- Email: [email protected]
Construction of a cell model for dynamically detecting process of autophagy in alveolar macrophages
e56
being the main parameter used to guide upper limb and trunk exercise prescription and progression. 44% of the respondents reported screening for sternal instability, with 10% using an objective measure (i.e. sternal instability scale (SIS), or imaging) as part of their assessment. If sternal instability was detected, the majority of respondents based the prescription of sternal precautions and exercise guidelines on clinical experience. Discussion: There is significant variation in practice with respect to sternal precautions as well as the prescription and progression of upper limb and trunk exercises in outpatient cardiac rehabilitation programs in Australia. Further research is warranted in order to establish evidence-based upper limb and trunk exercise guidelines for patients following cardiac surgery via a median sternotomy in order to facilitate recovery and attain optimal patient outcomes. http://dx.doi.org/10.1016/j.hlc.2014.12.117 Construction of a cell model for dynamically detecting process of autophagy in alveolar macrophages Qing Geng *, Tao Fan Department of Thoracic Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, China * Corresponding author. Introduction: Autophagy is a constitutive regulatory means of cellular homeostasis involved in diverse physiological and pathological events. Autophagy in macrophages is important for maturation, activation, polarisation, and regulation of cytokine production. Alveolar macrophages play a critical role in chronic obstructive pulmonary disease (COPD), ischaemia/reperfusion injury, infectious diseases and neurodegenerative disorders. The numbers of alveolar macrophages are markedly increased in the lungs of patients with COPD as a result of increased recruitment, proliferation and survival. Alveolar macrophages secrete many inflammatory mediators, oxidants, proteins and proteinases in response to cigarette smoke extract and other stimuli. We developed a new cell model for dynamically detecting process of autophagy in alveolar macrophages. Methods: The PAsGreen2 - N1 - LC3 plasmid was constructed and transfected into alveolar macrophagyes line (NR8383 cells) with transfection reagent. The stable transfectants were screened by G418. The PAsGreen2 - N1 - LC3 expression was analyzed by fluorescence microscope, Western blot and PCR technology. Results: Selected by G418, 4 transfected cell lines showed high level of AsGreen2-N1-LC3 expression, as demonstrated by Western blot analysis. More than 92% cells showed positive fluorescent signals under inverted fluorescence microscope. The result of Western blot confirmed that the amount of protein expressed by LC3 genes in NR8383 PAsGreen2 N1 - LC3 cells is 3.11628 times than that of normal NR8383 cells. The transcription of LC3 in NR8383 PAsGreen2 - N1 LC3 cells was relatively 5.188611 times than that of normal NR8383 cells detected by PCR technique. Discussion: An alveolar macrophage line (NR8383 cell line) stably expressing PAsGreen2 - N1 - LC3 was constructed successfully, which can be used for dynamic detection of
Abstracts
autophagy process in alveolar macrophagyes. Besides, as a new cell model, it can be used for further study of autophagy in the pathological processes of alveolar macrophages. http://dx.doi.org/10.1016/j.hlc.2014.12.118 Extracorporeal membrane oxygenation (ECMO) in post-cardiotomy cardiogenic shock (PCCS) in adults: a systematic review Alireza Kashani *, Con Manganas St. George Hospital, Sydney, NSW, Australia Corresponding author.
*
Background: Post-Cardiotomy Cardiogenic Shock (PCCS) is a rare but a significant cause of death in patients undergoing cardiac surgery. Venous-Arterial ECMO (VA-ECMO) is potentially life- saving. However, there are no clear selection criteria or prognostic predictors to direct patient selection and conduct of VA-ECMO in this specific subset of patients. Method: A systematic search was conducted in Ovid Medline, Cochrane Register of Systematic Reviews, and Embase for articles from Jan 2000 to July 2014 using the keywords “Cardiogenic Shock”, “Cardiotomy” or “Cardiac Surgery”, and “Extracorporeal Membrane Oxygenation” or “ECMO” or “Mechanical Circulatory Support”. Additional papers were identified by manual search of the references from the key articles. Those covering paediatric cases, case reports, or containing mixed data with other causes of refractory cardiogenic shock were excluded. Results: Indications, demographic data, statistically significant prognostic markers, and outcomes of VA-ECMO in PCCS were analysed. There were no randomised controlled trials. 11 case series were identified, in which 1328 patients received VA-ECMO for PCCS. 56.12% (31.27%-67.74%) of the patients could be weaned off. However, the overall survival to hospital discharge was 31.48% (15.69%-58.06%). Age was the commonest prognostic factor, followed by serum lactate levels 48 hours after initiation of ECMO, and dialysis. Conclusion: Due to continuing improvements in VAECMO over the last decade, increased numbers of patients supported can be successfully weaned off ECMO. However, this may not translate to successful discharge from the hospital and overall survival rate post cardiotomy shock has remained unchanged. This prompts our literature review of important selection criteria and prognostic predictors in this patient subset. http://dx.doi.org/10.1016/j.hlc.2014.12.119 A simple sternal closure technique to reduce the incidence of sternal complications in high risk patients Sean D. Galvin 1, Adam El Gamel 1,2* 1
Department of Cardiothoracic Surgery, Waikato Hospital, Hamilton, New Zealand 2 Department of Cardiothoracic Surgery, King’s College Hospital, London, UK * Corresponding author. Introduction: Sternal dehiscence after cardiac surgery may lead to mediastinitis and is associated with significant
