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Biomedical subjects

S J Neill

Publications and source records attributed to S J Neill.

At least 19 recordsLinked to original sources
PubMed2001

Does the redox status of cytochrome C act as a fail-safe mechanism in the regulation of programmed cell death?

It has now become recognized that one of the key events in the induction of apoptosis, or programmed cell death, in both plants and animals is the release of cytochrome c from mitochondria. It is also known that oxidative stress imposed on cells can have a profound effect on the onset or progression of apoptosis. Here, we discuss how the redox status of cytochrome c, and thus its structure, can be altered by the presence of reactive oxygen species (ROS) and reduced glutathione (GSH). We suggest that cytochrome c will only induce programmed cell death if present in the cytoplasm in the oxidized state, and that the presence of high levels of cytoplasmic GSH maintain cytochrome c in an inactive (reduced) state, thus behaving as a fail-safe mechanism if cytochrome c is released by mitochondria when programmed cell death is not the required outcome. If the redox status of the cell is disturbed however, perhaps in the presence of hydrogen peroxide, GSH concentrations will drop, the cellular E(h) will rise, and cytochrome c will tend towards the oxidized state, allowing programmed cell death to proceed. Therefore, we propose that the redox state of cytoplasmic cytochrome c may be a key regulator of programmed cell death.

Animals↗
PubMed2001

Role of reactive oxygen species in cell signalling pathways.

Reactive oxygen species (ROS) were originally thought to only be released by phagocytic cells during their role in host defence. It is now clear that ROS have a cell signalling role in many biological systems, both in animals and in plants. ROS induce programmed cell death or necrosis, induce or suppress the expression of many genes, and activate cell signalling cascades, such as those involving mitogen-activated protein kinases.

Animals↗
PubMed2001

Harpin induces activation of the Arabidopsis mitogen-activated protein kinases AtMPK4 and AtMPK6.

Mitogen-activated protein kinases (MAPKs) are key enzymes that mediate adaptive responses to various abiotic and biotic stresses, including pathogen challenge. The proteinaceous bacterial elicitor harpin (secreted by Pseudomonas syringae pv syringae) activates two MAPKs in suspension cultures of Arabidopsis var. Landsberg erecta. In this study, we show that harpin and exogenous hydrogen peroxide (H(2)O(2)) activate myelin basic protein kinases in Arabidopsis leaves. Using anti-AtMPK4 and anti-AtMPK6 antibodies, we identify the harpin-activated MAPKs in both leaves and suspension cultures as AtMPK4 and AtMPK6, and show that H(2)O(2), generated by Arabidopsis cells in response to challenge with harpin, activates only AtMPK6. However, treatments with catalase, which removes H(2)O(2), or diphenylene iodonium, which inhibits superoxide and H(2)O(2) production, do not inhibit harpin-induced activation of AtMPK4 or AtMPK6. In addition, activation of AtMPK4 but not AtMPK6 is inhibited by the MAPK kinase inhibitor PD98059. Neither harpin nor H(2)O(2) has any effect on AtMPK4 or AtMPK6 gene expression. In addition, the expression of AtMEKK1, AtMEK1, or AtMKK2, previously shown to be potential functional partners of AtMPK4, were not affected by either harpin or H(2)O(2) treatments. These data suggest that harpin activates several signaling pathways, one leading to stimulation of the oxidative burst and others leading to the activation of AtMPK4 or AtMPK6.

Arabidopsis↗
PubMed2001

Regulation of the Arabidopsis transcriptome by oxidative stress.

Oxidative stress, resulting from an imbalance in the accumulation and removal of reactive oxygen species such as hydrogen peroxide (H(2)O(2)), is a challenge faced by all aerobic organisms. In plants, exposure to various abiotic and biotic stresses results in accumulation of H(2)O(2) and oxidative stress. Increasing evidence indicates that H(2)O(2) functions as a stress signal in plants, mediating adaptive responses to various stresses. To analyze cellular responses to H(2)O(2), we have undertaken a large-scale analysis of the Arabidopsis transcriptome during oxidative stress. Using cDNA microarray technology, we identified 175 non-redundant expressed sequence tags that are regulated by H(2)O(2). Of these, 113 are induced and 62 are repressed by H(2)O(2). A substantial proportion of these expressed sequence tags have predicted functions in cell rescue and defense processes. RNA-blot analyses of selected genes were used to verify the microarray data and extend them to demonstrate that other stresses such as wilting, UV irradiation, and elicitor challenge also induce the expression of many of these genes, both independently of, and, in some cases, via H(2)O(2).

