National Ambulance Surveillance System: A novel method using coded Australian ambulance clinical records to monitor self-harm and mental health-related morbidity.

Self-harm and mental health are inter-related issues that substantially contribute to the global burden of disease. However, measurement of these issues at the population level is problematic. Statistics on suicide can be captured in national cause of death data collected as part of the coroner's review process, however, there is a significant time-lag in the availability of such data, and by definition, these sources do not include non-fatal incidents. Although survey, emergency department, and hospitalisation data present alternative information sources to measure self-harm, such data do not include the richness of information available at the point of incident. This paper describes the mental health and self-harm modules within the National Ambulance Surveillance System (NASS), a unique Australian system for monitoring and mapping mental health and self-harm. Data are sourced from paramedic electronic patient care records provided by Australian state and territory-based ambulance services. A team of specialised research assistants use a purpose-built system to manually scrutinise and code these records. Specific details of each incident are coded, including mental health symptoms and relevant risk indicators, as well as the type, intent, and method of self-harm. NASS provides almost 90 output variables related to self-harm (i.e., type of behaviour, self-injurious intent, and method) and mental health (e.g., mental health symptoms) in the 24 hours preceding each attendance, as well as demographics, temporal and geospatial characteristics, clinical outcomes, co-occurring substance use, and self-reported medical and psychiatric history. NASS provides internationally unique data on self-harm and mental health, with direct implications for translational research, public policy, and clinical practice. This methodology could be replicated in other countries with universal ambulance service provision to inform health policy and service planning.

'Extreme personal stress' and 'a lot of pain': Exploring the physical, mental and social contexts of extramedical pharmaceutical opioid use in Australian paramedic case descriptions.

INTRODUCTION AND AIMS: Despite growing concerns over the extramedical use of pharmaceutical opioids, few studies have focused on the contexts that shape this use or how it might lead to harm. There is also limited research examining how contexts might vary across different opioids, such as those with well-established abuse liabilities (e.g. oxycodone) and newer 'atypical' opioids (e.g. tapentadol). We aimed to address these gaps. DESIGN AND METHODS: We analysed Australian paramedic case descriptions for tapentadol cases (n = 82) and a representative sample of oxycodone cases (n = 82) from a 6-year period (2013-2018). We used framework analysis to identify 'contexts' shaping extramedical use from cases where use of tapentadol or oxycodone was assessed to have significantly contributed to the attendances. RESULTS: Demographically, case descriptions from both drug types were broadly similar (approximately 55% female, 44% were 35-54 years old), as were the contextual factors. The most prevalent contexts common to both oxycodone and tapentadol cases were psychological distress, physical pain and social stressors. Suicidal intent was present across multiple contexts. DISCUSSION AND CONCLUSIONS: This study is one of the first to explore the contexts of extramedical pharmaceutical opioid use leading to acute harm. Our analysis found patients in complex, emergency situations, seeking rapid relief from physical pain, psychological distress, social issues and/or suicidal thoughts. These data highlight the complex needs of those experiencing harm from extramedical pharmaceutical opioid use, regardless of opioid type, and the importance of contextual factors shaping both use and subsequent harm.

The impact of legislation on acute synthetic cannabinoid harms resulting in ambulance attendance.

BACKGROUND: Synthetic cannabinoid receptor agonists (SCRAs) have been challenging current drug policy due to the rapid emergence of new variants, and their propensity for acute harm. In Australia, as in other parts of the world, multiple regulatory changes have occurred in response to these new psychoactive compounds, and population surveys indicate use is declining. This suggests that related harms would also be declining. We examined the impact of drug legislative changes on acute SCRA-related harms resulting in ambulance attendance. Secondary aims were to describe patient and attendance characteristics. METHODS: A retrospective analysis of coded ambulance attendance data from Victoria, Australia (January 2014-December 2018). Interrupted time-series was used to analyse the trajectories of SCRA-related attendances relative to legislative changes. RESULTS: During the study period, 3727 SCRA-related ambulance attendances were identified. There was an upward trend in attendances following legislation scheduling specific SCRAs in Victoria in October 2016 (slope = 1.31, 95% CI 1.17, 1.45). A downward trend in attendances followed 'blanket' legislation targeting all new psychoactive substances, implemented in Victoria in November 2017 (slope = -1.87, 95% CI -2.27, -1.46). Patient median age was 33 years, 80.5% were male, co-occurring substance use was identified in 30.4% cases, and 15.2% had >1 SCRA-related attendance. Overall, 69.4% cases were transported to hospital, with the odds of transport to hospital increasing each year from 2016. CONCLUSION: This study represents a population-level examination of the impact of drug policy on acute SCRA-related harms resulting in ambulance attendance. Scheduling of specific SCRAs was associated with a spike in attendances, likely due to the introduction of more harmful variants in the drug market. Blanket legislation was associated with a reduction in SCRA-related attendances, however, a corresponding increase in cases transported to hospital indicates a greater severity of harm that may have been inadvertently promoted by this policy.

