Fracture in porous bone analysed with a numerical phase-field dynamical model.

A dynamic phase-field fracture finite element model is applied to discretized high-resolution three-dimensional computed tomography images of human trabecular bone to analyse rapid bone fracture. The model is contrasted to quasi-static experimental results and a quasi-static phase-field finite element model. The experiment revealed complex stepwise crack evolution with multiple crack fronts, and crack arrests, as the global tensile displacement load was incrementally increased. The quasi-static phase-field fracture model captures the fractures in the experiment reasonably well, and the dynamic model converges towards the quasi-static model when mechanically loaded at low rates. At higher load rates, i.e., at larger impulses, inertia effects significantly contribute to an increased initial global stiffness, higher peak forces and a larger number of cracks spread over a larger volume. Since the fracture process clearly is different at large impulses compared to small impulses, it is concluded that dynamic fracture models are necessary when simulating rapid bone fracture.

Concordance of cerebrospinal fluid real-time quaking-induced conversion across the European Creutzfeldt-Jakob Disease Surveillance Network.

BACKGROUND AND PURPOSE: Cerebrospinal fluid (CSF) real-time quaking-induced conversion (RT-QuIC) has a high degree of sensitivity and specificity for the diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) and this has led to its being included in revised European CJD Surveillance Network diagnostic criteria for sCJD. As CSF RT-QuIC becomes more widely established, it is crucial that the analytical performance of individual laboratories is consistent. The aim of this ring-trial was to ascertain the degree of concordance between European countries undertaking CSF RT-QuIC. METHODS: Ten identical CSF samples, seven from probable or neuropathologically confirmed sCJD and three from non-CJD cases, were sent to 13 laboratories from 11 countries for RT-QuIC analysis. A range of instrumentation and different recombinant prion protein substrates were used. Each laboratory analysed the CSF samples blinded to the diagnosis and reported the results as positive or negative. RESULTS: All 13 laboratories correctly identified five of the seven sCJD cases and the remaining two sCJD cases were identified by 92% of laboratories. Of the two sCJD cases that were not identified by all laboratories, one had a disease duration >26 months with a negative 14-3-3, whilst the remaining case had a 4-month disease duration and a positive 14-3-3. A single false positive CSF RT-QuIC result was observed in this study. CONCLUSIONS: This study shows that CSF RT-QuIC demonstrates an excellent concordance between centres, even when using a variety of instrumentation, recombinant prion protein substrates and CSF volumes. The adoption of CSF RT-QuIC by all CJD surveillance centres is recommended.

Evaluation of 11 SARS-CoV-2 antibody tests by using samples from patients with defined IgG antibody titers.

We evaluated the performance of 11 SARS-CoV-2 antibody tests using a reference set of heat-inactivated samples from 278 unexposed persons and 258 COVID-19 patients, some of whom contributed serial samples. The reference set included samples with a variation in SARS-CoV-2 IgG antibody titers, as determined by an in-house immunofluorescence assay (IFA). The five evaluated rapid diagnostic tests had a specificity of 99.0% and a sensitivity that ranged from 56.3 to 81.6% and decreased with low IFA IgG titers. The specificity was > 99% for five out of six platform-based tests, and when assessed using samples collected ≥ 22 days after symptom onset, two assays had a sensitivity of > 96%. These two assays also detected samples with low IFA titers more frequently than the other assays. In conclusion, the evaluated antibody tests showed a heterogeneity in their performances and only a few tests performed well with samples having low IFA IgG titers, an important aspect for diagnostics and epidemiological investigations.

Retrospective meta-transcriptomic identification of severe dengue in a traveller returning from Africa to Sweden, 1990.

Pathogens associated with haemorrhagic fever commonly have zoonotic origins. The first documented imported case of likely viral severe haemorrhagic fever in Sweden occurred in 1990. Despite extensive study, no aetiological agent was identified. Following retrospective investigation with total RNA-sequencing of samples collected between 7 and 36 days from onset of symptoms we identified dengue virus 3 (DENV-3) and a human pegivirus (HPgV). We conclude that the patient likely suffered from haemorrhagic symptoms due to an atypical severe and undiagnosed dengue infection.

Use of complementary and alternative medicine (CAM) among emergency department (ED) patients in Sweden.

