Paul McNeil - Academia.edu (original) (raw)
Papers by Paul McNeil
Experimental Eye Research, Apr 1, 1999
The nature and the extent of acute injury to corneal endothelial cells caused by exposure to ultr... more The nature and the extent of acute injury to corneal endothelial cells caused by exposure to ultrasound radiation were characterized, as well as the long-term reaction of these cells to this form of injury. It was found that the degree of lethal cell injury induced by ultrasound scaled with exposure intensity and duration. Immediate changes in plasma membrane permeability were induced by ultrasound exposure. This ultrasound-induced permeability change was, however, transient in many cells, allowing them to trap and retain a normally impermeant tracer, fluorescein dextran, in cytosol. Microvilli were present on ultrasound treated cells in far greater density than on control cells, characteristic of exocytosis-based resealing. Cultures containing a majority of transiently permeabilized endothelial cells were morphologically indistinguishable from untreated control cultures, and the fluorescein dextran-labeled cells in these populations locomoted and divided normally. We conclude that cell death due to ultrasound exposure can occur rapidly via a necrotic mechanism that can be attributed to mechanically induced damage to the plasma membrane. However, not all cells injured become necrotic : some survive and appear to behave normally after exposure. Conditions that favor plasma membrane disruption resealing, e.g. that result in sub-lethal rather than lethal cell injury, may mitigate the reduction in corneal endothelial cell density consequent on phacoemulsification and aspiration surgery.
Journal of Cell Science, Mar 1, 2002
Journal of Cell Biology, Aug 1, 1989
Growth factors may be required at sites of mechanical injury and normal wear and tear in vivo, su... more Growth factors may be required at sites of mechanical injury and normal wear and tear in vivo, suggesting that the direct action of mechanical forces on cells could lead to growth factor release. Scraping of cells from the tissue culture substratum at 37°C was used to test this possibility. We show that scraping closely mimics in vitro both the transient plasma membrane wounds observed in cells subject to mechanical forces in vivo (McNeil, P. L., and S. Ito. 1989. Gastroenterology. 96:1238-1248) and the transient plasma membrane wounds shown here to occur in endothelial cells under normal culturing conditions. Scraping of endothelial cells from the culturing substratum released into the culture medium a potent growth-promoting activity for Swiss 3T3 fibroblasts. Growth-promoting activity was released rapidly (within 5 min) after scraping but was not subsequently degraded by the endothelial cells for at least 24 h thereafter. A greater quantity of growth-promoting activity was released by cells scraped 4 h after plating than by those scraped 4 or 7 d afterwards. Thus release is not due to scraping-induced disruption of extracellular matrix. Release was only partially cold inhibitable, was poorly correlated with the level of cell death induced by scraping, and did not occur when cells were killed with metabolic poisons. These results suggest that mechanical disruption of plasma membrane, either transient or permanent, is the essential event leading to release. A basic fibroblast growth factor-like molecule and not platelet-derived growth factor appears to be partially responsible for the growthpromoting activity. We conclude that one biologically relevant route of release of basic fibroblast growth factor, a molecule which lacks the signal peptide sequence for transport into the endoplasmic reticulum, could be directly through mechanically induced membrane disruptions of endothelial cells growing in vivo and in vitro.
An edition of 26 posters, in A3, A2 and A1 sizes, concerning the ThreeSix optical/geometric typef... more An edition of 26 posters, in A3, A2 and A1 sizes, concerning the ThreeSix optical/geometric typeface system developed by MuirMcNeil. Designed by Paul McNeil and Hamish Muir. Output: 26 posters published online at www.print-process.com.
The visual grammar poster was designed and produced at the invitation of Modern Theory in 2012 as... more The visual grammar poster was designed and produced at the invitation of Modern Theory in 2012 as part of an international show of work from 20 design groups. The Visual Grammar show took place at MAD Brussels in September 2012. A specific briefing was provided and a fixed body of text was supplied to all participants who were asked to visually describe or define 'visual grammar' in the form of a poster. We took advantage of the project to prototype two new typeface designs. The intention for the poster was to use only the text provided in a way that distilled the words to visual forms without linguistic content. It was anticipated that separating form from meaning would speak about visual grammar in a direct and immediate way. Having undertaken a design development process involving around 50 alternative iterations, the final poster is a completely typeset composition where the boundary between what can and what can't be read is blurred. The body text is set in a single weight and size of ThreeTwo 00, an extensive modular type system which is currently in development. Here, letterforms are composed from dot grids at fixed increments. As type sizes become smaller, resolution decreases within the dot pitch, which remains constant. Although bodies of text appear to retain typographic features, characters are unrecognisable individually, testing the limits of legibility in context.
