Real-Time Whole-Genome Sequencing for Routine Typing,Surveillance, and Outbreak Detection of Verotoxigenic Escherichia coli

Fast and accurate identification and typing of pathogens are essential for effective surveillance and outbreak detection. The current routine procedure is based on a variety of techniques, making the procedure laborious, time-consuming, and expensive. With whole-genome sequencing (WGS) becoming cheaper, it has huge potential in both diagnostics and routine surveillance. The aim of this study was to perform a real-time evaluation of WGS for routine typing and surveillance of verocytotoxin-producing Escherichia coli (VTEC). In Denmark, the Statens Serum Institut (SSI) routinely receives all suspected VTEC isolates. During a 7-week period in the fall of 2012, all incoming isolates were concurrently subjected to WGS using IonTorrent PGM. Real-time bioinformatics analysis was performed using web-tools (www.genomicepidemiology.org) for species determination, multilocus sequence type (MLST) typing, and determination of phylogenetic relationship, and a specific VirulenceFinder for detection of E. coli virulence genes was developed as part of this study. In total, 46 suspected VTEC isolates were characterized in parallel during the study. VirulenceFinder proved successful in detecting virulence genes included in routine typing, explicitly verocytotoxin 1 (vtx1), verocytotoxin 2 (vtx2), and intimin (eae), and also detected additional virulence genes. VirulenceFinder is also a robust method for assigning verocytotoxin (vtx) subtypes. A real-time clustering of isolates in agreement with the epidemiology was established from WGS, enabling discrimination between sporadic and outbreak isolates. Overall, WGS typing produced results faster and at a lower cost than the current routine. Therefore, WGS typing is a superior alternative to conventional typing strategies. This approach may also be applied to typing and surveillance of other pathogens.     

PET/CT-Based Response Evaluation in Cancer—a Systematic Review of Design Issues

Molecular Imaging and Biology - Positron emission tomography/x-ray computed tomography (PET/CT) has long been discussed as a promising modality for response evaluation in cancer. When designing...     

Cardiovascular,hormonal and body fluid changes during prolonged exercise

Summary During prolonged heavy exercise a gradual upward drift in heart rate (HR) is seen after the first 10 min of exercise. This secondary rise might be caused by a reduction in stroke volume due to reduced filling of the heart, which is dependent upon both hemodynamic pressure and blood volume. Swimming and bicycling differ with respect to hydrostatic pressure and to water loss, due to sweating. Five subjects were studied during 90 min of bicycle exercise, and swimming the leg kick of free style. The horizontal position during swimming resulted in a larger cardiac output and stroke volume. After the initial rise in heart rate the secondary rise followed parallel courses in the two situations. The rises were positively related to the measured increments in plasma catecholamine concentrations, which continued to increase as exercise progresssed. The secondary rise in HR could not be explained by changes in plasma volume or in water balance, nor by changes in plasma [K]. The plasma volume decreased 5–6% (225–250 ml) within the first 5 to 10 min of exercise both in bicycling and swimming, but thereafter remained virtually unchanged. The sweat loss during bicycling was four times greater than during swimming; but during swimming the hydrostatic conditions induced a diuresis, so that the total water loss was only 25% less than during bicycling.     

Lack of association between cystatin C and different coronary atherosclerotic manifestations

Cystatin C (CysC) is known to be related to cardiovascular disease (CVD), including the presence and severity of coronary artery disease (CAD) and future clinical events. In this study, the association between CysC levels and (1) coronary artery calcification (CAC) in asymptomatic individuals from the general population as well as (2) different subgroups of patients with suspected or definite acute myocardial infarction (MI) was investigated. CysC levels were measured in serum from asymptomatic individuals as part of a screening study for CAC using non-contrast cardiac CT scan (N?=?1039) as well as in subgroups of hospitalized patients with a suspected MI (N?=?769). CysC was not associated with CAC in asymptomatic individuals after adjusting for relevant risk factors. No difference in CysC levels was observed between patients with type 1?MI (1.07?mg/L) and patients with normal troponin (with or without prior CAD: 1.14 and 1.01?mg/L, respectively). However, patients with type 2?MI and patient subgroups with elevated troponin but without MI had significantly higher CysC levels (1.24, 1.23 and 1.31?mg/L), even after adjusting for other risk factors. CysC was not associated with CAC in middle-aged asymptomatic individuals from the general population. Furthermore, CysC levels were found to be significantly lower in patients with type 1?MI compared to patients with type 2?MI and patients with elevated troponins but without MI. Thus, in two independent and clinically different populations, no association between CysC and coronary atherosclerotic manifestations could be demonstrated.     

Effects of sodium fluoride,vitamin D,and calcium on cortical bone remodeling in osteoporotic patients

Summary The purpose of this histomorphometric study of iliac bone biopsies from 10 postmenopausal osteoporotic patients was to describe the effects of sodium fluoride (combined with calcium and vitamin D) on remodeling in cortical bone after 6 months and after 5 years of tretment. Biopsies had been fixed in absolute methanol, embedded undecalcified in methylmetacrylate, and cut on a heavy-duty microtome. The therapy had no effect on the thickness of cortical bone in the iliac crest but increased the porosity slightly. It had no statistically significant effect on depth of resorption or thickness of new walls formed at remodeling sites but treatment increased the fraction of osteons undergoing remodeling in cortical bone. After 6 months of treatment, the increase was due to an enhanced activation of new remodeling sites, but in biopsies taken after 5 years of treatment, some degree of mineralization defect was observed and the duration of the remodeling cycle appeared to be prolonged. The mechanism underlying this qualitative change in the response to treatment is unknown, and it is unclear whether the mineralization defect may be prevented by, e.g., an altered supplementation of vitamin D or calcium.     

The anticonvulsant levetiracetam for the treatment of pain in polyneuropathy: A randomized,placebo‐controlled,cross‐over trial

Levetiracetam is an anticonvulsant which is assumed to act by modulating neurotransmitter release via binding to the vesicle protein SV2A. This could have an impact on signaling in the nociceptive system, and a pilot study indicated relief of neuropathic pain with levetiracetam. Objectives: The aim of this study was to test the analgesic effect of levetiracetam in painful polyneuropathy. Methods: This was a randomized, double‐blind, placebo‐controlled, cross‐over trial with levetiracetam 3000 mg/day versus placebo (6‐week treatment periods). Patients with diagnosed polyneuropathy and symptoms for more than 6 months, age between 20 and 80 years, pain intensity of more than 4 on a 0–10‐point numeric rating scale, and pain at least 4 days a week were included in the study. The primary outcome measure was pain relief at the end of each treatment period as measured on a 6‐point verbal scale. Results: Ninety‐three patients were screened for participation and 39 patients entered the study. Thirty‐five patients were included in the data analysis. There were no differences in the ratings of pain relief (levetiracetam 2.29 versus placebo 2.28, p =0.979), total pain intensity (levetiracetam 5.5 versus placebo 5.3, p =0.293) or any of the other outcome measures (p =0.147–1.00). Conclusion: This study indicates that the anticonvulsant levetiracetam has no clinically relevant effect on painful polyneuropathy.