Suggested new breakpoints of anti-MERS-CoV antibody ELISA titers: performance analysis of serologic tests.

To provide optimal cut-off values of anti-Middle East respiratory syndrome coronavirus (MERS-CoV) serologic tests, we evaluated performance of ELISA IgG, ELISA IgA, IFA IgM, and IFA IgG using 138 serum samples of 49 MERS-CoV-infected patients and 219 serum samples of 219 rRT-PCR-negative MERS-CoV-exposed healthcare personnel and patients. The performance analysis was conducted for two different purposes: (1) prediction of neutralization activity in MERS-CoV-infected patients, and (2) epidemiologic surveillance of MERS-CoV infections among MERS-CoV-exposed individuals. To evaluate performance according to serum collection time, we used 'days post onset of illness (dpoi)' and 'days post exposure (dpex)' assessing neutralization activity and infection diagnosis, respectively. Performance of serologic tests improved with delayed sampling time, being maximized after a seroconversion period. In predicting neutralization activity, ELISA IgG tests showed optimal performance using sera collected after 21 dpoi at cut-off values of OD ratio 0.4 (sensitivity 100% and specificity 100%), and ELISA IgA showed optimal performance using sera collected after 14 dpoi at cut-off value of OD ratio 0.2 (sensitivity 85.2% and specificity 100%). In diagnosis of MERS-CoV infection, ELISA IgG exhibited optimal performance using sera collected after 28 dpex, at a cut-off value of OD ratio 0.2 (sensitivity 97.3% and specificity 92.9%). These new breakpoints are markedly lower than previously suggested values (ELISA IgG OD ratio 1.1, sensitivity 34.8% and specificity 100% in the present data set), and the performance data help serologic tests to be practically used in the field of MERS management.

A patient with severe respiratory failure caused by novel human coronavirus.

We report a case of a 45-year-old patient who developed severe acute respiratory distress syndrome accompanied by renal failure. An infection with a novel human coronavirus was confirmed and found to be the reason for rapidly progressive respiratory failure of our patient.

Difficulties and challenges experienced by nurses in eldercare institutions in Albania: A qualitative content analysis.

INTRODUCTION: The global and Albanian populations of elderly people are steadily increasing. It is estimated that the number of elderly adults requiring care in Albania will rise from 90.9 thousand to 130.4 thousand by 2030. Despite the envisaged increase in the number and life expectancy of the elderly population in Albania, which will result in an increased demand for nursing care, little is known about the difficulties and challenges that nurses face while providing care for elderly Albanian individuals. AIM: To explore the difficulties and challenges nurses experience while caring for elderly people in Albanian eldercare institutions. METHODS: The study employed a qualitative design using purposive sampling of 20 nurses in 8 eldercare institutions who participated in face-to-face semi-structured interviews. The audio-recorded interviews were transcribed and subsequently subjected to analysis using Graneheim and Lundman's qualitative conventional content analysis. Data analysis was supported by the qualitative data analysis software MAXQDA 2020. The reporting of this study followed the consolidated criteria for reporting qualitative research (COREQ) checklist. RESULTS: Five key categories emerged from data analysis: (1) professional difficulties, (2) educational difficulties, (3) relationship challenges, (4) increased mental stress, and (5) participation in advocacy. This study showed that nursing staff experienced many barriers, challenges, and unmet needs when implementing care for elderly people in long-term care facilities. CONCLUSION: The findings indicate that nurses working in eldercare institutions faced significant challenges in caring for elderly people. Nurses need more legal, financial, educational, and emotional support. The study indicates that more organizational and national support is necessary for nursing staff to care for elderly people in eldercare Albanian institutions properly. Eldercare institution leaders need to recognize the importance of their role in overcoming the barriers and providing adequate support for their staff in caring for elderly people.

Contact investigation of a case of human novel coronavirus infection treated in a German hospital, October-November 2012.

