- Email: [email protected]
O065 Exposing the Tobacco Industry influence in the Middle East
O063 Is a Heart attack really a wake up call? Rates and correlates of smoking cessation following PCI for Acute Coronary Syndrome (ACS)
communities. An endgame vision of ‘No More Tobacco in the 21st’ can be realized through youth-led tobacco-free generations in future. Methods: NMT 21C or ‘No More Tobacco in the 21st Century’, a global, youth-led campaign was adopted as the global symbol of a movement for the elimination of tobacco in the Conference Declaration of the International Conference on Public Health Priorities in the 21st Century in Sept. 2013, in India in presence of 600 delegates from 52 countries. The campaign envisions a global movement empowering youth with skills to counter tobacco industry tactics and advocacy skills to affect radical tobacco control policies. Youth groups will be mobilized globally to unite through innovative social media strategies, to speak up against the tobacco industry and rally for tobacco elimination. Results: Policy initiatives around tobacco-free youth will be explored and tested. Evidencebased advocacy campaigns around tobacco control will be led by youth as messengers of tobacco-free society.
Arul Baradi*1, Nick Andrianopoulos2, Angela L. Brennan2, Andrew Teh1, Alex Huang3, Stephen J. Duffy3, David J. Clark4, Louise Roberts1, Gishel New1, Melanie Freeman1 1 Cardiology, Eastern Health, Box Hill, 2Cardiology, CCRET - Monash University, 3Cardiology, Alfred Hospital, Melbourne, 4Cardiology, Austin Hospital, Heidelberg, Australia Introduction: Up to 50% of smokers continue smoking following presentation with ACS. Predictors of successful smoking cessation have not been well described in this setting. Objectives: We sought to describe smoking cessation rates as well as predictors correlated with successful smoking cessation in a large multicentre Australian registry. Methods: We identified 2804 patients undergoing PCI for ACS who were currently smoking from the Melbourne Interventional Group registry between 2005 and 2011. After exclusion of patients who did not survive to 12 months and patients in whom data was incomplete, 1888 patients were included. Smoking cessation status was identified for these patients at 30-day and 12-month follow up. Successful cessation was defined as cessation at 30 days, persisting to 12 months; Delayed cessation was defined as persistent smoking at 30 days, but cessation at 12 months; Temporary cessation was defined as cessation at 30 days but relapse at 12 months; Unsuccessful cessation was persistent smoking at 30 days and 12 months. Results: Successful cessation was achieved in 45% of patients, delayed cessation in 10%, temporary cessation in 13%, while 32% were unsuccessful. Acute presentations with ST elevation MI, cardiogenic shock and out of hospital cardiac arrest (OHCA) are associated with higher rates of successful smoking cessation. Correlates of lower rates of cessation were lower socioeconomic status (SES), diabetes, peripheral vascular disease (PVD), and history of previous myocardial infarction (MI) or PCI. Conclusion: In current tobacco use trends continue, one billion people could die in the 21st century. Youth must take this battle forward to ensure a tobacco-free society within this century. Disclosure of Interest: None Declared O065 Exposing the Tobacco Industry influence in the Middle East Wael Abd El Meguid* Technical lead, Coalition for Tobacco Control, Cairo, Egypt
Conclusion: Smoking cessation continues to be a difficult component of secondary prevention to implement following acute coronary syndrome with nearly half of all patients relapsing within the first 12 months. Disclosure of Interest: None Declared O064 Mobilizing Youth for a Tobacco-Free Society: NMT 21C (No More Tobacco in the 21st Century) Manjusha Chatterjee*1, Monika Arora2, Amit Yadav1, Radhika Shrivastav3, Nikunj Sharma3, K. Srinath Reddy4 1 Health Promotion, 2Director-Health Promotion, Public Health Foundation of India(PHFI), 3 HRIDAY, 4President, Public Health Foundation of India(PHFI), New Delhi, India Introduction: A quarter of the world’s population is youth, 10-24 years of age. Everyday 80,000 to 100,000 young people initiate smoking, most of them in the developing countries. Of 1000 teenagers who smoke today, 500 will eventually die of tobacco related diseases- 250 in their middle age and 250 in their old age. The tobacco industry spends millions on marketing strategies to reach youth and capture potential users and studies have shown that tobacco advertising is positively related to tobacco use amongst youth, especially girls. Objectives: Youth involvement is critical to the vision to de-normalize tobacco use and the industry. Globally, children and adolescents contribute energy and creativity to catalyze a movement like tobacco control, that can save lives. With 10 years of the WHO FCTC, several countries adopting targets for minimal prevalence and availability of tobacco use (Norway, Finland) and others contemplating innovative and radical strategies: tobacco-free future generations (Singapore, Tasmania), plain packaging (Australia, New Zealand), tobacco-free films, gutkha ban (India), it is imperative to build youth interest in tobacco control. Young people have been instrumental in promoting policies, programs and attitudes to reduce the use of tobacco among their peers and within their families, schools and