Berg eBooks, 2009
A well-established history of costume has argued that a dynamic fashion system emerged within a c... more A well-established history of costume has argued that a dynamic fashion system emerged within a conjunction of the competing polities of France, England and Burgundy in the la te Middle Ages, and also that, around 1350, men began to ap? pear very different in their dress from women. The textiles and Other components of which fashion was manufactured were the focus of the medieval economy, an ancient and expanding trade spreadi ng innovation and promoting new aesthetics. Already by the fou rteenth century the majority of guilds in towns were occupied with providing luxury dress and trading in its components. Workshops began ro sep? arate from princely courtS and disseminated high levels of specialization. The most skilful makers moved from place to place, sometimes avoid ing corporate constraint and spreading new aesthetics within portable objects such as tailored clothes, textiles, embroideries, illuminated books, sta tuettes and jewels. Fashions in dress were centra l to technological innovation and the culture of social legibi liry and hierarchical display that characterized late medieval and Rena issance Europe. Although the idea of the 'Dark Ages' remains tenacious, alternative views of medieval life argue that the loss of certain hiera rch ies and the rise of print culture permitted Western Europe to develop a certain 'openness' to competing sets of ideas which produced new social forms
Journal of the American Society of Mining and Reclamation, 2003
Revegetation of mine residues (tailings) is an important aspect of rehabilitation after mining, a... more Revegetation of mine residues (tailings) is an important aspect of rehabilitation after mining, and represents a substantial investment. Therefore it is important to increase our understanding of all aspects of re-establishing a sustainable vegetation community on these materials. We report here on a multidisciplinary, collaborative research program, focusing on vegetation establishment, soil development, and water-balance modeling on a large residue revegetation experiment. The study focused on gold residues produced at the Boddington Gold Mine (BGM) and Hedges Gold Mine (HGM), in the southwest of Australia. The residue storage areas will be rehabilitated once no longer required, but revegetation may be hampered by the alkaline, saline, and sodic properties of the residue. A large field experiment was established to examine soil amendments and capping strategies. The treatments were three depths of gravel-rich subsoil (0 cm, 15 cm, and 30 cm) overlying residue treated with gypsum (30 t/ha). All plots subsequently received an application of topsoil (10 cm). The plots were established in 1999 with species from the local jarrah (Eucalyptus marginata Donn. ex Smith) forest, or salt-and waterlogging-tolerant native species. Ten months after application, gypsum had contributed to a decrease in residue pH and salinity. By March 2000, approximately 90% of the directly seeded species had emerged and survived, and 100% of transplanted seedlings had survived. Applying gravel subsoil in addition to topsoil did not improve plant growth in the first two years. In fact, aboveground biomass production was higher, from 4 to 8.5 t/ha/yr, in the absence of a gravel subsoil. However, in the third growing season, this trend was less apparent.
Current protocols in cell biology, May 1, 2001
Techniques for introducing normally impermeant macromolecules into the living cell—referred to as... more Techniques for introducing normally impermeant macromolecules into the living cell—referred to as “cell‐loading techniques”—are useful in a variety of settings for the cell biologist. Microinjection is probably the most commonly used technique for introducing fluorescent probes, fluorescently tagged proteins, and antibodies into living cells for short‐term studies of cell physiology and protein location and function. It is, however, not the only technique available, nor the easiest or least expensive to implement. Among the alternatives are several closely related techniques that, like microinjection, rely on the cell's ability to reseal a mechanically induced plasma membrane disruption created in order to gain temporary access to cell cytosol. Four such techniques are described in this unit: scrape loading, scratch loading, bead loading, and syringe loading. Unlike microinjection, these techniques allow one to rapidly load (in a matter of minutes) thousands or even many millions of many types of mammalian cells with normally impermeant molecules, and so to facilitate quantitative analyses of the effect of loading.
