Molecular dissection of PI3Kß synergistic activation by receptor tyrosine kinases, GßGγ, and Rho-family GTPases.

Phosphoinositide 3-kinase (PI3K) beta (PI3Kß) is functionally unique in the ability to integrate signals derived from receptor tyrosine kinases (RTKs), G-protein coupled receptors, and Rho-family GTPases. The mechanism by which PI3Kß prioritizes interactions with various membrane-tethered signaling inputs, however, remains unclear. Previous experiments did not determine whether interactions with membrane-tethered proteins primarily control PI3Kß localization versus directly modulate lipid kinase activity. To address this gap in our knowledge, we established an assay to directly visualize how three distinct protein interactions regulate PI3Kß when presented to the kinase in a biologically relevant configuration on supported lipid bilayers. Using single molecule Total Internal Reflection Fluorescence (TIRF) Microscopy, we determined the mechanism controlling PI3Kß membrane localization, prioritization of signaling inputs, and lipid kinase activation. We find that auto-inhibited PI3Kß prioritizes interactions with RTK-derived tyrosine phosphorylated (pY) peptides before engaging either GßGγ or Rac1(GTP). Although pY peptides strongly localize PI3Kß to membranes, stimulation of lipid kinase activity is modest. In the presence of either pY/GßGγ or pY/Rac1(GTP), PI3Kß activity is dramatically enhanced beyond what can be explained by simply increasing membrane localization. Instead, PI3Kß is synergistically activated by pY/GßGγ and pY/Rac1 (GTP) through a mechanism consistent with allosteric regulation.

The gut metagenome harbors metabolic and antibiotic resistance signatures of moderate-to-severe asthma.

Asthma is a common allergic airway disease that has been associated with the development of the human microbiome early in life. Both the composition and function of the infant gut microbiota have been linked to asthma risk, but functional alterations in the gut microbiota of older patients with established asthma remain an important knowledge gap. Here, we performed whole metagenomic shotgun sequencing of 95 stool samples from a cross-sectional cohort of 59 healthy and 36 subjects with moderate-to-severe asthma to characterize the metagenomes of gut microbiota in adults and children 6 years and older. Mapping of functional orthologs revealed that asthma contributes to 2.9% of the variation in metagenomic content even when accounting for other important clinical demographics. Differential abundance analysis showed an enrichment of long-chain fatty acid (LCFA) metabolism pathways, which have been previously implicated in airway smooth muscle and immune responses in asthma. We also observed increased richness of antibiotic resistance genes (ARGs) in people with asthma. Several differentially abundant ARGs in the asthma cohort encode resistance to macrolide antibiotics, which are often prescribed to patients with asthma. Lastly, we found that ARG and virulence factor (VF) richness in the microbiome were correlated in both cohorts. ARG and VF pairs co-occurred in both cohorts suggesting that virulence and antibiotic resistance traits are coselected and maintained in the fecal microbiota of people with asthma. Overall, our results show functional alterations via LCFA biosynthetic genes and increases in antibiotic resistance genes in the gut microbiota of subjects with moderate-to-severe asthma and could have implications for asthma management and treatment.

A pilot study of trauma-sensitive yoga and Breathe2Relax among service members in an intensive outpatient program.

Emerging research indicates that yoga is a promising adjunct to psychological trauma treatment. The current pilot study examined the associations between psychophysiological stress, diaphragmatic breathing (DB), and a trauma-sensitive yoga (TSY) regimen developed specifically for trauma-exposed service members in alignment with recent calls for precision in reporting therapeutic yoga protocols. Participants were 31 service members enrolled in a trauma-focused intensive outpatient program (IOP). Service members participated in a brief diaphragmatic breathing (DB) session using the Breathe2Relax (B2R) app followed by the TSY session. Heart rate (HR) and perceived stress were measured at baseline and after both the DB practice and the TSY session. We assessed Yoga and DB expectancies at baseline and post TSY. Participants also rated the acceptability and usability of the B2R app. Results of linear mixed effects regression models showed decreases in HR and perceived stress, compared to baseline, following DB (HR, b = -8.68, CI 95% = -13.34, -4.02; perceived stress, b = -1.77, CI 95% = -2.35, -1.18) and TSY (HR, b = -12.44, CI 95% = -17.15, -7.73; perceived stress b = -3.69, CI 95% = -4.29, -3.08). Higher levels of expectancies, compared to lower levels, related to stronger decreases in HR and perceived stress, particularly after TSY. Overall, participants rated the B2R usability as high; virtually all participants reported that "most would learn to use the app quickly," and 76.6% reported that they would use it frequently.

