PRMT1 promotes the tumor suppressor function of p14ARF and is indicative for pancreatic cancer prognosis.

The p14ARF protein is a well-known regulator of p53-dependent and p53-independent tumor-suppressive activities. In unstressed cells, p14ARF is predominantly sequestered in the nucleoli, bound to its nucleolar interaction partner NPM. Upon genotoxic stress, p14ARF undergoes an immediate redistribution to the nucleo- and cytoplasm, where it promotes activation of cell cycle arrest and apoptosis. Here, we identify p14ARF as a novel interaction partner and substrate of PRMT1 (protein arginine methyltransferase 1). PRMT1 methylates several arginine residues in the C-terminal nuclear/nucleolar localization sequence (NLS/NoLS) of p14ARF . In the absence of cellular stress, these arginines are crucial for nucleolar localization of p14ARF . Genotoxic stress causes augmented interaction between PRMT1 and p14ARF , accompanied by arginine methylation of p14ARF . PRMT1-dependent NLS/NoLS methylation promotes the release of p14ARF from NPM and nucleolar sequestration, subsequently leading to p53-independent apoptosis. This PRMT1-p14ARF cooperation is cancer-relevant and indicative for PDAC (pancreatic ductal adenocarcinoma) prognosis and chemotherapy response of pancreatic tumor cells. Our data reveal that PRMT1-mediated arginine methylation is an important trigger for p14ARF 's stress-induced tumor-suppressive function.

Assaying epigenome functions of PRMTs and their substrates.

Among the widespread and increasing number of identified post-translational modifications (PTMs), arginine methylation is catalyzed by the protein arginine methyltransferases (PRMTs) and regulates fundamental processes in cells, such as gene regulation, RNA processing, translation, and signal transduction. As epigenetic regulators, PRMTs play key roles in pluripotency, differentiation, proliferation, survival, and apoptosis, which are essential biological programs leading to development, adult homeostasis but also pathological conditions including cancer. A full understanding of the molecular mechanisms that underlie PRMT-mediated gene regulation requires the genome wide mapping of each player, i.e., PRMTs, their substrates and epigenetic marks, methyl-marks readers as well as interaction partners, in a thorough and unambiguous manner. However, despite the tremendous advances in high throughput sequencing technologies and the numerous efforts from the scientific community, the epigenomic profiling of PRMTs as well as their histone and non-histone substrates still remains a big challenge owing to obvious limitations in tools and methodologies. This review will summarize the present knowledge about the genome wide mapping of PRMTs and their substrates as well as the technical approaches currently in use. The limitations and pitfalls of the technical tools along with conventional approaches will be then discussed in detail. Finally, potential new strategies for chromatin profiling of PRMTs and histone substrates will be proposed and described.

PRMT4 is a novel coactivator of c-Myb-dependent transcription in haematopoietic cell lines.

Protein arginine methyltransferase 4 (PRMT4)-dependent methylation of arginine residues in histones and other chromatin-associated proteins plays an important role in the regulation of gene expression. However, the exact mechanism of how PRMT4 activates transcription remains elusive. Here, we identify the chromatin remodeller Mi2α as a novel interaction partner of PRMT4. PRMT4 binds Mi2α and its close relative Mi2ß, but not the other components of the repressive Mi2-containing NuRD complex. In the search for the biological role of this interaction, we find that PRMT4 and Mi2α/ß interact with the transcription factor c-Myb and cooperatively coactivate c-Myb target gene expression in haematopoietic cell lines. This coactivation requires the methyltransferase and ATPase activity of PRMT4 and Mi2, respectively. Chromatin immunoprecipitation analysis shows that c-Myb target genes are direct transcriptional targets of PRMT4 and Mi2. Knockdown of PRMT4 or Mi2α/ß in haematopoietic cells of the erythroid lineage results in diminished transcriptional induction of c-Myb target genes, attenuated cell growth and survival, and deregulated differentiation resembling the effects caused by c-Myb depletion. These findings reveal an important and so far unknown connection between PRMT4 and the chromatin remodeller Mi2 in c-Myb signalling.

FoxO3A promotes metabolic adaptation to hypoxia by antagonizing Myc function.

