Evolution, Enzymaktivität und Struktur-Funktions-Analyse der Norovirus-Enzyme Protease (NS6pro) und Polymerase (NS7pol)

Die humanen Noroviren (Familie Caliciviridae, Genus Norovirus) bilden den Haupterreger der nichtbakteriellen Gastroenterotiden. Seit Beginn der 1990er Jahre – besonders seit dem Herbst 2002 – kann weltweit eine sehr starke Zunahme der Norovirus-Infektionen beobachtet werden. Die Mehrheit der Erkrankungen (ca. 80%) ist dabei durch den Genotyp II.4 verursacht, wobei innerhalb dieses Genotyps kontinuierlich „neue“ genetisch veränderte Norovirus-Stämme auftreten, die kontinuierlich in „neue“ Subgenotypen eingeordnet werden. Bis dato wird vermutet, dass die hohe Zunahme der Infektionen durch die subgenotypspezifische Evolution des strukturellen Proteins VP1 bewirkt wurde, da dadurch die Bindung der Viruspartikel an die Rezeptoren der Wirtszelle – die HBGAs – verbessert sein könnte bzw. ein breiteres Spektrum an Rezeptoren erkannt werden könnte. Bisher ist aber weitgehend unbekannt, ob im Bereich der nichtstrukturellen Proteine auch eine subgenotypspezifische Evolution stattgefunden hat und ob diese durch eine verbesserte Enzymaktivität zu der Zunahme der Infektionen beigetragen haben könnte. Im Rahmen dieser Arbeit wurden demnach aus unterschiedlichen Stuhlproben die beiden Enzyme NS6pro und NS7pol von jeweils einem Stamm der epidemiologisch relevanten Subgenotypen II.4-1995, II.4-2002, II.4-2004, II.4-2006a und II.4-2006b rekombinant hergestellt und betreffend ihrer Evolution und Enzymaktivität untersucht. Anhand der durchgeführten Untersuchungen konnte festgestellt werden, dass bei den epidemiologisch relevanten Subgenotypen des Genotyps II.4 im Bereich der NS6pro und der NS7pol keine subgenotypspezifische Evolution stattgefunden hat und daher keine subgenotypspezifische Enzymaktivität vorliegt. Somit hatte die Enzymaktivität der NS6pro und der NS7pol keinen Einfluss auf die Zunahme der Infektionen. Dennoch konnte aber beobachtet werden, dass durch die Mutation einer einzelnen Aminosäure – entsprechend der Position in der räumlichen Struktur der NS6pro bzw. der NS7pol – die Enzymaktivität um bis zu 100 Prozent erhöht bzw. reduziert werden kann. Bei diesen bedeutenden Mutationen ist entweder die Substratbindung oder die Koordination bzw. die Zufuhr der Reaktionskomponenten betroffen. Somit konnte bestätigt werden, dass die Zunahme der Infektionen vermutlich doch durch die Evolution des VP1 bedingt ist. Dies ist auch nachvollziehbar, da das VP1 den Eintritt der Viruspartikel in das Zellplasma erlaubt, in dem danach erst die Virusvermehrung durch die Enzyme stattfindet. Hierbei muss aber erwähnt werden, dass die Evolution des VP1 durch die Mutationsrate der NS7pol bestimmt ist, sodass die NS7pol letztendlich doch für die Zunahme der Infektionen verantwortlich ist. Um aber definitiv abzuklären, ob durch die Evolution des VP1 tatsächlich die Bindung der Viruspartikel an die HBGAs verbessert bzw. ein breiteres Spektrum an Rezeptoren erkannt wird, müssten bspw. Bindungs-Assays zwischen den entsprechenden VLPs (virus-like particles) und bereits typisierten Serum- oder Speichelproben – ähnlich den Untersuchungen von Lindesmith et al. (2008) – durchgeführt werden. Darüber hinaus müsste die Evolution und die Funktionalität der restlichen nichtstrukturellen Proteine untersucht bzw. deren Einfluss auf die Zunahme der Infektionen überprüft werden