On 24 October 2012, a patient with acute respiratory distress syndrome of unknown origin and symptom onset on 5 October was transferred from Qatar to a specialist lung clinic in Germany. Late diagnosis on 20 November of an infection with the novel Coronavirus (NCoV) resulted in potential exposure of a considerable number of healthcare workers. Using a questionnaire we asked 123 identified contacts (120 hospital and three out-of-hospital contacts) about exposure to the patient. Eighty-five contacts provided blood for a serological test using a two-stage approach with an initial immunofluorescence assay as screening test, followed by recombinant immunofluorescence assays and a NCoV-specific serum neutralisation test. Of 123 identified contacts nine had performed aerosol-generating procedures within the third or fourth week of illness, using personal protective equipment rarely or never, and two of these developed acute respiratory illness. Serology was negative for all nine. Further 76 hospital contacts also tested negative, including two sera initially reactive in the screening test. The contact investigation ruled out transmission to contacts after illness day 20. Our two-stage approach for serological testing may be used as a template for similar situations.

Specific serology for emerging human coronaviruses by protein microarray.

We present a serological assay for the specific detection of IgM and IgG antibodies against the emerging human coronavirus hCoV-EMC and the SARS-CoV based on protein microarray technology. The assay uses the S1 receptor-binding subunit of the spike protein of hCoV-EMC and SARS-CoV as antigens. The assay has been validated extensively using putative cross-reacting sera of patient cohorts exposed to the four common hCoVs and sera from convalescent patients infected with hCoV-EMC or SARS-CoV.

Middle East Respiratory Syndrome coronavirus (MERS-CoV) serology in major livestock species in an affected region in Jordan, June to September 2013.

Between June and September 2013, sera from 11 dromedary camels, 150 goats, 126 sheep and 91 cows were collected in Jordan, where the first human Middle-East respiratory syndrome (MERS) cluster appeared in 2012. All sera were tested for MERS-coronavirus (MERS-CoV) specific antibodies by protein microarray with confirmation by virus neutralisation. Neutralising antibodies were found in all camel sera while sera from goats and cattle tested negative. Although six sheep sera reacted with MERS-CoV antigen, neutralising antibodies were not detected.

Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction.

We present two real-time reverse-transcription polymerase chain reaction assays for a novel human coronavirus (CoV), targeting regions upstream of the E gene (upE) or within open reading frame (ORF)1b, respectively. Sensitivity for upE is 3.4 copies per reaction (95% confidence interval (CI): 2.5­6.9 copies) or 291 copies/mL of sample. No cross-reactivity was observed with coronaviruses OC43, NL63, 229E, SARS-CoV, nor with 92 clinical specimens containing common human respiratory viruses. We recommend using upE for screening and ORF1b for confirmation.

Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infections.

We present a rigorously validated and highly sensitive confirmatory real-time RT-PCR assay (1A assay) that can be used in combination with the previously reported upE assay. Two additional RT-PCR assays for sequencing are described, targeting the RdRp gene (RdRpSeq assay) and N gene (NSeq assay), where an insertion/deletion polymorphism might exist among different hCoV-EMC strains. Finally, a simplified and biologically safe protocol for detection of antibody response by immunofluorescence microscopy was developed using convalescent patient serum.

Mn-induced Fermi-surface reconstruction in the SmFeAsO parent compound.

The electronic ground state of iron-based materials is unusually sensitive to electronic correlations. Among others, its delicate balance is profoundly affected by the insertion of magnetic impurities in the FeAs layers. Here, we address the effects of Fe-to-Mn substitution in the non-superconducting Sm-1111 pnictide parent compound via a comparative study of SmFe[Formula: see text]Mn[Formula: see text]AsO samples with [Formula: see text] 0.05 and 0.10. Magnetization, Hall effect, and muon-spin spectroscopy data provide a coherent picture, indicating a weakening of the commensurate Fe spin-density-wave (SDW) order, as shown by the lowering of the SDW transition temperature [Formula: see text] with increasing Mn content, and the unexpected appearance of another magnetic order, occurring at [Formula: see text] and 20 K for [Formula: see text] and 0.10, respectively. We attribute the new magnetic transition at [Formula: see text], occurring well inside the SDW phase, to a reorganization of the Fermi surface due to Fe-to-Mn substitutions. These give rise to enhanced magnetic fluctuations along the incommensurate wavevector [Formula: see text], further increased by the RKKY interactions among Mn impurities.

A simple method to approximate liver size on cross-sectional images using living liver models.