Correction: The hallmarks of childhood abuse and neglect: A systematic review.

[This corrects the article DOI: 10.1371/journal.pone.0243639.].

Molecular dissection of PI3Kß synergistic activation by receptor tyrosine kinases, GßGγ, and Rho-family GTPases.

The class 1A phosphoinositide 3-kinase (PI3K) beta (PI3Kß) is functionally unique in the ability to integrate signals derived from receptor tyrosine kinases (RTKs), heterotrimeric guanine nucleotide-binding protein (G-protein)-coupled receptors (GPCRs), and Rho-family GTPases. The mechanism by which PI3Kß prioritizes interactions with various membrane tethered signaling inputs, however, remains unclear. Previous experiments have not been able to elucidate whether interactions with membrane-tethered proteins primarily control PI3Kß localization versus directly modulate lipid kinase activity. To address this gap in our understanding of PI3Kß regulation, we established an assay to directly visualize and decipher how three distinct protein interactions regulate PI3Kß when presented to the kinase in a biologically relevant configuration on supported lipid bilayers. Using single molecule Total Internal Reflection Fluorescence (TIRF) Microscopy, we determined the mechanism controlling membrane localization of PI3Kß, prioritization of signaling inputs, and lipid kinase activation. We find that auto-inhibited PI3Kß prioritizes interactions with RTK-derived tyrosine phosphorylated (pY) peptides before engaging either GßGγ or Rac1(GTP). Although pY peptides strongly localize PI3Kß to membranes, stimulation of lipid kinase activity is modest. In the presence of either pY/GßGγ or pY/Rac1(GTP), PI3Kß activity is dramatically enhanced beyond what can be explained by simply increasing the strength of membrane localization. Instead, PI3Kß is synergistically activated by pY/GßGγ and pY/Rac1(GTP) through a mechanism consistent with allosteric regulation.

The gut microbiota of people with asthma influences lung inflammation in gnotobiotic mice.

The gut microbiota in early childhood is linked to asthma risk, but may continue to affect older patients with asthma. Here, we profile the gut microbiota of 38 children (19 asthma, median age 8) and 57 adults (17 asthma, median age 28) by 16S rRNA sequencing and find individuals with asthma harbored compositional differences from healthy controls in both adults and children. We develop a model to aid the design of mechanistic experiments in gnotobiotic mice and show enterotoxigenic Bacteroides fragilis (ETBF) is more prevalent in the gut microbiota of patients with asthma compared to healthy controls. In mice, ETBF, modulated by community context, can increase oxidative stress in the lungs during allergic airway inflammation (AAI). Our results provide evidence that ETBF affects the phenotype of airway inflammation in a subset of patients with asthma which suggests that therapies targeting the gut microbiota may be helpful tools for asthma control.

The gut metagenome harbors metabolic and antibiotic resistance signatures of moderate-to-severe asthma.

Asthma is a common allergic airway disease that develops in association with the human microbiome early in life. Both the composition and function of the infant gut microbiota have been linked to asthma risk, but functional alterations in the gut microbiota of older patients with established asthma remain an important knowledge gap. Here, we performed whole metagenomic shotgun sequencing of 95 stool samples from 59 healthy and 36 subjects with moderate-to-severe asthma to characterize the metagenomes of gut microbiota in children and adults 6 years and older. Mapping of functional orthologs revealed that asthma contributes to 2.9% of the variation in metagenomic content even when accounting for other important clinical demographics. Differential abundance analysis showed an enrichment of long-chain fatty acid (LCFA) metabolism pathways which have been previously implicated in airway smooth muscle and immune responses in asthma. We also observed increased richness of antibiotic resistance genes (ARGs) in people with asthma. One differentially abundant ARG was a macrolide resistance marker, ermF, which significantly co-occurred with the Bacteroides fragilis toxin, suggesting a possible relationship between enterotoxigenic B. fragilis, antibiotic resistance, and asthma. Lastly, we found multiple virulence factor (VF) and ARG pairs that co-occurred in both cohorts suggesting that virulence and antibiotic resistance traits are co-selected and maintained in the fecal microbiota of people with asthma. Overall, our results show functional alterations via LCFA biosynthetic genes and increases in antibiotic resistance genes in the gut microbiota of subjects with moderate-to-severe asthma and could have implications for asthma management and treatment.

