABA, SABA-DC, Simon Tam, Slurs, The Slants, Trademark Rights, USPTO

redpajama_set_name: RedPajamaCommonCrawl


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Home » TV & Movies » 'Never Have I Ever': Jeff Garlin Joins Fourth & Final Season
'Never Have I Ever': Jeff Garlin Joins Fourth & Final Season
EXCLUSIVE: Jeff Garlin (The Goldbergs, Curb Your Enthusiasm) has joined the cast of Netflix's Never Have I Ever for the show's fourth and final season.
Garlin will portray Len, a sweet man who fixes a mean sandwich and who makes Nirmala (Ranjita Chakravarty) reconsider if she really is done with relationships forever.
With a wedding confirmed to take place next season, could Devi Vishwakumar's (Maitreyi Ramakrishnan) grandmother be the one to walk down the aisle? She is just one of multiple potential brides to consider, however. Devi's mom Dr. Nalini Vishwakumar (Poorna Jagannathan) has a new love interest coming in this season portrayed by Ivan Hernandez, and her cousin Kamala Nandiwadal has been seriously dating English teacher Manish Kulkarni (Utkarsh Ambudkar) for a hot minute. And that's before considering wild cards like Eleanor Wong (Ramona Young) and Trent Harrison (Benjamin Norris). The blushing bride will be revealed when the final season drops in 2023.
Details Of New Netflix Studios Monmouth At Former New Jersey Army Base Unveiled: $850 Million Investment, 12 Sound Stages
From Mindy Kaling and Lang Fisher, Never Have I Ever is a coming-of-age comedy about the complicated life of first-generation Indian American teenager Devi, an overachieving high school sophomore who has a short fuse that gets her into difficult situations. Season 4 will focus on Devi's senior year in high school as she's really coming into her own.
The series is produced by Universal Television, in association with 3 Arts Entertainment, Original Langster, and Kaling International. Co-creator Fisher is the showrunner and executive produces alongside Kaling and 3 Arts Entertainment's Howard Klein and David Miner.
This is Garlin's first casting since he exited from ABC's The Goldberg's following HR investigations. He starred as Murray Goldberg in the comedy series for 9 seasons. Additionally, he plays Jeff Greene in HBO's Curb Your Enthusiasm. His TV credits also include Arrested Development, Everybody Loves Raymond, Mad About You, and Wizards of Waverly Place.
Investigation: How The BBC Is Making Money Producing Ads For China
2022: The Year Broadcast Networks Started To Wave The White Flag
Tops $150M Domestic After 2nd-Best Monday Of 2022; $500M+ Global With China Lower
Details On Netflix Studios Monmouth At Former NJ Army Base: $850M Investment, 12 Stages
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Barcelona 18 SECONDS late to submit paperwork for LA Galaxy star and face nervous wait to see if they can sign him | The Sun


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Missing neutron star found 30 years after supernova explosion
By Michael Irving
Missing neutron star found 30 ...
An artist's impression of a neutron star, obscured by a dust cloud
On February 23, 1987, a supernova lit up the night sky, visible to the naked eye. As the closest such event in almost 400 years, it provided the perfect opportunity to study supernovae, but one predicted piece has been missing ever since. Now, more than 30 years on astronomers say they've finally found the neutron star that was produced in the explosion.
Supernova 1987A was a once-in-several-lifetimes event, as the first supernova to be visible to the naked eye since 1604. That's because it was relatively close by – just 168,000 light-years away in the Large Magellanic Cloud, a dwarf galaxy that orbits the Milky Way. It flared up with the power of 100 million Suns, and glowed in the sky for several months. Even after it faded from view, the remnant of the explosion has remained as a cloud of dust and gas that astronomers have continued to study ever since.
But there was still one lingering question. Supernova theory said that a star of that mass should have collapsed into a neutron star, an incredibly small, dense, energetic object that should have been hard to miss. Yet missed it was, for 32 years. Some astronomers insisted it must have been hiding there, behind a thick veil of dust. Others suggested it may have collapsed into a black hole or a quark star – or maybe our theories are completely wrong, and there was nothing left at all.