AIM: To assess whether a simple. diameter-based formula applicable to cross-sectional images can be used to calculate the total liver volume. MATERIALS AND METHODS: On 119 cross-sectional examinations (62 computed tomography and 57 magnetic resonance imaging) a simple, formula-based method to approximate the liver volume was evaluated. The total liver volume was approximated measuring the largest craniocaudal (cc), ventrodorsal (vd), and coronal (cor) diameters by two readers and implementing the equation: Vol(estimated)=cc x vd x cor x 0.31. Inter-rater reliability, agreement, and correlation between liver volume calculation and virtual liver volumetry were analysed. RESULTS: No significant disagreement between the two readers was found. The formula correlated significantly with the volumetric data (r>0.85, p<0.0001). In 81% of cases the error of the approximated volume was <10% and in 92% of cases <15% compared to the volumetric data. CONCLUSION: Total liver volume can be accurately estimated on cross-sectional images using a simple, diameter-based equation.

Anti-cancer actions of a recombinant antibody (R6313/G2) against the angiotensin II AT1 receptor.

Although several tumour types express both AT1 and AT2 angiotensin II receptors, and angiotensin II stimulates cell proliferation, angiotensin-converting enzyme inhibitors and angiotensin receptor blockers are not effective anti-cancer agents. Development of a biologically active monoclonal antibody (6313/G2) against the AT1 receptor prompted the testing of a recombinant short-chain variable fragment form (R6313/G2) against breast cancer cells in vitro and in vivo. Cell lines MCF-7, MDA-MB-231 and T47D all expressed both receptor subtypes. In vitro, R6313/G2 suppressed cell proliferation in the presence of 100 nM angiotensin II, with IC50s of 30 nM, 153 nM and 2.8 microM for the three cell types respectively; in contrast, the AT1 receptor blocker losartan was effective only in T47D cells, at 25 microM. Studies on MCF-7 and T47D cells showed R6313/G2 also opposed the angiotensin II-induced inhibition of caspase-3/7 activity. In vivo, hollow fibres containing the cell lines were implanted in nu/nu balb-c mice at two sites, s.c. and i.p. Treatments of R6313/G2 at 2.5 nmol/kg and 25 nmol/kg twice per day for 7 days dose dependently reduced cell numbers for all three cell lines, but here MCF-7 cells responded most sensitively and MDA-MB-231 cells least. Although T47D cells were refractory at the s.c. site, growth was inhibited at the i.p. location, and otherwise results were similar at the two sites. In xenografts, MCF-7 cell tumours were dose dependently reduced by R6313/G2, and 13 and 27 nmol/kg R6313/G2 twice/day gave means of 74 and 76% tumour regression after 7 days. The data suggest that the anti-cancer action of R6313/G2 is considerably more effective than AT1 antagonists.

Neuromuscular characterisation in Cerebral Palsy using hybrid Hill-type models on isometric contractions.

BACKGROUND: Muscles of individuals with Cerebral Palsy (CP) undergo structural changes over their lifespan including an increase in muscle stiffness, decreased strength and coordination. Being able to identify these changes non-invasively would be beneficial to improve understanding of CP and assess therapy effectiveness over time. This study aims to adapt an existing EMG-driven Hill-type muscle model for neuromuscular characterisation during isometric contractions of the elbow joint. METHODS: Participants with (n = 2) and without CP (n = 8) performed isometric force ramps with contraction levels ranging between 15 and 70% of their maximum torque. During these contractions, high-density EMG data were collected from the M. Biceps and Triceps brachii with 64 electrodes on each muscle. The EMG-driven Hill-type muscle model was used to predict torques around the elbow joint, and muscle characterisation was performed by applying a genetic algorithm that tuned individuals' parameters to reduce the RMS error between observed and predicted torque data. RESULTS: Observed torques could be predicted accurately with an overall mean error of 1.24Nm ± 0.53Nm when modelling individual force ramps. The first four parameters of the model could be identified relatively reliably across different experimental protocols with a full-scale variation of below 20%. CONCLUSION: An HD-EMG muscle modelling approach to evaluating neuromuscular properties in participants with and without CP has been presented. This pilot study confirms the feasibility of the experimental protocol and demonstrates some parameters can be identified robustly using the isometric contraction force ramps.