Exposure of metazoan organisms to hypoxia engages a metabolic switch orchestrated by the hypoxia-inducible factor 1 (HIF-1). HIF-1 mediates induction of glycolysis and active repression of mitochondrial respiration that reduces oxygen consumption and inhibits the production of potentially harmful reactive oxygen species (ROS). Here, we show that FoxO3A is activated in hypoxia downstream of HIF-1 and mediates the hypoxic repression of a set of nuclear-encoded mitochondrial genes. FoxO3A is required for hypoxic suppression of mitochondrial mass, oxygen consumption, and ROS production and promotes cell survival in hypoxia. FoxO3A is recruited to the promoters of nuclear-encoded mitochondrial genes where it directly antagonizes c-Myc function via a mechanism that does not require binding to the consensus FoxO recognition element. Furthermore, we show that FoxO3A is activated in human hypoxic tumour tissue in vivo and that FoxO3A short-hairpin RNA (shRNA)-expressing xenograft tumours are decreased in size and metabolically changed. Our findings define a novel mechanism by which FoxO3A promotes metabolic adaptation and stress resistance in hypoxia.

Inferential comprehension of 3-6 year olds within the context of story grammar: a scoping review.

BACKGROUND: The ability to make inferences plays a crucial role in reading comprehension and the educational success of school-aged children. However, it starts to unfold much earlier than school entry and literacy. Given that it is likely to be targeted in speech language therapy, it would be useful for clinicians to have access to information about a developmental sequence of inferential comprehension. Yet, at this time, there is no clear proposition of the way in which this ability develops in young children prior to school entry. AIMS: To reduce the knowledge gap with regards to inferential comprehension in young children by conducting a scoping review of the literature. The two objectives of this research are: (1) to describe typically developing children's comprehension of causal inferences targeting elements of story grammar, with the goal of proposing milestones in the development of this ability; and (2) to highlight key elements of the methodology used to gather this information in a paediatric population. METHODS & PROCEDURES: A total of 16 studies from six databases that met the inclusion criteria were qualitatively analysed in the context of a scoping review. This methodological approach was used to identify common themes and gaps in the knowledge base to achieve the intended objectives. MAIN CONTRIBUTION: Results permit the description of key elements in the development of six types of causal inference targeting elements of story grammar in children between 3 and 6 years old. Results also demonstrate the various methods used to assess this ability in young children and highlight particularly interesting procedures for use with this younger population. CONCLUSIONS: These findings point to the need for additional studies to understand this ability better and to develop strategies to stimulate an evidence-based developmental sequence in children from an early age.

Uninterrupted in vivo cerebral microdialysis measures of the acute neurochemical response to a single or repeated concussion in a rat model combining force and rotation.

Altered extracellular amino acid concentrations following concussion or mild traumatic brain injury can result in delayed neuronal damage through overactivation of NMDA glutamatergic receptors. However, the consequences of repeated concussions prior to complete recovery are not well understood. In this study, we utilized in vivo cerebral microdialysis and a weight-drop model to investigate the acute neurochemical response to single and repeated concussions in adult rats that were fully conscious. A microdialysis probe was inserted into the hippocampus and remained in place during impact. Primary outcomes included concentrations of glutamate, GABA, taurine, glycine, glutamine, and serine, while secondary outcomes were righting times and excitotoxic indices. Compared to sham injury, the first concussion resulted in significant increases in glutamate, GABA, taurine, and glycine levels, longer righting times, and higher excitotoxic indices. Following the second concussion, righting times were significantly longer, suggesting cumulative effects of repeated concussion while only partial increases were observed in glutamate and taurine levels. GABA and glycine levels, and excitotoxic indices were comparable to sham injury. These findings suggest that single and repeated concussions may induce acute increases in several amino acids, while repeated concussions could exacerbate neurological symptoms despite less pronounced neurochemical changes.

Inferential Comprehension Abilities in French-Speaking Preschoolers Exposed to Neglect in the Early Longitudinal Language and Neglect Study.