GHEART Vol 9/1S/2014
j
March, 2014
j
ORAL/2014 WCC Orals
Introduction: There are greater efforts to make antismoking efforts more effective especially after adoption of the international treaty FCTC. Tobacco companies recognize the impact of these strong tobacco control policies and globally they spend billions of dollars working to defeat such measures in order to maintain their corporate profits. The industry is constantly interfering in tobacco control policies through a continuous, systematic, and highly sophisticated multi-layer strategy that is constantly innovated. It resorts to a variety of tactics, employ them at different levels and violate several number of FCTC provisions. The type and combination of interference practices vary depending upon how comprehensive the legislation is and if it is effectively enforced. TI tactics can be observed as either; 1 practices aiming to advertise, promote and market tobacco or, 2 part of a strategy to weaken a government plan to improve or amend tobacco control legislation or hold back/slow down effective initiatives. Objectives: the coalition of Tobacco control and other regional partners conducted monitoring for the TI in order to screen tobacco industry vested interests that hamper development of tobacco control and health policies in order to help in planning to face them to save lives. Methods: Common tactics known to be conducted by the TI has been identified in a checklist and customized into 6 main categories mainly legislative, strategies against the following policies; Ban of Tobacco Advertisement, Promotion and Sponsorship, packaging and labeling, Smoke Free places and Taxation rise as well as using Public relations. other 3 partners has been identified to participate in that regional effort. Results:
Country
Legislative
Packaging and Labeling
Advertisement, Promotion and Sponsorship
Smoke Free implementation
Raise Taxation
Public relation
Egypt
2
6
18
9
3
4
Lebanon
4
6
22
5
2
7
Turkey
4
3
13
10
5
7
Pakistan
2
9
22
11
4
7
e17
ORAL ABSTRACTS
who discontinued on the day of surgery. Fifty-one percent of current smokers resumed smoking in the year after surgery, with 80% of these resuming within 2 weeks. Conclusion: The timing of smoking cessation prior to surgery does not impact on cardiovascular, major medical outcomes or death in the perioperative period. Therefore, physicians seeing a patient even within 8 weeks of surgery should have no reservations about encouraging abstinence. Resumption of smoking after surgery is common and indicates that there are opportunities to encourage more sustained abstinence. Disclosure of Interest: None Declared
ORAL ABSTRACTS
Conclusion: the TI harms the public health globaly by influencing the health policies. Monitoring of the intererences and exposing these tactics helps a lot in facing that danger. Monitoring of these identified TI tactics on a regular basis is an important step as tactics change or increase in intensity depending on the TI’s goal and local context. Disclosure of Interest: None Declared O066 Mechanisms of BNP interactions with neutrophil superoxide release: attenuation in acute and chronic heart failure Saifei Liu*1,2, Doan Ngo3, Cher-rin Chong1, Yuliy Chirkov3, Simon Stewart4, John Horowitz3 Cardiology, University of Adelaide, 2Cardiology, Basil Hetzel Institute, 3Cardiology, The Queen Elizabeth Hospital, Adelaide, 4Preventative Cardiology, Baker IDI Heart & Diabetes Institute, Melbourne, Australia 1
Introduction: B-type natriuretic peptide (BNP) is upregulated in heart failure (HF). Although BNP is considered cardioprotective, the net response to endogenous BNP is insufficient to restore cardiovascular hemeostasis in HF . Equivocal clinical benefit associated with administration of a recombinant BNP, nesiritide, raises the possibility of BNP resistance in HF patients. Objectives: To test the hypothesis that tissue responsiveness to BNP is impaired in patients with HF. Methods: HF patients admitted to hospital were studied both at acute presentation and after 3 weeks’ HF therapy. Isolated neutrophils were pre-incubated with either BNP (1mmol/L) or the cell permeable cGMP analog 8-pCPT-cGMP (500mmol/L). Superoxide (O2-) generated in response to N-Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) (1mmol/ L) and phorbol 12-myristate 13-acetate (PMA) (100nmol/L) was quantitated by EPR spectroscopy in both healthy and HF patients. BNP effects on fMLP induced phosphorylation of p47phox was measured via western blot. Results: In healthy subjects both PMA and fMLP -associated O2- production was inhibited by BNP by 234% (n¼32, P<0.001) and 299% (n¼23, P<0.01) respectively. This effect was not age dependent. Also, BNP suppressed fMLP induced phosphorylation of the NAD(P)H oxidase subunit p47phox on serine345 by 7617% (n¼6, P<0.01). In acute HF patients, there was significant attenuation of BNP effects (P<0.05 versus healthy subjects: Figure A) for both PMA- and fMLP-induced O2- release. Treated HF patients showed no significant sensitization of neutrophils to BNP (Figure B). Importantly, 8-pCPT-cGMP inhibited neutrophil O2- release in healthy subjects, with no attenuation of this effect in acute HF patients.