Influence of the immune system on the course of infection with murine coronavirus JHM in suckling mice.

The course of infection with murine corona virus JHM in C3H mice depends on the age of the animals. Mice up to 20 days of age are fully susceptible while mice older than 23 days resist the infection. Protection of suckling mice from death of infection can be provided by intraperitoneal administration of immune spleen cells but not by non-immune spleen cells from adult mice. The immune spleen cells can be generated by priming adult mice, or by priming non-immune spleen cells from adult mice in baby mice with inactivated JHM virus. Thus the immune system might well be involved in the different outcome of infection with JHM-virus in suckling and adult mice, but it does not seem to be the exclusive factor responsible for the achievement of natural resistance.

[Ultrasound screening for Down's syndrome early in pregnancy: nuchal translucency thickness]. / Echoscopische screening op het syndroom van Down vroeg in de zwangerschap: de nekplooimeting.

Screening for Down's syndrome (DS) in the Netherlands is based on maternal age. Women aged 36 years or above are offered amniocentesis or chorionic villus sampling to determine if the foetus has Down's syndrome or other chromosomal abnormalities. Disadvantages of this method are a low detection rate of affected pregnancies (no more than 15%) and iatrogenic miscarriages (0.5-1%). Pregnant women under the age of 36 are not eligible for screening, even though 70% of the children with Down's syndrome are born in this age group. Screening by measuring nuchal translucency thickness, an early ultrasound marker for Down's syndrome carried out in the first trimester of pregnancy, has a higher detection rate with a less invasive method and therefore fewer iatrogenic miscarriages. However, Dutch law prohibits the offer of screening to women with a low individual risk of DS (Population Screening Act), because of the anxiety this may provoke in these women. The Dutch Health Council's Prenatal Screening Committee has advised the minister of Public Health, Welfare and Sport that risk-assessment screening, in particular the triple test, should no longer be postponed and that research into (other) screening options should be stimulated.

Characterization of SM201, an anti-hFcγRIIB antibody not interfering with ligand binding that mediates immune complex dependent inhibition of B cells.

The monoclonal antibody SM201 specifically recognizes the human inhibitory FcγRIIB without showing cross-reactivity to the related but activating FcγRIIA. The epitope recognized by SM201 is located outside the IgG-binding site of FcγRIIB. As a result, the antibody does not interfere with hIgG binding to the receptor. It was therefore hypothesized that SM201 may amplify the inhibitory signaling of FcγRIIB after coligation of B cell receptor (BCR) and FcγRIIB by immune complexes (ICs). Mechanistic and functional studies were conducted in established B cell lines as well as in primary B cells from healthy donors to substantiate the anticipated working principle. Using an experimental setup mimicking IC binding, we were able to demonstrate that SM201 enhances the extent of ITIM phosphorylation of FcγRIIB. The antibody works synergistically with ICs and the mode of action is strictly dependent on their presence. Additionally, SM201 did not induce apoptosis, cellular depletion or NK cell activation, which indicates an advantageous safety profile. This establishes an innovative approach for the treatment of antibody-mediated autoimmune diseases.

[How to Read a Chest-X-ray?]. / Wie lese ich eine Thoraxaufnahme?

Only a standartized, systematic approach allows a complete review of a chest x-ray and the collection of all relevant informations. In detail, alterations of the following structures should be sought: soft tissue, bones, pleura, mediastinum, lung, heart, pulmonary circulation and hili. Technical factors and the position of the patient should also be considered.

State of the art 3D imaging of abdominal organs.

Since its introduction in the late 70's 3D imaging has assumed growing importance for the visualization of abdominal pathologies. Main clinical applications include the preoperative planning for resection of liver, pancreas and kidney tumors, living-donor liver and renal transplantation, and tumor radiofrequency ablation of liver and kidney. The advantages of 3D imaging are the exact visualization of vessels in areas with complex and variable vascular anatomy, demonstration of possible resection margins, and predicting operative risks, especially when dealing with complex or minimal invasive surgery. 3D imaging is an important visual aid for the surgeon in order to facilitate crucial decisions and to lower the operative risk. In future, 3D planning and simulation of operations will be become standard procedures. Therefore an efficient use of 3D imaging is not possible without a close collaboration between radiologist and surgeons.