The nucleocapsid of hepatitis B virus (HBV), or HBcAg, is a highly symmetric structure formed by multiple dimers of a single core protein that contains potent T helper epitopes in its 183-aa ...sequence. Both factors make HBcAg an unusually strong immunogen and an attractive candidate as a carrier for foreign epitopes. The immunodominant c/e1 epitope on the capsid has been suggested as a superior location to convey high immunogenicity to a heterologous sequence. Because of its central position, however, any c/e1 insert disrupts the core protein's primary sequence; hence, only peptides, or rather small protein fragments seemed to be compatible with particle formation. According to recent structural data, the epitope is located at the tips of prominent surface spikes formed by the very stable dimer interfaces. We therefore reasoned that much larger inserts might be tolerated, provided the individual parts of a corresponding fusion protein could fold independently. Using the green fluorescent protein (GFP) as a model insert, we show that the chimeric protein efficiently forms fluorescent particles; hence, all of its structurally important parts must be properly folded. We also demonstrate that the GFP domains are surface-exposed and that the chimeric particles elicit a potent humoral response against native GFP. Hence, proteins of at least up to 238 aa can be natively displayed on the surface of HBV core particles. Such chimeras may not only be useful as vaccines but may also open the way for high resolution structural analyses of non-assembling proteins by electron microscopy.
Purpose Huntington disease (HD) is a chronic, debilitating genetic disease that affects physical, emotional, cognitive, and social health. Existing patient-reported outcomes (PROs) of health-related ...quality of life (HRQOL) used in HD are neither comprehensive, nor do they adequately account for clinically meaningful changes in function. While new PROs examining HRQOL (i.e., Neuro-QoL— Quality of Life in Neurological Disorders and PROMIS— Patient-Reported Outcomes Measurement Information System) offer solutions to many of these shortcomings, they do not include HD-specific content, nor have they been validated in HD. HDQLIFE addresses this by validating 12 PROMIS/Neuro-QoL domains in individuals with HD and by using established PROMIS methodology to develop new, HD-specific content. Methods New item pools were developed using cognitive debriefing with individuals with HD, and expert, literacy, and translatability reviews. Existing item banks and new item pools were field tested in 536 individuals with prodromal, early-, or late-stage HD. Results Moderate to strong relationships between Neuro-QoL/PROMIS measures and generic self-report measures of HRQOL, and moderate relationships between NeuroQoL/PROMIS and clinician-rated measures of similar constructs supported the validity of Neuro-QoL/PROMIS in individuals with HD. Exploratory and confirmatory factor analysis, item response theory, and differential item functioning analyses were utilized to develop new item banks for Chorea, Speech Difficulties, Swallowing Difficulties, and Concern with Death and Dying, with corresponding six-item short forms. A four-item short form was developed for Meaning and Purpose. Conclusions HDQLIFE encompasses both validated Neuro-QoL/PROMIS measures, as well as five new scales in order to provide a comprehensive assessment of HRQOL in HD.
Time-of-flight (ToF) performances of large area diamond detectors based on polycrystalline samples of dimensions 20 mm × 20 mm and thickness of 0.3 mm are presented. The devices that feature ...segmented Cr/Au electrodes in a sandwich configuration were irradiated with a heavy ions 197Au beam of 1 GeV u−1 showing, when mounted with a separation distance of 2 cm, a ToF resolution σ¯ of 37.5 ps averaged on 16 strip pairs. When ToF measurements were performed over a particle path of 30 m, a resolution σ of 45 ps was achieved. The detectors were mounted on high-vacuum compatible printed circuit boards (PCBs) with integrated processing electronics. This is the first time that ToF measurements have been performed using integrated electronics with such a large separation distance between the diamond detectors.
Only few studies have focused on the cognitive processes of the respondents that are involved when answering physical activity questionnaires (PAQs). This study aimed at examining whether two PAQs ...work as intended with different segments of the survey population in different cultural settings in Europe.
The International Physical Activity Questionnaire - Short Form (IPAQ-SF) and the US National Health Interview Survey - Adult Core Physical Activity Questionnaire (NHIS-PAQ) were tested in Belgium, Estonia, Germany and the UK using a standardized cognitive interviewing procedure. IPAQ-SF measures total vigorous physical activity (PA), moderate PA, walking and sitting. NHIS-PAQ measures leisure-time vigorous PA, light and moderate PA and muscle-strengthening PA. In total 62 persons completed cognitive interviews, at least 15 interviews were conducted in each country.
Both PAQs performed as intended with young and high-skilled persons and those having a regular exercise schedule. For the others, however, the testing revealed that problems occurred with both PAQs relating to understanding the concepts of '(light and) moderate' and 'vigorous' PA, classifying activities into the provided answer options of different PA intensities, recalling instances of 'normal' activities such as walking and sitting, and calculating the total duration of more than one activity or instance of an activity. The revealed problems with the questionnaires were quite similar in different countries; profound cultural differences were not observed.