being the main parameter used to guide upper limb and trunk exercise prescription and progression. 44% of the respondents reported screening for sternal instability, with 10% using an objective measure (i.e. sternal instability scale (SIS), or imaging) as part of their assessment. If sternal instability was detected, the majority of respondents based the prescription of sternal precautions and exercise guidelines on clinical experience. Discussion: There is significant variation in practice with respect to sternal precautions as well as the prescription and progression of upper limb and trunk exercises in outpatient cardiac rehabilitation programs in Australia. Further research is warranted in order to establish evidence-based upper limb and trunk exercise guidelines for patients following cardiac surgery via a median sternotomy in order to facilitate recovery and attain optimal patient outcomes. http://dx.doi.org/10.1016/j.hlc.2014.12.117 Construction of a cell model for dynamically detecting process of autophagy in alveolar macrophages Qing Geng *, Tao Fan Department of Thoracic Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, China * Corresponding author. Introduction: Autophagy is a constitutive regulatory means of cellular homeostasis involved in diverse physiological and pathological events. Autophagy in macrophages is important for maturation, activation, polarisation, and regulation of cytokine production. Alveolar macrophages play a critical role in chronic obstructive pulmonary disease (COPD), ischaemia/reperfusion injury, infectious diseases and neurodegenerative disorders. The numbers of alveolar macrophages are markedly increased in the lungs of patients with COPD as a result of increased recruitment, proliferation and survival. Alveolar macrophages secrete many inflammatory mediators, oxidants, proteins and proteinases in response to cigarette smoke extract and other stimuli. We developed a new cell model for dynamically detecting process of autophagy in alveolar macrophages. Methods: The PAsGreen2 - N1 - LC3 plasmid was constructed and transfected into alveolar macrophagyes line (NR8383 cells) with transfection reagent. The stable transfectants were screened by G418. The PAsGreen2 - N1 - LC3 expression was analyzed by fluorescence microscope, Western blot and PCR technology. Results: Selected by G418, 4 transfected cell lines showed high level of AsGreen2-N1-LC3 expression, as demonstrated by Western blot analysis. More than 92% cells showed positive fluorescent signals under inverted fluorescence microscope. The result of Western blot confirmed that the amount of protein expressed by LC3 genes in NR8383 PAsGreen2 N1 - LC3 cells is 3.11628 times than that of normal NR8383 cells. The transcription of LC3 in NR8383 PAsGreen2 - N1 LC3 cells was relatively 5.188611 times than that of normal NR8383 cells detected by PCR technique. Discussion: An alveolar macrophage line (NR8383 cell line) stably expressing PAsGreen2 - N1 - LC3 was constructed successfully, which can be used for dynamic detection of
Abstracts
autophagy process in alveolar macrophagyes. Besides, as a new cell model, it can be used for further study of autophagy in the pathological processes of alveolar macrophages. http://dx.doi.org/10.1016/j.hlc.2014.12.118 Extracorporeal membrane oxygenation (ECMO) in post-cardiotomy cardiogenic shock (PCCS) in adults: a systematic review Alireza Kashani *, Con Manganas St. George Hospital, Sydney, NSW, Australia Corresponding author.
*
Background: Post-Cardiotomy Cardiogenic Shock (PCCS) is a rare but a significant cause of death in patients undergoing cardiac surgery. Venous-Arterial ECMO (VA-ECMO) is potentially life- saving. However, there are no clear selection criteria or prognostic predictors to direct patient selection and conduct of VA-ECMO in this specific subset of patients. Method: A systematic search was conducted in Ovid Medline, Cochrane Register of Systematic Reviews, and Embase for articles from Jan 2000 to July 2014 using the keywords “Cardiogenic Shock”, “Cardiotomy” or “Cardiac Surgery”, and “Extracorporeal Membrane Oxygenation” or “ECMO” or “Mechanical Circulatory Support”. Additional papers were identified by manual search of the references from the key articles. Those covering paediatric cases, case reports, or containing mixed data with other causes of refractory cardiogenic shock were excluded. Results: Indications, demographic data, statistically significant prognostic markers, and outcomes of VA-ECMO in PCCS were analysed. There were no randomised controlled trials. 11 case series were identified, in which 1328 patients received VA-ECMO for PCCS. 56.12% (31.27%-67.74%) of the patients could be weaned off. However, the overall survival to hospital discharge was 31.48% (15.69%-58.06%). Age was the commonest prognostic factor, followed by serum lactate levels 48 hours after initiation of ECMO, and dialysis. Conclusion: Due to continuing improvements in VAECMO over the last decade, increased numbers of patients supported can be successfully weaned off ECMO. However, this may not translate to successful discharge from the hospital and overall survival rate post cardiotomy shock has remained unchanged. This prompts our literature review of important selection criteria and prognostic predictors in this patient subset. http://dx.doi.org/10.1016/j.hlc.2014.12.119 A simple sternal closure technique to reduce the incidence of sternal complications in high risk patients Sean D. Galvin 1, Adam El Gamel 1,2* 1
Department of Cardiothoracic Surgery, Waikato Hospital, Hamilton, New Zealand 2 Department of Cardiothoracic Surgery, King’s College Hospital, London, UK * Corresponding author. Introduction: Sternal dehiscence after cardiac surgery may lead to mediastinitis and is associated with significant