Detection of enteroviruses in stools precedes islet autoimmunity by several months: possible evidence for slowly operating mechanisms in virus-induced autoimmunity

Aims/hypothesis

This case–control study was nested in a prospective birth cohort to evaluate whether the presence of enteroviruses in stools was associated with the appearance of islet autoimmunity in the Type 1 Diabetes Prediction and Prevention study in Finland.

Methods

Altogether, 1673 longitudinal stool samples from 129 case children who turned positive for multiple islet autoantibodies and 3108 stool samples from 282 matched control children were screened for the presence of enterovirus RNA using RT-PCR. Viral genotype was detected by sequencing.

Results

Case children had more enterovirus infections than control children (0.8 vs 0.6 infections per child). Time-dependent analysis indicated that this excess of infections occurred more than 1 year before the first detection of islet autoantibodies (6.3 vs 2.1 infections per 10 follow-up years). No such difference was seen in infections occurring less than 1 year before islet autoantibody seroconversion or after seroconversion. The most frequent enterovirus types included coxsackievirus A4 (28% of genotyped viruses), coxsackievirus A2 (14%) and coxsackievirus A16 (11%).

Conclusions/interpretation

The results suggest that enterovirus infections diagnosed by detecting viral RNA in stools are associated with the development of islet autoimmunity with a time lag of several months.

     

Under-notification of cryptosporidiosis by routine clinical and laboratory practices among non-hospitalised children with acute diarrhoea in Southern Spain

Introduction  

In a prevalence study of 400 stool samples from non-hospitalised children under 7 years of age with diarrhoea, the presence of Cryptosporidium was evaluated.     

Analysis of pancreatic volume in acute‐onset,slowly‐progressive and fulminant type 1 diabetes in a Japanese population

Aims/Introduction

A decrease in the size of the pancreas is observed in islet autoantibody‐positive non‐diabetic donors and acute‐onset type 1 diabetes irrespective of the diabetes duration. Little is known, however, about the relationship between the size of the pancreas and type 1 diabetes subtypes, including fulminant type 1 diabetes.

Materials and Methods

We examined the pancreatic volume (PV) in 44 adult patients with type 1 diabetes (16 acute‐onset type 1 diabetes, 18 slowly progressive type 1 diabetes and 10 fulminant type 1 diabetes) and 39 age‐ and body mass index‐matched non‐diabetic controls. PV was measured by computed tomography. The ability to secrete insulin was assessed by stimulated C‐peptide after intravenous glucagon administration.

Results

PV was significantly correlated with bodyweight in both control participants and type 1 diabetes patients. The PV index (PVI; PV/bodyweight) was decreased by 39% in type 1 diabetes compared with that in controls. PVI was significantly decreased in acute‐onset type 1 diabetes patients and slowly progressive type 1 diabetes patients (both P < 0.0001), but not in fulminant type 1 diabetes patients (P = 0.10), compared with control participants. In cases patients with recent‐onset type 1 diabetes (0–7 days post‐diagnosis), PVI was significantly decreased in acute‐onset type 1 diabetes patients (n = 8, P = 0.0005), but not in fulminant type 1 diabetes patients (n = 7, P = 0.44), compared with controls. PVI showed no correlations with the diabetes duration, C‐peptide levels, glycated hemoglobin, glutamic acid decarboxylase autoantibody levels, serum amylase or daily total insulin dose in type 1 diabetes subtypes.

Conclusions

The present results show that patients with acute‐onset type 1 diabetes and slowly progressive type 1 diabetes have small pancreases irrespective of the diabetes duration or C‐peptide levels. In contrast to earlier findings on acute‐onset type 1 diabetes, we found no reduction of PVI at the onset of fulminant type 1 diabetes.     

Evaluation of a Non-invasive Method to Detect Cytomegalovirus (CMV)-DNA in Stool Samples of Patients with Inflammatory Bowel Disease (IBD): A Pilot Study

Background  

A severe flare of colitis in patients with IBD treated with immunosuppressive therapy may be complicated by an underlying CMV infection. The aim of this pilot study was to investigate the diagnostic efficacy of quantitative polymerase chain reaction (PCR) to detect CMV DNA in stool samples of IBD patients.     

High frequency of enterovirus D68 in children hospitalised with respiratory illness in Norway,autumn 2014

Objectives

An unexpectedly high proportion of children were admitted for severe respiratory infections at the Oslo University Hospital, Ullevål, Norway, during September and October, 2014. In light of the ongoing outbreak of enterovirus-D68 (EV-D68) in North America a real-time RT-PCR for screening of enterovirus and enterovirus D68 was established.