Conclusion: BNP inhibits the phosphorylation of p47phox-Ser345 and neutrophil O2release. This effect is impaired at the level of cGMP-formation in both acute and treated HF, potentially contributing to increased redox stress in HF. Disclosure of Interest: None Declared O067 STARS SiRNA knockdown altered cardiac hypertrophy associated gene expression in cardiac myoblasts and rendered cells susceptible to hydrogen peroxide induced cell death Daniel P. Fothergill, Nelson W. Chong* Department of Cardiovascular Sciences, University of Leicester, Leicester, United Kingdom Introduction: Changes in calcium signalling and reactivation of foetal genes is frequently associated with maladaptive cardiac hypertrophy and heart failure. Striated muscle activator of Rho signalling (STARS) is highly enriched in muscles. Ventricular STARS is transiently upregulated (1 hr) after pressure overload by aortic banding, prior to detectable increase in left ventricular mass. Transgenic mouse studies indicated STARS is involved in the transition from left ventricular hypertrophy to heart failure. STARS knockdown in zebrafish resulted in severe cardiac abnormalities and ectopic over-expression of STARS led to cellular hypertrophy in cardiac myoblasts. However, the mechanism of STARS is still unclear. Objectives: To examine the effect of STARS knockdown on gene expression and cytoprotection in cardiac myoblasts (H9c2) using two established cell model systems of oxidative stress [simulated ischemia-reperfusion and hydrogen peroxide (H2O2)]. Methods: Simulated ischemia-reperfusion (I/R) was done by preincubating H9c2 cells with "ischemic buffer" (which includes sodium dithionite and 2-deoxyglucose in PBS) for 1 hr. Thereafter cells were washed and incubated with complete DMEM for the indicated times. Gene expression was measured using quantitative PCR. STARS SiRNA were transfected into cells and left for 72 hrs. Non-targeting SiRNA was used as the negative control.
e18
SiRNA targeted cells were exposed to H2O2 (16 hrs) and cell viability measured using a MTT assay. Results: Simulated I/R induced STARS expression with high levels at 1-2 hr post-reperfusion (80.8 fold, n¼4) and decreased to 2.50.2 fold at 4 hr. Elimination of sodium dithionite abolished STARS induction. Hypertrophy associated genes were significantly increased during reperfusion: Anp (2.51 fold), Bnp (3.50.8 fold) and Rcan1 (5.81.1 fold). STARS SiRNA reduced STARS mRNA expression by w90% and decreased expression of b-MHC, Anp, Bnp and the anti-apoptotic gene Bcl2 by w35% (p<0.05, n¼3). STARS knockdown attenuated Rcan1 induction during I/R (5.8 to 2.5 fold, p<0.05, n¼3) but had no effect on Anp and Bnp expression. Hydrogen peroxide had no effect on cell viability at 400 mM but at 600 mM reduced viability to 76.24.3% (p<0.05, n¼3). However, STARS knockdown further reduced cell viability with H2O2 treatment (400mM, 72.71.8%; 600mM, 58.93.6%, p<0.05, n¼3). Conclusion: STARS could play a role in cardiac hypertrophy and protection against oxidant stress via the regulation of hypertrophic associated and anti-apoptotic genes. Disclosure of Interest: None Declared O068 Loss of myocardial protection against myocardial infarction in middle-aged transgenic mice overexpressing cardiac thioredoxin-1 Virginia Perez, Verónica D’Annunzio, Cristian Garmendia, Clara Llamosas, Anabella Gómez, Natalia Wulf, Eliana Cicale, Verónica Casanova, Ricardo J. Gelpi* Departament of Pathology, Institute of Cardiovascular Physiopathology, Faculty of Medicine, University of Buenos Aires, Buenos Aires, Argentina Introduction: Thioredoxin-1 (Trx1) protects the heart from ischemia/reperfusion (I/R) injury but this protection is abolished in elderly mice. However, it’s not clear whether this also occurs in middle-age, when the deleterious effects of aging are already taking place. This absence of studies in middle-aged patients is striking, since it’s known that ischemic episodes in patients start manifesting themselves during middle-age and they are not exclusive of advanced age. Objectives: The present study was designed to determine whether Trx1 expression and activity, as well as p-Akt, are altered in young and middle-aged (MA) transgenic mice overexpressing cardiac Trx1, in a way that may contribute to increased susceptibility to myocardial I/R. Methods: Langendorff-perfused hearts were subjected to 30 minutes of global ischemia