Adaptation, Physiological↗
PubMed2000

Hydrogen peroxide-induced gene expression in Arabidopsis thaliana.

Hydrogen peroxide (H(2)O(2)) is generated in plants after exposure to a variety of biotic and abiotic stresses, and has been shown to induce a number of cellular responses. Previously, we showed that H(2)O(2) generated during plant-elicitor interactions acts as a signaling molecule to induce the expression of defense genes and initiate programmed cell death in Arabidopsis thaliana suspension cultures. Here, we report for the first time the identification by RNA differential display of four genes whose expression is induced by H(2)O(2). These include genes that have sequence homology to previously identified Arabidopsis genes encoding a late embryogenesis-abundant protein, a DNA-damage repair protein, and a serine/threonine kinase. Their putative roles in H(2)O(2)-induced defense responses are discussed.

Amino Acid Sequence↗
PubMed2000

Acute childhood illness at home: the parents' perspective.

This critical review of British literature explores the phenomenon of acute childhood illness at home from the parents' perspective. The Literature was searched using four CD-ROM databases: CINAHL, MEDLINE, ASSIA and PSYCHLIT, augmented by hand searching of current journal issues. Sandelowski's (1995) stages of qualitative data analysis were used to develop a rigorous approach to conducting qualitative overviews. The majority of childhood illness is of short duration and takes place in the child's own home. Yet the research which addresses this area is limited. Mothers constantly monitor their child's health, identifying illness as a change in behaviour of the child from their perception of normal. Their usual response to illness is nonprofessional care, including general nursing care and 'over the counter' medicines. When mothers do seek help this is usually from the family doctor. Mothers are often dissatisfied with the help and advice they receive. This is partially explained by the mismatches which occur concerning the perceptions of mothers and of health care professionals regarding appropriate use of services and the needs of these mothers and their children. The literature reviewed indicates a need to address the service users perceptions of the services provided. Areas where further research is required are identified.

Acute Disease↗
PubMed2000

NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures.

Recent research has implicated nitric oxide (NO) in the induction of the hypersensitive response (HR) during plant-pathogen interactions. Here we demonstrate that Arabidopsis suspension cultures generate elevated levels of NO in response to challenge by avirulent bacteria, and, using NO donors, show that these elevated levels of NO are sufficient to induce cell death in Arabidopsis cells independently of reactive oxygen species (ROS). We also provide evidence that NO-induced cell death is a form of programmed cell death (PCD), requiring gene expression, and has a number of characteristics of PCD of mammalian cells: NO induced chromatin condensation and caspase-like activity in Arabidopsis cells, while the caspase-1 inhibitor, Ac-YVAD-CMK, blocked NO-induced cell death. A well-established second messenger mediating NO responses in mammalian cells is cGMP, produced by the enzyme guanylate cyclase. A specific inhibitor of guanylate cyclase blocked NO-induced cell death in Arabidopsis cells, and this inhibition was reversed by the cell-permeable cGMP analogue, 8Br-cGMP, although 8Br-cGMP alone did not induce cell death or potentiate NO-induced cell death. This suggests that cGMP synthesis is required but not sufficient for NO-induced cell death in Arabidopsis. In-gel protein kinase assays showed that NO activates a potential mitogen-activated protein kinase (MAPK), although a specific inhibitor of mammalian MAPK activation, PD98059, which blocked H2O2-induced cell death, did not inhibit the effects of NO.

Amino Acid Chloromethyl Ketones↗
PubMed2000

ABA activation of an MBP kinase in Pisum sativum epidermal peels correlates with stomatal responses to ABA.

