Clinical significance of Stratifin,ERα and PR promoter methylation in tumor and serum DNA in Indian breast cancer patients

Objectives:The objective of this study was to determine the concordance of promoter methylation of stratifin, ERα and PR in tumor and circulating DNA in breast cancer patients and their association with clinicopathological parameters and disease prognosis.Design and methods:Methylation specific PCR were carried out to investigate the promoter methylation status of stratifin, ERα and PR in tumor and circulating DNA in 100 breast cancer patients in a prospective study. The effect of promoter methylation on protein expression was evaluated by immunohistochemistry.Results:Significant association was observed between promoter methylation of stratifin in tumors (61%) and paired sera (56%) (r = 0.78; p ≤ 0.001). Loss of stratifin expression was observed in 47% tumors and was associated with poor overall survival (p = 0.05). Significant correlation was observed between methylation status of ERα with PRB (p < 0.0001, OR = 20.8, 95% CI = 7.4–58.0) and stratifin (p = 0.003, OR = 2.0, 95% CI = 0.8–4.4).Conclusion:This study underscores the potential utility of serum DNA methylation of these genes as surrogate for tumor DNA methylation as a promising tool for cancer diagnosis.     

Hypomethylation of LINE-1 but not Alu in lymphocyte subsets of systemic lupus erythematosus patients

BackgroundT lymphocytes from SLE patients have a global decrease in the 5-methylcytosine content. Previous studies have identified hypomethylation in the promoter of several genes but there is limited study in the interspersed repetitive sequences (IRSs).MethodsWe examined and compared the methylation levels of long interspersed nuclear element 1 s (LINE-1) and Alu elements in normal and SLE CD4+ T lymphocytes, CD8+ T lymphocytes and B lymphocytes by the combined bisulfite restriction analysis-interspersed repetitive sequences (COBRA-IRS).ResultsHypomethylation of LINE-1 but not Alu was found in CD4+ T lymphocytes, CD8+ T lymphocytes, and B lymphocytes of SLE patient (P = 0.005, 0.002, and 0.007, respectively). Moreover, when the SLE patients were divided into active and inactive groups, LINE-1 hypomethylation was more significantly distinguished in both CD4+ and CD8+ T lymphocytes of patients from the active SLE group when compared to the controls. Surprisingly, Alu hypomethylation was also observed in CD8+ T lymphocytes from the inactive SLE group when compared to the normal controls (P = 0.0056).ConclusionsThe hypomethylation in each lymphocyte subset of SLE was IRSs specific, mainly found in LINE-1 rather than Alu.     

Prognostic significance of RASSF1A promoter methylation in operable breast cancer

ObjectivesThe aim of our study was to evaluate the prognostic significance of RASSF1A promoter methylation status in operable breast cancer.Design and methodsBy using Methylation Specific PCR, we evaluated the specificity of RASSF1A promoter methylation in 10 breast tumors and matching normal tissues, 10 breast fibroadenomas and 11 normal breast tissues. The prognostic significance of RASSF1A methylation was validated in 93 formalin fixed paraffin-embedded (FFPE) tissues obtained from patients with operable breast cancer.ResultsMethylation of RASSF1A promoter was observed in 1/31 (3.2%) non-cancerous breast tissues and 53/93 (57.0%) early stage breast tumors. The only positive sample in the non-cancerous breast tissues group was found in a histological normal tissue surrounding the tumor. During the follow-up period, 24/93 (25.8%) patients relapsed and 19/93 (20.4%) died. Disease-Free-Interval (DFI) was significantly associated with RASSF1A methylation (p = 0.028).ConclusionsRASSF1A promoter methylation provides important prognostic information in early stage breast cancer patients.     

RAS mutation analysis in a large cohort of Chinese patients with acute myeloid leukemia