PURPOSE: Using a longitudinal design, this study aimed to describe inferential comprehension abilities of neglected French-speaking preschool children from 42 to 66 months of age in comparison to non-neglected peers, to examine the association with receptive vocabulary, and to determine whether rates of change in inferential abilities over time was stable between the two group conditions. METHOD: An inferential comprehension task and the French version of the Peabody Picture Vocabulary Test-Fourth Edition were administered to a group of neglected children (n = 37-40) and to a group of same-age non-neglected children (n = 71-91) at 42, 54, and 66 months old, as part of the Early Longitudinal Language and Neglect study. RESULTS: Results show that children exposed to neglect obtain significantly lower scores compared to their same-age peers on inferential comprehension and receptive vocabulary measures at all three time points (p < .001) with large to very large effect sizes and indicate moderate to strong correlations between the two variables. Children from the neglected group present difficulties in inferencing compared to same-age non-neglected peers, a disadvantage that remains stable over time. CONCLUSIONS: This study demonstrates the significant gap in inferential comprehension abilities between neglected and non-neglected preschool children. These results reiterate the importance of early detection of language comprehension difficulties in young children coming from vulnerable environments.

Developmental trajectories of speech and language in neglected children aged 3 to 5 years: Results of the ELLAN study.

BACKGROUND: Neglected children are at high risk for significant difficulties in speech and language development. Because no longitudinal study has been conducted to date, the dynamic description of development during the preschool period is unknown. OBJECTIVES: Establish the developmental trajectories of speech sounds, receptive and expressive vocabulary, and morphosyntax among neglected children during the preschool years and compare them with those of non-neglected children. PARTICIPANTS AND SETTING: Participants are 69 neglected children and 99 same age non-neglected peers (37 and 46 males respectively) recruited at 36 months of age. Data were collected at home. METHODS: Data were collected at six-month intervals between the ages of 3 and 5.5 years using psychometrically robust tools. Neglected and control groups were compared according to age using repeated measures ANOVAs on all variables. A discrete mixture model for clustering longitudinal data was used for testing the heterogeneity of the language trajectories among neglected children. RESULTS: The language development of the neglected children as a whole group is lower than that of the control group for all variables. Two subgroups are identified within the neglected group: one with a developmental trajectory similar to that of the non-neglected children, and another whose trajectory is far below that of the control group. The effect sizes of these differences vary between 1.4 and 3 standard deviations under the mean. CONCLUSIONS: A large proportion of neglected children present significant speech and language difficulties from the age of 3, but some of them catch up and develop similarly to non-neglected children.

Efficacy of prophylactic versus therapeutic administration of the NMDA receptor antagonist MK-801 on the acute neurochemical response to a concussion in a rat model combining force and rotation.

OBJECTIVE: Alterations in amino acid concentrations are a major contributor to the persistent neurological and behavioral effects induced by concussions and mild traumatic brain injuries (TBIs). Glutamate, the most abundant excitatory amino acid in the CNS, has a major role in the pathophysiological process of concussion. The indiscriminate liberation of glutamate immediately after a concussion triggers an excitotoxic response that leads to cell death, neuronal damage, and the dysfunction of surviving neurons, largely by overactivation of N-methyl-d-aspartate (NMDA) glutamatergic receptors. The aim of the present study was to investigate the efficacy of prophylactic versus therapeutic administration of MK-801, a promising NMDA receptor antagonist, on the acute changes in amino acid extracellular concentrations involved in excitotoxicity resulting from a concussive trauma. METHODS: The immediate neurochemical response to a concussion cannot be characterized in humans. Therefore, the authors used their previously validated combination of a weight-drop concussion rat model and in vivo cerebral microdialysis. The microdialysis probe was inserted inside the hippocampus and left inserted at impact to allow uninterrupted sampling of amino acids of interest immediately after concussion. The primary outcome included amino acid concentrations and the secondary outcome included righting time. Samples were taken in 10-minute increments for 60 minutes before, during, and 60 minutes after impact, and analyzed for glutamate, gamma-aminobutyric acid, taurine, glycine, glutamine, and serine using high-performance liquid chromatography. Righting time was acquired as a neurological restoration indicator. Physiological saline or 10 mg/kg MK-801 was administrated intraperitoneally 60 minutes before or immediately following induction of sham injury or concussion. RESULTS: Following induction of concussion, glutamate, taurine, and glycine levels as well as righting times in cases from the MK-801 treatment group were comparable to those of vehicle-treated animals. In contrast, righting times and amino acid concentrations observed within the first 10 minutes after induction of concussion in cases assigned to the MK-801 prophylaxis group were comparable to those of sham-injured animals. CONCLUSIONS: These results suggest that presynaptic actions and peak availability of MK-801 following prophylactic administration significantly inhibit the immediate and indiscriminate release of glutamate, taurine, and glycine in extracellular fluid after a concussion.