Now, we may finally have our answer. As part of a new study, researchers from Cardiff University claim to have found that predicted neutron star, hiding in the dust. To do so, the team examined the supernova remnant using the Atacama Large Millimeter/submillimeter Array (ALMA) telescope in Chile.
The astronomers found that one certain patch of the dust looked brighter than the rest at the submillimeter wavelengths that ALMA measures. The neutron star was already suspected to be in this particular area.
"Although the light from the neutron star is absorbed by the dust cloud that surrounds it, this in turn makes the cloud shine in sub-millimeter light, which we can now see with the extremely sensitive ALMA telescope," says Mikako Matsuura, an author of the study.
The team says that the finding helps confirm that our theories about supernovae are on the right track after all. In future, the researchers say that the dust cloud may start to clear up, making the neutron star directly visible.
The research was published in the Astrophysical Journal.
Source: Cardiff University
SpaceCardiff UniversitySupernovaStarsNeutron GeneratorsCosmic dustALMANew Atlas Audio
Michael Irving
Michael has always been fascinated by space, technology, dinosaurs, and the weirder mysteries of the universe. With a Bachelor of Arts in Professional Writing and several years experience under his belt, he joined New Atlas as a staff writer in 2016.
Douglas Rogers November 21, 2019 11:11 AM
1987A actually proves the existence of neutrinos, as the required neutrino pulse was observed at the right time.
Pmeon March 2, 2020 08:36 PM
Neutrons have a half life of 10.3 minutes.
That is one short lived star.


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RELIGION-INFO.COM
A Guide to Religions, Religious Information and Help in Search for God
Spiritualism (Spiritism or Spiritualist Church)
Spiritualism is a religion with an estimated 13,000,000 followers or Spiritualists. Spiritualism is of ancient origin; The modern origin of Spiritualism can be traced back to either scientist and medium Emanuel Swedenborg (1688-1772) of Sweden in 1744 -or- Catherine and Margaretta Fox a.k.a.. the Fox sisters in Hydesville, New York in 1848. The first Spiritualism Church was founded in 1853.
[NOTE: In 1888, the sisters admitted to be fraudulent mediums when paid $1,500 (now equivalent to over $2,500,000.00) by the New York Herald newspaper. However, a year later, they retracted their confessions.]
Spiritualism is a religion marked by the belief that each and every person has a spirit and spirits of the dead can communicate with spirits of the living. This happens under the "right" conditions, usually through mediums (who go into a "trance" during "séances") and other types of psychic individuals, who may occasionally demonstrate a variety of psychic power including clairvoyance, clairaudience, telepathy and telekinesis.
Some mediums and other types of psychic individuals claim that they actually see dead spirits, who still wander around our material world. In some cases, the dead spirits may be deceased relatives of individuals, who they hang around and try to serve as spirit guides. Furthermore, some mediums claim to have the ability to "channel" dead spirits (or allow dead spirits to temporarily use their bodies to communicate to the living).
[NOTE: In more modern terminology, mediums may be referred to "channelers" and what they do is referred to as "channeling".]
Most Spiritualism churches are independently organized and small, but there are many of them scattered throughout the world. Some of the larger Spiritualism churches include the Church of the New Jerusalem (formed by followers of Emanuel Swedenborg), The Swedenborgian Church (also, formed by followers of Emanuel Swedenborg) and the Spiritualists' National Union based in the United Kingdom.
Furthermore, there are many followers of Spiritualism who are not members of any Spiritualism church, but use the services of mediums and other types of psychics to communicate with dead spirits.
Unfortunately, Spiritualism suffers greatly from an ongoing history of many fraudulent participants. However, there are very likely good apples in the basket along with the bad apples. In recent decades, the police have successfully used medium/psychics to help solve some "otherwise, unsolvable" criminal cases.
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Site created on 12/21/2004 | This page was last updated on 05/21/2007
COPYRIGHT © 2004-2007 • John Chin • ALL RIGHTS RESERVED
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This post comes to you from my kitchen. We haven't been birding for a while due to my Dad being pretty ill and in the ICU at Holy Redeemer Hospital and the really cold, rainy weather that we've had in December. For example, I went outside this morning to fill the feeders and needed ice skates! There is a layer of ice covering everything and it's still raining. Yuck.