ObjectiveWe examined RAS mutational status and correlated this with presenting morphology, cytogenetics, clinical outcome and other gene aberrations in a large cohort of Chinese acute myeloid leukemia (AML) patients.Designs and methodsN-RAS and K-RAS were screened for mutations at hot-spot codons 12, 13 and 61 using high resolution melting analysis (HRMA) and direct DNA sequencing in 504 Chinese AML patients and their clinical relevance was analyzed.ResultsThe frequencies of mutations of N-RAS and K-RAS were 9.7% (49/504) and 2.9% (15/504), respectively. c.35 G > A (rs121913237: G > A; p.Gly12Asp and rs121913529: G > A; p.Gly12Asp) and c.38 G > A (rs121434596: G > A; p.Gly13Asp and rs112445441: G > A; p.Gly13Asp) were the most common base substitutions (46% in N-RAS and 60% in K-RAS, respectively). AML patients with RAS mutations presented significantly higher white blood cell count (WBC) at diagnosis than those without mutations (p < 0.001). RAS mutations were underrepresented in patients with t(15;17) (2.9%, p = 0.01), while overrepresented in cases with abn11q23 (50%, p = 0.002) and inv(16) (66.6%, p = 0.04). In the FAB subtypes M4 and M5, RAS mutations were more frequent (21.6% and 20.6%, respectively) than they were in other subtypes (7.5%, p = 0.006 and 0.005, respectively). FLT3-ITD and RAS mutation were rarely coexistent (p = 0.03). RAS mutation didn't influence overall survival (OS) either in the entire cohort or within some defined subgroups.ConclusionsRAS mutations are associated with some biologically specific subtypes of AML but don't impact clinical outcome in Chinese patients.     

Identification of a novel IRGM promoter single nucleotide polymorphism associated with tuberculosis

BackgroundHuman immunity-related GTPase M (IRGM) is found to play an important role in defense against intracellular pathogen Mycobacterium tuberculosis in vitro by regulating autophagy. To verify whether single nucleotide polymorphisms (SNPs) in the promoter region of IRGM gene are associated with tuberculosis (TB) 1.7 kb IRGM promoter region was sequenced and SNP analysis was conducted in TB patients and healthy controls.MethodsA simple and rapid procedure for extracting DNA from clotted-blood was developed in this study. A 1.7 kb IRGM promoter region was amplified and sequenced for nucleotide polymorphism search. Then, 3 SNPs were selected and analyzed in 216 TB patients and 275 healthy subjects by ligase detection reaction technique.ResultsDNA extracted by our method was of high quality and suitable for PCR, sequencing, and genotyping. We identified 29 polymorphisms in the 1.7 kb IRGM promoter region, including 11 novel polymorphisms not yet reported. Large population analysis showed that frequencies of ? 1208A allele (P = 0.031), ? 1208AA genotype (P = 0.042), and ? 1208A/?1161C/?947C (P = 0.035) and ? 1208G/?1161C/?947C (P = 0.030) haplotypes in cases were significantly different from those in controls.ConclusionsIn 1.7 kb IRGM promoter region, only ? 1208A/G polymorphism is associated with susceptibility to TB.     

Rapid and quantitative detection of CpG-methylation status using TaqMan PCR combined with methyl-binding-domain polypeptide

ObjectivesTo assess the medical applicability of CpG methylation as molecular markers for cancer diagnosis, we established a new system to determine DNA methylation based on TaqMan PCR combined with a methyl-binding-domain polypeptide 2.Design and methodsWe evaluated the diagnostic applicability of this approach by examining the methylation status of two tumor suppressor genes, RASSF1A and APC, in 10 paired hepatocellular carcinoma (HCC) and the corresponding non-tumor liver tissues.ResultsMethylation levels of total 20 clinical samples measured by the TaqMan PCR assay showed a significantly positive correlation (R = 0.814, P < 0.0005 for RASSF1A, R = 0.736, P < 0.00001 for APC) with those calculated by bisulfite sequencing. The methylated DNA amount measured by our TaqMan PCR system precisely replicated the methylation status estimated by direct sequencing.ConclusionsThis suggests our method may serve as a reliable and easy-to-use tool for cancer diagnosis using methylated genes as biomarkers.     

TNF-α −308G > A and IL-6 −174G > C polymorphisms in Tunisian patients with coronary artery disease

ObjectiveOur aim was to evaluate the contribution of tumor necrosis factor (TNF)-α ?308G > A and interleukin (IL)-6 ?174G > C gene promoter variants to the presence of coronary artery disease (CAD) in Tunisians.Design and methodsStudy subjects comprised 418 angiographically proven CAD patients and 406 age-, gender-, and ethnic origin-matched controls. Genotyping was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis.ResultsThere were no significant differences in the allelic distribution of TNF-α ?308A (19.6% vs. 19.0%, P = 0.73), and IL-6 ?174C (15.6% vs. 14.3%, P = 0.47) promoter polymorphisms between CAD patients and control subjects, respectively. In addition, single locus analysis revealed no differences in genotype frequencies between the two study groups, and the combined distribution of both genotypes did not differ significantly between controls and CAD patients (P > 0.05).ConclusionThere is no allelic or genotypic association of TNF-α ?308G > A and IL-6 ?174G > C promoter polymorphisms with CAD in Tunisians, thereby confirming an ethnic-selective contribution of both gene variants to CAD presence.     