Design

We developed a duplex real-time RT-PCR for rapid screening of enterovirus D68. The method target the 5′ non-translated region (NTR) of the HEV genome at a location generally used for enterovirus detection.

Sample

Nasopharyngeal samples (n = 354), from children <15 years of age, received for respiratory virus analysis in OUH during September 1st and October 31nd, 2014, were tested for enterovirus and screened for enterovirus D68.

Main outcome measures and results

The duplex real-time RT-PCR method was an efficient tool for rapid screening for EV-D68 in respiratory specimens. Enterovirus was detected in 66 (22%) of 303 pediatric nasopharyngeal samples collected from children hospitalised with acute respiratory infection within the two-month period. Out of these, 33 (50%) were EV-D68. EV-D68 was associated with acute flaccid paralysis in one child.

Conclusions

An unexpectedly high proportion of children admitted for severe respiratory infections at the Oslo University Hospital, Ullevål, Norway, were diagnosed with EV- D68 during September 1st and October 31nd, 2014. These results emphasise that greater vigilance is required throughout Europe as enteroviruses are cause of severe respiratory disease.     

Efficacy of pulmonary vein isolation with a novel hot balloon ablation catheter

Background  

Due to the variable anatomy of the human pulmonary vein (PV), PV isolation for atrial fibrillation ablation is challenging. A novel hot balloon ablation catheter has been developed to improve ablation outcomes. The efficacy, histopathologic lesion characteristics, and thermodynamics of this technology were examined.     

Significance of hepatic resection and adjuvant hepatic arterial infusion chemotherapy for hepatocellular carcinoma with portal vein tumor thrombus in the first branch of portal vein and the main portal trunk: a project study for hepatic surgery of the Japanese Society of Hepato‐Biliary‐Pancreatic Surgery

Background

The prognosis of hepatocellular carcinoma (HCC) with tumor thrombus in the major portal vein (PV) is extremely poor. The purpose of this study was to clarify the impact of hepatic resection for HCC with tumor thrombus in the major PV.

Patients

Four hundred patients undergoing macroscopic curative resection for HCC involving the first branch or trunk of the PV between 2001 and 2010 at the 22 institutions were enrolled. We examined the effect of adjuvant hepatic arterial infusion chemotherapy (HAIC) on prognosis and validated the prognostic index consisting of ascites, prothrombin activity, and maximal tumor diameter.

Results

Median survival time (MST) and 5‐year overall survival rate were 21.5 months and 25.7%. MST of HAIC group was longer than that of non‐HAIC group (28.1 months vs. 18.7 months, P = 0.0024). Significant prognostic factors for overall survival were PIVKA‐II, tumor diameter, and adjuvant HAIC. MST for patients with prognostic index 0, 1, 2, and 3 was 39.0, 21.1, 18.9, and 5.7 months, respectively (P = 0.005).

Conclusions

Macroscopic curative resection with adjuvant HAIC might provide better survival outcome. Furthermore, the prognostic index was useful to select adequate treatment modalities for patients with HCC with tumor thrombosis in the major PV.     

Enterovirus RNA in longitudinal blood samples and risk of islet autoimmunity in children with a high genetic risk of type 1 diabetes: the MIDIA study

Aims/hypothesis

Only a few longitudinal molecular studies of enterovirus and islet autoimmunity have been reported, and positive results seem to be limited to Finland. We aimed to investigate an association between enterovirus RNA in blood and islet autoimmunity in the MIDIA study from Norway, a country which largely shares environmental and economic features with Finland.

Methods

We analysed serial blood samples collected at ages 3, 6, and 9 months and then annually from 45 children who developed confirmed positivity for at least two autoantibodies (against insulin, GAD65 and IA-2) and 92 matched controls, all from a cohort of children with a single high-risk HLA-DQ-DR genotype. Enterovirus was tested in RNA extracted from frozen blood cell pellets, using real-time RT-PCR with stringent performance control.

Results

Out of 807 blood samples, 72 (8.9%) were positive for enterovirus. There was no association between enterovirus RNA and islet autoimmunity in samples obtained strictly before (7.6% cases, 10.0% controls, OR 0.75 [95% CI 0.36, 1.57]), or strictly after the first detection of islet autoantibodies (10.5% case, 5.8% controls, OR 2.00 [95% CI 0.64, 6.27]). However, there was a tendency towards a higher frequency of enterovirus detection in the first islet autoantibody-positive sample (15.8%) compared with the corresponding time point in matched controls (3.2%, OR 8.7 [95% CI 0.97, 77]). Neither of these results was changed by adjusting for potential confounders, restricting to various time intervals or employing various definitions of enterovirus positivity.