The National Ambulance Surveillance System: A novel method for monitoring acute alcohol, illicit and pharmaceutical drug related-harms using coded Australian ambulance clinical records.

Although harmful consumption of alcohol and other drugs (both illicit and pharmaceutical) significantly contribute to global burden of disease, not all harms are captured within existing morbidity data sources. Indeed, harms occurring in the community may be missed or under-reported. This paper describes the National Ambulance Surveillance System, a unique Australian system for monitoring and mapping acute harms related to alcohol and other drug consumption. Data are sourced from paramedic electronic patient care records provided by ambulance services from across Australia. Coding occurs in a purpose-built system, by a team of specialised research assistants. Alcohol, and specific illicit and pharmaceutical drugs, rather than broad drug classes, are manually coded and the dataset is reviewed and cleaned prior to analysis. The National Ambulance Surveillance System is an ongoing, dynamic surveillance system of alcohol and other drug-related harms across Australia. The data includes more than 140 output variables per attendance, including individual substances, demographics, temporal, geospatial, and clinical data (e.g., Glasgow Coma Scale score, naloxone provision and response, outcome of attendance). The National Ambulance Surveillance System is an internationally unique population-level surveillance system of acute harms arising from alcohol and other drug consumption. Dissemination of National Ambulance Surveillance System data has been used to inform and evaluate policy approaches and potential points of intervention, as well as guide workforce development needs and clinical practice at the local and national level. This methodology could be replicated in other countries.

Aggression and violence at ambulance attendances where alcohol, illicit and/or pharmaceutical drugs were recorded: A 5-year study of ambulance records in Victoria, Australia.

BACKGROUND: This study describes the frequency and characteristics of aggression and/or violence in ambulance attendances involving alcohol, illicit and/or pharmaceutical drug use in Victoria, Australia between January 2012 and January 2017. METHODS: Patient characteristics, context, and substance use involvement in ambulance attendances were examined to determine associations with attendances where aggression and/or violence was recorded. RESULTS: There were 205,178 ambulance attendances where use of alcohol, pharmaceutical drugs or illicit substances contributed to the reason for the attendance. Paramedics recorded acts of aggression and/or violence in 11,813 (5.76 %) of these attendances. Aggression/violence was more likely to be recorded in certain contexts. Compared with attendances where aggression/violence was not recorded, attendances where aggression/violence was recorded were significantly more likely to involve younger and male patients, and occur on Friday and Saturday nights. Alcohol intoxication was involved in more than half of attendances where aggression/violence was recorded, and was almost twice as prevalent as those involving illicit drug use where aggression/violence was recorded. This pattern was consistent across all hours, high-alcohol hours only, by metropolitan/regional location, and by police co-attendance. CONCLUSIONS: Aggression and violence are frequently recorded in ambulance attendances involving alcohol, pharmaceutical drugs or illicit substances, and, most often involve alcohol. This violence poses a recurring threat to the health and safety of paramedics, bystanders, and patients. Greater priority should be given to reducing alcohol-related violence through evidence-based policy measures targeting high-risk groups (e.g. young adult males) and contexts (e.g. weekends, late at night) where harm is most likely to occur.

Queensland Alcohol-related violence and Night Time Economy Monitoring project (QUANTEM): a study protocol.