followed by 120 minutes of reperfusion. We used 3 and 12 month-old male wild type (WT) mice (WT-Y, n¼8 and WT-MA, n¼7 respectively) and 3 and 12 month-old mice overexpressing cardiac Trx1 (Trx1-Y, n¼7 and Trx1-MA, n¼6 respectively). Infarct size (TTC) was measured. Trx1, Akt and p-Akt (Ser473) expression (western blot), Trx1 activity (insulin reduction assay), and nitration (western blot) were also measured. Results: Infarct size was 46.84.1% in WT-Y, 52.65.2% in WT-MA, 27.33.5% in Trx1-Y (p<0.05), and 49.16.3% in Trx1-MA. Although Trx1 expression in Trx1-Y and Trx1-MA was the same, Trx1 activity was significantly reduced by 57.05.2% (P< 0.05) in Trx1-MA. The reduction in activity in Trx1-MA was accompanied by an increase in nitration by 17.50.9% (p<0.05). The expression of p-Akt showed a significant increase during reperfusion, with respect to the pre-ischemic values in Trx1-Y (1.550.05 AU vs. 0.960.06 AU, p< 0.05), while it did not increase during reperfusion in Trx1-MA (0.900.08 AU vs. 0.870.10 AU). Conclusion: As expected, Trx1 cardiac overexpression reduced infarct size in young but not in middle-aged mice. The lack of protection in transgenic middle-aged mice even with normal Trx1 expression may be associated to decreased Trx1 activity, increased nitration and inhibition of p-Akt (Ser473). Disclosure of Interest: None Declared O069 New Zealand’s Unique Experiment: Primary prevention of rheumatic fever with a focus on schools Diana Lennon1, on behalf of Rheumatic Fever Epidemiology Group: Te Aro Moxon*1, Clair Mills2, John Malcolm3, Vicki Pennock4, Margot McLean5, Peter Reed6, Neil Poskitt7, Sue Crengle8, Catherine Jackson9, Rheumatic Fever Epidemiology Group 1 Department of Paediatrics: Child & Youth Health, The University of Auckland, Auckland, 2 Public Health, Northland District Health Board, Whangarei, 3Paediatrics, Whakatane Hospital, Whakatane, 4Paediatrics, Waikato Hospital, Waikato, 5Public Health, Hutt Valley District Health Board, Wellington, 6Children’s Research Centre, Starship Children’s Hospital, Auckland, 7 Lakes General Practice, Rotorua, 8Waitemata District Health Board, 9Public Health, Auckland District Health Board, Auckland, New Zealand Introduction: Acute rheumatic fever (ARF) incidence data is scarce. Reduction of ARF is a NZ government priority aiming for 2/3’s reduction by 2017 using hospital discharge data. Prospective surveillance process (Auckland Regional RF Register (ARRFR) provides free nurse-delivered benzathine penicillin. Objectives: An audit of ARF surveillance processes in Auckland, NZ. Application of best surveillance method in ARF endemic areas for 10 years of retrospective ARF epidemiology as baseline prior to primary prevention. Methods: Cases for 1998-2010 for children resident in Auckland aged <15 years, was compared from the ARRFR register, hospital admissions (ICD codes I00-I02), and EpiSurv (national notifiable disease database). Clinical records were reviewed; a consistent definition was applied to all data sources including grading of diagnostic certainty (www.heartfoundation. org.nz). The same methodology was applied retrospectively to other endemic areas. Results: A total of 555 confirmed (definite and probable) Auckland ARF cases aged <15 years (mean age 9.9y) were identified from the ARRFR, including 7 recurrences. Hospital
GHEART Vol 9/1S/2014
j
March, 2014
j
ORAL/2014 WCC Orals
communities. An endgame vision of ‘No More Tobacco in the 21st’ can be realized through youth-led tobacco-free generations in future. Methods: NMT 21C or ‘No More Tobacco in the 21st Century’, a global, youth-led campaign was adopted as the global symbol of a movement for the elimination of tobacco in the Conference Declaration of the International Conference on Public Health Priorities in the 21st Century in Sept. 2013, in India in presence of 600 delegates from 52 countries. The campaign envisions a global movement empowering youth with skills to counter tobacco industry tactics and advocacy skills to affect radical tobacco control policies. Youth groups will be mobilized globally to unite through innovative social media strategies, to speak up against the tobacco industry and rally for tobacco elimination. Results: Policy initiatives around tobacco-free youth will be explored and tested. Evidencebased advocacy campaigns around tobacco control will be led by youth as messengers of tobacco-free society.