Pretty good for a backyard.


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Dirt Road by James Kelman: A Theatrical Exploration
James Kelman, adapted by David Greig
Edinburgh International Book Festival and Royal Lyceum Theatre Edinburgh
Dirt Road was one of a series of "theatrical explorations" at this year's Book Festival that locked a writer, a director and some actors into a rehearsal room for three days with a book to see what they came up with.
The resulting one-off performances, in collaboration with Edinburgh's Royal Lyceum Theatre, were adaptations of selected sections of the novel lasting up to an hour followed by a Q&A session with the actors, director, novelist and adapter.
James Kelman's Dirt Road was scripted by Lyceum artistic director David Greig, directed by Wils Wilson and featured actor-musicians Moyo Akandé, Brian James O'Sullivan, Harry Ward and Aly Macrae, the latter also composing the music. Some of the selected text will be familiar to anyone who attended Kelman's own event at last year's Book Festival when he read from this book.
The story we saw involved young Scottish musician—accordion player to be precise—Murdo travelling to America with his father, who has no appreciation of music at all, after the death of his mother to see family.
They end up in the middle of nowhere, where Murdo comes across a blues band playing in the open air. He joins in on the squeezebox, and then is invited to joint them on stage at a festival in Lafayette in two weeks' time. Murdo and his father move on, but the idea of playing in a festival begins to obsess him more and more.
This was an interesting experiment that produced a captivating performance, well-acted and with some great music. However the question arises, what exactly were we watching? It was an entertaining and informative hour and a half, but it felt like the middle of a process rather than an end product. It would be great if these particular pieces were developed into something more substantial rather than being abandoned after one experimental showing.
As an experimental scratch piece, this was very well put together and well performed and certainly worth seeing.
Reviewer: David Chadderton


SUNDERLAND'S Seb Larsson cannot wait to take on league leaders Manchester City on New Year's Day and is hoping a bumper crowd on Wearside will help inflict only a second top-flight defeat of the season on Roberto Mancini's title chasers.
Swede Larsson was terrific in the 1-1 Boxing Day home draw with Everton, which Sunderland should have won had referee Howard Webb not handed David Moyes' side a controversial second-half penalty leveller. City are next and Larsson said: " Hopefully we can produce a good display at the Stadium of Light, with the backing of a big crowd.
"It is not an easy place to come and we want to get it back to the fortress it has been in the past. Everton looked very happy to get a point from here .
Webb's penalty award blunder, after Everton's Leon Osman kicked the ground with his own foot, hurt Martin O'Neill's men.
Larsson said: "It was the wrong decision but I don't think the player was trying to fall down intentionally.


Collin Smith, a fourth grade student at Champ Cooper visited Mrs. April's class to encourage the new First grade readers to participate in the Accelerated Reader Program. Collin currently has 100 points for this school year and was eager to help. After listening to the read aloud, the students worked together to take a quiz on the story. It was a great addition to the Reading lesson. Accelerated Reader is a great tool to help build reading flucency and comprehension. The students look forward to more guest readers and more AR points!
Jordan Cooper, a Kindergartner at O.W. Dillon Elementary School, has read over 100 Accelerated Reader books and earned over 50 A.R. points!! She is shown above with a stack of books she has read just in computer lab... the stack doesn't even include any that she has read in library. Congratulations go out to Jordan on this amazing accomplishment!
Liam Tallo - Tucker 2nd grader in Sherry Majeau's class is the first student to reach 100 AR points this year! Students attended an Accelerated Reader Party in the Library at Tucker, where they received prizes for all their hard work.
Tucker Elementary students take Accelerated Reader quizzes at school to test their comprehension on books they are reading. All of the students at Tucker combined took 7,446 quizzes! The following students earned over 100 points each. Top: Kaitlyn Hall, Zairana Jordan; Bottom: Madeline Falgout (top point earner), Maddie Ridgedell, John Cranford, Kollin Delatte, Karrlea Haist (not pictured.) These students, along with all students who earned 50 or more points came to the library for indoor bounce time, a movie and popcorn. Great job AR Stars!