In-gel protein kinase assays using myelin basic protein (MBP) as substrate have been used to demonstrate that abscisic acid (ABA) activates an MBP kinase (AMBP kinase) in epidermal peels prepared from leaves of the Argenteum mutant of pea, Pisum sativum L. AMBP kinase has the characteristics of a mitogen-activated protein kinase (MAPK): it utilizes MBP preferentially as an artificial substrate, it is rapidly and transiently activated, it is of the appropriate size (molecular weight c. 45 kDa), requires tyrosine phosphorylation for activity and is tyrosine phosphorylated upon activation. Reverse transcription-PCR was used to generate a previously-cloned MAPK from guard cells, epidermis and mesophyll and immunoblotting using an antibody raised against a mammalian MAPK detected MAPK-related proteins, including one of 45 kDa, in epidermal peels, mesophyll and guard cells. Inhibition of AMBP kinase activation by PD98059, a specific inhibitor of MAPK kinase, and thus MAPK activation, correlated with PD98059-inhibition of ABA-induced stomatal closure and dehydrin gene expression, suggesting that ABA effects in pea epidermal peels require MAPK activation. AMBP kinase was not activated by ABA in guard cells isolated by enzyme treatment. However, a protein kinase of c. 43 kDa was activated by ABA in isolated guard cells, but not in mesophyll or epidermal tissue.

Abscisic Acid↗
PubMed1999

Flow cytometry and surface plasmon resonance analyses demonstrate that the monoclonal antibody JIM19 interacts with a rice cell surface component involved in abscisic acid signalling in protoplasts.

Abscisic acid (ABA) is a plant hormone involved in many developmental and physiological processes, but as yet, no ABA receptor has been identified. Flow cytometry of rice protoplasts and immunoblotting of purified plasma membranes (PMs) have been used to demonstrate that the monoclonal antibody JIM19 recognizes carbohydrate epitopes of cell surface glycoproteins. Using surface plasmon resonance technology specific binding of PMs to JIM19 was observed. Such interaction was antagonized significantly by ABA, but not by the biologically inactive ABA catabolite phaseic acid. These in vitro interactions were correlated with the biological activities of JIM19, ABA and phaseic acid on activation of the ABA-inducible Em promoter using two different transient reporter gene assays, beta-glucuronidase/luciferase and quantitative flow cytometry of Aequoria green fluorescent protein. Pre-treatment with JIM19 resulted in significant inhibition of ABA-inducible gene expression. Taken together, these data suggest that JIM19 interacts with a functional PM complex involved in ABA signalling.

Abscisic Acid↗
PubMed1999

Harpin induces mitogen-activated protein kinase activity during defence responses in Arabidopsis thaliana suspension cultures.

Elicitation of Arabidopsis thaliana (L.) Heynh. suspension cultures with the bacterial protein harpin (from Pseudomonas syringae pv. syringae) induced the activation of two kinases of 39 and 44 kDa, as demonstrated by in-gel kinase assays using myelin basic protein (MBP) as a substrate. Both these kinases appeared to be tyrosine-phosphorylated upon activation, as demonstrated by treatment with tyrosine phosphatase and immunoprecipitation using an anti-phosphotyrosine monoclonal antibody. An inhibitor of mammalian mitogen-activated protein kinase (MAPK) activation, PD98059, inhibited harpin-induced MBPK activation, but did not inhibit the activity of these kinases. PD98059 also inhibited harpin-induced programmed cell death and defence gene expression, suggesting the involvement of harpin-induced MAPKs in defence responses in Arabidopsis thaliana.

Apoptosis↗
PubMed1998

Harpin and hydrogen peroxide both initiate programmed cell death but have differential effects on defence gene expression in Arabidopsis suspension cultures.

Programmed cell death is increasingly viewed as a key component of the hypersensitive disease resistance response of plants. The generation of reactive oxygen species (ROS) such as H2O2 triggers a cell death programme in Arabidopsis suspension cultures following challenge with the bacterial elicitor harpin. Both harpin and exogenous H2O2 initiate a cell death pathway that requires gene expression, and also act as signalling molecules to induce the expression of plant defence genes encoding enzymes such as phenylalanine ammonia-lyase (PAL), glutathione S-transferase (GST) and anthranilate synthase (ASA1), an enzyme of phytoalexin biosynthesis in Arabidopsis. H2O2 induces the expression of PAL1 and GST but not that of ASA1. Harpin initiates two signalling pathways, one leading to increased ROS generation and expression of PAL1 and GST mRNA, and another leading to increased GST and ASA1 expression, independent of H2O2.