BACKGROUND: It has been suggested that the combination of complementary and alternative medicine (CAM) with conventional medicine carries a risk of adverse effects. The prevalence of CAM usage among patients in the Swedish emergency department (ED) is unknown. Hence, the aim of this study was to investigate CAM use among visiting patients at a Swedish ED. METHOD: A cross-sectional descriptive study was performed between August and October 2016 at an ED in Sweden. The questionnaire included 16 items regarding CAM use, factors associated with CAM use and patient healthcare communication and was distributed to 1600 patients. RESULTS: A total of 1029 questionnaires was returned (RR 64.3%). Current CAM use was reported by 7.9%, during the last year by 38.0%, and within lifetime by 72.9%. Factors associated with CAM use were: being a woman, middle-aged, in full-time employment, with secondary education level, higher use of non-prescription drugs and lower use of prescription drugs. Patient healthcare personnel communication about CAM was found to be approximately 5%. CONCLUSION: CAM usage exists among patients visiting Swedish EDs and almost one in ten uses CAM on the same day as the ED visit. CAM usage is associated with demographic factors. However, communication about CAM usage with ED personnel is poor.

Genetic analyses of Seoul hantavirus genome recovered from rats (Rattus norvegicus) in the Netherlands unveils diverse routes of spread into Europe.

Seoul virus (SEOV) is the etiologic agent of hemorrhagic fever with renal syndrome. It is carried by brown rats (Rattus norvegicus), a commensal rodent that closely cohabitates with humans in urban environments. SEOV has a worldwide distribution, and in Europe, it has been found in rats in UK, France, Sweden, and Belgium, and human cases of SEOV infection have been reported in Germany, UK, France, and Belgium. In the search of hantaviruses in brown rats from the Netherlands, we found both serological and genetic evidence for the presence of SEOV in the local wild rat population. To further decipher the relationship with other SEOV variants globally, the complete genome of SEOV in the Netherlands was recovered. SEOV sequences obtained from three positive rats (captured at close trapping locations at the same time) were found highly similar. Phylogenetic analyses demonstrated that two lineages of SEOV circulate in Europe. Strains from the Netherlands and UK, together with the Baxter strain from US, constitute one of these two, while the second includes strains from Europe and Asia. Our results support a hypothesis of diverse routes of SEOV spread into Europe. These findings, combined with other indications on the expansion of the spatial European range of SEOV, suggest an increased risk of this virus for the public health, highlighting the need for increased surveillance.

Discovery of Novel Viruses in Mosquitoes from the Zambezi Valley of Mozambique.

Mosquitoes carry a wide variety of viruses that can cause vector-borne infectious diseases and affect both human and veterinary public health. Although Mozambique can be considered a hot spot for emerging infectious diseases due to factors such as a rich vector population and a close vector/human/wildlife interface, the viral flora in mosquitoes have not previously been investigated. In this study, viral metagenomics was employed to analyze the viral communities in Culex and Mansonia mosquitoes in the Zambezia province of Mozambique. Among the 1.7 and 2.6 million sequences produced from the Culex and Mansonia samples, respectively, 3269 and 983 reads were classified as viral sequences. Viruses belonging to the Flaviviridae, Rhabdoviridae and Iflaviridae families were detected, and different unclassified single- and double-stranded RNA viruses were also identified. A near complete genome of a flavivirus, tentatively named Cuacua virus, was obtained from the Mansonia mosquitoes. Phylogenetic analysis of this flavivirus, using the NS5 amino acid sequence, showed that it grouped with 'insect-specific' viruses and was most closely related to Nakiwogo virus previously identified in Uganda. Both mosquito genera had viral sequences related to Rhabdoviruses, and these were most closely related to Culex tritaeniorhynchus rhabdovirus (CTRV). The results from this study suggest that several viruses specific for insects belonging to, for example, the Flaviviridae and Rhabdoviridae families, as well as a number of unclassified RNA viruses, are present in mosquitoes in Mozambique.

Hantavirus in new geographic regions, Sweden.

In Sweden, human cases of Puumala hantavirus (PUUV) infections are reported from the northern endemic regions. We found hantavirus-specific antibodies in yellow-necked mice (Apodemus flavicollis) trapped in human dwellings in the surroundings of the cities of Uppsala and Stockholm, which are situated far south from the traditional endemic areas of PUUV. Because the yellow-necked mouse is the most common rodent in human dwellings, hantaviruses in this rodent species may be important for the public health.