Profiles of teacher-child interaction quality in groups of 3-year-old children in Quebec and France.

Theory and studies support that educational quality may differ according to socio-political context even in states with similar cultures. Based on a secondary analysis of data, this study aims at identifying latent profiles of adult-child interaction quality in groups of three-year-old children in Quebec's (Canada) early childhood centers and France's kindergarten classrooms using the CLASS Pre-K. This study also aims to explore existing associations between identified profiles, socio-political contexts, and structural characteristics (staff qualifications, ages, group size). Latent profile analyses showed four interaction quality profiles, namely a high-quality profile (HQ), a medium-high-quality profile (MHQ), a medium quality profile (MQ), and a medium-low-quality profile (MLQ). The scores of the three CLASS Pre-K domains associated with identified profiles show a higher average interaction quality in Quebec compared with France, suggesting a more favorable sociocultural context for interaction quality in Quebec. As for characteristics of structural quality, analyses suggest that the group size variable is significantly associated with scores of interaction quality, with the HQ and the MHQ profiles showing a significantly lower group size than the MQ and MLQ profiles. Age is also significantly associated with profiles, exhibiting a general trend of younger participants found in higher quality profiles. Courses of action to enhance French policies are discussed.

Systematic investigation of PRMT6 substrate recognition reveals broad specificity with a preference for an RG motif or basic and bulky residues.

Protein arginine methyltransferase 6 (PRMT6) catalyses the asymmetric dimethylation of arginines on numerous substrate proteins within the human cell. In particular, PRMT6 methylates histone H3 arginine 2 (H3R2) which affects both gene repression and activation. However, the substrate specificity of PRMT6 has not been comprehensively analysed. Here, we systematically characterise the substrate recognition motif of PRMT6, finding that it has broad specificity and recognises the RG motif. Working with a H3 tail peptide as a template, on which we made 204 amino acid substitutions, we use targeted mass spectrometry to measure their effect on PRMT6 in vitro activity. We first show that PRMT6 methylates R2 and R8 in the H3 peptide, although H3R8 is methylated with lower efficiency and is not an in vivo PRMT6 substrate. We then quantify the effect of 194 of these amino acid substitutions on methylation at both H3R2 and H3R8. In both cases, we find that PRMT6 tolerates essentially any amino acid substitution in the H3 peptide, but that positively charged and bulky residues are preferred near the target arginine. We show that PRMT6 also has preference for glycine, but only in the position immediately following the target arginine. This indicates that PRMT6 recognises the RG motif rather than the RGG motif. We further confirm this preference for the RG motif on another PRMT6 substrate, histone H4R3. This broad specificity and recognition of RG rather than RGG are distinctive among the PRMT family and has implications for the development of drugs to selectively target PRMT6. DATABASES: Panorama Public (https://panoramaweb.org/PRMT6motif.url); ProteomeXchange (PXD016711).

Targeting Uric Acid Prevents Brain Injury and Anxiety in a Rat Model of Hemorrhagic Shock.

ABSTRACT: Secondary brain injury following hemorrhagic shock (HS) is a frequent complication in patients, even in the absence of direct brain trauma, leading to behavioral changes and more specifically anxiety and depression. Despite preclinical studies showing inflammation and apoptosis in the brain after HS, none have addressed the impact of circulating mediators. Our group demonstrated an increased uric acid (UA) circulation in rats following HS. Since UA is implicated in endothelial dysfunction and inflammatory response, we hypothesized UA could alter the blood-brain barrier (BBB) and impact the brain. Male Wistar rats were randomly assigned to: SHAM, HS (hemorrhagic shock) and HS + U (hemorrhagic shock + 1.5 mg/kg of uricase). The uricase intervention, specifically targeting UA, was administered during fluid resuscitation. It prevented BBB dysfunction (fluorescein sodium salt permeability and expression of intercellular adhesion molecule-1) following HS. As for neuroinflammation, all of the results obtained (MPO activity; Iba1 and GFAP expression) showed a significant increase after HS, also prevented by the uricase. The same pattern was observed after quantification of apoptosis (caspase-3 activity and TUNEL) and neurodegeneration (Fluoro-Jade). Finally, the forced swim, elevated plus maze, and social interaction tests detected anxiety-like behavior after HS, which was blunted in rats treated with the uricase. In conclusion, we have identified UA as a new circulatory inflammatory mediator, responsible for brain alterations and anxious behavior after HS in a murine model. The ability to target UA holds the potential of an adjunctive therapeutic solution to reduce brain dysfunction related to hemorrhagic shock in human.