European public mental health responses to the COVID-19 pandemic.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic increased multiple risk factors for mental health. Evidence-based, intersectoral public mental health responses are therefore critical. The primary aim of this study was to collate public mental health responses from across Europe. METHODS: We conducted a cross-sectional survey in March 2021. Participants were public and mental health professionals from across Europe. We developed an online instrument exploring five domains: changes in mental health supports during the pandemic; mental health support for vulnerable groups; multi-sectoral and service-user involvement; published mental health response plans; and perceived quality of overall country response. RESULTS: Fifty-two individuals from 20 European nations responded. Reported changes in mental health supports included an increase in online mental health supports (n = 18); but no change in long-term mental health funding (n = 13); and a decrease in access to early interventions (n = 9). Responses indicated mental health support for vulnerable groups was limited, as was multi-sectoral and service-user involvement. Few national mental health response plans existed (n = 9) and 48% of respondents felt their countries mental health response had been 'poor' or 'very poor'. CONCLUSIONS: Our results give insights into the changes in mental health support at a country level across Europe during the COVID-19 pandemic. They indicate countries were not prepared to respond and people with existing vulnerabilities were often neglected in response planning. To be prepared for future pandemics and environmental disasters Public Mental Health preparedness plans are highly needed. These must be developed cross-departmentally, and through the meaningful inclusion of vulnerable groups.

Recruiting and exploring vulnerabilities among young people at risk, or in the early stages of serious mental illness (borderline personality disorder and first episode psychosis).

Introduction: Many gaps exist in our understanding of the developmental pathways to severe mental illness (SMI), including borderline personality disorder (BPD) and psychosis. However, those who have experienced adverse childhood experiences (ACEs) are at an increased risk and there is evidence to suggest that one of the earliest markers is emotional dysregulation. An area which has received relatively less research attention is the role neurodevelopmental disorders (NDDs) play. The aim of this feasibility study was therefore to explore the clinical profiles of young people early in the course of SMI, including their profiles of ACEs, emotional regulation difficulties, borderline personality traits and NDDs. Methods: A cross-sectional study of young people (aged 15-25) at risk of SMI, currently being seen within NHS mental health services, was conducted. This included those with early symptoms of psychosis and/or BPD as assessed by diagnostic interview. Eligible participants self-completed a battery of sociodemographic, clinical, and psychological measures in the company of a researcher. This included assessments of: symptoms of NDDs; borderline pathology traits; ACEs; and difficulties in emotional regulation. Statistical analyses included Mann-Whitney U tests and multiple regression. Results: Of the 118 potentially eligible participants who were referred, 48 were ultimately included in the study. Young people early in the course of SMI reported a high prevalence of ACEs and deficits in emotional regulation. In total, 79% met criteria for attention deficit hyperactivity disorder (ADHD) and/or autism spectrum disorder (ASD). Emotional dysregulation was found to significantly mediate the association between both ACEs and the frequency of NDDs and borderline personality traits, however given the small sample size these results are preliminary in nature. Conclusion: Young people early in the course of SMI are at an increased risk of experiencing multiple childhood adversities and our results indicate a high prevalence of NDDs amongst them. Emotional dysregulation emerged as a potentially significant early marker of future clinical severity. We suggest that the clinical implications of our findings include routine screening for NDDs and ACEs and an increased recognition of the significance of emotional dysregulation. However, larger scale longitudinal studies are needed to investigate these preliminary findings further.

The mental health effects of a Universal Basic Income: A synthesis of the evidence from previous pilots.