Quality of intra-partum care at a university hospital in Nepal: A prospective cross-sectional survey.

OBJECTIVE: To investigate the quality of intra-partum care provided to women with an expected normal birth at a university hospital in Nepal. METHODS: A prospective cross-sectional study was conducted during three weeks in November 2013. Nurses at the labor ward collected data from 292 consecutive births. Of these, 164 women of low risk were expected to have a normal birth and were included in the study; 107 (65%) were nulliparous. The self-administered questionnaire covered maternal characteristics, previous pregnancies and births, current pregnancy, labor and birth. Nine items assessed care management, five of which comprised the Bologna score with a total possible score of 5: presence of a companion, use of partograph, non-use of augmentation, non-supine position, and skin-to-skin contact. RESULTS: The women were assisted by physicians (56%), nurses (42%) or students under supervision (2%). All were in good health after birth. Two had a postpartum hemorrhage exceeding 500 ml and 49% had an episiotomy. Apgar score in all neonates was ≥ 7 at five minutes. Mean Bologna score was 1.43 (variance 0-3). CONCLUSIONS: The management of care in normal birth could be improved in the studied setting, and there is a need for more research to support such improvement.

Two clinical cases of renal syndrome caused by Dobrava/Saaremaa hantaviruses imported to the Netherlands from Poland and Belarus, 2012-2014.

We report the rare event of two imported cases in the Netherlands presenting with renal syndrome caused by Dobrava (DOBV)/Saaremaa (SAAV) hantaviruses. DOBV/SAAV hantaviruses are not circulating in the Netherlands and their clinical manifestation is typically more severe than that of the endemic Puumala virus (PUUV). This report aims to increase awareness among healthcare professionals and diagnostic laboratories to consider different hantaviruses as a cause of renal failure.

Emerging Viruses in the Republic of Suriname: Retrospective and Prospective Study into Chikungunya Circulation and Suspicion of Human Hantavirus Infections, 2008-2012 and 2014.

INTRODUCTION: Suriname is a country on the northeastern Atlantic coast of South America. It is unique in the sense that different ethnic cultures live together within the country, resulting in high levels of transport of both humans and products between the Asian, African, and European continents as well as the Caribbean. Travel is only one of the many factors present in Suriname contributing to the risk for the emergence or introduction of any infectious disease. Recently, circulation of both chikungunya virus (CHIKV) and hantavirus was reported in areas neighboring Suriname. Here we report a retrospective and prospective study into chikungunya and hantavirus circulation. METHODS: A chikungunya and hantavirus retrospective serological study was conducted on samples submitted for dengue, leptospirosis, and/or influenza virus diagnostics between 2008 and 2012 to the Bureau of Public Health in Suriname. This was followed by a prospective CHIKV serological and molecular surveillance study until the detection of the first autochthonous CHIKV cases in Suriname in May and June of 2014. RESULTS: None of the tested samples showed the presence of CHIKV antibodies in the retrospective serological study. Prospective testing of CHIKV-suspected patients resulted in the detection of the first autochthonous CHIKV cases in Suriname in May, 2015. In one sample, we were able to isolate and sequence the virus. Retrospective testing for the presence of hantavirus antibodies showed a relative high response in both pan-hantavirus enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA). However, neutralization tests did not yield any evidence for infection with either Seoul or Andes hantavirus. CONCLUSION: Here we report the presence of CHIKV in the republic of Suriname and the first serological indication of hantavirus infections in symptomatic patients.

Culex torrentium Mosquito Role as Major Enzootic Vector Defined by Rate of Sindbis Virus Infection, Sweden, 2009.

We isolated Sindbis virus (SINV) from the enzootic mosquito vectors Culex torrentium, Cx. pipiens, and Culiseta morsitans collected in an area of Sweden where SINV disease is endemic. The infection rate in Cx. torrentium mosquitoes was exceptionally high (36 infections/1,000 mosquitoes), defining Cx. torrentium as the main enzootic vector of SINV in Scandinavia.

First evidence of Seoul hantavirus in the wild rat population in the Netherlands.