Between the dates of January 7th and April 26, 2013, one hundred and thirty nine 1st and 2nd graders at Midway Elementary read 3,026 AR books. Students were given a ticket for every 5 points earned and were entered into a drawing to win a special t-shirt. The winner of the t-shirt was 1st grader, Ma'kiya Jackson from Ms. Peggy Frederick's class. Ma'kiya read 45 books and earned 21 AR points. The top 3 readers were 1st graders TyQuan Jackson and Madison Walker from Ms. Cheryl Donakey's class, and 1st grader Ali Crowe from Ms. Peggy Frederick's class. TyQuan read 95 books and earned 39 AR points. Madison read 84 books and earned 29 AR points. Ali read 73 books and earned 37 AR points. Each student received a $10.00 gift card from Books A Million donated by Master Teacher Robyn Hutchinson. Pictured above Ali Crowe, Ma'Kiya Jackson, Madison Walker, and TyQuan Jackson.
Ambria Cook, 6th grader in Mrs. Spears' Class at O.W. Dillon Elementary has already exceeded her Accelerated Reader Goal for the 2012-2013 school year. Her goal was 25 A.R. points for the year, but Ambria has already made 59.2 points. Students at O.W. Dillon are not allowed to get points by reading books at just any level. Ambria had to read books that were at least 6th grade level or higher in order to earn her points. We are very proud of her!!
In celebration of Sumner High students' success with the Accelerated Reader Program in the first semester of the school year, Mrs. Lesley Bridges, SHS Librarian, took top point earners to a reward lunch at the Boston Restaurant. Students invited to this lunch each earned 250+ points each in the first semester. Two of these students, Christy Miller and Randi Shaidee, earned more than 500 points each.
Overall, Sumner High has had a wonderful school year with AR. Grades 9-12 have earned 11,423.5 points and Mrs. Delatte's English II Honors class earned the most points as a class with 2358 points total. We would like to congratulate all of our students on their accomplishments and remind them that reading is the foundation of all learning. Congrats SHS!
Second graders Sara Addison and Saylor Baham in Mrs. Walker's class at Loranger Elementary have been very busy reading. Sara Addison has read over 114 books with an average Accelerated Reader score of 90%. Saylor Baham has read over 185 books with an average AR score of 95%. For earning "super sized" AR points, Mrs. Walker gave each a "Super Sized" candy bar.
This year at Hammond Westside Montessori the 6th grade English Language Arts teachers are focusing on Accelerated Reader. Accelerated Reader is an online program where students are assigned to read books, keep journals on what they read, and finally take tests on the books that they read. The students in Mrs. Tongia Keith's 6th grade class finished their first class novel Hiroshima on Friday, September 7th. Every student was assigned to take a test on Hiroshima. Almost every student in all three classes passed the AR test. The teachers are extremely proud of their students' accomplishment. Already, students have started their second class novel Sadako. We look forward to even more student achievement from the 6th grade students at Hammond Westside Montessori. Pictured front row from left are Kalen Alexander, Jimmiesha Felder, Teniya Pines, Tyveion Franklin, Atilah Tircuit, and Ymmot Jackson. Middle row from left are Chelsea Watson, Amari Wilford, Ashante Travis, Jazmine Donahue, and Erin Bell. Back row from left are Shyterrica Stalls, Keshon Adams, Troy Buggage, Amani Gilmore, Trey Schillace, Kendel Moore, and Cadarious Barton.


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Teaching how to feel safe with risk
November 27, 2015 Jamie MacAlister	Leave a comment
This week an Ashridge team won a Training Journal award for our leadership development work with one of our clients, Heineken. https://www.ashridge.org.uk/executive-organisation-development/custom-programmes/case-studies/Heineken/ The essence of the programme is helping participants to experiment with different leadership styles and interventions in high stress situations, which they can do in the relatively "safe" environment away from the workplace. This is done through a classroom simulation of an organisation in crisis, where actors play roles which create the stress. Participants wear heart-rate monitors, and are regularly evaluating their own approaches and physiological responses, as well as those of each other. The idea is they develop a kind of muscle memory for the impact of different kinds of interventions and responses in situations involving significant personal risk.