The importance of Calanus glacialis for the feeding success of young polar cod: a circumpolar synthesis.

Understanding the feeding ecology of polar cod (Boreogadus saida) during its first year of life is crucial to forecasting its response to the ongoing borealization of Arctic seas. We investigated the relationships between diet composition and feeding success in 1797 polar cod larvae and juveniles 4.5-55.6 mm standard length (SL) collected in five Arctic seas from 1993 to 2009. Prey were identified to species and developmental stages when possible, measured, and their carbon content was estimated using taxon-specific allometric equations. Feeding success was defined as the ratio of ingested carbon to fish weight. Carbon uptake in polar cod larvae < 15 mm was sourced primarily from calanoid copepods eggs and nauplii which were positively selected from the plankton. With increasing length, carbon sources shifted from eggs and nauplii to the copepodites of Calanus glacialis, Calanus hyperboreus and Pseudocalanus spp. Calanus glacialis copepodites were the main carbon source in polar cod > 25 mm and the only copepodite positively selected for. Pseudocalanus spp. copepodites became important replacement prey when C. glacialis left the epipelagic layer at the end of summer. Calanus glacialis was the preferred prey of polar cod, contributing from 23 to 84% of carbon uptake at any stage in the early development. Feeding success was determined by the number of prey captured in larvae < 15 mm and by the size of prey in juveniles > 30 mm. As Arctic seas warm, the progressive displacement of C. glacialis by the smaller Calanus finmarchicus could accelerate the replacement of polar cod, the dominant Arctic forage fish, by boreal species.

Parental behaviors associated with the level of pragmatic language ability among 42-month-old neglected children.

BACKGROUND: Exposure to neglect can severely compromise children's pragmatic skills (social language use). The disruptions of parent-child interactions that typically occur in context of neglect may compromise several parental behaviors which are known to foster language skills such as pragmatics. OBJECTIVES: 1- Compare the behaviors of neglectful and non-neglectful parents in four domains which are of interest for pragmatic language development, namely, responsive, supportive, affective, and control behaviors, and 2- Identify parental behaviors associated with the levels of pragmatic ability of 42-month-old neglected children. PARTICIPANTS: Study sample consisted of 21 neglected children living in their biological family, recruited in four Youth Centers in the province of Québec (Canada) and 95 non-neglected children recruited in child-care centers. METHOD: Parental behaviors were video recorded in context of free-play with the child at the participants' homes between 2015 and 2017, and subsequently analyzed using the Coding Observations of Parent-Child Interactions (COPI), developed to observe ten parental behaviors associated with early language development. The level of pragmatic ability of children was established using the Language Use Inventory: French, a standardized questionnaire completed with parents of both groups. RESULTS: Parents in situation of neglect scored lower than parents in the control group on eight of the ten behaviors (p < .001). Parental reciprocity was associated with the level of pragmatic ability of 42-month-old neglected children (p = .04). CONCLUSIONS: The results of this exploratory study provide insight on the associations between parental behaviors and the level of pragmatic language skills of children experiencing neglect.

Impact of uric acid on liver injury and intestinal permeability following resuscitated hemorrhagic shock in rats.

BACKGROUND: Multiorgan failure is a consequence of severe ischemia-reperfusion injury after traumatic hemorrhagic shock (HS), a major cause of mortality in trauma patients. Circulating uric acid (UA), released from cell lysis, is known to activate proinflammatory and proapoptotic pathways and has been associated with poor clinical outcomes among critically ill patients. Our group has recently shown a mediator role for UA in kidney and lung injury, but its role in liver and enteric damage after HS remains undefined. Therefore, the objective of this study was to evaluate the role of UA on liver and enteric injury after resuscitated HS. METHODS: A murine model of resuscitated HS was treated during resuscitation with a recombinant uricase, a urate oxidase enzyme (rasburicase; Sanofi-Aventis, Canada Inc, Laval, Canada), to metabolize and reduce circulating UA. Biochemical analyses (liver enzymes, liver apoptotic, and inflammatory markers) were performed at 24 hours and 72 hours after HS. Physiological testing for enteric permeability and gut bacterial product translocation measurement (plasma endotoxin) were performed 72 hours after HS. In vitro, HT-29 cells were exposed to UA, and the expression of intercellular adhesion proteins (ZO-1, E-cadherin) was measured to evaluate the influence of UA on enteric permeability. RESULTS: The addition of uricase to resuscitation significantly reduced circulating and liver UA levels after HS. It also prevented HS-induced hepatolysis and liver apoptotic/inflammatory mediators at 24 hours and 72 hours. Hemorrhagic shock-induced enteric hyperpermeability and endotoxemia were prevented with uricase. CONCLUSIONS: After resuscitated HS, UA is an important mediator in liver and enteric injury. Uric acid represents a therapeutic target to minimize organ damage in polytrauma patients sustaining HS.