PLOS Biology, Aug 22, 2006
When a cell suffers a plasma membrane disruption, extracellular Ca 2þ rapidly diffuses into its c... more When a cell suffers a plasma membrane disruption, extracellular Ca 2þ rapidly diffuses into its cytosol, triggering there local homotypic and exocytotic membrane fusion events. One role of this emergency exocytotic response is to promote cell survival: the internal membrane thus added to the plasma membrane acts as a reparative ''patch.'' Another, unexplored consequence of disruption-induced exocytosis is secretion. Many of the cells lining the gastrointestinal tract secrete mucus via a compound exocytotic mechanism, and these and other epithelial cell types lining the digestive tract are normally subject to plasma membrane disruption injury in vivo. Here we show that plasma membrane disruption triggers a potent mucus secretory response from stomach mucous cells wounded in vitro by shear stress or by laser irradiation. This disruption-induced secretory response is Ca 2þ dependent, and coupled to cell resealing: disruption in the absence of Ca 2þ does not trigger mucus release, but results instead in cell death due to failure to reseal. Ca 2þ-dependent, disruption-induced mucus secretion and resealing were also demonstrable in segments of intact rat large intestine. We propose that, in addition to promoting cell survival of membrane disruptions, disruption-induced exocytosis serves also the important protective function of liberating lubricating mucus at sites of mechanical wear and tear. This mode of mechanotransduction can, we propose, explain how lubrication in the gastrointestinal tract is rapidly and precisely adjusted to widely fluctuating, diet-dependent levels of mechanical stress.
Journal of Cellular Physiology, Jul 1, 1991
Cells of gut and skin frequently suffer mechanically-induced plasma membrane disruptions in vivo,... more Cells of gut and skin frequently suffer mechanically-induced plasma membrane disruptions in vivo, and bioactive molecules, including basic fibroblast growth factor (bFGF), could enter and leave cytoplasm through these disruptions. We here provide three lines of evidence that bFGF is released with surprising efficiency through plasma membrane disruptions, resembling those known to occur in vivo, produced by scraping endothelial cells from their culturing substratum. First, 41 O h of the total of bFGF extractable in 1 M NaCl by freeze-thaw and sonication was released simply by scraping the endothelial cells. Second, relative to release of lactate dehydrogenase, cells wounded by scraping under conditions promoting >60% cell survival released a significantly larger amount (up to twofold more) of growth promoting activity than did cells uniformly killed and irreversibly permeabilized by scraping in the cold or by freezing and thawing. Last, cells that survived membrane disruptions released, and contained, less bFGF on each subsequent wounding, consistent with release of bFCF through transient (i.e., survivable) membrane disruptions. A polyclonal antibody against bFGF completely neutralized the growth promoting activity released by scraping, confirming that bFGF is released through endothelial cell plasma membrane disruptions. Cell fractionation and immunolocalization, including a novel per
Journal of Surgical Research, Feb 1, 2006
Introduction: Burn injury results in early T cell apoptosis, lymphopenia and associated T cell dy... more Introduction: Burn injury results in early T cell apoptosis, lymphopenia and associated T cell dysfunction. Subsequent reconstitution of the T cell compartment late after burn injury via homeostatic proliferation generates hyper-responsive memory CD8ϩ T cells of unclear functional significance. In this study, we hypothesized that the hyper-responsive CD8ϩ population isolated late after burn injury 1) has enhanced activity in vivo and 2) that burn injury associated apoptosis is necessary for the generation and hyperresponsiveness of this unique T cell population. Methods: To test these hypotheses, we used a mouse model of allogeneic skin graft rejection to assess in vivo activity and the glucocortocoid receptor inhibitor mifepristone (RU486) to block burn associated apoptosis. Female C57Bl/6 (B6) mice transgenic for the T cell receptor recognizing the male minor histocompatibility antigen HY (HY TCR mice) were anesthetized and treated as follows (nϭ6 per group): 1) 20% TBSA burn; 2) 20% TBSA burn and 3 daily injections (20g/g) of RU486; 3) sham and 4) sham with RU486. Mice were sacrificed 14 days after burn injury, splenic CD8ϩ T cells purified (Ͼ95% purity) by magnetic cell separation and CD8ϩ T cells from each group were adoptively transferred i/v into female wildtype B6 mice (1ϫ106 cells / mouse, nϭ6 per group). Transferred cells were allowed to redistribute for 48 hours, then each mouse received allogeneic male (HYϩ) skin and control isogeneic female (HYϪ) tail skin grafts. Grafts were monitored daily, scored for rejection and median survival times of allogeneic male grafts were calculated. Statistical analysis was performed by logrank test with significance defined as pϽ0.05. Results: Mice that received CD8ϩ HY TCR T cells isolated late after burn injury rejected male skin graft significantly faster than mice that received CD8ϩ T cells from sham mice (11 days versus 15 days, pϭ0.038), demonstrating that transferred cells from burn mice have enhanced activity in vivo. This effect was blocked when RU486 was administered to burn mice and subsequent skin graft rejection in adoptive transfer recipients was compared to the burn alone recipients (19 days versus 11 days, pϭ0.0013). Administration of RU486 to burn mice also resulted in similar graft rejection when compared to sham (19 days versus 15 days, pϭ0.30), sham with RU486 (19 days versus 15 days, pϭ0.55) and no cells (19 days versus 17 days, pϭ0.75) adoptive transfer recipients. Conclusions: This study demonstrates for the first time the functional significance in vivo of hyper-responsive CD8ϩ T cells late after burn injury. While blocking apoptosis may have beneficial effects on immune function early after burn injury it also appears to prevent the development of these
... [Art/Design Item] [Creative Arts and Design > Graphic Design]. Details. Creators: Mc N... more ... [Art/Design Item] [Creative Arts and Design > Graphic Design]. Details. Creators: Mc Neil, Paul. Description/Abstract: Design of printed promotional material commissioned by the Energy Saving Trust, 2002. This design research ...