Both questionnaires were difficult to answer for many respondents and rather user-unfriendly. They are designed to measure an exactness of PA quantity (frequency and duration) and intensity which would be desirable to obtain from a scientific point of view; however, respondents can hardly provide this information for cognitive reasons. Studies investigating the respondents' perspective are useful for improving physical activity information based on self-reports.
The present work reports on a long-term irradiation test performed on a 0.3-mm thick polycrystalline diamond detector prototype. The device, biased at 300 V, was continuously irradiated for about ...60 h by using a 12C beam at 62 MeV/nucl. at the LNS-INFN Cyclotron facility in Catania. An ionization chamber, calibrated using a single crystal diamond detector, was used to evaluate the total absorbed dose in the diamond detector under study. Data analysis carried out on recorded oscilloscope waveforms shows no significant variation of the signal properties in terms of amplitude, slope and charge after exposure to an integrated flux of 6.2⋅ 1013 ions/cm2. The results indicate that a polycrystalline diamond detector can be used for calibrating intensity monitors at the future superconducting fragment separator Super-FRS at FAIR (Darmstadt).
•No change observed in the signal proprieties of an irradiated pcCVD diamond material•Diamond detectors as precise intensity monitors with heavy ion beams•Survival of a pcCVD diamond detector after long-term beam current calibrations
Hydrogen peroxide is a reactive oxygen species (ROS) naturally occurring at low levels in aquatic environments and production varies widely across different ecosystems. Oxygenic photosynthesis ...generates hydrogen peroxide as a byproduct, of which some portion can be released to ambient water. However, few studies have examined hydrogen peroxide dynamics in relation to cyanobacterial harmful algal blooms (cHABs). A year-long investigation of algal succession and hydrogen peroxide dynamics was conducted at the Caloosahatchee River, Florida, USA. We aimed to identify potential biological mechanisms responsible for elevated hydrogen peroxide production during cHAB events through the exploration of the freshwater microbial metatranscriptome. Hydrogen peroxide concentrations were elevated from February to September of 2021 when cyanobacteria were active and abundant. We observed one Microcystis cHAB event in spring and one in winter. Both had distinct nutrient uptake and cyanotoxin gene expression patterns. While meaningful levels of microcystin were only detected during periods of elevated hydrogen peroxide, cyanopeptolin was by far the most expressed cyanotoxin during the spring bloom when hydrogen peroxide was at its yearly maxima. Gene expressions of five microbial enzymes (Rubisco, superoxide dismutase, cytochrome b559, pyruvate oxidase, and NADH dehydrogenase) positively correlated to hydrogen peroxide concentrations. Additionally, there was higher nitrogen-fixing gene (nifDKH) expression by filamentous cyanobacteria after the spring bloom but no secondary bloom formation occurred. Overall, elevated environmental hydrogen peroxide concentrations were linked to cyanobacterial dominance and greater expression of specific enzymes in the photosynthesis of cyanobacteria. This implicates cyanobacterial photosynthesis and growth results in increased hydrogen peroxide generation as reflected in measured environmental concentrations.
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•Hydrogen peroxide dynamics coincided with annual cyanobacterial growth pattern.•Hydrogen peroxide surge during cHAB events coincided with high Rubisco expression.•Microcystis gene expression decreased by 98% after bloom formation.•Highest cyanobacterial nitrogen fixation expressions occurred after cHAB event.•High cyanopeptolin gene expressions by Microcystis during hydrogen peroxide maxima.
Few field trials examining hydrogen peroxide as a cyanobacterial harmful algal bloom (cHAB) treatment have been conducted in subtropical and tropical regions. None have been tested in Florida, home ...to Lake Okeechobee and downstream waterways which periodically experience Microcystis bloom events. To investigate treatment effects in Florida, we applied a 490 μM (16.7 mg/L; 0.0015%) hydrogen peroxide spray to a minor bloom of Microcystis aeruginosa on the downstream side of Franklin Lock and Dam in the Caloosahatchee River. Although hydrogen peroxide decreased to background level one day post-treatment, succession was observed in phytoplankton community amplicon sequencing. The relative abundance of Microcystis decreased on day 3 by 86%, whereas the picocyanobacteria Synechococcus became dominant, increasing by 77% on day 3 and by 173% on day 14 to 57% of the phytoplankton community. Metatranscriptomics revealed Synechococcus likely benefitted from the antioxidant defense of upregulated peroxiredoxin, peroxidase/catalase, and rubrerythrin expressions immediately after treatment, and upregulated nitrate transport and urease to take advantage of available nitrogen. Our results indicated hydrogen peroxide induces succession of the phytoplankton community from Microcystis to non-toxic picocyanobacteria and could be used for selective suppression of harmful cyanobacteria.
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•490 μM hydrogen peroxide treatment selectively suppressed dominant bloom species.•Succession to picocyanobacteria within 3 days of hydrogen peroxide treatment.•Microcystis phylotypes exhibited different sensitivity to hydrogen peroxide.
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