Quantitative Proteomics and Phosphoproteomics Support a Role for Mut9-Like Kinases in Multiple Metabolic and Signaling Pathways in Arabidopsis.

Protein phosphorylation is one of the most prevalent posttranslational modifications found in eukaryotic systems. It serves as a key molecular mechanism that regulates protein function in response to environmental stimuli. The Mut9-like kinases (MLKs) are a plant-specific family of Ser/Thr kinases linked to light, circadian, and abiotic stress signaling. Here we use quantitative phosphoproteomics in conjunction with global proteomic analysis to explore the role of the MLKs in daily protein dynamics. Proteins involved in light, circadian, and hormone signaling, as well as several chromatin-modifying enzymes and DNA damage response factors, were found to have altered phosphorylation profiles in the absence of MLK family kinases. In addition to altered phosphorylation levels, mlk mutant seedlings have an increase in glucosinolate metabolism enzymes. Subsequently, we show that a functional consequence of the changes to the proteome and phosphoproteome in mlk mutant plants is elevated glucosinolate accumulation and increased sensitivity to DNA damaging agents. Combined with previous reports, this work supports the involvement of MLKs in a diverse set of stress responses and developmental processes, suggesting that the MLKs serve as key regulators linking environmental inputs to developmental outputs.

Plastid osmotic stress influences cell differentiation at the plant shoot apex.

The balance between proliferation and differentiation in the plant shoot apical meristem is controlled by regulatory loops involving the phytohormone cytokinin and stem cell identity genes. Concurrently, cellular differentiation in the developing shoot is coordinated with the environmental and developmental status of plastids within those cells. Here, we employ an Arabidopsis thaliana mutant exhibiting constitutive plastid osmotic stress to investigate the molecular and genetic pathways connecting plastid osmotic stress with cell differentiation at the shoot apex. msl2 msl3 mutants exhibit dramatically enlarged and deformed plastids in the shoot apical meristem, and develop a mass of callus tissue at the shoot apex. Callus production in this mutant requires the cytokinin receptor AHK2 and is characterized by increased cytokinin levels, downregulation of cytokinin signaling inhibitors ARR7 and ARR15, and induction of the stem cell identity gene WUSCHEL Furthermore, plastid stress-induced apical callus production requires elevated plastidic reactive oxygen species, ABA biosynthesis, the retrograde signaling protein GUN1, and ABI4. These results are consistent with a model wherein the cytokinin/WUS pathway and retrograde signaling control cell differentiation at the shoot apex.

MSL1 is a mechanosensitive ion channel that dissipates mitochondrial membrane potential and maintains redox homeostasis in mitochondria during abiotic stress.

Mitochondria must maintain tight control over the electrochemical gradient across their inner membrane to allow ATP synthesis while maintaining a redox-balanced electron transport chain and avoiding excessive reactive oxygen species production. However, there is a scarcity of knowledge about the ion transporters in the inner mitochondrial membrane that contribute to control of membrane potential. We show that loss of MSL1, a member of a family of mechanosensitive ion channels related to the bacterial channel MscS, leads to increased membrane potential of Arabidopsis mitochondria under specific bioenergetic states. We demonstrate that MSL1 localises to the inner mitochondrial membrane. When expressed in Escherichia coli, MSL1 forms a stretch-activated ion channel with a slight preference for anions and provides protection against hypo-osmotic shock. In contrast, loss of MSL1 in Arabidopsis did not prevent swelling of isolated mitochondria in hypo-osmotic conditions. Instead, our data suggest that ion transport by MSL1 leads to dissipation of mitochondrial membrane potential when it becomes too high. The importance of MSL1 function was demonstrated by the observation of a higher oxidation state of the mitochondrial glutathione pool in msl1-1 mutants under moderate heat- and heavy-metal-stress. Furthermore, we show that MSL1 function is not directly implicated in mitochondrial membrane potential pulsing, but is complementary and appears to be important under similar conditions.

Plastid osmotic stress activates cellular stress responses in Arabidopsis.