Journal of Surgical Research, Feb 1, 2006
Introduction: In the developmental stage, pancreas derives from the endodermal cells where the tr... more Introduction: In the developmental stage, pancreas derives from the endodermal cells where the transcription factor, pancreatic duodenal homeobox gene-1 (pdx-1) is expressed. In the adulthood, pdx-1 expression is limited to pancreatic  cells and pdx-1 is necessary for maintaining the function. Recent studies have shown that ectopic expression of pdx-1 like liver successfully induced insulin production and ameliorated hyperglycemia. The purpose of this study was to investigate the effects of enforced expression by the adenovirus-mediated gene transfer of pdx-1 in the jejunal epithelia where pdx-1 is not expressed. Methods: The recombinant replication-deficient adenovirus carrying pdx-1 gene was constructed using the COS-TPC method. ICR mice were treated with STZ (220 mg/kg i.p.) After determining the hyperglycemia, a loop-jejunostomy was constructed and the adenovirus solutions (Ad-pdx-1 and Ad-lacZ 1ϫ108 PFU/body) were injected into the lumen of the loop. (1) Blood glucose concentrations were measured by drawing blood from ocular veins. (2) Immunohistochemical or fluorescent analyses of PDX-1 and insulin in the adenovirus-infected jejunal epithelia were performed. (3) RT-PCR of pdx-1 and other pancreatic markers were investigated. (4) Immunoreactinve insulin (IRI) extracted from the adenovirus-infected jejunum were measured. Results: (1) Hyperglycemia was improved in STZ-treated mice infected with Ad-pdx-1. (2) Ad-pdx-1 extensively induced ectopic PDX-1 expression in the jejunum and the scattered cells in the jejunal epithelia were positive for insulin expression. (3) mRNA of insulin Ϫ1, Ϫ2 and pdx-1 were expressed in mice with Ad-pdx-1. (4) IRI level of the jejunum extract was significantly increased in mice with Ad-pdx-1. Conclusions: Gene transfer of pdx-1 in intestinal epithelia could be a promising strategy for diabetes mellitus by inducing ectopic insulin producing cells. 244. THE PERITONEAL CAVITY IS A DISTINCT COMPARTMENT OF ANGIOGENIC MOLECULAR MEDIATORS.
Agro Food Industry Hi-tech, Jul 1, 2012
Research, design and production of Generator typeface and poster on behalf of Dr Charley Peters a... more Research, design and production of Generator typeface and poster on behalf of Dr Charley Peters and Saturation Point Projects — an LCC funded research project. Generator, curated by Saturation Point Projects (saturationpoint.org.uk), was an exhibition exploring the language of generative art that took place at the Kaleidoscope Gallery in 2015. Artworks on display, by Katrina Blannin, Christina France, Hanz Hancock, James Irwin, Patrick Morrissey, Andy Parkinson, Charley Peters and Mary Yacoob, employed analogue systems to define visual outputs. MuirMcNeil were invited to design and produce a poster to accompany the event. Featuring a text by Laura Davidson, the 70x50cm poster is printed in orange and violet neon inks in two bespoke typefaces generated exclusively for the project.
Inclusion of ThreeSix Typeface and Unit Editions U:D/R03 journal in Wim Crouwel exhibition at Des... more Inclusion of ThreeSix Typeface and Unit Editions U:D/R03 journal in Wim Crouwel exhibition at Design Museum, London, 30 March – 03 July 2011.
A special edition of Unit Editions design research journal, concerning the ThreeSix optical/geome... more A special edition of Unit Editions design research journal, concerning the ThreeSix optical/geometric typeface system developed by MuirMcNeil. Written and designed by Paul McNeil and Hamish Muir, with an introduction by Wim Crouwel, U:D/R 03 examines ThreeSix in relation to its historical, ideological and typographic contexts. The publication also describes the design process in detail. This edition was produced in newsprint, single colour litho printing.
Contribution of article to publication and limited edition poster to Team Impression’s X Ten proj... more Contribution of article to publication and limited edition poster to Team Impression’s X Ten project.