In the introduction to my book, I state that the book is about the journey towards feeling safe with risk, which is the key to effective strategy.
One of my stated aims from this book is to help leaders feel safe with risk. This is of course a paradox, yet I believe the reality is that leaders will not take the risks they need to take if they don't feel safe in doing so. This is looking under the bonnet of the leader, to understand what might be going on behind the scenes to support right risk taking. Up to now we have been looking at the outward evidence of risk-taking decisions in organisations, with some exploration of what psychological factors might influence those decisions. But the deeper issue is how risk makes you feel when you take it, particularly personal risk.
I believe at some level, right risk takers do have a mechanism for helping them to feel safe. Part of this may indeed be the hormonal effect which not only prepares us with the capabilities to engage more effectively with risk, as in the case of testosterone, but also provides an anaesthetic to numb the fear. This makes me think of adrenaline, and reminds me of the day my Achilles tendon snapped while playing indoor soccer. I can still remember the excruciating pain in my ankle the moment it happened, as if I had been struck there by the sharp edge of a brick that had been thrown at my foot. Within only five or so seconds, the pain was already starting to reduce. The adrenalin was kicking in. Then I thought I had just been taken out with a tackle from behind. The first aider who took care of me accurately diagnosed it as an Achilles tendon going after a bit of research – no one had been anywhere near me. But the numbing effect of the adrenalin meant I drove myself to the hospital, even though I was only able to use my heel and not the front of my foot to depress the clutch.
The idea of feeling safe with risk is one of the bases on which we have tried to offer Executive Education at Ashridge.
There is a conundrum. In traditional teaching environments, away from the buzz of everyday working life, how do we replicate the real risks that leaders face, in the relatively "safe" environment of the classroom? Conceptualising risk for learning purposes appears to have an anaesthetising impact on our actual experience of it. We may think our way through a case study involving risk in an abstract way, using suitable models to help us; but the emotion of risk is generally missing in a "safe" environment. Virtually nothing we do or say in a classroom setting will put the company finances at risk, nor the health of our stakeholders, nor our own careers. To some extent this is exactly why the "classroom" is designed to be a "safe" environment – it's confidential, it's safe. "What's in the room stays in the room?"
Indeed, the unique selling proposition for "away-from-the-workplace" learning is that it is a safe environment in which to experiment, in which to take some 'risks' that participants wouldn't normally take in the workplace. These risks can be about exploring and articulating new ideas to colleagues, about having conversations that they wouldn't normally have, about interacting with others in a way that may not feel comfortable. So for some aspects of working with risk, the "classroom" offers an advantage, even if it may feel a little artificial or clinical.
But, paradoxically, if we are creating safety to work with risk, how can this be authentic?
We have successfully worked around this dilemma in our management development practice by using simulations that generate the emotion of risk. These take the form either of team-based competitive decision making in a computerised market-place simulation over a series of rounds, or of working through specific artificial challenges working with professional actors, whose role it is to re-create some of the emotion of difficult, risky, situations. Participants have also been asked to wear heart rate monitors over one or two days, as they work together to resolve simulated problems which may be creating tensions between individuals. Participants are encouraged to experiment with roles which are different from those they would normally have, and with types of interventions which they would typically not employ. Their emotional responses are monitored and measured.
These programmes demonstrate that they can have a significant residual learning impact – i.e. this learning manifests itself after, not during, the experience of the programme. Participants start to apply certain aspects of what they have experimented with on the programme back at work and they are encouraged, post programme, to reflect and to continue learning from these reflections. They effectively build on the risk that they have already taken during the programme by applying the experiment in a real work environment. There is effectively a physiological stress memory, reflected in the heart rate print-out, which gets replayed in the workplace. This has been called the development of emotional muscle memory, in the same way that a tennis player only really learns how to hit an effective tennis shot when he does it automatically without having to think about it – i.e. when he has developed muscle memory.
