Gene delivery to dendritic cells by orally administered recombinant Saccharomyces cerevisiae in mice

DNA vaccination has caught the attention of many for triggering humoral as well as cellular immune responses. And delivering DNA into the antigen presenting cells (APCs) in order to induce efficient immunoresponse has become the backbone of this field. It has been confirmed that Saccharomyces cerevisiae, though non-pathogenic, is being engulfed by the dendritic cells and macrophages and delivers not only proteins, but also DNA materials (already confirmed in vitro). In this research, S. cerevisiae is used to deliver green fluorescent protein (GFP) reporter gene controlled under cytomegalovirus (CMV) promoter in living organism (mice). The recombinant yeast, transfected with the plasmid containing the GFP gene, was heat killed and orally administered to mice. After 60 h of yeast administration, mice were sacrificed and intestine was separated, washed and frozen in liquid nitrogen. Tissues were cut at the size of 10 μm using Cryostat machine, and GFP expression was successfully detected under a fluorescence microscope. After 45 days Western blot was able to detect GFP antibody in the blood of mice. These results imply that S. cerevisiae, being non-pathogenic, cheap, and easy to culture could be a good candidate to deliver DNA materials to the immune cells for vaccination.     

Clinical signs of pulmonary congestion predict outcome in patients with acute chest pain

BACKGROUND: Pulmonary congestion is associated with poor outcome in patients with acute coronary syndromes. In consecutive patients presenting with acute unexplained chest pain to a primary care facility, the prognostic impact of pulmonary congestion is indeterminate. Therefore, we assessed the predictive value of clinical signs of pulmonary congestion in patients presenting with acute chest pain to an emergency department with regard to the origin of the symptoms. METHODS: 1288 consecutive patients with acute chest pain were prospectively assessed for clinical signs of pulmonary congestion. The diagnosis was confirmed by chest radiography. The association of pulmonary congestion and short- and intermediate-term mortality in patients with coronary (n = 381) and non-coronary (n = 907) causes of chest pain was determined using multivariate Cox regression analysis. RESULTS: 108 (8%) patients had clinical signs of pulmonary congestion. Within the mean follow-up period of 23 months (SD 4) 67 patients died, mainly within the first 6 months. Of 108 patients with pulmonary congestion, 82 (76%) had coronary and 26 (24%) had non-coronary chest pain. Pulmonary congestion was independently associated with mortality in patients with coronary chest pain (hazard ratio 6.4, 95% confidence interval 2.5 to 16.1, p < 0.0001), both in patients with acute coronary syndromes or angina pectoris. However, in patients with non-coronary chest pain we observed no independent association of pulmonary congestion with outcome. CONCLUSION: Clinical signs of pulmonary congestion indicate an increased risk for poor outcome in patients with chest pain due to myocardial ischemia. Mortality of these patients is high, particularly in the first months after presentation. Therefore, hospital admission is warranted, including patients with angina pectoris, who otherwise may be candidates for early discharge.     

Acute chest pain — identification of patients at low risk for coronary events. The impact of symptoms, medical history and risk factors

BACKGROUND: The evaluation of patients with acute chest pain remains challenging, as it implies the risk of fatal misdiagnosis. It is well recognized that typical angina does not specifically identify patients at high risk. We investigated the predictive value of characteristics atypical for myocardial ischemia for exclusion of acute or subacute coronary events, focusing on patients' symptoms, medical history and risk factors. METHODS: We prospectively studied 1288 consecutive patients presenting with acute chest pain at a non-trauma emergency department. Patients' symptoms, history and risk factors were evaluated using seven predefined criteria and assigned as typical or atypical for ischemic coronary chest pain. Positive predictive value (PPV) and 95% confidence intervals (95% CI) were calculated to predict or exclude acute myocardial infarction (AMI) and major adverse cardiac events (MACE: cardiovascular death, percutaneous coronary interventions, bypass surgery, or myocardial infarction) within six months. RESULTS: AMI occurred in 168 patients (13%), and 6-months MACE (including AMI) overall in 240 patients (19%). Presence of four or more criteria typical for myocardial ischemia was associated with a PPV of 0.21 (0.17 to 0.25) for predicting AMI and 0.30 (0.25 to 0.35) for 6-months MACE. Presence of four or more criteria atypical for coronary ischemia was associated with a PPV of 0.94 (0.91 to 0.96) for excluding AMI and 0.93 (0.90 to 0.96) for excluding 6-months MACE. In 165 of 476 patients under 40 years of age (35%), four or more atypical criteria excluded AMI and 6-months MACE with PPVs of 0.98 (0.96 to 1.0). CONCLUSION: Evaluation of criteria atypical for myocardial ischemia with acute chest pain may help to identify candidates for early discharge, whereas typical characteristics have very little diagnostic value.     

Cardiopulmonary resuscitation-associated major liver injury

AIM: To evaluate the frequency, presentation, treatment and outcome of cardiopulmonary resuscitation-associated major liver injury in patients after non-traumatic in- or out-of-hospital cardiac arrest. MATERIALS AND METHODS: Retrospective analysis of a cardiac arrest registry in a tertiary care hospital emergency department. We reviewed patients charts, laboratory data, diagnostic imaging studies and autoptic findings. RESULTS: Within 14.5 years, major liver injury (rupture/laceration, haemorrhage/haematoma) was found in 15 of 2558 cardiac arrest victims (0.6%). Eleven were male (73%), median age was 58 (IQR 53-67). In seven, resuscitation was started out-of-hospital. In 9 of the 15 patients, liver injury was correctly diagnosed intra vitam. In 5, haematocrit level was low on admission, in 8 haematocrit dropped significantly during observation; haemostasis was compromised in 13 patients, 4 of them receiving thrombolytic therapy. Bedside abdominal sonography showed free intra-peritoneal fluid in 8 of 9 cases examined. In 11 patients, we found liver rupture/laceration, in 4 liver haemorrhage/haematoma. The site of injury was the left liver lobe in 11, six underwent emergent surgery. Two of 15 patients survived to 6 months in good neurological condition, 1 after emergency surgery. No patient died from bleeding due to liver injury. CONCLUSION: Our single centre observation confirms that resuscitation-associated major liver injury is infrequent and shows that most patients had compromised haemostasis. Low or dropping haematocrit should trigger suspicion. Bedside sonography reveals intra-peritoneal fluid or liver injury. A conservative therapeutic approach or emergency surgery may be warranted. Major liver injury alone scarcely appears to influence overall outcome.