Little is known about cytoplasmic osmoregulatory mechanisms in plants, and even less is understood about how the osmotic properties of the cytoplasm and organelles are coordinately regulated. We have previously shown that Arabidopsis (Arabidopsis thaliana) plants lacking functional versions of the plastid-localized mechanosensitive ion channels Mechanosensitive Channel of Small Conductance-Like2 (MSL2) and MSL3 contain leaf epidermal plastids under hypoosmotic stress, even during normal growth and development. Here, we use the msl2 msl3 mutant as a model to investigate the cellular response to constitutive plastid osmotic stress. Under unstressed conditions, msl2 msl3 seedlings exhibited several hallmarks of drought or environmental osmotic stress, including solute accumulation, elevated levels of the compatible osmolyte proline (Pro), and accumulation of the stress hormone abscisic acid (ABA). Furthermore, msl2 msl3 mutants expressed Pro and ABA metabolism genes in a pattern normally seen under drought or osmotic stress. Pro accumulation in the msl2 msl3 mutant was suppressed by conditions that reduce plastid osmotic stress or inhibition of ABA biosynthesis. Finally, treatment of unstressed msl2 msl3 plants with exogenous ABA elicited a much greater Pro accumulation response than in the wild type, similar to that observed in plants under drought or osmotic stress. These results suggest that osmotic imbalance across the plastid envelope can elicit a response similar to that elicited by osmotic imbalance across the plasma membrane and provide evidence for the integration of the osmotic state of an organelle into that of the cell in which it resides.

Autonomy in place of birth: a concept analysis.

This article examines one of the relevant concepts in the current debate on home birth-autonomy in place of birth-and its uses in general language, ethics, and childbirth health care literature. International discussion on childbirth services. A concept analysis guided by the model of Walker and Avant. The authors suggest that autonomy in the context of choosing place of birth is defined by three main attributes: information, capacity and freedom; given the antecedent of not harming others, and the consequences of accountability for the outcome. Model, borderline and contrary cases of autonomy in place of birth are presented. A woman choosing place of birth is autonomous if she receives all relevant information on available choices, risks and benefits, is capable of understanding and processing the information and choosing place of birth in the absence of coercion, provided she intends no harm to others and is accountable for the outcome. The attributes of the definition can serve as a useful tool for pregnant women, midwives, and other health professionals in contemplating their moral status and discussing place of birth.

Two mechanosensitive channel homologs influence division ring placement in Arabidopsis chloroplasts.

Chloroplasts must divide repeatedly to maintain their population during plant growth and development. A number of proteins required for chloroplast division have been identified, and the functional relationships between them are beginning to be elucidated. In both chloroplasts and bacteria, the future site of division is specified by placement of the Filamentous temperature sensitive Z (FtsZ) ring, and the Min system serves to restrict FtsZ ring formation to mid-chloroplast or mid-cell. How the Min system is regulated in response to environmental and developmental factors is largely unstudied. Here, we investigated the role in chloroplast division played by two Arabidopsis thaliana homologs of the bacterial mechanosensitive (MS) channel MscS: MscS-Like 2 (MSL2) and MSL3. Immunofluorescence microscopy and live imaging approaches demonstrated that msl2 msl3 double mutants have enlarged chloroplasts containing multiple FtsZ rings. Genetic analyses indicate that MSL2, MSL3, and components of the Min system function in the same pathway to regulate chloroplast size and FtsZ ring formation. In addition, an Escherichia coli strain lacking MS channels also showed aberrant FtsZ ring assembly. These results establish MS channels as components of the chloroplast division machinery and suggest that their role is evolutionarily conserved.

A randomized controlled trial of the effectiveness of educating parents about distraction to decrease postoperative pain in children at home after tonsillectomy.