Apolipoprotein M promoter polymorphisms alter promoter activity and confer the susceptibility to the development of type 1 diabetes

ObjectivesApolipoprotein M plays an important role in the formation of preβ-HDL and cholesterol efflux to HDL. In the present study, we investigate the potential association between the ApoM promoter polymorphisms and type 1 diabetes.Design and methodsThe study was conducted in Peking Union Medical College, Beijing, China and Karolinska Institutet, Stockholm, Sweden. Two populations, including 493 Han Chinese subjects (177 T1D patients/316 controls) and 225 Swedish (124/101), are enrolled in the present study. Three single nucleotide polymorphisms (SNP) C-1065A, T-855C and T-778C in the promoter region of the ApoM gene are genotyped using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol. Promoter activity was measured by reporter gene assay.ResultsSNP T-778C was strongly associated with T1D in both Han Chinese (p = 0.002, OR = 2.188, CI 95% = 1.338–3.581) and Swedish (p = 0.021, OR = 2.865, CI 95% = 1.128–7.278) populations. The luciferase activity of ?778C promoter was 1.41 times as high as that of ?778T promoter (9.90 ± 1.92 vs. 7.04 ± 0.76, p = 0.001).ConclusionsAllele C of SNP T-778C may increase promoter activity and confer the risk susceptibility to the development of T1D.     

Rapid determination of AKAP12 promoter methylation levels in peripheral blood using methylation-sensitive high resolution melting (MS-HRM) analysis: Application in colorectal cancer

BackgroundColorectal cancer is the third most common form of cancer and hypermethylation has been shown to increase the risk of developing this disease. DNA hypermethylation in the A kinase anchor protein 12 (AKAP12/Gravin) promoter region and the accompanied underexpression of it has been noted in a variety of human cancers.MethodsWe applied methylation-specific high resolution melting (MS-HRM) technology to detect quantitatively A kinase anchor protein 12 (AKAP12/Gravin) methylation in peripheral blood from 100 colorectal cancer patients and 50 healthy volunteers and in 3 colorectal cancer cell lines.ResultsIn this study 48 of the 100 colorectal cancer samples (48%) were found to be methylated at the AKAP12 promoter region. AKAP12 methylation was significantly higher in the colorectal cancer samples with differentiation (p = 0.03). We also compared the results generated by MS-HRM with a traditional methylation-specific PCR (MSP) assay. We found that intra-assay variability ranged from 6.14 to 9.90% and inter-assay variability ranged from 14.5 to 17.2%. The AKAP12 MS-HRM assay was able to reproducibly detect 1% methylated DNA, whereas the MSP method was unable to detect less than 5% methylation.ConclusionsWe demonstrate the utility of quantitative AKAP12 MS-HRM analysis of promoter methylation in peripheral blood samples. AKAP12 MS-HRM quantitative methods with excellent detection capabilities have many promising applications in the research and diagnosis of colorectal cancer.     

Management of Moderate-to-Severe Dyspnea in Hospitalized Patients Receiving Palliative Care

ContextBenzodiazepines (BZDs) are commonly prescribed for relief of dyspnea in palliative care, yet few data describe their efficacy.ObjectivesTo describe the management of moderate-to-severe dyspnea in palliative care patients.MethodsChart review of inpatients with moderate or severe dyspnea on initial evaluation by a palliative care service. We recorded dyspnea scores at follow-up (24 hours later) and use of BZDs and opioids.ResultsThe records of 115 patients were reviewed. The mean age of patients was 64 years and primary diagnoses included cancer (64%, n = 73), heart failure (8%, n = 9), and chronic obstructive pulmonary disease (5%, n = 6). At initial assessment, 73% (n = 84) of the patients had moderate and 27% (n = 31) had severe dyspnea. At follow-up, 74% (n = 85) of patients reported an improvement in their dyspnea, of which 42% (n = 36) had received opioids alone, 37% (n = 31) had BZDs concurrent with opioids, 2% (n = 2) had BZDs alone, and 19% (n = 16) had received neither opioids nor BZDs. Logistic regression analysis identified that patients who received BZDs and opioids had increased odds of improved dyspnea (odds ratio 5.5, 95% CI 1.4, 21.3) compared with those receiving no medications.ConclusionMost patients reported improvement in dyspnea at 24 hours after palliative care service consultation. Consistent with existing evidence, most patients with dyspnea received opioids but only the combination of opioids and BZDs was independently associated with improvement in dyspnea. Further research on the role of BZDs alone and in combination with opioids may lead to better treatments for this distressing symptom.     