Morphosyntactic Development and Severe Parental Neglect in 4-Year-Old French-Speaking Children: ELLAN study.

Language is the most frequently compromised area of development in English-speaking neglected children, particularly the morphosyntactic component of language. This is very worrisome given its central role in academic success and social participation. No previous study has examined the morphosyntactic skills of French-speaking neglected children, despite the morphological richness of French. This study aimed to fill this gap. Forty-four neglected (mean age = 48.32 months, SD = 0.45) and 92 non-neglected (mean age = 48.07 months, SD = 0.24) French-speaking children participated. Measures of morphosyntactic skills were derived from a sample of spontaneous language collected during standardized semistructured play and analyzed using Systematic Analysis of Language Transcripts software (2012) . Four morphosyntactic indicators were compared using analyses of variance and Kolmogorov-Smirnov tests: the mean length of utterances (MLU), verbal inflections, word-level errors, and omission errors. The results indicate that 25.6% of the neglected children presented clinically significant morphosyntactic difficulties, as evidenced by a significantly shorter MLU (M = 5.60, SD = 1.13; M = 6.90, SD = 1.30), fewer verbal inflections, and more frequent word omission errors compared to their non-neglected peers. The results confirm that French-speaking neglected children present many morphosyntactic difficulties. This study argues for sustained speech-language services for these children.

The Pragmatic Language Skills of Severely Neglected 42-Month-Old Children: Results of the ELLAN Study.

The goals of this study were twofold: (1) to compare the pragmatic language skills (i.e., social communication skills) of 42-month-old neglected children with those of same-aged non-neglected children and (2) to measure the prevalence of pragmatic difficulties among the neglected children. The study sample was composed of 45 neglected and 95 non-neglected 42-month-old French-speaking children. The Language Use Inventory: French (LUI-French) was completed with all parents. This measure, comprised of 159 scored items divided into 10 subscales, was used to assess the children's pragmatic skills. The 10th percentile on the LUI-French (95% confidence interval ) was used to identify children with pragmatic difficulties. The neglected children had lower scores than the non-neglected children on all 10 dimensions of pragmatics evaluated (p < .01), as well as lower LUI-French Total Scores (p < .001). The effect sizes of these differences varied between 0.84 and 2.78. Forty-four percent of the neglected children presented significant pragmatic difficulties compared to 4.2% of their non-neglected peers (p < .001). It can be concluded that exposure to neglect significantly compromises children's pragmatic skills. These results support the need for interventions geared toward neglected children and their families to support the early development of their pragmatic skills.

Caspase-(8/3) activation and organ inflammation in a rat model of resuscitated hemorrhagic shock: A role for uric acid.

BACKGROUND: Multiple organ failure can develop after hemorrhagic shock (HS). Uric acid (UA) is released from dying cells and can be proinflammatory. We hypothesized that UA could be an alternative mediator of organ apoptosis and inflammation after HS. METHODS: Ventilated male Wistar rats were used for the HS model. Two durations of shock (5 minutes vs. 60 minutes) were compared, and shams were instrumented only; animals were resuscitated and observed for 24 hours/72 hours. Caspases-(8/3), myeloperoxidase (MPO), TNF-α were measured in lungs and kidneys. Plasma UA and cytokine (IL-1ß, IL-18, TNF-α) were measured. A second set of animals were randomized to vehicle versus Rasburicase intraperitoneal intervention (to degrade UA) during resuscitation. Another group received exogenous UA intraperitoneally without HS. Measures mentioned above, in addition to organs UA, were performed at 24 hours. In vitro, caspases-(8/3) activity was tested in epithelial cells exposed to UA. RESULTS: Hemorrhagic shock increased organ (kidney and lung) TNF-α, MPO, and caspases activity in various patterns while caspase-8 remained elevated over time. Hemorrhagic shock led to increased plasma UA at 2 hours, which remained high until 72 hours; TNF-α and IL-18 were elevated at 24 hours. The exogenous UA administration in sham animals reproduced the activation of caspase-8 and MPO in organs, and TNF-α in the lung. The increased plasma and organ UA levels, plasma and lung TNF-α, as well as organ caspase-(8/3) and MPO, observed at 24 hours after HS, were prevented by the administration of Rasburicase during resuscitation. In vitro, soluble UA induced caspases-(3/8) activity in epithelial cells. CONCLUSION: Uric acid is persistently high after HS and leads to the activation of caspases-8 and organ inflammation; these can be prevented by an intervention to degrade UA. Therefore, UA is an important biomarker and mediator that could be considered a therapeutic target during HS resuscitation in human.