BACKGROUND: Alcohol-related harm is a substantial burden on the community in Australia and internationally, particularly harm related to risky drinking practices of young people in the night-time economy. This protocol paper describes a study that will report on the changes in a wide range of health and justice outcome measures associated with major policy changes in the state of Queensland, Australia. A key element includes trading hours restrictions for licensed premises to 2 am for the state and 3 am in Safe Night Precincts (SNPs). Other measures introduced include drinks restrictions after midnight, increased patron banning measures for repeat offenders, mandatory ID scanning of patrons in late-night venues, and education campaigns. METHODS: The primary aim of the study is to evaluate change in the levels of harm due to these policy changes using administrative data (e.g., police, hospital, ambulance, and court data). Other study elements will investigate the impact of the Policy by measuring foot traffic volume in SNPs, using ID scanner data to quantify the volume of people entering venues and measure the effectiveness of banning notices, using patron interviews to quantify the levels of pre-drinking, intoxication and illicit drug use within night-time economy districts, and to explore the impacts of the Policy on business and live music, and costs to the community. DISCUSSION: The information gathered through this project aims to evaluate the effectiveness of the Policy and to draw on these findings to inform future prevention and enforcement approaches by policy makers, police, and venue staff.

Long-term use of a 4% sodium cromoglicate cutaneous emulsion in the treatment of moderate to severe atopic dermatitis in children.

INTRODUCTION: Sodium cromoglicate (SCG), a chromone with anti-inflammatory, anti-itch and anti-allergic properties. We report a long-term study of a 4% aqueous solution of SCG in children with moderate to severe atopic dermatitis (AD). MATERIALS AND METHODS: Children aged 1 to 12 years with AD were entered into a 12-week randomised clinical trial (RCT), followed by 12 months open treatment with known 4% SCG emulsion (Altocrom®). Primary endpoint was change in SCORAD score. Secondary endpoints included symptom severity, Quality of Life, concomitant treatment usage, global assessments. RESULTS: One hundred and seventy-seven subjects entered, 118 treated with 4% SCG emulsion and 59 with vehicle: 128 completed 12 months in open study. SCORAD score reduced during RCT by -15.3 (-33%) on 4% SCG emulsion and -18.0 (-39%) on vehicle: p = 0.2331. After 12 months reduction was 56%. No secondary endpoint showed differences between treatments during RCT. Thirty-two subjects reported treatment related events during RCT and open trial. Eleven (7%) reported application site discomfort. Most were reported as mild and most resolved without intervention and the study drug was stopped in one case only. CONCLUSIONS: SCG 4% cutaneous emulsion was well tolerated in children treated for 15 months.

Alcohol and burden of disease in Australia: the challenge in assessing consumption.

OBJECTIVE: Alcohol consumption is one of the major avoidable risk factors for disease, illness and injury in the Australian community. Population health scientists and economists use estimates of alcohol consumption in burden of disease frameworks to estimate the impact of alcohol on disease, illness and injury. This article highlights challenges associated with estimating alcohol consumption in these models and provides a series of recommendations to improve estimates. METHODS: Key challenges in measuring alcohol consumption at the population level are identified and discussed with respect to how they apply to burden of disease frameworks. RESULTS: Methodological advances and limitations in the estimation of alcohol consumption are presented with respect to use of survey data, population distributions of alcohol consumption, consideration of 'patterns' of alcohol use including 'bingeing', and capping exposure. Key recommendations for overcoming these limitations are provided. Implications and conclusion: Alcohol-related burden has a significant impact on the health of the Australian population. Improving estimates of alcohol related consumption will enable more accurate estimates of this burden to be determined to inform future alcohol policy by legislators.

Trends in alcohol-attributable morbidity and mortality for Victoria, Australia from 2000/01 to 2009/10.