Today, parents are more involved with postoperative pain management, because children are discharged as early as possible after surgery. The purpose of this randomized controlled trial was to determine the effectiveness of educating parents to provide distraction in addition to pharmacologic pain management in decreasing postoperative pain at home for children ages 3 to 7 years. Ninety-three children aged 3-7 years having tonsillectomy at Landspitali, The National University Hospital of Iceland, and their parents were randomized to one of two intervention groups. The interventions were pharmacologic pain management education (control group) and pharmacologic pain management education with distraction (experimental group). Pain was measured by parent's report of pain behavior during the evening on postoperative days 1 and 2 and child's report of pain intensity on eight time points on postoperative days 1 and 2. Sixty-nine children had complete data. The results from RM-ANCOVA showed that when adjusting for pain intensity in the morning on postoperative day 1 that educating parents about distraction in addition to educating them about pain medication management decreases mean pain behavior scores (p < .001). There was no difference in pain intensity between the groups using RM-ANCOVA. However, there was association between pain intensity in the morning on postoperative day 1 and follow-up pain intensity (p < .001). The results from this study support the importance of educating parents of children having tonsillectomy about distraction in addition to educating them about pain medication management.

MscS-like mechanosensitive channels in plants and microbes.

The challenge of osmotic stress is something all living organisms must face as a result of environmental dynamics. Over the past three decades, innovative research and cooperation across disciplines have irrefutably established that cells utilize mechanically gated ion channels to release osmolytes and prevent cell lysis during hypoosmotic stress. Early electrophysiological analysis of the inner membrane of Escherichia coli identified the presence of three distinct mechanosensitive activities. The subsequent discoveries of the genes responsible for two of these activities, the mechanosensitive channels of large (MscL) and small (MscS) conductance, led to the identification of two diverse families of mechanosensitive channels. The latter of these two families, the MscS family, consists of members from bacteria, archaea, fungi, and plants. Genetic and electrophysiological analysis of these family members has provided insight into how organisms use mechanosensitive channels for osmotic regulation in response to changing environmental and developmental circumstances. Furthermore, determining the crystal structure of E. coli MscS and several homologues in several conformational states has contributed to our understanding of the gating mechanisms of these channels. Here we summarize our current knowledge of MscS homologues from all three domains of life and address their structure, proposed physiological functions, electrophysiological behaviors, and topological diversity.

Relations between feeding intolerance and stress biomarkers in preterm infants.

OBJECTIVES: Feeding intolerance (FI) in preterm infants is common but the etiology remains unclear. This study examined FI as a stress-related disease involving brain-gut interactions and tested the model of allostatic load and complications of prematurity. Specific aims were to describe demographic/medical variables and biomarker levels at each time and over time for the sample; describe/compare variables and biomarker levels at each time for infants with/without FI; and compare biomarker interquartile/interpercentile distributions between infants with/without FI. METHODS: Preterm infants <32 weeks' gestation were recruited. The primary outcome was FI by day 7 defined as a feeding withheld, discontinued, or decreased because the infant was not tolerating enteral feedings. Allostatic load was operationalized using cortisol and 8-hydroxydeoxyguanosine (8-OHdG) from cord blood and from saliva and urine on days 1, 7, and 14. Descriptive statistics and comparative analyses were performed. RESULTS: Seven of 31 infants enrolled met criteria for FI. Infants with FI had lower median urinary cortisol on day 1 (P = 0.007) and trended to have lower cortisol in the cord blood (P = 0.056). Interquartile distributions were significantly different between infants with/without FI for urinary cortisol on day 1 (P = 0.034) and trended for differences in 8-OHdG on day 14 (P = 0.087). Interpercentile distributions were significantly different in salivary cortisol on day 14 (P = 0.034) and trended for differences in 8-OHdG on day 1 (P = 0.079). CONCLUSIONS: Results support further testing of the model in a larger sample; investigation of the cellular mechanisms associated with the stress and the free radical/antioxidant systems; and inclusion of prenatal factors.

Social regulation of the stress response in the transitional newborn: a pilot study.