We report the first detection of Seoul hantavirus (SEOV)-specific antibodies in the wild brown rat population in the Netherlands. SEOV-reactive antibodies were found in three rats out of 16 in a repeated series of tests including immunofluorescence assay, immunoblot, and enzyme-linked immunosorbent assay. Focus reduction neutralization test confirmed the presence of SEOV-specific antibodies, and reverse-transcription polymerase chain reaction (RT-PCR) confirmed the presence of hantaviral RNA. This discovery follows the recent findings of SEOV infections in wild and pet brown rats and humans in England, Wales, France, Belgium, and Sweden, indicating an even higher importance of this hantavirus for public health in large areas of Europe.

Clinical and radiological findings in methadone-induced delayed leukoencephalopathy.

OBJECTIVE: To increase awareness of the incidence of delayed leukoencephalopathy in rehabilitation medicine. SUBJECT: A 34-year-old male patient in an inpatient neuro-rehabilitation clinic who developed cognitive, psychological and physical deterioration 33 days after methadone intake. METHODS: Clinical follow-up for 7 months, brain imaging with magnetic resonance imaging and computed tomography, electroencephalography, multidisciplinary team evaluation and rehabilitation, pharmacological treatment, and examination of medical records. RESULTS: Clinical findings showed neuropsychological and motor deterioration. Brain images demonstrated that previous white matter infarctions had developed to cystic substance defects, and that abnormally high signals developed in the white matter of most cerebral lobes, with the exception of the grey matter and the cerebellum. Clinical improvement coincided with a modification in pharmacological treatment (increase in sertraline and introduction of baclofen). Brain images at 3 and 6 months after the methadone overdose showed reduced intensity of signal abnormalities and complete normalization of diffusion weighted images. Evaluation 7 months after injury estimated moderate brain injury with moderate disability and partial recovery of the patient's capacity for previous activities of daily living. CONCLUSION: Delayed leukoencephalopathy should be suspected in patients who deteriorate after methadone overdose. Drugs such as sertraline and baclofen may be of use in treating delayed leukoencephalopathy.

Direct regulation of abiotic responses by the Arabidopsis circadian clock component PRR7.

Up to 30% of the plant transcriptome is circadian clock-regulated in different species; however, we still lack a good understanding of the mechanisms involved in these genome-wide oscillations in gene expression. Here, we show that PSEUDO-RESPONSE REGULATOR 7 (PRR7), a central component of the Arabidopsis clock, is directly involved in the repression of master regulators of plant growth, light signaling and stress responses. The expression levels of most PRR7 target genes peak around dawn, in an antiphasic manner to PRR7 protein levels, and were repressed by PRR7. These findings indicate that PRR7 is important for cyclic gene expression by repressing the transcription of morning-expressed genes. In particular we found an enrichment of the genes involved in abiotic stress responses, and in accordance we observed that PRR7 is involved in the oxidative stress response and the regulation of stomata conductance.

Drug experts of the future, today?--depiction of the pharmacist profession in Swedish professional and lay print media.

BACKGROUND: The Swedish pharmacy market is presently being reregulated. The state-owned pharmacy monopoly company was divided and sold during 2009, and certain nonprescription medicines are now allowed in nonpharmacy settings. The changes will likely affect the pharmacist's role and the image of the community pharmacist in society. This change may affect how pharmacists are seen by society at large, and therefore, a baseline showing how pharmacists are depicted before the reregulation is of great value. OBJECTIVES: The aim of this study was to describe how the pharmacist profession is depicted in print media in Sweden, with a focus on community pharmacy. METHODS: A deductive qualitative content analysis with material from print media was conducted, using professional criteria as a framework. Swedish print media from October 2005 to October 2008 were searched and all relevant articles included. RESULTS: A total of 139 articles were included. Most articles came from professional journals, that is, journals directed toward pharmacist or related professions. The results show that the pharmacist profession is not highly visible and that this lack of visibility is disappointing to pharmacists. CONCLUSIONS: Society, as reflected in print media, does not display an awareness of the pharmacist role in Sweden. Although this is disappointing for the profession, it allows pharmacists to influence the depiction and hence their position in society.

Non-radioactive in situ hybridization protocol applicable for Norway spruce and a range of plant species.