Conclusions/interpretation

Positivity for enterovirus RNA in blood did not predict the later induction of islet autoantibodies, but enterovirus tended to be detected more often at the islet autoantibody seroconversion stage.     

Pulmonary vein isolation by duty-cycled bipolar and unipolar antrum ablation using a novel multielectrode ablation catheter system: first clinical results

Background  

The electrical disconnection of the pulmonary veins (PV) plays an important role in the ablation strategy of paroxysmal atrial fibrillation (PAF). Circumferential antral ablation with a conventional ablation technique using a steerable ablation catheter is sometimes difficult to perform and does not always result in isolation of the targeted PV.     

Enterovirus Infections in Germany: Comparative Evaluation of Different Laboratory Diagnostic Methods

Background: The diagnosis of an enterovirus infection may be achieved through direct virus detection from fecal or cerebrospinal fluid (CSF) samples by virus isolation or PCR. Serologically, a significant rise in antibody titer may be detected and different enteroviral types can be differentiated using the neutralization assay. Patients and Methods: We investigated the contribution of these different laboratory parameters to the diagnosis of enterovirus infections occurring in the Frankfurt am Main area during the years 1997 to 1999, including an echovirus 30 outbreak in a group of children with aseptic meningitis in 1997. Samples were referred from 1,013 patients; virus isolation was attempted from 579 CSF specimens and from 400 stool samples. 208 CSF samples were tested by PCR. Results: During the echorivus 30 outbreak we identified 22.3% of samples as positive, almost exclusively echovirus 30. In 1998 only 7.1% of samples were positive and a rather broad range of agents was isolated. In 1999 10.4% were positive, predominantly coxsackie B5 and echovirus 11. We could show that in acute enterovirus infections, virus detection by cell culture and PCR is superior to serological methods (neutralization assay and IgM assay). For virus isolation, there was a higher rate of positives from stool compared to CSF (1997: 27.8% versus 25%; 1998: 14.4% versus 3%; 1999: 17.9% versus 8.5%). When comparing PCR and virus isolation from the CSF, the former yielded a higher rate of positive results but was not clearly superior to virus isolation from CSF. Conclusion: The recommended method for the diagnosis of acute enterovirus infections is virus isolation from feces. In cases of suspected aseptic meningitis virus isolation and PCR are valuable for the direct detection of virus in CSF. Received: March 31, 2000 · Revision accepted: March 5, 2001     

Pulmonary venous structural remodeling in a canine model of chronic atrial dilation due to mitral regurgitation

OBJECTIVE:

Structural remodelling plays an important role in the genesis and maintenance of atrial fibrillation (AF). Although some studies that associate structural remodelling with atrial dilation have been reported, structural pulmonary venous (PV) remodelling due to chronic atrial dilation remains unclear.

METHODS:

Six sham dogs and five mitral regurgitation (MR) dogs (three months after partial mitral valve avulsion) were studied. Separate cryosections from the PV and left atrium (LA) were immunolabelled with antibodies against connexin (Cx) 40 and Cx43 and analyzed by confocal laser scanning microscopy. Tissue samples from the PV and LA were stained with hematoxylin and eosin, and Masson’s trichrome.

RESULTS:

In MR models, a decrease in Cx40 (0.57±0.2% versus 1.18±0.3%, P<0.05) and Cx43 (0.48±0.2% versus 1.56±0.5%, P<0.05) expression was observed compared with sham dogs. The distribution pattern of Cx40 and Cx43 changed from homogeneous to heterogeneous. Gap junction remodelling was not observed in the LA. In Masson’s trichrome-stained sections from MR dogs, regions with increased interstitial fibrosis were present in the PV. Thickness in the PV and the PV-LA junction did not change in the MR group.

CONCLUSION:

The present study demonstrated a decrease in Cx40 and Cx43 expression and increased interstitial fibrosis in PV due to MR. These changes may potentially be a mechanism that renders the dilated atria more susceptible to AF.     

Introduction of an expert system for the discrimination of local pulmonary vein and atrial far field signals

Background  

Discrimination of local and far field potentials during sinus rhythm and atrial fibrillation (AF) is essential for successful pulmonary vein (PV) isolation. We sought to introduce an expert system for the classification of electrophysiologic PV signals.     

Clostridium difficile as a cause of acute diarrhea: a prospective study in a tertiary care center

Introduction

Clostridium difficile-associated diarrhea (CDAD) is an increasing problem. Recent reports suggest presence of community acquired CDAD (CA CDAD). Studies in India have shown varied results.