The purpose of the study was to explore relationships between caregiver holding and feeding behaviors and the transitional newborn infant's cortisol response. Behaviors of 46 mothers, fathers, and their term transitional newborn infants were measured with the Index of Mother-Infant Separation (IMIS). Repeated measures of infant salivary cortisol were used to calculate area under the curve. A higher percentage of observations in which mother was holding infant was related to lower infant total cortisol during the first 6 hours after birth (r = -.24, p = .05, one-tailed).

Feeding intolerance: a concept analysis.

Feeding intolerance is a well-known phenomenon in the NICU and is linked to morbidity and mortality in the premature infant. However, a universal definition for this concept is lacking. Properly defining a concept is a key step in a successful research project. A concept analysis is an examination of the fundamental elements of a concept to bring clarification and definition to the topic of interest. The purpose of this concept analysis was to clarify the phenomenon of feeding intolerance in the premature infant and to provide a universal conceptual and operational definition for researchers and clinicians to use in practice theory.

Genetic and physical interactions between the organellar mechanosensitive ion channel homologs MSL1, MSL2, and MSL3 reveal a role for inter-organellar communication in plant development.

Plant development requires communication on many levels, including between cells and between organelles within a cell. For example, mitochondria and plastids have been proposed to be sensors of environmental stress and to coordinate their responses. Here we present evidence for communication between mitochondria and chloroplasts during leaf and root development, based on genetic and physical interactions between three Mechanosensitive channel of Small conductance-Like (MSL) proteins from Arabidopsis thaliana. MSL proteins are Arabidopsis homologs of the bacterial Mechanosensitive channel of Small conductance (MscS), which relieves cellular osmotic pressure to protect against lysis during hypoosmotic shock. MSL1 localizes to the inner mitochondrial membrane, while MSL2 and MSL3 localize to the inner plastid membrane and are required to maintain plastid osmotic homeostasis during normal growth and development. In this study, we characterized the phenotypic effect of a genetic lesion in MSL1, both in wild type and in msl2 msl3 mutant backgrounds. msl1 single mutants appear wild type for all phenotypes examined. The characteristic leaf rumpling in msl2 msl3 double mutants was exacerbated in the msl1 msl2 msl3 triple mutant. However, the introduction of the msl1 lesion into the msl2 msl3 mutant background suppressed other msl2 msl3 mutant phenotypes, including ectopic callus formation, accumulation of superoxide and hydrogen peroxide in the shoot apical meristem, decreased root length, and reduced number of lateral roots. All these phenotypes could be recovered by molecular complementation with a transgene containing a wild type version of MSL1. In yeast-based interaction studies, MSL1 interacted with itself, but not with MSL2 or MSL3. These results establish that the abnormalities observed in msl2 msl3 double mutants is partially dependent on the presence of functional MSL1 and suggest a possible role for communication between plastid and mitochondria in seedling development.

Quality Pain Management in Adult Hospitalized Patients: A Concept Evaluation.

AIM: To explore the concept of quality pain management (QPM) in adult hospitalized patients. BACKGROUND: Pain is common in hospitalized patients, and pain management remains suboptimal in some settings. DESIGN: A concept evaluation based on Morse et al.'s method. DATA SOURCE: Of more than 5,000 articles found, data were restricted to 37 selected key articles published in peer-reviewed journals. REVIEW METHODS: Data were extracted from the selected articles and then synthesized according to the following: definition, characteristics, boundaries, preconditions, and outcomes. RESULTS: QPM relates to the Structure: organizationally supported evidence-based policies, competent staff, interprofessional and specialized care, and staff accountability; PROCESS: screening, assessment/reassessment and communication of pain and its treatment, patient/family education, individualized evidence-based treatment, embedded in safe, effective, patient-centered, timely, efficient, and equitable services; and OUTCOMES: reduced pain severity and functional interference, decreased prevalence/severity of adverse consequences from pain or pain treatment, and increase in patient satisfaction. CONCLUSIONS: QPM is a multifaceted concept that remains poorly defined in the literature. Studies should aim to develop valid, reliable, and operational measures of the pillars of QPM and to look at the relationship among these factors. Authors need to state how they define and what aspects of QPM they are measuring.