Arul Baradi*1, Nick Andrianopoulos2, Angela L. Brennan2, Andrew Teh1, Alex Huang3, Stephen J. Duffy3, David J. Clark4, Louise Roberts1, Gishel New1, Melanie Freeman1 1 Cardiology, Eastern Health, Box Hill, 2Cardiology, CCRET - Monash University, 3Cardiology, Alfred Hospital, Melbourne, 4Cardiology, Austin Hospital, Heidelberg, Australia Introduction: Up to 50% of smokers continue smoking following presentation with ACS. Predictors of successful smoking cessation have not been well described in this setting. Objectives: We sought to describe smoking cessation rates as well as predictors correlated with successful smoking cessation in a large multicentre Australian registry. Methods: We identified 2804 patients undergoing PCI for ACS who were currently smoking from the Melbourne Interventional Group registry between 2005 and 2011. After exclusion of patients who did not survive to 12 months and patients in whom data was incomplete, 1888 patients were included. Smoking cessation status was identified for these patients at 30-day and 12-month follow up. Successful cessation was defined as cessation at 30 days, persisting to 12 months; Delayed cessation was defined as persistent smoking at 30 days, but cessation at 12 months; Temporary cessation was defined as cessation at 30 days but relapse at 12 months; Unsuccessful cessation was persistent smoking at 30 days and 12 months. Results: Successful cessation was achieved in 45% of patients, delayed cessation in 10%, temporary cessation in 13%, while 32% were unsuccessful. Acute presentations with ST elevation MI, cardiogenic shock and out of hospital cardiac arrest (OHCA) are associated with higher rates of successful smoking cessation. Correlates of lower rates of cessation were lower socioeconomic status (SES), diabetes, peripheral vascular disease (PVD), and history of previous myocardial infarction (MI) or PCI. Conclusion: In current tobacco use trends continue, one billion people could die in the 21st century. Youth must take this battle forward to ensure a tobacco-free society within this century. Disclosure of Interest: None Declared O065 Exposing the Tobacco Industry influence in the Middle East Wael Abd El Meguid* Technical lead, Coalition for Tobacco Control, Cairo, Egypt
Conclusion: Smoking cessation continues to be a difficult component of secondary prevention to implement following acute coronary syndrome with nearly half of all patients relapsing within the first 12 months. Disclosure of Interest: None Declared O064 Mobilizing Youth for a Tobacco-Free Society: NMT 21C (No More Tobacco in the 21st Century) Manjusha Chatterjee*1, Monika Arora2, Amit Yadav1, Radhika Shrivastav3, Nikunj Sharma3, K. Srinath Reddy4 1 Health Promotion, 2Director-Health Promotion, Public Health Foundation of India(PHFI), 3 HRIDAY, 4President, Public Health Foundation of India(PHFI), New Delhi, India Introduction: A quarter of the world’s population is youth, 10-24 years of age. Everyday 80,000 to 100,000 young people initiate smoking, most of them in the developing countries. Of 1000 teenagers who smoke today, 500 will eventually die of tobacco related diseases- 250 in their middle age and 250 in their old age. The tobacco industry spends millions on marketing strategies to reach youth and capture potential users and studies have shown that tobacco advertising is positively related to tobacco use amongst youth, especially girls. Objectives: Youth involvement is critical to the vision to de-normalize tobacco use and the industry. Globally, children and adolescents contribute energy and creativity to catalyze a movement like tobacco control, that can save lives. With 10 years of the WHO FCTC, several countries adopting targets for minimal prevalence and availability of tobacco use (Norway, Finland) and others contemplating innovative and radical strategies: tobacco-free future generations (Singapore, Tasmania), plain packaging (Australia, New Zealand), tobacco-free films, gutkha ban (India), it is imperative to build youth interest in tobacco control. Young people have been instrumental in promoting policies, programs and attitudes to reduce the use of tobacco among their peers and within their families, schools and
GHEART Vol 9/1S/2014
j
March, 2014
j
ORAL/2014 WCC Orals