Fluid sparing and norepinephrine use in a rat model of resuscitated haemorrhagic shock: end-organ impact.

BACKGROUND: Haemostasis and correction of hypovolemia are the pillars of early haemorrhage shock (HS) management. Vasopressors, which are not recommended as first-line therapy, are an alternative to aggressive fluid resuscitation, but data informing the risks and benefits of vasopressor therapy as fluid-sparing strategy is lacking. We aimed to study its impact on end organs, in the setting of a haemodynamic response to the initial volume resuscitation. METHODS: Following controlled HS (60 min) induced by blood withdrawal, under anaesthesia and ventilation, male Wistar rats (N = 10 per group) were randomly assigned to (1) sham, (2) HS with fluid resuscitation only [FR] and (3) HS with fluid resuscitation to restore haemodynamic (MAP: mean arterial pressure) then norepinephrine [FR+NE]. After a reperfusion time (60 min) during which MAP was maintained with fluid or norepinephrine, equipment was removed and animals were observed for 24 h (N = 5) or 72 h (N = 5) before euthanasia. Besides haemodynamic parameters, physiological markers (creatinine, lactate, pH, PaO2) and one potential contributor to vasoplegia (xanthine oxidase activity) were measured. Apoptosis induction (caspase 3), tissue neutrophil infiltration (MPO: myeloperoxidase) and illustrative protein markers were measured in the lung (Claudin-4), kidney (KIM-1) and brain amygdala (Iba1). RESULTS: No difference was present in MAP levels during HS or reperfusion between the two resuscitation strategies. FR required significantly more fluid than FR+NE (183% vs 106% of bleed-out volume; p = 0.003), when plasma lactate increased similarly. Xanthine oxidase was equally activated in both HS groups. After FR+NE, creatinine peaked higher but was similar in all groups at later time points. FR+NE enhanced MPO in the lung, when Claudin-4 increased significantly after FR. In the brain amygdala, FR provoked more caspase 3 activity, MPO and microglial activation (Iba1 expression). CONCLUSION: Organ resuscitation after controlled HS can be assured with lesser fluid administration followed by vasopressors administration, without signs of dysoxia or worse evolution. Limiting fluid administration could benefit the brain and seems not to have a negative impact on the lung or kidney.

MYB induces the expression of the oncogenic corepressor SKI in acute myeloid leukemia.

Acute myeloid leukemia (AML) arises through clonal expansion of transformed myeloid progenitor cells. The SKI proto-oncogene is highly upregulated in different solid tumors and leukemic cells, but little is known about its transcriptional regulation during leukemogenesis. MYB is an important hematopoietic transcription factor involved in proliferation as well as differentiation and upregulated in most human acute leukemias. Here, we find that MYB protein binds within the regulatory region of the SKI gene in AML cells. Reporter gene assays using MYB binding sites present in the SKI gene locus show MYB-dependent transcriptional activation. SiRNA-mediated depletion of MYB in leukemic cell lines reveals that MYB is crucial for SKI gene expression. Consistently, we observed a positive correlation of MYB and SKI expression in leukemic cell lines and in samples of AML patients. Moreover, MYB and SKI both were downregulated by treatment with histone deacetylase inhibitors. Strikingly, differentiation of AML cells induced by depletion of MYB is attenuated by overexpression of SKI. Our findings identify SKI as a novel MYB target gene, relevant for the MYB-induced differentiation block in leukemic cells.