Could Federer have won by taking more risk?
Yesterday, Djokovic won the ATP World Tour Finals tennis at the O2, beating Federer by two sets to zero. I was disappointed because Federer had indicated he would pursue a more aggressive style of tennis, in particular by coming to the net to volley more often. Overall, in 116 rallies in all, Federer lost because he only won 46% of them. When he came to the net in rallies, he won 65% of the time. But he only came to the net for 17 of those rallies! The statistics suggest that if he had come to the net more often, he might have won. Why didn't he? Part of the reason could be summarised in this extract from my book: "Risky Strategy"
My game is tennis, and I notice that an approach to risk is being played out on a point-by-point basis. An approach can be planned in advance, in the form of an overall match or set strategy, but in reality you have a point strategy, or even an in-the-moment shot strategy. So how does risk manifest itself in tennis? Well, one way is in the approaches to the net as shown in the following diagram. This is based on some fairly rough and ready research of rally length and outcomes in professional games. The average rally length in a game is around a surprisingly low five to six shots total, so less than three per player. In terms of winning or losing shots, at the top professional level of the game, this is split very close to 50:50 – it's surprising how close a lot of professional games are in terms of points won or lost.
So what is this chart showing us? This is an equivalent to a normal distribution curve for two types of tennis shot: the blue bars represent the distribution for shots played from the back of the court, and the red from the net area. There are just three points in the distribution for each shot type, as we are only interested in three possible outcomes from the shot; either it's a winning shot, it's a losing shot, or the rally continues so that the player gets another shot.
So first to note that our flatter 'curve' is our more risky shot, remembering the way I demonstrated risk and variability in the earlier chapter. In other words, there is more variability in possible outcome with our flatter (brown) net shot – which is what we would expect. The net shot is more risky; there is more chance of the extreme outcomes, either a win or a loss.
A back of the court shot is less risky; it has a more peaked profile – the most likely outcome from the shot by a good margin is a continuation of the rally.
So what we see from this is that the shot which has a higher chance of winning the point is the shot at the net – ie the more risky shot.
What is interesting is that tennis players know this, and yet the shot at the net is a relatively rare part of the game at the professional level today. It used to be more regularly played; up until the late 1970s and early 1980s, the serve and volley at the net was a regular way for players to win points, particularly on grass courts. Of course, there are a number of reasons why that has changed. Tennis balls are slower and make it harder to hit winners. The perfected top spin ground shot, pioneered by Bjorn Borg, changed everything, making it easier to hit dipping passing shots. And longer rallies and games means energy is a bigger factor – coming to the net expends more energy.
However, the most interesting feature of this analysis is one other reason why us tennis players are more reluctant to come to the net. Yes there may be a higher chance that the shot will be a winner. But there is also a higher chance it will be a loser. And, in the tiger moment, away from rational elephant analysis, our loss aversion kicks in!
Choosing risks and winning aspirations ; the Race to the Pole
[An excerpt from the book "Risky Strategy" to be published in 2016]
We see an intriguing relationship between "winning" and "risk" in the race to the South Pole in 1910, between the English Robert Falcon Scott and the Norwegian Roald Admundsen. For both explorers, winning was in part at least about being the first to reach the South Pole, one of the last unexplored areas on Earth. For both, this undoubtedly involved a high level of personal risk, to themselves and their chosen teams. But this is where the similarities end.
Admundsen reached the Pole first, and returned to their ship with a team completely in tact. Scott by contrast arrived at the Pole weeks later; he and his entire team failed to return alive.
What else was different? Firstly, their specified goals were different. Admundsen had a single goal: to be the first to reach the South Pole. It was probably the best example of a clearly defined and focused winning aspiration – easy to confirm or measure success. Scott had two goals: to get to the Pole first, like Admundsen, and to gather scientific information about the Antarctic. As a result of this, they each took different types of risks.