Association of the MS4A2 gene promoter C-109T or the 7th exon E237G polymorphisms with asthma risk: A meta-analysis

Background and objectiveA large number of studies have examined the association between the Membrane-spanning 4 domains, superfamily A, number 2 (MS4A2) gene C-109T (rs1441586) or E237G (rs569108) variants and asthma risk. However, the results are inconsistent and inconclusive. To derive a more precise estimation, a meta-analysis was performed.MethodsMeta-analyses were conducted with the data from case–control association studies (24 studies with 4496 asthmatics and 4571 controls for E237G variant and 9 studies including 2005 cases and 1868 control for C-109T polymorphisms, respectively). Random-effects model was used to calculate summary odds ratios (ORs).ResultsFor the MS4A2 gene E237G variant, no significant associations with asthma were found in overall population; we observed an elevated risk of atopic asthma among subjects with the 237G allele (OR = 1.341, 95% CI: 1.039–1.732 for G versus E and OR = 1.374, 95% CI: 1.032–1.828 for EG + GG versus EE) in the stratified meta-analysis. As for the MS4A2 gene C-109T polymorphism, no significant associations with asthma risk were observed in the total population; in subgroup analysis by ethnicity of subjects we found increased asthma risk among Asians carrying T allele (OR = 1.140, 95% CI: 1.019–1.276 for T versus C and OR = 1.359, 95% CI: 1.029–1.794 for TT versus CC).ConclusionsData indicated that the MS4A2 gene E237G variant may be a risk factor for developing atopic asthma and the promoter -109T allele is a potential risk factor of asthma in Asians.     

Neuropeptide Y polymorphisms and ischemic stroke in Chinese population

BackgroundStroke is the second most common cause of death in developed countries and a major cause of adult disability and mortality worldwide. New data strongly suggest that neuropeptide Y (NPY) may be a candidate gene for ischemic stroke.MethodsWe investigated 450 ischemic stroke patients and 423 healthy controls matched for sex and age in a Han Chinese population. Three functional polymorphisms (? 883TGins/del, ? 602G/T and ? 399 T/C) located in NPY gene promoter were genotyped using DNA sequencing methods.ResultsOf 3 NPY polymorphisms investigated in our study, the ? 399CC genotype (OR: 1.699, 95% CI: 1.124–2.567, P = 0.011) and the ? 399C allele (OR: 1.254, 95% CI: 1.031–1.524, P = 0.023) were more frequent among ischemic stroke patients than in controls, especially in the small vessel disease (SVD) subtype patients. The similar results were observed in multivariable logistic regression analysis. Haplotype analysis revealed that the ? 883ins/?399C haplotype was a risk marker for ischemic stroke (P = 0.008).ConclusionsThe C allele of ? 399 T/C polymorphism in the promoter regions of NPY is an independent risk factor for ischemic stroke, suggesting that NYP system may involve in the mechanisms of stroke pathology.     

Two minutes CPR versus five cycles CPR prior to reanalysis of the cardiac rhythm: A prospective,randomized simulator-based trial

Aim of the studyWhile the 2005 cardiopulmonary resuscitation (CPR) guidelines recommended to provide CPR for five cycles before the next cardiac rhythm check, the current 2010 guideline now recommend to provide CPR for 2 min. Our aim was to compare adherence to both targets in a simulator-based randomized trial.Methods119 teams, consisting of three to four physicians each, were randomized to receive a graphical display of the simplified circular adult BLS algorithm with the instruction to perform CPR for either 2 min or five cycles 30:2. Subsequently teams had to treat a simulated unwitnessed cardiac arrest. Data analysis was performed using video-recordings obtained during simulations. The primary endpoint was adherence, defined as being within ±20% of the instructed target (i.e. 96–144 s in the 2 min teams and 4–6 cycles in the fivex30:2 teams).Results22/62 (35%) of the “two minutes” teams and 48/57 (84%) of the “five × 30:2″ teams provided CPR within a range of ± 20% of their instructed target (P < 0.0001). The median time of CPR prior to rhythm check was 91 s and 87 s, respectively, (P = 0.59) with a significant larger variance (P = 0.023) in the “two minutes” group.ConclusionsThis randomized simulator-based trial found better adherence and less variance to an instruction to continue CPR for five cycles before the next cardiac rhythm check compared to continuing CPR for 2 min. Avoiding temporal targets whenever possible in guidelines relating to stressful events appears advisable.     