INTRODUCTION: Amongst its advocates, one of the key arguments for a Universal Basic Income (UBI) is its potential to improve population mental health. However, while previous authors have variously examined the potential effects of UBI on income, employment and labour market demand, the direct mental health consequences of previous pilots have been less frequently examined. The purpose of this paper is therefore to conduct a review of the literature on UBI and to re-examine the existing research with a mental health focus. METHODS: Six electronic databases were used to conduct a review of the literature. We searched for empirical research studies of any design, conducted since the year 2000 in High Income Countries, exploring the effects of interventions similar to a UBI on the mental health of children or working-age adults. Grey literature and government reports were also included. RESULTS: A total of 1566 articles were screened of which seven peer reviewed studies and eight governmental reports were ultimately selected for inclusion. None of the identified studies directly compared the impact of individual payments with those made on a household basis, or the effects of payments which were truly universal. However, several studies evaluated the mental health outcomes associated with payments provided unconditionally, and consistently reported clear and significant improvements in mental wellbeing. Potential mediating factors included improved time with family and friends, a reduction in perceived stigma and a renewed sense of hope for the future. CONCLUSIONS: Our review has produced evidence to suggest that prophesies surrounding the mental health benefits of a UBI are at least partially justified. However, future studies should aim to be conducted at an area level, with an adequately powered sample size, and investigate interventions of a considerable duration using a longitudinal design.

Nature and nurture? A review of the literature on childhood maltreatment and genetic factors in the pathogenesis of borderline personality disorder.

BACKGROUND: Borderline Personality Disorder (BPD) is a psychiatric disorder associated with significant morbidity and mortality. However, the neurobiological alterations underlying the condition remain poorly understood. As a result, existing treatments remain inadequate. One of the main risk factors for the development of BPD is a history of childhood maltreatment. However, it is considered neither causative nor specific to the condition. Current theory is therefore increasingly moving toward a 'Gene x Environment' (GxE) model of the condition. The purpose of the current work was to conduct a systematic literature review, which comprehensively identifies all published molecular level GxE studies that have explored the role of specific genetic loci, in influencing the risk of BPD following exposure to childhood abuse or neglect. METHODS: Four electronic databases were used to systematically search for molecular level GxE studies of any design, which focused on the development of BPD following exposure to childhood abuse or neglect, without language or date restrictions. Articles were screened independently by two reviewers and results were synthesized narratively. RESULTS: A total of 473 articles were screened of which sixteen were selected for inclusion in our review. Implicated genes were categorised according to their influence on; Neurotransmitter Systems, Neurodevelopment and Neuroendocrine Systems. CONCLUSIONS: The identified studies have produced several relevant and statistically significant results. Of particular note, is the repeated finding that genes involved in HPA axis regulation, may be altered by exposure to childhood maltreatment, influencing subsequent susceptibility to BPD. This is both biologically plausible and of potential clinical significance.

Escaping the inescapable: Risk of mental health disorder, somatic symptoms and resilience in Palestinian refugee children.

Exposure to war, conflict and forced migration puts children at risk of mental health problems. The present study examined the levels of psychological distress and resilience factors among 106 Palestinian refugee children aged 11 to 17 in the West Bank. In a cross-sectional, mixed method design along with qualitative interviews, three questionnaires were administered: the Strength and Difficulties Questionnaire and Patient Health Questionnaire-15, assessed the risk of mental health disorders and psychosomatic complaints, and the Child and Youth Resilience Measure assessed the availability of resilience-enhancing factors. Palestinian refugee children were found to be at greater risk for mental disorders and psychosomatic complaints than were children living in non-conflict affected settings. In addition, resilience-enhancing resources were significantly reduced and were negatively correlated with both symptom outcomes. Risk factors identified included poverty, violence and marginalisation. Key protective factors were youth education, supportive relationships and social participation. Our findings support interventions that address the identified protective factors, which may promote the mental health of this vulnerable population.

The hallmarks of childhood abuse and neglect: A systematic review.

BACKGROUND: Studies on the impacts of child maltreatment (CM) have been conducted in diverse areas. Mechanistic understanding of the complex interplay between factors is lacking. Hallmarking is an approach which identifies common factors across studies and highlights the most robust findings. OBJECTIVES: In a review of systematic reviews and meta-analyses, we addressed the following questions: 1) What are the hallmarks associated with exposure to CM across the bio-ecological spectrum? 2) What is the strength of evidence to support each hallmark? 3) What are the gaps that future research should address? METHODS: A comprehensive literature search was carried out to find relevant systematic reviews or meta-analyses. 269 articles were read in full and 178 articles, encompassing more than 6000 original papers, were included in the final synthesis. All reviews were independently rated for quality by at least 2 reviewers using AMSTAR-2. RESULTS: Of 178 review articles, 6 were rated as high quality (all meta-analyses) and 46 were rated as medium quality. Most were from high income countries. CONCLUSIONS: Based on the most commonly reported high-quality research findings we propose that the hallmarks of exposure to child maltreatment are: Increased risk of psychopathology; Increased risk of obesity; Increased risk of high- risk sexual behaviours, Increased risk of smoking; and Increased risk of child maltreatment in children with disabilities. Research gaps include a lack of focus on complexity and resilience. Little can be concluded about directions of causality or mechanisms. Adequately powered prospective studies are required to move the field forward.