For Scott with his dual goals, in a situation where time was a critical factor,he delayed on his return trip to collect geological samples, which also added weight to his sledges. Admundsen, on the other hand, took a different kind of risk, which was consistent with his winning goal. While Scott chose a route from McMurdo Sound, which had already been partly explored by Ernest Shackleton in 2007, Admundsen chose an unchartered route, from the Bay of Whales, which was on the edge of the Great Ice Barrier, where explorers had feared that the ice could fracture and send you floating away. But Admundsen chose this because it was 60 miles closer to the Pole. In addition, because Scott has assumed Admundsen would choose the same route as him, he assumed that he was ahead of him, as there were no signs of Admundsen's tracks. With this assumption, he had no compelling driver to go faster.
However, in this amazing story, even though the single-minded winning goal that Admundsen led to certain risks, he compensated for this by reducing risk in other ways. He had investigated the records of other explorers who had been in the area, and noted that the ice had remained unchanged for decades. So this gave him more confidence in his choice of routes.
Admundsen had also researched the best type of transport to use extensively by spending time with the Inuit Eskimos in the Arctic. He used dogs which were very well suited to the extreme conditions. Scott, on the other hand, for most of the journey used a combination of ponies, motor sledges and man-hauling. The motor sledges were untested and quickly broke down. The ponies although they could haul heavier loads than dogs were also ill suited – they had no natural vegetation to feed on, sweated through their hides which led heavy ice to form and caused them to sink more deeply into the snow – they all had to be put down. This left man-hauling for most of the journey, recommended by Shackleton to be the best and most noble approach. But a noble but ill-conceived approach didn't get the job done.
What this famous piece of history tells us about winning and risk, is that while a winning aspiration creates the need and appetite for risk, leaders need to be choiceful about which risks they take, which ones they work hard to avoid.
For such a time as this – students seek new "capitalism"
I was struck by Paul Polman's comments in this interview that students that he visits around the world are seeing the need to "evolve capitalism and create a more equitable and sustainable form of growth. They get it, they really get this need for change!" http://www.theleadershipvanguard.com/#/paul-polman/
Last week, I ran two sessions on "Risks in the global supply chain" with undergraduates at Hult Business School – representing a fairly balanced mix of students from Europe, The Americas, Asia and Africa. We talked about global strategy making, referring to my book, "Risky Strategy" – there was some interest. I then introduced a case study based on the research we had just completed on modern slavery in the global supply chain, in which we had talked to a number of the UK's top retailers. The engagement levels increased dramatically.
The class discussions battled with the challenges presented by a capitalist system that requires retailers to compete in the interests of the consumers, and the need for those same retailers to collaborate to truly be able to tackle this pernicious issue of slavery. Has the mantra the "customer is king" in reality been translated to mean the "worker is slave"? Is this particularly the case because the "customer" in value terms is primarily from the wealthy developed nations, and the "worker" predominantly from the poorer developing nations?
Is this a time for business leaders like Polman to challenge these basic assumptions? Its risky… yes and ..? Is this a time for business to become part of the solution rather than part of the problem. The students seem to think so!


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Vampire Action Novel 'Blood Oath' Adaptation in the Works
Vampire Action Novel Blood Oath Adaptation in the Works
Cat Parker
— May 17, 2010
Warp Films' Lucas Foster (Law Abiding Citizen) has picked up rights to Blood Oath, a vampire action novel written by Christopher Farnsworth, and plans to fast-track the feature franchise about a vampire secret agent, according to Heat Vision.
Blood Oath centers on a vampire captured by the U.S. Army just after the Civil War who is forced to swear an oath to President Andrew Johnson and has protected the presidency in the Secret Service for nearly 150 years.
In the story, the vampire, named Nathaniel Cade, is paired with a new human handler and finds himself embroiled in a plot involving a monstrous new form of biological weapon with a connection to the White House.
The deal comes just as the book, from Penguin/Putnam, hits stores Tuesday.
Foster has picked up the rights for "Oath" and its follow-up, "Black Site," which Farnworth is still writing, and will develop scripts for them simultaneously. Farnsworth will work with a screenwriter to help guide the look and feel of the scripts and receive an exec producer credit.
"I'm using my own money so I'm pretty careful with the things I develop, and I tend to make the things I develop," Foster said. "I want to get a fast start, and I'm hoping to get this in production next year."