Experimental Eye Research, Apr 1, 1999
The nature and the extent of acute injury to corneal endothelial cells caused by exposure to ultr... more The nature and the extent of acute injury to corneal endothelial cells caused by exposure to ultrasound radiation were characterized, as well as the long-term reaction of these cells to this form of injury. It was found that the degree of lethal cell injury induced by ultrasound scaled with exposure intensity and duration. Immediate changes in plasma membrane permeability were induced by ultrasound exposure. This ultrasound-induced permeability change was, however, transient in many cells, allowing them to trap and retain a normally impermeant tracer, fluorescein dextran, in cytosol. Microvilli were present on ultrasound treated cells in far greater density than on control cells, characteristic of exocytosis-based resealing. Cultures containing a majority of transiently permeabilized endothelial cells were morphologically indistinguishable from untreated control cultures, and the fluorescein dextran-labeled cells in these populations locomoted and divided normally. We conclude that cell death due to ultrasound exposure can occur rapidly via a necrotic mechanism that can be attributed to mechanically induced damage to the plasma membrane. However, not all cells injured become necrotic : some survive and appear to behave normally after exposure. Conditions that favor plasma membrane disruption resealing, e.g. that result in sub-lethal rather than lethal cell injury, may mitigate the reduction in corneal endothelial cell density consequent on phacoemulsification and aspiration surgery.
Journal of Cell Science, Mar 1, 2002
Journal of Cell Biology, Aug 1, 1989
Growth factors may be required at sites of mechanical injury and normal wear and tear in vivo, su... more Growth factors may be required at sites of mechanical injury and normal wear and tear in vivo, suggesting that the direct action of mechanical forces on cells could lead to growth factor release. Scraping of cells from the tissue culture substratum at 37°C was used to test this possibility. We show that scraping closely mimics in vitro both the transient plasma membrane wounds observed in cells subject to mechanical forces in vivo (McNeil, P. L., and S. Ito. 1989. Gastroenterology. 96:1238-1248) and the transient plasma membrane wounds shown here to occur in endothelial cells under normal culturing conditions. Scraping of endothelial cells from the culturing substratum released into the culture medium a potent growth-promoting activity for Swiss 3T3 fibroblasts. Growth-promoting activity was released rapidly (within 5 min) after scraping but was not subsequently degraded by the endothelial cells for at least 24 h thereafter. A greater quantity of growth-promoting activity was released by cells scraped 4 h after plating than by those scraped 4 or 7 d afterwards. Thus release is not due to scraping-induced disruption of extracellular matrix. Release was only partially cold inhibitable, was poorly correlated with the level of cell death induced by scraping, and did not occur when cells were killed with metabolic poisons. These results suggest that mechanical disruption of plasma membrane, either transient or permanent, is the essential event leading to release. A basic fibroblast growth factor-like molecule and not platelet-derived growth factor appears to be partially responsible for the growthpromoting activity. We conclude that one biologically relevant route of release of basic fibroblast growth factor, a molecule which lacks the signal peptide sequence for transport into the endoplasmic reticulum, could be directly through mechanically induced membrane disruptions of endothelial cells growing in vivo and in vitro.
An edition of 26 posters, in A3, A2 and A1 sizes, concerning the ThreeSix optical/geometric typef... more An edition of 26 posters, in A3, A2 and A1 sizes, concerning the ThreeSix optical/geometric typeface system developed by MuirMcNeil. Designed by Paul McNeil and Hamish Muir. Output: 26 posters published online at www.print-process.com.
The visual grammar poster was designed and produced at the invitation of Modern Theory in 2012 as... more The visual grammar poster was designed and produced at the invitation of Modern Theory in 2012 as part of an international show of work from 20 design groups. The Visual Grammar show took place at MAD Brussels in September 2012. A specific briefing was provided and a fixed body of text was supplied to all participants who were asked to visually describe or define 'visual grammar' in the form of a poster. We took advantage of the project to prototype two new typeface designs. The intention for the poster was to use only the text provided in a way that distilled the words to visual forms without linguistic content. It was anticipated that separating form from meaning would speak about visual grammar in a direct and immediate way. Having undertaken a design development process involving around 50 alternative iterations, the final poster is a completely typeset composition where the boundary between what can and what can't be read is blurred. The body text is set in a single weight and size of ThreeTwo 00, an extensive modular type system which is currently in development. Here, letterforms are composed from dot grids at fixed increments. As type sizes become smaller, resolution decreases within the dot pitch, which remains constant. Although bodies of text appear to retain typographic features, characters are unrecognisable individually, testing the limits of legibility in context.