Evaluation of the PotoClean® decontamination technology for reprocessing of water supply lines in dental units during routine work

Background: A frequent problem in dental units is the microbial contamination of water and biofilm formation in the water supply lines. After random identification of a bacterial contaminated dental unit (310 cfu/ml) in a practise with 3 dental units we implemented the present study to evaluate the efficacy of the PotoClean® technology, based on anodic oxidation

Pupil light reflex evoked by light-emitting diode and computer screen: methodology and association with need for recovery in daily life

Objectives: Pupil light reflex (PLR) has been widely used as a method for evaluating parasympathetic activity. The first aim of the present study is to develop a PLR measurement using a computer screen set-up and compare its results with the PLR generated by a more conventional setup using light-emitting diode (LED). The parasympathetic nervous system, which is known to control the ‘rest and digest’ response of the human body, is considered to be associated with daily life fatigue. However, only few studies have attempted to test the relationship between self-reported daily fatigue and physiological measurement of the parasympathetic nervous system. Therefore, the second aim of this study was to investigate the relationship between daily-life fatigue, assessed using the Need for Recovery scale, and parasympathetic activity, as indicated by the PLR parameters. Design: A pilot study was conducted first to develop a PLR measurement set-up using a computer screen. PLRs evoked by light stimuli with different characteristics were recorded to confirm the influence of light intensity, flash duration, and color on the PLRs evoked by the system. In the subsequent experimental study, we recorded the PLR of 25 adult participants to light flashes generated by the screen set-up as well as by a conventional LED set-up. PLR parameters relating to parasympathetic and sympathetic activity were calculated from the pupil responses. We tested the split-half reliability across two consecutive blocks of trials, and the relationships between the parameters of PLRs evoked by the two set-ups. Participants rated their need for recovery prior to the PLR recordings. Results: PLR parameters acquired in the screen and LED set-ups showed good reliability for amplitude related parameters. The PLRs evoked by both set-ups were consistent, but showed systematic differences in absolute values of all parameters. Additionally, higher need for recovery was associated with faster and larger constriction of the PLR. Conclusions: This study assessed the PLR generated by a computer screen and the PLR generated by a LED. The good reliability within set-ups and the consistency between the PLRs evoked by the set-ups indicate that both systems provides a valid way to evoke the PLR. A higher need for recovery was associated with faster and larger constricting PLRs, suggesting increased levels of parasympathetic nervous system activity in people experiencing higher levels of need for recovery on a daily basis

Exhaled and nasal nitric oxide in laryngectomized patients

<p>Abstract</p> <p>Background</p> <p>Nitric oxide (NO) shows differing concentrations in lower and upper airways. Patients after total laryngectomy are the only individuals, in whom a complete separation of upper and lower airways is guaranteed. Thus the objective of our study was to assess exhaled and nasal NO in these patients.</p> <p>Methods</p> <p>Exhaled bronchial NO (FE_NO) and nasal nitric oxide (nNO) were measured in patients after total laryngectomy (n = 14) and healthy controls (n = 24). To assess lung function we additionally performed spirometry. Co-factors possibly influencing NO, such as smoking, infections, and atopy were excluded.</p> <p>Results</p> <p>There was a markedly (p < 0.001) lower FE_NOin patients after total laryngectomy (median (range): 4 (1-22) ppb) compared to healthy controls 21 (9-41) ppb). In contrast, nNO was comparable between groups (1368 <it>versus </it>1380 in controls) but showed higher variability in subjects after laryngectomy.</p> <p>Conclusions</p> <p>Our data suggest that either bronchial NO production in patients who underwent laryngectomy is very low, possibly due to alterations of the mucosa or oxidant production/inflammation, or that substantial contributions to FE_NOarise from the larynx, pharynx and mouth, raising FE_NOdespite velum closure. The data fit to those indicating a substantial contribution to FE_NOby the mouth in healthy subjects. The broader range of nNO values found in subjects after laryngectomy may indicate chronic alteration or oligo-symptomatic inflammation of nasal mucosa, as frequently found after total laryngectomy.</p