VEGF-A gene promoter polymorphisms and microvascular complications in patients with essential hypertension

ObjectivesWe investigated the possible involvement of vascular endothelial growth factor (VEGF-A) gene promoter polymorphisms in essential hypertension (EH).Design and methods1225 bp of the VEGF-A gene promoter were screened for polymorphisms using PCR amplification and direct DNA sequence analysis in 62 EH and 62 normotensive (HS) individuals. Circulating VEGF-A levels were determined by immunoassay.Results?152G/A (p = 0.009) and ?116G/A (p = 0.016) polymorphisms were correlated to hypertension (p < 0.05). Median platelet VEGF-A load in EH was 2.10 fg/plt. Patients with microvascular complications (MC) had higher platelet VEGF-A load than those without (p = 0.005). Multivariate analyses showed that ?116 A allele was an independent predictor of microalbuminuria (p = 0.014) and increased platelet VEGF-A load (p = 0.009) in EH. Platelet VEGF-A load independently predicted MC (p = 0.049) in addition to ?116G/A polymorphism (p = 0.035).ConclusionsAbnormal regulation of VEGF-A due to polymorphism at position ?116 might represent a genetic factor for increased VEGF-A production and MC in EH.     

Association of alpha 2A adrenergic receptor gene (ADRΑ2A) polymorphism with irritable bowel syndrome,microscopic and ulcerative colitis

BackgroundAlpha 2 adrenergic receptors (α2 ARs) play a central role in the regulation of systemic sympathetic activity. Prejunctional alpha 2A adrenoceptor regulates through negative feedback at presynaptic nerve ending. A-1291 C > G polymorphism located in α2-adrenergic receptor gene (ADRΑ2A) has been identified. We investigated the possible association between 1291 C > G polymorphism in the promoter region of ADRΑ2A in clinical subtypes of IBS, ulcerative and microscopic colitis patients.MethodsThis prospective case control study included 92 patients with diarrhea predominant IBS (D-IBS), 44 with constipation predominant IBS (C-IBS), 15 with alternating diarrhea and constipation IBS (M-IBS), 75 ulcerative colitis (UC), 41 microscopic colitis (MC) and 100 healthy controls. The subjects were genotyped by using PCR amplification of the promoter region of ADRΑ2A gene followed by digestion with the restriction enzyme MspI. The study was approved by the institute ethical committee.ResultsA strong genotypic association was observed between α2A-1291 C > G polymorphism and D-IBS (χ² = 6.38, df = 2, p < 0.05). There was no significant difference in α2A-1291 C > G genotype and allele frequency between C-IBS, M-IBS, UC, MC cases and control subjects.ConclusionsA significant association was observed between α2A-1291C > G polymorphism and D-IBS. Thus, α2 AR gene may be a potential candidate involved in the pathophysiology of D-IBS.     

Dysregulation of miR-124-1 predicts favorable prognosis in acute myeloid leukemia

ObjectiveMicroRNA miR-124 has been suggested as a tumor suppressor for its role in inhibiting cell growth, inducing differentiation and promoting apoptosis. The present study was aimed to investigate the expression status of miR-124-1 and its clinical relevance in patients with acute myeloid leukemia (AML).Designs and methodsReal-time quantitative PCR was performed to detect the expression level of miR-124-1 in AML patients. The clinical significance of miR-124-1 expression in AML was investigated.ResultsmiR-124-1 underexpression was identified in 30 (36%) of 83 AML patients. No significant difference could be observed in sex, age and blood parameters between the patients with and without miR-124-1 underexpression. The frequency of miR-124-1 underexpression was higher in the patients with t(15;17) than in others (62% versus 30%, P = 0.040). The status of miR-124-1 expression was not correlated with the mutations of nine genes (FLT3-ITD, NPM1, C-KIT, IDH1/IDH2, DNMT3A, N/K-RAS and C/EBPA). The patients with miR-124-1 underexpression had borderline longer overall survival and relapse-free survival than those without miR-124-1 underexpression (P = 0.052 and 0.045, respectively).ConclusionsThese findings suggest that miR-124-1 underexpression is a common event and might have a favorable impact on prognosis in AML.     