Berg eBooks, 2009
A well-established history of costume has argued that a dynamic fashion system emerged within a c... more A well-established history of costume has argued that a dynamic fashion system emerged within a conjunction of the competing polities of France, England and Burgundy in the la te Middle Ages, and also that, around 1350, men began to ap? pear very different in their dress from women. The textiles and Other components of which fashion was manufactured were the focus of the medieval economy, an ancient and expanding trade spreadi ng innovation and promoting new aesthetics. Already by the fou rteenth century the majority of guilds in towns were occupied with providing luxury dress and trading in its components. Workshops began ro sep? arate from princely courtS and disseminated high levels of specialization. The most skilful makers moved from place to place, sometimes avoid ing corporate constraint and spreading new aesthetics within portable objects such as tailored clothes, textiles, embroideries, illuminated books, sta tuettes and jewels. Fashions in dress were centra l to technological innovation and the culture of social legibi liry and hierarchical display that characterized late medieval and Rena issance Europe. Although the idea of the 'Dark Ages' remains tenacious, alternative views of medieval life argue that the loss of certain hiera rch ies and the rise of print culture permitted Western Europe to develop a certain 'openness' to competing sets of ideas which produced new social forms
Journal of the American Society of Mining and Reclamation, 2003
Revegetation of mine residues (tailings) is an important aspect of rehabilitation after mining, a... more Revegetation of mine residues (tailings) is an important aspect of rehabilitation after mining, and represents a substantial investment. Therefore it is important to increase our understanding of all aspects of re-establishing a sustainable vegetation community on these materials. We report here on a multidisciplinary, collaborative research program, focusing on vegetation establishment, soil development, and water-balance modeling on a large residue revegetation experiment. The study focused on gold residues produced at the Boddington Gold Mine (BGM) and Hedges Gold Mine (HGM), in the southwest of Australia. The residue storage areas will be rehabilitated once no longer required, but revegetation may be hampered by the alkaline, saline, and sodic properties of the residue. A large field experiment was established to examine soil amendments and capping strategies. The treatments were three depths of gravel-rich subsoil (0 cm, 15 cm, and 30 cm) overlying residue treated with gypsum (30 t/ha). All plots subsequently received an application of topsoil (10 cm). The plots were established in 1999 with species from the local jarrah (Eucalyptus marginata Donn. ex Smith) forest, or salt-and waterlogging-tolerant native species. Ten months after application, gypsum had contributed to a decrease in residue pH and salinity. By March 2000, approximately 90% of the directly seeded species had emerged and survived, and 100% of transplanted seedlings had survived. Applying gravel subsoil in addition to topsoil did not improve plant growth in the first two years. In fact, aboveground biomass production was higher, from 4 to 8.5 t/ha/yr, in the absence of a gravel subsoil. However, in the third growing season, this trend was less apparent.
Current protocols in cell biology, May 1, 2001
Techniques for introducing normally impermeant macromolecules into the living cell—referred to as... more Techniques for introducing normally impermeant macromolecules into the living cell—referred to as “cell‐loading techniques”—are useful in a variety of settings for the cell biologist. Microinjection is probably the most commonly used technique for introducing fluorescent probes, fluorescently tagged proteins, and antibodies into living cells for short‐term studies of cell physiology and protein location and function. It is, however, not the only technique available, nor the easiest or least expensive to implement. Among the alternatives are several closely related techniques that, like microinjection, rely on the cell's ability to reseal a mechanically induced plasma membrane disruption created in order to gain temporary access to cell cytosol. Four such techniques are described in this unit: scrape loading, scratch loading, bead loading, and syringe loading. Unlike microinjection, these techniques allow one to rapidly load (in a matter of minutes) thousands or even many millions of many types of mammalian cells with normally impermeant molecules, and so to facilitate quantitative analyses of the effect of loading.
PLOS Biology, Aug 22, 2006
When a cell suffers a plasma membrane disruption, extracellular Ca 2þ rapidly diffuses into its c... more When a cell suffers a plasma membrane disruption, extracellular Ca 2þ rapidly diffuses into its cytosol, triggering there local homotypic and exocytotic membrane fusion events. One role of this emergency exocytotic response is to promote cell survival: the internal membrane thus added to the plasma membrane acts as a reparative ''patch.'' Another, unexplored consequence of disruption-induced exocytosis is secretion. Many of the cells lining the gastrointestinal tract secrete mucus via a compound exocytotic mechanism, and these and other epithelial cell types lining the digestive tract are normally subject to plasma membrane disruption injury in vivo. Here we show that plasma membrane disruption triggers a potent mucus secretory response from stomach mucous cells wounded in vitro by shear stress or by laser irradiation. This disruption-induced secretory response is Ca 2þ dependent, and coupled to cell resealing: disruption in the absence of Ca 2þ does not trigger mucus release, but results instead in cell death due to failure to reseal. Ca 2þ-dependent, disruption-induced mucus secretion and resealing were also demonstrable in segments of intact rat large intestine. We propose that, in addition to promoting cell survival of membrane disruptions, disruption-induced exocytosis serves also the important protective function of liberating lubricating mucus at sites of mechanical wear and tear. This mode of mechanotransduction can, we propose, explain how lubrication in the gastrointestinal tract is rapidly and precisely adjusted to widely fluctuating, diet-dependent levels of mechanical stress.