Airway Microbiota-Host Interactions Regulate Secretory Leukocyte Protease Inhibitor Levels and Influence Allergic Airway Inflammation.

Homeostatic mucosal immune responses are fine-tuned by naturally evolved interactions with native microbes, and integrating these relationships into experimental models can provide new insights into human diseases. Here, we leverage a murine-adapted airway microbe, Bordetella pseudohinzii (Bph), to investigate how chronic colonization impacts mucosal immunity and the development of allergic airway inflammation (AAI). Colonization with Bph induces the differentiation of interleukin-17A (IL-17A)-secreting T-helper cells that aid in controlling bacterial abundance. Bph colonization protects from AAI and is associated with increased production of secretory leukocyte protease inhibitor (SLPI), an antimicrobial peptide with anti-inflammatory properties. These findings are additionally supported by clinical data showing that higher levels of upper respiratory SLPI correlate both with greater asthma control and the presence of Haemophilus, a bacterial genus associated with AAI. We propose that SLPI could be used as a biomarker of beneficial host-commensal relationships in the airway.

Financial hardship and risk of suicide among U.S. Army personnel.

Financial stress has been frequently identified as a risk factor for suicidal behavior, both in military and civilian groups. However, it remains unclear to what degree financial stress may be associated independently with suicide behavior when accounting for other risk factors. This study examined data on suicide and suicide attempt cases in the Department of Defense Suicide Event Report compared with service members who did not have recent suicide behavior. The resulting multinomial regression analysis found that financial distress had a weak association with suicide, and its relationship to suicide attempts was not statistically significant. Compared with financial distress, relationship problems and substance abuse history appeared to have much stronger associations with suicidal behavior, as did having a diagnosis of a mood disorder, such as major depressive disorder. The major conclusion from these data are that although financial distress may be a risk factor for suicidal behavior, the relationship is likely indirect and considerably less substantial than previously suspected. In addition, its relative influence is significantly less than other identified risk factors. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Breathe Soft, What Bugs through Early Windows Break?

In this issue of Cell Host & Microbe, Teo et al. (2018) explore the development of the upper airway microbiota over the first 5 years of life and provide evidence for a "critical window" of microbial exposure that contributes to chronic wheezing, a precursor to asthma.

Prevalence and predictive factors of psychological morbidity following facial injury: a prospective study of patients attending a maxillofacial outpatient clinic within a major UK city.

Adults presenting to maxillofacial surgery services are at high risk of psychological morbidity. This study examined the prevalence of depression, post-traumatic stress disorder (PTSD), anxiety, drug and alcohol use, and appearance-related distress among maxillofacial trauma outpatients over medium-term follow-up. It also explored socio-demographic and injury-related variables associated with psychological distress to inform targeted psychological screening protocols for maxillofacial trauma services. Significant associations were found between level of distress at time of injury and number of traumatic life events with levels of depression at 3 months. No significant associations were found between predictor variables and PTSD at 3 months, or with any psychiatric diagnosis at 6 months. The lack of evidence for an identifiable subgroup of patients who were at higher risk of psychological distress indicated that routine screening of all maxillofacial trauma outpatients should be offered in order to best respond to their mental health needs. The feasibility of the medical team facilitating this is challenging and should ideally be undertaken by psychologists integrated within the MDT. This study led to the funding of a clinical psychologist to provide collaborative care with the maxillofacial surgeons, resulting in brief assessment and treatment to over 600 patients in the first year of the service.