Mothers in the NICU: outsider to partner.

The emerging care delivery model for Neonatal Intensive Care Units (NICU) is family-focused, developmentally supportive care. The purpose of this study was to explore and describe mothers' experience of becoming a mother while their infants were receiving care in the NICU. A qualitative research design was used. Interviews with 15 mothers whose infants were in a Level III NICU were analyzed using Spradley's domain analysis approach. Mothers developed from outsider to engaged parent along four continua: (1) focus: from NICU to baby; (2) ownership: from their baby to my baby; (3) caregiving: from passive to active; and (4) voice: from silence to advocacy. Mothers entered the continua at different points and moved at different rates toward "engaged parenting." The final stage, partnering, required active participation of nurses. Mothers' development evolved in predictable patterns. The results of this study can be considered in implementation and evaluation plans for NICUs moving to family-focused developmental care.

RNA Sequencing Analysis of the msl2msl3, crl, and ggps1 Mutants Indicates that Diverse Sources of Plastid Dysfunction Do Not Alter Leaf Morphology Through a Common Signaling Pathway.

Determining whether individual genes function in the same or in different pathways is an important aspect of genetic analysis. As an alternative to the construction of higher-order mutants, we used contemporary expression profiling methods to perform pathway analysis on several Arabidopsis thaliana mutants, including the mscS-like (msl)2msl3 double mutant. MSL2 and MSL3 are implicated in plastid ion homeostasis, and msl2msl3 double mutants exhibit leaves with a lobed periphery, a rumpled surface, and disturbed mesophyll cell organization. Similar developmental phenotypes are also observed in other mutants with defects in a range of other chloroplast or mitochondrial functions, including biogenesis, gene expression, and metabolism. We wished to test the hypothesis that the common leaf morphology phenotypes of these mutants are the result of a characteristic nuclear expression pattern that is generated in response to organelle dysfunction. RNA-Sequencing was performed on aerial tissue of msl2msl3 geranylgeranyl diphosphate synthase 1 (ggps1), and crumpled leaf (crl) mutants. While large groups of co-expressed genes were identified in pairwise comparisons between genotypes, we were only able to identify a small set of genes that showed similar expression profiles in all three genotypes. Subsequent comparison to the previously published gene expression profiles of two other mutants, yellow variegated 2 (var2) and scabra3 (sca3), failed to reveal a common pattern of gene expression associated with superficially similar leaf morphology defects. Nor did we observe overlap between genes differentially expressed in msl2msl3, crl, and ggps1 and a previously identified retrograde core response module. These data suggest that a common retrograde signaling pathway initiated by organelle dysfunction either does not exist in these mutants or cannot be identified through transcriptomic methods. Instead, the leaf developmental defects observed in these mutants may be achieved through a number of independent pathways.

A new way of thinking about complications of prematurity.

The morbidity and mortality of preterm infants are impacted by their ability to maintain physiologic homeostasis using metabolic, endocrine, and immunologic mechanisms independent of the mother's placenta. Exploring McEwen's allostatic load model in preterm infants provides a new way to understand the altered physiologic processes associated with frequently occurring complications of prematurity such as bronchopulmonary dysplasia, intraventricular hemorrhage, necrotizing enterocolitis, and retinopathy of prematurity. The purpose of this article is to present a new model to enhance understanding of the altered physiologic processes associated with complications of prematurity. The model of allostatic load and complications of prematurity was derived to explore the relationship between general stress of prematurity and complications of prematurity. The proposed model uses the concepts of general stress of prematurity, allostasis, physiologic response patterns (adaptive-maladaptive), allostatic load, and complications of prematurity. These concepts are defined and theoretical relationships in the proposed model are interpreted using the four maladaptive response patterns of repeated hits, lack of adaptation, prolonged response, and inadequate response. Empirical evidence for cortisol, inflammation, and oxidative stress responses are used to support the theoretical relationships. The proposed model provides a new way of thinking about physiologic dysregulation in preterm infants. The ability to describe and understand complex physiologic mechanisms involved in complications of prematurity is essential for research. Advancing the knowledge of complications of prematurity will advance clinical practice and research and lead to testing of interventions to reduce negative outcomes in preterm infants.