Impact of stimulus-related factors and hearing impairment on listening effort as indicated by pupil dilation

Previous research has reported effects of masker type and signal-to-noise ratio (SNR) on listening effort, as indicated by the peak pupil dilation (PPD) relative to baseline during speech recognition. At about 50% correct sentence recognition performance, increasing SNRs generally results in declining PPDs, indicating reduced effort. However, the decline in PPD over SNRs has been observed to be less pronounced for hearing-impaired (HI) compared to normal-hearing (NH) listeners. The presence of a competing talker during speech recognition generally resulted in larger PPDs as compared to the presence of a fluctuating or stationary background noise. The aim of the present study was to examine the interplay between hearing-status, a broad range of SNRs corresponding to sentence recognition performance varying from 0 to 100% correct, and different masker types (stationary noise and single-talker masker) on the PPD during speech perception. Twenty-five HI and 32 age-matched NH participants listened to sentences across a broad range of SNRs, masked with speech from a single talker (−25 dB to +15 dB SNR) or with stationary noise (−12 dB to +16 dB). Correct sentence recognition scores and pupil responses were recorded during stimulus presentation. With a stationary masker, NH listeners show maximum PPD across a relatively narrow range of low SNRs, while HI listeners show relatively large PPD across a wide range of ecological SNRs. With the single-talker masker, maximum PPD was observed in the mid-range of SNRs around 50% correct sentence recognition performance, while smaller PPDs were observed at lower and higher SNRs. Mixed-model ANOVAs revealed significant interactions between hearing-status and SNR on the PPD for both masker types. Our data show a different pattern of PPDs across SNRs between groups, which indicates that listening and the allocation of effort during listening in daily life environments may be different for NH and HI listeners

Toward a more comprehensive understanding of the impact of masker type and signal-to-noise ratio on the pupillary response while performing a speech-in-noise test

Difficulties arising in everyday speech communication often result from the acoustical environment, which may contain interfering background noise or competing speakers. Thus, listening and understanding speech in noise can be exhausting. Two experiments are presented in the current study that further explored the impact of masker type and Signal-to-Noise Ratio (SNR) on listening effort by means of pupillometry. In both studies, pupillary responses of participants were measured while performing the Danish Hearing in Noise Test (HINT; Nielsen and Dau, 2011). The first experiment aimed to replicate and extend earlier observed effects of noise type and semantic interference on listening effort (Koelewijn et al., 2012). The impact of three different masker types, i.e. a fluctuating noise, a 1-talker masker and a 4-talker masker on listening effort was examined at a fixed speech intelligibility. In a second experiment, effects of SNR on listening effort were examined while presenting the HINT sentences across a broad range of fixed SNRs corresponding to intelligibility scores ranging from 100% to 0% correct performance. A peak pupil dilation (PPD) was calculated and a Growth Curve Analysis (GCA) was performed to examine listening effort involved in speech recognition as a function of SNR. The results of two experiments showed that the pupil dilation response is highly affected by both masker type and SNR when performing the HINT. The PPD was highest, suggesting the highest level of effort, for speech recognition in the presence of the 1-talker masker in comparison to the 4-talker babble and the fluctuating noise masker. However, the disrupting effect of one competing talker disappeared for intelligibly levels around 50%. Furthermore, it was demonstrated that the pupillary response strongly varied as a function of SNRs. Listening effort was highest for intermediate SNRs with performance accuracies ranging between 30% and 70% correct. GCA revealed time-dependent effects of the SNR on the pupillary response that were not reflected in the PPD

Impact of SNR, masker type and noise reduction processing on sentence recognition performance and listening effort as indicated by the pupil dilation response