Introduction: There are greater efforts to make antismoking efforts more effective especially after adoption of the international treaty FCTC. Tobacco companies recognize the impact of these strong tobacco control policies and globally they spend billions of dollars working to defeat such measures in order to maintain their corporate profits. The industry is constantly interfering in tobacco control policies through a continuous, systematic, and highly sophisticated multi-layer strategy that is constantly innovated. It resorts to a variety of tactics, employ them at different levels and violate several number of FCTC provisions. The type and combination of interference practices vary depending upon how comprehensive the legislation is and if it is effectively enforced. TI tactics can be observed as either; 1 practices aiming to advertise, promote and market tobacco or, 2 part of a strategy to weaken a government plan to improve or amend tobacco control legislation or hold back/slow down effective initiatives. Objectives: the coalition of Tobacco control and other regional partners conducted monitoring for the TI in order to screen tobacco industry vested interests that hamper development of tobacco control and health policies in order to help in planning to face them to save lives. Methods: Common tactics known to be conducted by the TI has been identified in a checklist and customized into 6 main categories mainly legislative, strategies against the following policies; Ban of Tobacco Advertisement, Promotion and Sponsorship, packaging and labeling, Smoke Free places and Taxation rise as well as using Public relations. other 3 partners has been identified to participate in that regional effort. Results:
Country
Legislative
Packaging and Labeling
Advertisement, Promotion and Sponsorship
Smoke Free implementation
Raise Taxation
Public relation
Egypt
2
6
18
9
3
4
Lebanon
4
6
22
5
2
7
Turkey
4
3
13
10
5
7
Pakistan
2
9
22
11
4
7
e17
ORAL ABSTRACTS
who discontinued on the day of surgery. Fifty-one percent of current smokers resumed smoking in the year after surgery, with 80% of these resuming within 2 weeks. Conclusion: The timing of smoking cessation prior to surgery does not impact on cardiovascular, major medical outcomes or death in the perioperative period. Therefore, physicians seeing a patient even within 8 weeks of surgery should have no reservations about encouraging abstinence. Resumption of smoking after surgery is common and indicates that there are opportunities to encourage more sustained abstinence. Disclosure of Interest: None Declared
ORAL ABSTRACTS
Conclusion: the TI harms the public health globaly by influencing the health policies. Monitoring of the intererences and exposing these tactics helps a lot in facing that danger. Monitoring of these identified TI tactics on a regular basis is an important step as tactics change or increase in intensity depending on the TI’s goal and local context. Disclosure of Interest: None Declared O066 Mechanisms of BNP interactions with neutrophil superoxide release: attenuation in acute and chronic heart failure Saifei Liu*1,2, Doan Ngo3, Cher-rin Chong1, Yuliy Chirkov3, Simon Stewart4, John Horowitz3 Cardiology, University of Adelaide, 2Cardiology, Basil Hetzel Institute, 3Cardiology, The Queen Elizabeth Hospital, Adelaide, 4Preventative Cardiology, Baker IDI Heart & Diabetes Institute, Melbourne, Australia 1
Introduction: B-type natriuretic peptide (BNP) is upregulated in heart failure (HF). Although BNP is considered cardioprotective, the net response to endogenous BNP is insufficient to restore cardiovascular hemeostasis in HF . Equivocal clinical benefit associated with administration of a recombinant BNP, nesiritide, raises the possibility of BNP resistance in HF patients. Objectives: To test the hypothesis that tissue responsiveness to BNP is impaired in patients with HF. Methods: HF patients admitted to hospital were studied both at acute presentation and after 3 weeks’ HF therapy. Isolated neutrophils were pre-incubated with either BNP (1mmol/L) or the cell permeable cGMP analog 8-pCPT-cGMP (500mmol/L). Superoxide (O2-) generated in response to N-Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) (1mmol/ L) and phorbol 12-myristate 13-acetate (PMA) (100nmol/L) was quantitated by EPR spectroscopy in both healthy and HF patients. BNP effects on fMLP induced phosphorylation of p47phox was measured via western blot. Results: In healthy subjects both PMA and fMLP -associated O2- production was inhibited by BNP by 234% (n¼32, P<0.001) and 299% (n¼23, P<0.01) respectively. This effect was not age dependent. Also, BNP suppressed fMLP induced phosphorylation of the NAD(P)H oxidase subunit p47phox on serine345 by 7617% (n¼6, P<0.01). In acute HF patients, there was significant attenuation of BNP effects (P<0.05 versus healthy subjects: Figure A) for both PMA- and fMLP-induced O2- release. Treated HF patients showed no significant sensitization of neutrophils to BNP (Figure B). Importantly, 8-pCPT-cGMP inhibited neutrophil O2- release in healthy subjects, with no attenuation of this effect in acute HF patients.