Recognizing new perspectives in eating difficulties following stroke: a concept analysis.

PURPOSE: To describe and identify various perspectives on eating difficulties following stroke to enable theory development and facilitate advancement of interventions. METHODS: Concept analysis in line with Morse's principles of exploring pragmatic utility was used throughout the data collection and analysis. Peer-reviewed research on eating difficulties was systematically and critically appraised. Literature included the bibliographic databases, Medline, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Scopus and PsycInfo, published up until November 2011. A total of 33 key articles were included in the final analysis. RESULTS: A conceptual model of eating difficulties is presented showing the multiple affecting factors, namely meaning-related, functional and contextual factors and the consequences of non-intervention, feeling of loss, social isolation, negative sensations and depression - all of which potentially increase the impact of disease severity, functional outcome and quality of life. Based upon the dimensions identified, the definition of eating difficulties was synthesized as "any activity and emotional requirement and relations, which alone or in combination interfere with the process of preparing food, transferring food into the mouth, chewing and swallowing". CONCLUSION: Eating difficulties have profound effects on people. The conceptual model serves to guide health care professionals to assess and help stroke patients in facing a life with eating difficulties.

Baby lost and found: mothers' experiences of infants who cry persistently.

INTRODUCTION: The purpose of this study was to describe mothers' experiences of parenting an irritable infant. Although "colic" is regarded as a "self-limiting" condition that usually disappears by 3 to 4 months of age, the entire family is affected by the infant's crying; no definitive cause or cure has been identified. SAMPLE AND SETTING: Twelve middle-class married mothers (mean age=27.6 years) of irritable infants were interviewed. The women responded to open-ended questions, beginning with a description of the "typical day" with the infant. All interviews were tape recorded, transcribed verbatim, and entered into the AtlasTi qualitative analysis program. Grounded theory methods were used to analyze the data. Transcripts were read repeatedly to verify coding and emerging concepts. RESULTS: The basic social psychological problem was the loss of the perceived baby and competence as a mother. The psychosocial process was the search for the baby and sense of self as mother. Processes involved cycles of hope and despair and trial and error as mothers became more isolated. DISCUSSION: Implications for practitioners include support and listening to mothers during this difficult period.

The voices of children: stories about hospitalization.

INTRODUCTION: The study explored children's views of hospitalization through their own voices. METHOD: In this secondary analysis, 93 children aged 5 to 9 years told stories about hospitalization using a set of drawings of children in the hospital. Children were recruited in the hospital and in the community. Themes were identified through qualitative analysis. RESULTS: Children's stories focused on being alone and feeling scared, mad, and sad. These children wanted protection. Children in the stories were not always facing scary events. They were simply not at home and feeling bored, lonely, and sad. They wanted companions. Children displayed awareness of both good and bad outcomes. The hospital was a unique environment that could be fun as well as threatening. DISCUSSION: Children's views of hospitalization were not invariably negative. The themes of hospitalized and never-hospitalized children were different only in degree of detail.

The experience of spouses as informal caregivers for recipients of hematopoietic stem cell transplants.

Spouses often are the primary caregiver for the hematopoietic stem cell transplant (HSCT) recipients and participate in the transplant recipients' care throughout the HSCT trajectory. The purpose of this qualitative study was to explore and describe the experiences of spouses of HSCT recipients during the acute phase of the transplant trajectory. The participants were spousal caregivers (8 wives and 3 husbands) of transplant recipients. Each participant was interviewed 1 to 6 times. Data were analyzed using Spradley's domain analysis. As couples entered the transplant experience, spouses described a sense of dislocation from normal life. They were now riding a rollercoaster in the dark as they lived the uncertainty of this risky therapy. Spouses structured the uncertainty with rituals, formed a positive perspective, and envisioned the future. They described a caregiving role but also needed to balance "me and my world" with "us and our world."