Recent studies have shown that activating the noise reduction scheme in hearing aids results in a smaller peak pupil dilation (PPD), indicating reduced listening effort, at 50% and 95% correct sentence recognition with a 4-talker masker. The objective of this study was to measure the effect of the noise reduction scheme (on or off) on PPD and sentence recognition across a wide range of signal-to-noise ratios (SNRs) from +16 dB to −12 dB and two masker types (4-talker and stationary noise). Relatively low PPDs were observed at very low (−12 dB) and very high (+16 dB to +8 dB) SNRs presumably due to ‘giving up’ and ‘easy listening’, respectively. The maximum PPD was observed with SNRs at approximately 50% correct sentence recognition. Sentence recognition with both masker types was significantly improved by the noise reduction scheme, which corresponds to the shift in performance from SNR function at approximately 5 dB toward a lower SNR. This intelligibility effect was accompanied by a corresponding effect on the PPD, shifting the peak by approximately 4 dB toward a lower SNR. In addition, with the 4-talker masker, when the noise reduction scheme was active, the PPD was smaller overall than that when the scheme was inactive. We conclude that with the 4-talker masker, noise reduction scheme processing provides a listening effort benefit in addition to any effect associated with improved intelligibility. Thus, the effect of the noise reduction scheme on listening effort incorporates more than can be explained by intelligibility alone, emphasizing the potential importance of measuring listening effort in addition to traditional speech reception measures

Sacred turf: the Wimbledon tennis championships and the changing politics of Englishness

© 2015 Taylor & Francis. This article is about ‘Wimbledon’, widely celebrated – not least in its own publicity material – as the world’s premier tennis tournament. It examines ‘Wimbledon’ essentially as a text (hence the inverted commas), viewed politically and historically. In this context, ‘Wimbledon’ is seen as a signifier of a certain kind of Englishness, carefully adapted to meet changing social and economic circumstance. Loose parallels are drawn between the cultural trajectory of ‘Wimbledon’ and that of the British royal family. The transmutations of ‘Wimbledon’ as a tennis championship are also seen as reflecting Britain’s decline as a world power during the twentieth century

Incidence and outcomes of kidney replacement therapy for end-stage kidney disease due to primary glomerular disease in Europe:Findings from the ERA Registry

Background and hypothesis: Primary glomerular disease (PGD) is a major cause of end-stage kidney disease (ESKD) leading to kidney replacement therapy (KRT). We aimed to describe incidence (trends) in individuals starting KRT for ESKD due to PGD and to examine their survival and causes of death.Methods: We used data from the European Renal Association (ERA) Registry on 69,854 patients who started KRT for ESKD due to PGD between 2000 and 2019. ERA primary renal disease codes were used to define six PGD subgroups. We examined age and sex standardized incidence, trend of the incidence, and survival.Results: The standardized incidence of KRT for ESKD due to PGD was 16.6 per million population (pmp), ranging from 8.6 pmp in Serbia to 20.0 pmp in France. IgA nephropathy (IgAN) and focal segmental glomerulosclerosis (FSGS) had the highest incidence of 4.6 pmp and 2.6 pmp, respectively. Histologically non-examined PGDs represented over 50% of cases in Serbia, Bosnia and Herzegovina, and Romania and were also common in Greece, Estonia, Belgium, and Sweden. The incidence declined from 18.6 pmp in 2000 to 14.5 pmp in 2013, after which it stabilized. All PGD subgroups had five-year survival probabilities above 50%, with crescentic glomerulonephritis having the highest risk of death (adjusted hazard ratio: 1.8 [95% confidence interval: 1.6-1.9]) compared with IgAN. Cardiovascular disease was the most common cause of death (33.9%).Conclusion: The incidence of KRT for ESKD due to PGD showed large differences between countries and was highest for IgAN and FSGS. Lack of kidney biopsy facilities in some countries may have affected accurate assignment of the cause of ESKD. The recognition of the incidence and outcomes of KRT among different PGD subgroups may contribute to a more individualized patient care approach