BACKGROUND: To examine trends in alcohol-attributable morbidity (AAMorb) (2000/01-2009/10) and mortality (AAMort) (2000-07) by age, sex and region. METHODS: Time-series analyses of population data for Victoria, Australia. We used joinpoint regression to quantify trends by estimating quarterly percent change (QPC) for rates of morbidity and mortality. We present the average QPC (AQPC) as a weighted average of QPCs. A test of parallelism was used to examine pairwise differences. RESULTS: AAMorb increased significantly over time for Victoria (AQPC = 1.0%, 95% confidence interval 0.8-1.2). While females (1.6, 1.1-2.0), age groups 25-44 (1.0, 0.9-1.1) and 45-64 (1.2, 0.2-2.2), and metropolitan population (1.2, 0.5-1.9) were broad subgroups more at risk, multivariate analysis detected specific increases for metropolitan females aged 15-44 (1.8, 1.0-2.6) and 45+ (1.6, 0.2-3.0). Relatively greater increases in morbidity among metropolitan subgroups were widespread. AAMort remained stable for Victoria and for most subgroups, although significant declines in mortality were specifically experienced by metropolitan 15-24 (-2.0, -2.9 to -1.0) and 25-44 (-1.0, -1.7 to -0.3) age groups, and by regional males aged 45+ (-0.8, -1.3 to -0.3). Metropolitan males aged 45+ were a special high-risk population. DISCUSSION: Our study has identified overlooked subgroups as being at increasing risk for alcohol-attributable chronic harm necessitating their inclusion in future policies for harm reduction.

Relationships between age of puberty onset and height at age 18 years in girls and boys.

BACKGROUND: Changes during puberty may influence final adult height. Height is related to multiple health conditions, including lung function. We investigated the association between the age of onset of five puberty events and height at age 18 years, analyzing boys and girls separately. METHODS: Of 1456 children recruited into the Isle of Wight birth cohort (1989-1990), 1313 were followed up at age 18 years. Height was measured, and age of pubertal onset was collected at age 18 years. Cluster analysis was performed on the five puberty events in boys and girls and linear regression was applied with the clusters predicting height at age 18 years. Individual linear regression analyses assessed the age of onset of each pubertal event as a potential predictor for height at age 18 years. RESULTS: Of the 1313 children followed up at age 18 years, 653 were males and 660 were females. All puberty variables had high internal consistency. In girls, earlier age of menarche, breast development, and growth spurt were related to shorter height. In boys, earlier age of growth spurt and slower progression through puberty were related to taller height at age 18 years. CONCLUSIONS: Given that boys and girls may have opposing associations between pubertal timing and adult height and that height is an important predictor of lung function, the effect of pubertal timing on respiratory health should be explored.

The social location of heavy episodic alcohol consumption in the Victorian population.

INTRODUCTION AND AIMS: To examine heavy episodic drinking across demographic subgroups to identify where heavy episodic drinking is socially located in an Australian state. DESIGN AND METHODS: Cross-sectional survey, 2483 adult Victorians using Computer Assisted Telephone Interviewing. Two measures of heavy drinking were used: (i) heavy episodic drinking-more than five standard drinks at least weekly; and (ii) typically heavy drinking-50% or more of all drinking occasions involving consumption of 5+ standard drinks. Associations between heavy episodic drinking and eight potential sociodemographic correlates (gender, age, education, income, marital status, ethnic origin, religion and geographical remoteness) were examined. RESULTS: There were few significant correlates of heavy episodic drinking apart from gender and age, once gender and age were controlled. Men were more likely to report heavy episodic drinking than women, but this was attenuated in the measure of typically heavy drinking, suggesting that women reporting heavy episodic drinking were more likely to typically drink that much when they drank. Younger people were more likely to report weekly heavy episodic drinking and more likely to report engaging in this pattern on at least half of their drinking occasions, and this was also true for those unmarried or in de facto relationships. Those of Asian background were less likely to report heavy drinking. In multivariate analysis, the remaining sociodemographic variables were largely unrelated to the drinking measures. DISCUSSION AND CONCLUSION: The study clearly shows that the prevalence of heavy episodic drinking varies particularly across gender and age groups in Victoria. These variations appear to hold across key sociodemographic variables such as income and education.

Alcohol intoxication in the context of major public holidays, sporting and social events: a time-series analysis in Melbourne, Australia, 2000-2009.