Journal of Cellular Physiology, Jul 1, 1991
Cells of gut and skin frequently suffer mechanically-induced plasma membrane disruptions in vivo,... more Cells of gut and skin frequently suffer mechanically-induced plasma membrane disruptions in vivo, and bioactive molecules, including basic fibroblast growth factor (bFGF), could enter and leave cytoplasm through these disruptions. We here provide three lines of evidence that bFGF is released with surprising efficiency through plasma membrane disruptions, resembling those known to occur in vivo, produced by scraping endothelial cells from their culturing substratum. First, 41 O h of the total of bFGF extractable in 1 M NaCl by freeze-thaw and sonication was released simply by scraping the endothelial cells. Second, relative to release of lactate dehydrogenase, cells wounded by scraping under conditions promoting >60% cell survival released a significantly larger amount (up to twofold more) of growth promoting activity than did cells uniformly killed and irreversibly permeabilized by scraping in the cold or by freezing and thawing. Last, cells that survived membrane disruptions released, and contained, less bFGF on each subsequent wounding, consistent with release of bFCF through transient (i.e., survivable) membrane disruptions. A polyclonal antibody against bFGF completely neutralized the growth promoting activity released by scraping, confirming that bFGF is released through endothelial cell plasma membrane disruptions. Cell fractionation and immunolocalization, including a novel per
Journal of Surgical Research, Feb 1, 2006
Introduction: Burn injury results in early T cell apoptosis, lymphopenia and associated T cell dy... more Introduction: Burn injury results in early T cell apoptosis, lymphopenia and associated T cell dysfunction. Subsequent reconstitution of the T cell compartment late after burn injury via homeostatic proliferation generates hyper-responsive memory CD8ϩ T cells of unclear functional significance. In this study, we hypothesized that the hyper-responsive CD8ϩ population isolated late after burn injury 1) has enhanced activity in vivo and 2) that burn injury associated apoptosis is necessary for the generation and hyperresponsiveness of this unique T cell population. Methods: To test these hypotheses, we used a mouse model of allogeneic skin graft rejection to assess in vivo activity and the glucocortocoid receptor inhibitor mifepristone (RU486) to block burn associated apoptosis. Female C57Bl/6 (B6) mice transgenic for the T cell receptor recognizing the male minor histocompatibility antigen HY (HY TCR mice) were anesthetized and treated as follows (nϭ6 per group): 1) 20% TBSA burn; 2) 20% TBSA burn and 3 daily injections (20g/g) of RU486; 3) sham and 4) sham with RU486. Mice were sacrificed 14 days after burn injury, splenic CD8ϩ T cells purified (Ͼ95% purity) by magnetic cell separation and CD8ϩ T cells from each group were adoptively transferred i/v into female wildtype B6 mice (1ϫ106 cells / mouse, nϭ6 per group). Transferred cells were allowed to redistribute for 48 hours, then each mouse received allogeneic male (HYϩ) skin and control isogeneic female (HYϪ) tail skin grafts. Grafts were monitored daily, scored for rejection and median survival times of allogeneic male grafts were calculated. Statistical analysis was performed by logrank test with significance defined as pϽ0.05. Results: Mice that received CD8ϩ HY TCR T cells isolated late after burn injury rejected male skin graft significantly faster than mice that received CD8ϩ T cells from sham mice (11 days versus 15 days, pϭ0.038), demonstrating that transferred cells from burn mice have enhanced activity in vivo. This effect was blocked when RU486 was administered to burn mice and subsequent skin graft rejection in adoptive transfer recipients was compared to the burn alone recipients (19 days versus 11 days, pϭ0.0013). Administration of RU486 to burn mice also resulted in similar graft rejection when compared to sham (19 days versus 15 days, pϭ0.30), sham with RU486 (19 days versus 15 days, pϭ0.55) and no cells (19 days versus 17 days, pϭ0.75) adoptive transfer recipients. Conclusions: This study demonstrates for the first time the functional significance in vivo of hyper-responsive CD8ϩ T cells late after burn injury. While blocking apoptosis may have beneficial effects on immune function early after burn injury it also appears to prevent the development of these
... [Art/Design Item] [Creative Arts and Design > Graphic Design]. Details. Creators: Mc N... more ... [Art/Design Item] [Creative Arts and Design > Graphic Design]. Details. Creators: Mc Neil, Paul. Description/Abstract: Design of printed promotional material commissioned by the Energy Saving Trust, 2002. This design research ...