Association between myeloperoxidase polymorphisms and its plasma levels with severity of coronary artery disease

ObjectivesMyeloperoxidase (MPO) polymorphism ?463 has been related to higher cardiovascular risk. This study was conducted to test whether the MPO promoter polymorphism ?463A/G and MPO plasma levels are associated with coronary artery disease (CAD) severity.Design and methodsPatients submitted to elective coronariography were enrolled, CAD severity was assessed and blood samples collected to identify the MPO polymorphism and its plasma levels.ResultsGenotypes were determined in 118 patients. Among these patients, 12 (10%) were homozygous for AA, 69 (58%) for GG and 37 (32%) were heterozygous. Mean MPO plasma levels were 8.6 ± 4.7 ng/mL for AA, 8.6 ± 7.0 ng/mL for AG and 9.4 ± 5.6 ng/mL for GG genotypes. The CAD severity was not associated with MPO genotypes (p = 0.43), however, patients with higher CAD score presented higher MPO levels (p = 0.02).ConclusionWe found no association between MPO polymorphism and CAD severity, although a relation was observed for MPO plasma levels and extension of CAD.     

Video laryngoscopy versus direct laryngoscopy for tracheal intubation during in-hospital cardiopulmonary resuscitation

AimTracheal intubation during cardiopulmonary resuscitation (CPR) is a high-risk procedure. Here, we investigated the efficacy of video laryngoscopy for tracheal intubation during CPR.MethodsData regarding tracheal intubation during CPR from in-hospital cardiac arrests occurring between January 2011 and December 2013 (n = 229) were prospectively collected and retrospectively analyzed.ResultsThe initial laryngoscopy method was video laryngoscopy in 121 patients (52.8%) and direct laryngoscopy in 108 patients (47.2%). The rate of successful intubation at the first attempt was higher with video laryngoscopy (71.9%; 87/121) than with direct laryngoscopy (52.8%; 57/108; p = 0.003). The rate of success at the first attempt was higher for experienced (73.0%; 84/115) than inexperienced operators, including residents (52.6%; 60/114; p = 0.001). Mortality at day 28 after CPR was not significantly different between patients with successful tracheal intubation at the first attempt and without (68.1% [98/144] vs. 67.1% [57/85]; p = 0.876). In multivariate logistic regression analysis, a predicted difficult airway (odds ratio [95% confidence interval] = 0.22 [0.10–0.49]; p < 0.001), intubation by an experienced operator (2.63 [1.42–4.87]; p = 0.002), and use of video laryngoscopy rather than direct laryngoscopy (2.42 [1.30–4.45]; p = 0.005) were independently associated with a successful tracheal intubation at the first attempt.ConclusionUse of video laryngoscopy during CPR from in-hospital cardiac arrest is independently associated with successful tracheal intubation at the first attempt.     

Impact of intensified postresuscitation treatment on outcome of comatose survivors of out-of-hospital cardiac arrest according to initial rhythm

AimWe investigated the impact of intensified postresuscitation treatment in comatose survivors of out-of-hospital cardiac arrest (OHCA) of presumed cardiac etiology according to the initial rhythm at the emergency medical team arrival.MethodsInterventions and survival with Cerebral Performance Category (CPC) 1–2 within each group were retrospectively compared between the periods of conservative (1995–2003) and intensified (2004–2012) postresuscitation treatment.ResultsIn shockable group, therapeutic hypothermia (TH) increased from 1 to 93%, immediate invasive coronary strategy from 28 to 78%, intraaortic balloon pump from 4 to 21%, vasopressors/inotropes from 47 to 81% and antimicrobial agents from 65 to 86% during the intensified period as compared to conservative period (p < 0.001). This was associated with increased survival with CPC 1–2 from 27 to 47% (p < 0.001). After adjusting for age, sex and prehospital confounders, TH (OR = 2.12, 95% CI 1.25–3.61), percutaneous coronary intervention (OR 1.77, 95% CI 1.15–2.73) and antimicrobial agents (OR = 12.21, 95% CI 5.13–29.08) remained associated with survival with CPC 1–2. In non-shockable patients, TH also significantly increased from 1 to 74%, immediate invasive coronary strategy from 8 to 51%, intraaortic balloon pump from 2 to 9% and vasopressors/inotropes from 56 to 84% during intensified period without concomitant increase in survival with CPC 1–2 (7% vs. 9%; p = 0.27). After adjustment, only antimicrobial agents (OR = 8.43, 95% CI: 1.05–67.72) remained associated with survival with CPC 1–2.ConclusionIntensified postresuscitation treatment was associated with doubled survival in comatose survivors of OHCA with shockable rhythm. Such association could not be demonstrated in patients with non-shockable rhythm.