AIMS: To assess the relationship between ambulance attendances, emergency department (ED) presentations and hospital admissions for acute alcohol intoxication and the timing of public holidays, sporting and social events. DESIGN: Time-series analysis was used to explore trends in intoxication in the context of major events. SETTING: Population of Melbourne, Victoria, Australia between 2000 and 2009. PARTICIPANTS: All patients attended by ambulance, presenting to hospital EDs, or admitted to hospital who were classified as acutely alcohol intoxicated. MEASUREMENT: Analysis of daily numbers of presentations for acute alcohol intoxication associated with major events were undertaken, including lead and lag effects. Analyses controlled for day of week and month of year to address temporal and seasonal variations. FINDINGS: Alcohol intoxication presentations were significantly elevated the day before all public holidays, with intoxication cases on the day of public holidays only higher on New Year's Day (ambulance 6.57, 95% confidence intervals (CI): 3.4-9.74; ED 3.34, 95% CI: 1.28-5.4) and ANZAC Day (ambulance 3.71, 95% CI: 0.68-6.75). The Australian Football League (AFL) Grand Final (ED 2.37, 95% CI: 0.55-4.19), Commonwealth Games (ED 2.45, 95% CI: 0.6-4.3) and Melbourne Cup Day (ambulance 6.14, 95% CI: 2.42-9.85) represented the sporting events with significant elevations in acute intoxication requiring medical attention. The last working day before Christmas was the only social event where a significant increase in acute intoxication occurred (ambulance 8.98, 95% CI: 6.8-11.15). CONCLUSIONS: Acute alcohol intoxication cases requiring ambulance, emergency department and hospital in-patient treatment increase substantially on the day preceding public holidays and other major social events.

Multifaceted allergen avoidance during infancy reduces asthma during childhood with the effect persisting until age 18 years.

BACKGROUND: Asthma is a chronic disease that often starts in childhood. The key risk factors are a child's environment and their genetic characteristics. The aim of this study was to evaluate the efficacy of environmental modification in the first 12 months of life on the prevalence of asthma in high-risk individuals. METHODS: Children (n=120) considered at high risk of allergic disorders (either dual heredity or single heredity and a high cord total IgE), were enrolled in a single-blinded, randomised controlled trial. Infants in the intervention arm were either breast fed with the mother on a low allergen diet or given an extensively hydrolysed formula. Exposure to house dust mite allergen was reduced. The control group followed standard advice. Children were assessed at ages 1, 2, 4, 8 and 18 years for the presence of asthma and atopy. RESULTS: At 18 years of age, there was a significantly lower prevalence of asthma in the prevention group compared with the control group (OR: 0.23, 95% CI 0.08 to 0.70, p=0.01), primarily due to asthma that developed during childhood but persisted until age 18 years. Repeated-measure analysis showed that there was an overall reduction in asthma prevalence from 1 to 18 years (OR: 0.51, CI 0.32 to 0.81, p=0.04). Prevalence of atopy was not significantly different between the two groups at age 18. CONCLUSION: Comprehensive allergen avoidance in the first year of life is effective in preventing asthma onset in individuals considered at high risk due to heredity. The effect occurs in the early years, but persists through to adulthood.

The effect of parental allergy on childhood allergic diseases depends on the sex of the child.

BACKGROUND: The parent-of-origin effect is important in understanding the genetic basis of childhood allergic diseases and improving our ability to identify high-risk children. OBJECTIVE: We sought to investigate the parent-of-origin effect in childhood allergic diseases. METHODS: The Isle of Wight Birth Cohort (n= 1456) has been examined at 1, 2, 4, 10, and 18 years of age. Information on the prevalence of asthma, eczema, rhinitis, and environmental factors was obtained by using validated questionnaires. Skin prick tests were carried out at ages 4, 10, and 18 years, and total IgE measurement was carried out at 10 and 18 years. Parental history of allergic disease was assessed soon after the birth of the child, when maternal IgE levels were also measured. Prevalence ratios (PRs) and their 95% CIs were estimated, applying log-linear models adjusted for confounding variables. RESULTS: When stratified for sex of the child, maternal asthma was associated with asthma in girls (PR, 1.91; 95% CI, 1.34-2.72; P= .0003) but not in boys (PR, 1.29; 95% CI, 0.85-1.96; P= .23), whereas paternal asthma was associated with asthma in boys (PR, 1.99; 95% CI, 1.42-2.79; P< .0001) but not in girls (PR, 1.03; 95% CI, 0.59-1.80; P= .92). Maternal eczema increased the risk of eczema in girls (PR, 1.92; 95% CI, 1.37-2.68; P= .0001) only, whereas paternal eczema did the same for boys (PR, 2.07; 95% CI, 1.32-3.25; P = .002). Similar trends were observed when the effect of maternal and paternal allergic disease was assessed for childhood atopy and when maternal total IgE levels were related to total IgE levels in children at ages 10 and 18 years. CONCLUSIONS: The current study indicates a sex-dependent association of parental allergic conditions with childhood allergies, with maternal allergy increasing the risk in girls and paternal allergy increasing the risk in boys. This has implications for childhood allergy prediction and prevention.