The high-throughput expression analysis technologies available today give scientists an overflow of expression profiles but their resolution in terms of tissue specific expression is limited because of problems in dissecting individual tissues. Expression data needs to be confirmed and complemented with expression patterns using e.g. in situ hybridization, a technique used to localize cell specific mRNA expression. The in situ hybridization method is laborious, time-consuming and often requires extensive optimization depending on species and tissue. In situ experiments are relatively more difficult to perform in woody species such as the conifer Norway spruce (Picea abies). Here we present a modified DIG in situ hybridization protocol, which is fast and applicable on a wide range of plant species including P. abies. With just a few adjustments, including altered RNase treatment and proteinase K concentration, we could use the protocol to study tissue specific expression of homologous genes in male reproductive organs of one gymnosperm and two angiosperm species; P. abies, Arabidopsis thaliana and Brassica napus. The protocol worked equally well for the species and genes studied. AtAP3 and BnAP3 were observed in second and third whorl floral organs in A. thaliana and B. napus and DAL13 in microsporophylls of male cones from P. abies. For P. abies the proteinase K concentration, used to permeablize the tissues, had to be increased to 3 g/ml instead of 1 g/ml, possibly due to more compact tissues and higher levels of phenolics and polysaccharides. For all species the RNase treatment was removed due to reduced signal strength without a corresponding increase in specificity. By comparing tissue specific expression patterns of homologous genes from both flowering plants and a coniferous tree we demonstrate that the DIG in situ protocol presented here, with only minute adjustments, can be applied to a wide range of plant species. Hence, the protocol avoids both extensive species specific optimization and the laborious use of radioactively labeled probes in favor of DIG labeled probes. We have chosen to illustrate the technically demanding steps of the protocol in our film.

An indirect competitive immunoassay for insulin autoantibodies based on surface plasmon resonance.

We have developed a sensitive and specific method based on surface plasmon resonance (SPR) for detection of insulin autoantibodies (IAA) in serum samples from individuals at high risk of developing type 1 diabetes (T1D). When measuring trace molecules in undiluted sera with label-free techniques like SPR, non-specific adsorption of matrix proteins to the sensor surface is often a problem, since it causes a signal that masks the analyte response. The developed method is an indirect competitive immunoassay designed to overcome these problems. Today, IAA is mainly measured in radio immunoassays (RIAs), which are time consuming and require radioactively labeled antigen. With our SPR-based immunoassay the overall assay time is reduced by a factor of >100 (4 days to 50min), while sensitivity is maintained at a level comparable to that offered by RIA.

Detection of global glycosylation changes of serum proteins in type 1 diabetes using a lectin panel and multivariate data analysis.

Global glycosylation changes of serum proteins in type 1 diabetic patients have in this paper been investigated based on the interaction of the saccharide moiety of serum proteins with different lectins. Lectins are proteins, which bind carbohydrates specifically and reversibly. Panels with lectins of various carbohydrate specificities were immobilized on gold surfaces. Sera from healthy individuals, newly diagnosed type 1 diabetes patients and type 1 diabetes patients having had the disease for 4-6 years, respectively, were applied to the lectin panel. The biorecognition was evaluated with null ellipsometry. Data obtained were related to an internal standard of lactoferrin. Multivariate data analysis (MVDA) techniques were used to analyze data. Principal component analysis showed that the lectin panel enabled discrimination between sera from the three different above-mentioned groups. Using an artificial neuronal net (ANN), it was possible to correctly categorize unknown serum samples into one of the three groups.

Mitochondrial regulation of flower development.

Flower development in plants depends not only on a set of nuclear genes but also on the coordinate action of the mitochondrion. Certain mitochondrial genomes in combination with certain nuclear genomes lead to the expression of cytoplasmic male-sterility (CMS). Both mitochondrial genes that determine male-sterility and nuclear Restorer-of-fertility genes that suppress the male-sterile phenotype have been cloned. Lately, the interactions between mitochondrial and nuclear genes through retrograde signalling in CMS-systems have been dissected. Of special interest are the altered expression patterns of floral homeotic genes in certain CMS-systems. Here, we review the mitochondrial influence on flower development and give examples from CMS-systems developed in Brassica, Daucus carota, Nicotiana tabacum and Triticum aestivum.