Anthranilate Synthase↗
PubMed1998

Developing children's nursing through action research.

Action research provides an ideal strategy for the integration of theory and practice, as it focuses on the development of theory in practice. Action research is gaining in popularity and acceptance. The collaborative approaches of action research parallel the dominant philosophies of participation and partnership in contemporary children's nursing. Taking the lead in identifying areas of practice for action research is a natural extension of the children's nurse's role as advocate for the child and family. The challenge for the future is to expand the use of action research in children's nursing and to include service users--the children and their families--in the research teams.

Child↗
PubMed1997

Educating the new community children's nurses: challenges and opportunities?

This paper aims to stimulate debate around the issue of educating nurses within the new Specialist Practitioner courses in areas where no Community Children's Nursing services currently exist. The United Kingdom Central Council (UKCC) (1994) have provided the impetus and the educational structure for the development of community services for children through the publication of learning outcomes for the new, long awaited, Community Children's Nursing qualification. In outlining learning outcomes for this and the other Specialist Community Nursing Practice courses, the UKCC is clearly communicating that the care of sick children in the community should be the province of Community Children's Nurses and not held to be within the remit of other community nurses. Consequently, areas which do not currently offer a Community Children's Nursing service to these children and their families are presented with several challenges: educating nurses for these new roles; developing a new service; and redefining the roles of practitioners currently caring for these children in the community. This paper addresses the first of these challenges and identifies three factors influencing the education of these new practitioners: funding, supervision and placements. A strategic model for the support of these new students in areas with no existing service is proposed.

Community Health Nursing↗
PubMed1996

Generation of active oxygen in elicited cells of Arabidopsis thaliana is mediated by a NADPH oxidase-like enzyme.

Suspension-cultured cells of Arabidopsis thaliana generated active oxygen species (AOS) (measured by luminol-dependent chemiluminescence) following challenge with the bacterial protein elicitor harpin or the protein kinase activator phorbol 12-myristate 13-acetate. These responses were blocked by inhibitors of superoxide dismutase (SOD), NADPH oxidase and protein kinase. Harpin treatment also resulted in an increase in cell death, a response reduced by inhibitors of AOS generation or AOS scavengers. Extracellular SOD activity was found to be present in cell culture medium. Immunoblotting of Arabidopsis extracts revealed the presence of proteins immunologically related to the human neutrophil NADPH oxidase complex, and cell-free reconstitution assays showed that human neutrophil cytosol combined with Arabidopsis membranes could initiate superoxide generation. These data suggest that the enzyme catalysing the generation of superoxide in elicited Arabidopsis cells is similar to the mammalian NADPH oxidase and that a signalling cascade leading to AOS generation involves protein phosphorylation.

Arabidopsis↗
PubMed1983

Biosynthesis of abscissic acid.

The fungus Cercospora rosicola has been studied as a model system for abscissic acid biosynthesis. 1'-dDeoxyabscissic acid and 4'-hydroxy-a-ionylidene acetic acid have been identified as endogenous compounds in this fungus. The results of feeding these and other putative intermediates suggest that abscissic acid biosynthesis proceeds via the successive oxidations of a 3-methyl-5-(2',6',6'-trimethylcyclohex-2'-en-1'-yl)-2,4-pentadienyl intermediate. Preliminary results suggest that a similar pathway may operate in plants.

Abscisic Acid↗
PubMedDate not supplied

Parent participation. 1: Literature review and methodology.

A qualitative exploratory study was undertaken to examine parents' views and experiences of participation in the care of their child in hospital and to identify factors that inhibit or facilitate participation. Parents of children aged 2-5 years of age were interviewed within 2 months of discharge, using tape-recorded, semi-structured interviews. The findings demonstrate that parents clearly wish to participate in their children's care at a level of their own choosing. However, they preferred professionals to be responsible for their children's clinical care, while they continue to be responsible for the children's normal day-to-day care. Problems were identified in the area of relationships between parents and professionals, centred on communication and the continuing paternalistic nature of these relationships. Parents of children who experienced single, short hospital admissions found involvement in their children's care particularly difficult. Guidelines were developed from the results, to facilitate parent participation in the care of children in this age group.

Child↗