Protein kinase D isoforms are dispensable for integrin-mediated lymphocyte adhesion and homing to lymphoid tissues.

Leukocyte function-associated antigen-1 (LFA-1) and very late antigen-4 (VLA-4) integrins are essential for lymphocyte adhesion, trafficking and effector functions. Protein kinase D (PKD) has previously been implicated in lymphocyte integrin regulation through regulation of Rap1 activity. However, the true role of PKD in integrin regulation in primary lymphocytes has not previously been investigated. The major PKD isoform in lymphocytes is PKD2. Here we employed PKD2-deficient mice, a specific PKD kinase inhibitor, as well as PKD-null DT40 B cells to investigate the role of PKD in integrin regulation in lymphocytes. We report that PKD2-deficient lymphocytes bound normally to integrin ligands in static and shear flow adhesion assays. They also homed normally to lymphoid organs after adoptive transfer into wild-type mice. DT40 B cells devoid of any PKD isoforms and primary lymphocytes pretreated with a specific PKD inhibitor bound normally to integrin ligands, indicating that multiple PKD isoforms do not redundantly regulate lymphocyte integrins. In addition, PKD2-deficient lymphocytes, as well as DT40 cells devoid of any PKD isoforms, could activate Rap1 in response to B-cell receptor ligation or phorbol ester treatment. Together, these results show that the PKD family does not play a critical role in lymphocyte integrin-mediated cell adhesion or lymphocyte trafficking in vivo.

Characterisation of asthma that develops during adolescence; findings from the Isle of Wight Birth Cohort.

BACKGROUND: Understanding of adolescent-onset asthma remains limited. We sought to characterise this state and identify associated factors within a longitudinal birth cohort study. METHODS: The Isle of Wight Whole Population Birth Cohort was recruited in 1989 (N=1456) and characterised at 1, 2, 4, 10 and 18-years. "Adolescent-onset asthma" was defined as asthma at age 18 without prior history of asthma, "persistent-adolescent asthma" as asthma at both 10 and 18 and "never-asthma" as those without asthma at any assessment. RESULTS: Adolescent-onset asthma accounted for 28.3% of asthma at 18-years and was of similar severity to persistent-adolescent asthma. Adolescent-onset asthmatics showed elevated bronchial hyper-responsiveness (BHR) and atopy at 10 and 18 years. BHR in this group at 10 was intermediate to that of never-asthmatics and persistent-adolescent asthma. By 18 their BHR, bronchodilator reversibility and sputum eosinophilia was greater than never-asthmatics and comparable to persistent-adolescent asthma. At 10, males who later developed adolescent-onset asthma had reduced FEV(1) and FEF(25-75), while females had normal lung function but then developed impaired FEV(1) and FEF(25-75) in parallel with adolescent asthma. Factors independently associated with adolescent-onset asthma included atopy at 10 (OR=2.35; 95% CI=1.08-5.09), BHR at 10 (3.42; 1.55-7.59), rhinitis at 10 (2.35; 1.11-5.01) and paracetamol use at 18-years (1.10; 1.01-1.19). CONCLUSIONS: Adolescent-onset asthma is associated with significant morbidity. Predisposing factors are atopy, rhinitis and BHR at age 10 while adolescent paracetamol use is also associated with this state. Awareness of potentially modifiable influences may offer avenues for mitigating this disease state.