Aims

The following are the aims of this study: (a) the prevalence of CDAD and CA CDAD in patients with acute diarrhea; (b) the incremental yield of second stool sample for the diagnosis of C. difficile infection (CDI); and (c) the risk factors for CDI.

Patients and Methods

Patients with acute diarrhea (<4 weeks) between April 2009 and December 2010 had two stool sample tested for C. difficile toxin (CDT) by enzyme-linked immunofluorescent assay. Demographic, clinical data, risk factors, clinical course, complications, treatment, and response were noted.

Results

Of 150 patients (mean age, 47.3 years; 76 males), 12 (8 %) had their first stool sample positive for CDT. Two patients (1.3 %) had community acquired CDI. The study group was compared with 138 patients (“control group”, stool samples negative for CDT). Compared to the controls, the study group were more likely to have had intensive care unit (ICU) stay (p?=?0.018) and tube feeding (p?=?0.035). Eleven patients were treated with metronidazole. One patient did not respond to metronidazole and was treated with vancomycin. No patient developed complications of CDAD.

Conclusions

The prevalence of CDAD in our population was 8 % and of CA CDAD was 1.3 %. There was no advantage of testing two samples. ICU stay and tube feeding were major risk factors for the CDAD. Metronidazole was an effective first-line therapy.     

Pulmonary vein isolation using a compliant endoscopic laser balloon ablation system in a swine model

Introduction  

The incidence of pulmonary vein reconnection and recurrent atrial fibrillation after a single catheter ablation procedure is unacceptably high. We studied the ability of a novel endoscopic laser balloon ablation system (EAS) to achieve acute and chronic pulmonary vein (PV) isolation in a swine model.     

Incorrect identification of recent Asian strains of Coxsackievirus A16 as human enterovirus 71: Improved primers for the specific detection of human enterovirus 71 by RT PCR

Background  

Human enterovirus 71 has emerged as an important pathogen in the Asia Pacific region and it is important to be able to make a rapid and specific diagnosis for outbreak control. Recent Asian strains of Coxsackievirus A16 have changes in the VP1 gene which causes mispriming of widely used primers for human enterovirus 71 specific identification.     

No evidence of enteroviruses in the intestine of patients with type 1 diabetes

Aims/hypothesis  

The purpose of this study was to investigate whether the gut mucosa is a reservoir for enterovirus persistence in patients with type 1 diabetes.     

Quantitative profiling of CpG island methylation in human stool for colorectal cancer detection

Purpose

The aims of this study were to investigate the use of quantitative CGI methylation data from stool DNA to classify colon cancer patients and to relate stool CGI methylation levels to those found in corresponding tissue samples.

Methods

We applied a quantitative methylation-specific PCR assay to determine CGI methylation levels of six genes, previously shown to be aberrantly methylated during colorectal carcinogenesis. Assays were performed on DNA from biopsies of “normal” mucosa and stool samples from 57 patients classified as disease-free, adenoma, or cancer by endoscopy, and in tumour tissue from cancer patients. Additionally, CGI methylation was analysed in stool DNA from an asymptomatic population of individuals covering a broad age range (mean?=?47?±?24 years)

Results

CGI methylation levels in stool DNA were significantly higher than in DNA from macroscopically normal mucosa, and a significant correlation between stool and mucosa was observed for ESR1 only. Multivariate statistical analyses using the methylation levels of each CGI in stool DNA as a continuous variable revealed a highly significant (p?=?0.003) classification of cancer vs. non-cancer (adenoma + disease-free) patients (sensitivity?=?65 %, specificity?=?81 %).

Conclusion

CGI methylation profiling of stool DNA successfully identified patients with cancer despite the methylation status of CGIs in stool DNA not generally reflecting those in DNA from the colonic mucosa.     

Rapid ventricular pacing to optimize rotational angiography in atrial fibrillation ablation

Background  

Conventional pulmonary vein (PV) angiography cannot precisely delineate the left atrium (LA)–PV anatomy, which is essential for the ablation of atrial fibrillation (AF). The aim of the study was to test the feasibility of a novel method of rotational angiography for the AF ablation.     

High-density biatrial pacing protects against atrial fibrillation by synchronizing left atrial tissue

Purpose  

The induction and sustainability of atrial fibrillation (AF) are most commonly due to pulmonary vein (PV) triggers and left atrial (LA) substrate. We hypothesized that simultaneous pacing of the PV ostia, LA, and right atrium (RA) reduces AF susceptibility by synchronizing atrial activity.