Conclusion: BNP inhibits the phosphorylation of p47phox-Ser345 and neutrophil O2release. This effect is impaired at the level of cGMP-formation in both acute and treated HF, potentially contributing to increased redox stress in HF. Disclosure of Interest: None Declared O067 STARS SiRNA knockdown altered cardiac hypertrophy associated gene expression in cardiac myoblasts and rendered cells susceptible to hydrogen peroxide induced cell death Daniel P. Fothergill, Nelson W. Chong* Department of Cardiovascular Sciences, University of Leicester, Leicester, United Kingdom Introduction: Changes in calcium signalling and reactivation of foetal genes is frequently associated with maladaptive cardiac hypertrophy and heart failure. Striated muscle activator of Rho signalling (STARS) is highly enriched in muscles. Ventricular STARS is transiently upregulated (1 hr) after pressure overload by aortic banding, prior to detectable increase in left ventricular mass. Transgenic mouse studies indicated STARS is involved in the transition from left ventricular hypertrophy to heart failure. STARS knockdown in zebrafish resulted in severe cardiac abnormalities and ectopic over-expression of STARS led to cellular hypertrophy in cardiac myoblasts. However, the mechanism of STARS is still unclear. Objectives: To examine the effect of STARS knockdown on gene expression and cytoprotection in cardiac myoblasts (H9c2) using two established cell model systems of oxidative stress [simulated ischemia-reperfusion and hydrogen peroxide (H2O2)]. Methods: Simulated ischemia-reperfusion (I/R) was done by preincubating H9c2 cells with "ischemic buffer" (which includes sodium dithionite and 2-deoxyglucose in PBS) for 1 hr. Thereafter cells were washed and incubated with complete DMEM for the indicated times. Gene expression was measured using quantitative PCR. STARS SiRNA were transfected into cells and left for 72 hrs. Non-targeting SiRNA was used as the negative control.
e18
SiRNA targeted cells were exposed to H2O2 (16 hrs) and cell viability measured using a MTT assay. Results: Simulated I/R induced STARS expression with high levels at 1-2 hr post-reperfusion (80.8 fold, n¼4) and decreased to 2.50.2 fold at 4 hr. Elimination of sodium dithionite abolished STARS induction. Hypertrophy associated genes were significantly increased during reperfusion: Anp (2.51 fold), Bnp (3.50.8 fold) and Rcan1 (5.81.1 fold). STARS SiRNA reduced STARS mRNA expression by w90% and decreased expression of b-MHC, Anp, Bnp and the anti-apoptotic gene Bcl2 by w35% (p<0.05, n¼3). STARS knockdown attenuated Rcan1 induction during I/R (5.8 to 2.5 fold, p<0.05, n¼3) but had no effect on Anp and Bnp expression. Hydrogen peroxide had no effect on cell viability at 400 mM but at 600 mM reduced viability to 76.24.3% (p<0.05, n¼3). However, STARS knockdown further reduced cell viability with H2O2 treatment (400mM, 72.71.8%; 600mM, 58.93.6%, p<0.05, n¼3). Conclusion: STARS could play a role in cardiac hypertrophy and protection against oxidant stress via the regulation of hypertrophic associated and anti-apoptotic genes. Disclosure of Interest: None Declared O068 Loss of myocardial protection against myocardial infarction in middle-aged transgenic mice overexpressing cardiac thioredoxin-1 Virginia Perez, Verónica D’Annunzio, Cristian Garmendia, Clara Llamosas, Anabella Gómez, Natalia Wulf, Eliana Cicale, Verónica Casanova, Ricardo J. Gelpi* Departament of Pathology, Institute of Cardiovascular Physiopathology, Faculty of Medicine, University of Buenos Aires, Buenos Aires, Argentina Introduction: Thioredoxin-1 (Trx1) protects the heart from ischemia/reperfusion (I/R) injury but this protection is abolished in elderly mice. However, it’s not clear whether this also occurs in middle-age, when the deleterious effects of aging are already taking place. This absence of studies in middle-aged patients is striking, since it’s known that ischemic episodes in