Protein kinase D2 has a restricted but critical role in T-cell antigen receptor signalling in mature T-cells.

PKD (protein kinase D) 2 is a serine/threonine kinase activated by diacylglycerol in response to engagement of antigen receptors in lymphocytes. To explore PKD2 regulation and function in TCR (T-cell antigen receptor) signal transduction we expressed TCR complexes with fixed affinity for self antigens in the T-cells of PKD2-null mice or mice deficient in PKD2 catalytic activity. We also developed a single cell assay to quantify PKD2 activation as T-cells respond to developmental stimuli or engagement of α/ß TCR complexes in vivo. Strikingly, PKD2 loss caused increases in thymic output, lymphadenopathy and splenomegaly in TCR transgenic mice. The precise magnitude and timing of PKD2 activation during T-cell development is thus critical to regulate thymic homoeostasis. PKD2-null T-cells that exit the thymus have a normal transcriptome, but show a limited and abnormal transcriptional response to antigen. Transcriptional profiling reveals the full consequences of PKD2 loss and maps in detail the selective, but critical, function for PKD2 in signalling by α/ß mature TCR complexes in peripheral T-cells.

Angiotensin II acts through the angiotensin 1a receptor to upregulate pendrin.

Pendrin is an anion exchanger expressed in the apical regions of B and non-A, non-B intercalated cells. Since angiotensin II increases pendrin-mediated Cl(-) absorption in vitro, we asked whether angiotensin II increases pendrin expression in vivo and whether angiotensin-induced hypertension is pendrin dependent. While blood pressure was similar in pendrin null and wild-type mice under basal conditions, following 2 wk of angiotensin II administration blood pressure was 31 mmHg lower in pendrin null than in wild-type mice. Thus pendrin null mice have a blunted pressor response to angiotensin II. Further experiments explored the effect of angiotensin on pendrin expression. Angiotensin II administration shifted pendrin label from the subapical space to the apical plasma membrane, independent of aldosterone. To explore the role of the angiotensin receptors in this response, pendrin abundance and subcellular distribution were examined in wild-type, angiotensin type 1a (Agtr1a) and type 2 receptor (Agtr2) null mice given 7 days of a NaCl-restricted diet (< 0.02% NaCl). Some mice received an Agtr1 inhibitor (candesartan) or vehicle. Both Agtr1a gene ablation and Agtr1 inhibitors shifted pendrin label from the apical plasma membrane to the subapical space, independent of the Agtr2 or nitric oxide (NO). However, Agtr1 ablation reduced pendrin protein abundance through the Agtr2 and NO. Thus angiotensin II-induced hypertension is pendrin dependent. Angiotensin II acts through the Agtr1a to shift pendrin from the subapical space to the apical plasma membrane. This Agtr1 action may be blunted by the Agtr2, which acts through NO to reduce pendrin protein abundance.

Protein kinase C mediates platelet secretion and thrombus formation through protein kinase D2.

Platelets are highly specialized blood cells critically involved in hemostasis and thrombosis. Members of the protein kinase C (PKC) family have established roles in regulating platelet function and thrombosis, but the molecular mechanisms are not clearly understood. In particular, the conventional PKC isoform, PKCα, is a major regulator of platelet granule secretion, but the molecular pathway from PKCα to secretion is not defined. Protein kinase D (PKD) is a family of 3 kinases activated by PKC, which may represent a step in the PKC signaling pathway to secretion. In the present study, we show that PKD2 is the sole PKD member regulated downstream of PKC in platelets, and that the conventional, but not novel, PKC isoforms provide the upstream signal. Platelets from a gene knock-in mouse in which 2 key phosphorylation sites in PKD2 have been mutated (Ser707Ala/Ser711Ala) show a significant reduction in agonist-induced dense granule secretion, but not in α-granule secretion. This deficiency in dense granule release was responsible for a reduced platelet aggregation and a marked reduction in thrombus formation. Our results show that in the molecular pathway to secretion, PKD2 is a key component of the PKC-mediated pathway to platelet activation and thrombus formation through its selective regulation of dense granule secretion.