Journal of Surgical Research, Feb 1, 2006
Introduction: In the developmental stage, pancreas derives from the endodermal cells where the tr... more Introduction: In the developmental stage, pancreas derives from the endodermal cells where the transcription factor, pancreatic duodenal homeobox gene-1 (pdx-1) is expressed. In the adulthood, pdx-1 expression is limited to pancreatic  cells and pdx-1 is necessary for maintaining the function. Recent studies have shown that ectopic expression of pdx-1 like liver successfully induced insulin production and ameliorated hyperglycemia. The purpose of this study was to investigate the effects of enforced expression by the adenovirus-mediated gene transfer of pdx-1 in the jejunal epithelia where pdx-1 is not expressed. Methods: The recombinant replication-deficient adenovirus carrying pdx-1 gene was constructed using the COS-TPC method. ICR mice were treated with STZ (220 mg/kg i.p.) After determining the hyperglycemia, a loop-jejunostomy was constructed and the adenovirus solutions (Ad-pdx-1 and Ad-lacZ 1ϫ108 PFU/body) were injected into the lumen of the loop. (1) Blood glucose concentrations were measured by drawing blood from ocular veins. (2) Immunohistochemical or fluorescent analyses of PDX-1 and insulin in the adenovirus-infected jejunal epithelia were performed. (3) RT-PCR of pdx-1 and other pancreatic markers were investigated. (4) Immunoreactinve insulin (IRI) extracted from the adenovirus-infected jejunum were measured. Results: (1) Hyperglycemia was improved in STZ-treated mice infected with Ad-pdx-1. (2) Ad-pdx-1 extensively induced ectopic PDX-1 expression in the jejunum and the scattered cells in the jejunal epithelia were positive for insulin expression. (3) mRNA of insulin Ϫ1, Ϫ2 and pdx-1 were expressed in mice with Ad-pdx-1. (4) IRI level of the jejunum extract was significantly increased in mice with Ad-pdx-1. Conclusions: Gene transfer of pdx-1 in intestinal epithelia could be a promising strategy for diabetes mellitus by inducing ectopic insulin producing cells. 244. THE PERITONEAL CAVITY IS A DISTINCT COMPARTMENT OF ANGIOGENIC MOLECULAR MEDIATORS.
Agro Food Industry Hi-tech, Jul 1, 2012
Research, design and production of Generator typeface and poster on behalf of Dr Charley Peters a... more Research, design and production of Generator typeface and poster on behalf of Dr Charley Peters and Saturation Point Projects — an LCC funded research project. Generator, curated by Saturation Point Projects (saturationpoint.org.uk), was an exhibition exploring the language of generative art that took place at the Kaleidoscope Gallery in 2015. Artworks on display, by Katrina Blannin, Christina France, Hanz Hancock, James Irwin, Patrick Morrissey, Andy Parkinson, Charley Peters and Mary Yacoob, employed analogue systems to define visual outputs. MuirMcNeil were invited to design and produce a poster to accompany the event. Featuring a text by Laura Davidson, the 70x50cm poster is printed in orange and violet neon inks in two bespoke typefaces generated exclusively for the project.
Inclusion of ThreeSix Typeface and Unit Editions U:D/R03 journal in Wim Crouwel exhibition at Des... more Inclusion of ThreeSix Typeface and Unit Editions U:D/R03 journal in Wim Crouwel exhibition at Design Museum, London, 30 March – 03 July 2011.
A special edition of Unit Editions design research journal, concerning the ThreeSix optical/geome... more A special edition of Unit Editions design research journal, concerning the ThreeSix optical/geometric typeface system developed by MuirMcNeil. Written and designed by Paul McNeil and Hamish Muir, with an introduction by Wim Crouwel, U:D/R 03 examines ThreeSix in relation to its historical, ideological and typographic contexts. The publication also describes the design process in detail. This edition was produced in newsprint, single colour litho printing.
Contribution of article to publication and limited edition poster to Team Impression’s X Ten proj... more Contribution of article to publication and limited edition poster to Team Impression’s X Ten project.