Los adultos que consultan en los servicios de cirugía máxilo-facial tienen un alto riesgo de presentar morbilidad psicológica. Este estudio examinó la prevalencia de depresión, trastorno por estrés postraumático (TEPT), ansiedad, uso de alcohol y drogas, y distrés relacionado con la apariencia entre los pacientes con trauma máxilofacial en un seguimiento ambulatorio de mediano plazo. También se exploraron variables socio-demográficas y otras relacionadas con las lesiones que se asocian con distrés psicológico para contar con protocolos de evaluación psicológica orientados a los servicios de trauma máxilo-facial. Se encontraron asociaciones significativas entre el nivel de distrés al momento de la lesión y el número de acontecimientos traumáticos con los niveles de depresión a los tres meses. En cambio, no hubo asociaciones significativas entre las variables predictoras y el TEPT a los tres meses, o con algún diagnóstico psiquiátrico a los seis meses. La falta de evidencia de un subgrupo identificable de pacientes que estuvieron en alto riesgo de distrés psicológico indicaron que se debe ofrecer la evaluación de rutina a todos los pacientes ambulatorios con trauma máxilo-facial para responder mejor a sus necesidades de salud mental. Constituye un desafío configurar un equipo médico que permita esto y lo ideal es que se forme un equipo multidisciplinario en que estén integrados psicólogos. Este estudio permitió el financiamiento de un psicólogo clínico, quien aportó atención en colaboración con los cirujanos máxilo-faciales, lo que se tradujo en una evaluación breve y el tratamiento de más de 600 pacientes durante el primer año de funcionamiento del servicio.

Les adultes hospitalisés des services de chirurgie maxillo-faciale sont à risque élevé de morbidité psychologique. Cette étude analyse la prévalence de la dépression, du syndrome de stress post-traumatique (SSPT), de l'anxiété, de la consommation de drogues et d'alcool ainsi que de la détresse liée à l'apparence chez des patients ayant subi un traumatisme maxillo-facial, avec un suivi à moyen terme en ambulatoire. Les variables socio-démographiques et liées à la lésion ainsi que la détresse psychologique sont également examinées afin de renseigner des protocoles ciblés de dépistage psychologique pour les services de chirurgie maxillo-faciale. Le niveau de détresse au moment de la lésion et le nombre d'événements traumatiques de la vie sont significativement associés aux niveaux de dépression à 3 mois. Aucune association significative n'a été trouvée entre les variables prédictives et le SSPT à 3 mois ou un diagnostic psychiatrique quel qu'il soit à 6 mois. L'identification d'un sous-groupe de patients à risque élevé de détresse psychologique est difficile : le dépistage de routine de tous les patients suivis en ambulatoire après chirurgie maxillo-faciale devrait donc être proposé afin de mieux répondre à leurs besoins en santé mentale. Créer l'équipe médicale qui le permettrait est compliqué ; idéalement, cette tâche devrait être confiée à des psychologues au sein d'une équipe pluridisciplinaire. Grâce à l'étude, le poste d'un psychologue clinicien a été financé, qui travaille en collaboration avec les chirurgiens maxillo-faciaux. C'est ainsi que plus de 600 patients ont été évalués et traités au cours de la première année.

Characterisation of gamma delta (γδ) T cell populations in patients with sepsis.

Gamma delta (γδ) T cells contribute to both innate and acquired immune responses during infection. In this pilot study, we measured the in vitro responses of γδT cell populations from patients with sepsis compared to cells from healthy subjects. We also measured production of interferon (IFN)γ. Mononuclear cells were isolated from 10 healthy control subjects and 20 patients with sepsis. Cells were cultured for 7 days with interleukin (IL)-2 plus the bisphosphonate zoledronic acid which results in indirect cell activation. Flow cytometry was used to characterise the γδT cells and enzyme immunoassay was used to measure IFNγ production. The median [range] proportion of γδT cells in healthy controls after activation was 19.2% [2.0-55.9%], compared to only 0.61% [0.1-3.6%] (P < 0.0001) in patients with sepsis. However, IFNγ levels in culture supernatants were similar in both the patients and healthy subjects. We therefore characterised the cells further by CD27 and CD45RA expression in a additional group of patients and found that the population of γδT cells was mainly CD27 negative which characterised these cells as non-proliferating effector cells. Our results suggest predominance of a non-proliferative effector subset of γδT cells in patients with sepsis, which retain functional activity and may contribute towards the host response to inflammation and infection.