patients start manifesting themselves during middle-age and they are not exclusive of advanced age. Objectives: The present study was designed to determine whether Trx1 expression and activity, as well as p-Akt, are altered in young and middle-aged (MA) transgenic mice overexpressing cardiac Trx1, in a way that may contribute to increased susceptibility to myocardial I/R. Methods: Langendorff-perfused hearts were subjected to 30 minutes of global ischemia followed by 120 minutes of reperfusion. We used 3 and 12 month-old male wild type (WT) mice (WT-Y, n¼8 and WT-MA, n¼7 respectively) and 3 and 12 month-old mice overexpressing cardiac Trx1 (Trx1-Y, n¼7 and Trx1-MA, n¼6 respectively). Infarct size (TTC) was measured. Trx1, Akt and p-Akt (Ser473) expression (western blot), Trx1 activity (insulin reduction assay), and nitration (western blot) were also measured. Results: Infarct size was 46.84.1% in WT-Y, 52.65.2% in WT-MA, 27.33.5% in Trx1-Y (p<0.05), and 49.16.3% in Trx1-MA. Although Trx1 expression in Trx1-Y and Trx1-MA was the same, Trx1 activity was significantly reduced by 57.05.2% (P< 0.05) in Trx1-MA. The reduction in activity in Trx1-MA was accompanied by an increase in nitration by 17.50.9% (p<0.05). The expression of p-Akt showed a significant increase during reperfusion, with respect to the pre-ischemic values in Trx1-Y (1.550.05 AU vs. 0.960.06 AU, p< 0.05), while it did not increase during reperfusion in Trx1-MA (0.900.08 AU vs. 0.870.10 AU). Conclusion: As expected, Trx1 cardiac overexpression reduced infarct size in young but not in middle-aged mice. The lack of protection in transgenic middle-aged mice even with normal Trx1 expression may be associated to decreased Trx1 activity, increased nitration and inhibition of p-Akt (Ser473). Disclosure of Interest: None Declared O069 New Zealand’s Unique Experiment: Primary prevention of rheumatic fever with a focus on schools Diana Lennon1, on behalf of Rheumatic Fever Epidemiology Group: Te Aro Moxon*1, Clair Mills2, John Malcolm3, Vicki Pennock4, Margot McLean5, Peter Reed6, Neil Poskitt7, Sue Crengle8, Catherine Jackson9, Rheumatic Fever Epidemiology Group 1 Department of Paediatrics: Child & Youth Health, The University of Auckland, Auckland, 2 Public Health, Northland District Health Board, Whangarei, 3Paediatrics, Whakatane Hospital, Whakatane, 4Paediatrics, Waikato Hospital, Waikato, 5Public Health, Hutt Valley District Health Board, Wellington, 6Children’s Research Centre, Starship Children’s Hospital, Auckland, 7 Lakes General Practice, Rotorua, 8Waitemata District Health Board, 9Public Health, Auckland District Health Board, Auckland, New Zealand Introduction: Acute rheumatic fever (ARF) incidence data is scarce. Reduction of ARF is a NZ government priority aiming for 2/3’s reduction by 2017 using hospital discharge data. Prospective surveillance process (Auckland Regional RF Register (ARRFR) provides free nurse-delivered benzathine penicillin. Objectives: An audit of ARF surveillance processes in Auckland, NZ. Application of best surveillance method in ARF endemic areas for 10 years of retrospective ARF epidemiology as baseline prior to primary prevention. Methods: Cases for 1998-2010 for children resident in Auckland aged <15 years, was compared from the ARRFR register, hospital admissions (ICD codes I00-I02), and EpiSurv (national notifiable disease database). Clinical records were reviewed; a consistent definition was applied to all data sources including grading of diagnostic certainty (www.heartfoundation. org.nz). The same methodology was applied retrospectively to other endemic areas. Results: A total of 555 confirmed (definite and probable) Auckland ARF cases aged <15 years (mean age 9.9y) were identified from the ARRFR, including 7 recurrences. Hospital
GHEART Vol 9/1S/2014
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March, 2014
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ORAL/2014 WCC Orals