Darkness inhibits autokinase activity of bacterial bathy phytochromes.

Bathy phytochromes are a subclass of bacterial biliprotein photoreceptors that carry a biliverdin IXα chromophore. In contrast to prototypical phytochromes that adopt a red-light-absorbing Pr ground state, the far-red light-absorbing Pfr-form is the thermally stable ground state of bathy phytochromes. Although the photobiology of bacterial phytochromes has been extensively studied since their discovery in the late 1990s, our understanding of the signal transduction process to the connected transmitter domains, which are often histidine kinases, remains insufficient. Initiated by the analysis of the bathy phytochrome PaBphP from Pseudomonas aeruginosa, we performed a systematic analysis of five different bathy phytochromes with the aim to derive a general statement on the correlation of photostate and autokinase output. While all proteins adopt different Pr/Pfr-fractions in response to red, blue, and far-red light, only darkness leads to a pure or highly enriched Pfr-form, directly correlated with the lowest level of autokinase activity. Using this information, we developed a method to quantitatively correlate the autokinase activity of phytochrome samples with well-defined stationary Pr/Pfr-fractions. We demonstrate that the off-state of the phytochromes is the Pfr-form and that different Pr/Pfr-fractions enable the organisms to fine-tune their kinase output in response to a certain light environment. Furthermore, the output response is regulated by the rate of dark reversion, which differs significantly from 5 s to 50 min half-life. Overall, our study indicates that bathy phytochromes function as sensors of light and darkness, rather than red and far-red light, as originally postulated.

In Vitro Mastication Simulation and Wear Test of Virgilite and Advanced Lithium Disilicate Ceramics.

PURPOSE: To compare wear behavior, durability during in vitro mastication simulation, and fracture force of an established and a novel lithium disilicate CAD/CAM material, as well as to examine the impact of cementation and reduced ceramic thickness on durability and fracture force. MATERIALS AND METHODS: Specimens (n = 8 per group) were prepared from lithium disilicate (LS2; IPS e.max, Ivoclar Vivadent) and advanced lithium disilicate (ALD; Cerec Tessera, Dentsply Sirona). Specimens were polished, and two-body wear test and thermocycling were performed (50 N, 120,000 cycles, 1.6 Hz, H2O dist., 5°C/55°C, 600 cycles). Maximum vertical loss, surface roughness, surface roughness depth, and antagonist wear were determined. Single crowns (n = 8 per group; thickness 1.5 mm/1.0 mm) were manufactured from LS2 and ALD and mounted on human molar teeth with adhesive resin (AB; CalibraCeram, Dentsply Sirona), glass-ionomer cement (GIC; Ketac Cem, 3M ESPE), and hybrid glass-ionomer cement (HGIC; Calibra Bio, Dentsply Sirona). Thermocycling and mechanical loading (2 × 3000 × 5°C/55°C, 2 minutes, H20 dist., 1.2 × 106 50 N) were performed. Fracture force was determined by a universal testing machine (1446, ZwickRoell), and one-way analysis and Bonferroni post hoc test (α = .05) were used for statistical analyses. RESULTS: Mean (ALD: 210 ± 42.4 µm; LS2: 264.3 ± 56.1 µm) and maximum (ALD: 391.1 ± 86.3 µm; LS2: 518.3 ± 113.2 µm) wear between groups were significantly different (P ≤ .047). Fracture force varied between 1,911.4 ± 468.4 N (ALD/AB 1 mm) and 2,995.3 ± 880.6 N (LS2/GIC), without significant differences (P ≥ .152). CONCLUSION: ALD showed better wear behavior than LS2, but provided similar fracture force. Cementation and reduction of ceramic thickness had only minor effects on fracture force.

American prejudice during the COVID-19 pandemic.

In the United States, anti-Asian sentiment has pervaded the Coronavirus 2019 (COVID-19) pandemic. Could Americans' fear of contracting the virus relate to prejudice against Asian individuals? According to intergroup threat theory, prejudice increases toward groups of people when they are perceived as a likely cause of symbolic and/or real threat, including disease threat. We tested this perspective in the context of the COVID-19 pandemic by investigating the relationship between Americans' concern about contracting COVID-19 and their feelings toward individuals from multiple countries. Between May 12-14 2020, participants residing in the United States (N = 932) completed an online survey assessing their (1) perceived threat of COVID-19 infection, (2) feelings of warmth and coldness toward people in America, China, Italy, Japan, and Greece, and (3) trait-level prejudice. Perceived threat of COVID-19 infection differentially related to feelings toward American and Chinese nationals and was unrelated to feelings toward people from other countries assessed. Specifically, greater threat of infection was associated with less warmth toward individuals from China, an effect moderated by trait-level prejudice. That is, participants high (but not medium or low) in trait prejudice showed a significant relationship between threat of COVID-19 infection and reduced warmth toward Chinese individuals. Threat of infection also related to greater warmth and less coldness toward American nationals, consistent with prior work indicating that disease threats amplify ethnocentrism. Collectively, results suggest that perceived threat of COVID-19 infection may correspond with prejudice toward the national outgroup associated with the disease's origin (i.e., China), as well as national ingroup favoritism, among Americans prone to prejudice.

Investigating the mechanical and optical properties of novel Urethandimethacrylate (UDMA) and Urethanmethacrylate (UMA) based rapid prototyping materials.

OBJECTIVE: This study is focused on testing experimental rapid prototyping materials for occlusal splints made from Urethandimethacrylate (UDMA) and Urethanmethacrylate (UMA). METHODS: Materials were mixed from UDMA and UMA in ratios of 1.0:0.0, 0.75:0.25, 0.5:0.5, 0.25:0.75 and 0.0:1.0. Specimens were printed using digital light processing (DLP). After post-processing, the specimens underwent testing on flexural strength, modulus of elasticity, hardness, wear behavior, surface roughness, gloss and color stability. All tests were performed after 24 h (baseline) and 10 days of water storage (aging). Splints underwent cyclic pull-off and insertion testing, which was alongside simulated using finite element analysis. RESULTS: The mechanical properties were significantly influenced by changes in the UDMA:UMA ratio. Statistical analysis revealed that increased amounts of UMA correlated with a decrease in flexural strength (92.0 to 30.7 MPa), modulus of elasticity (2.4 to 0.6 GPa), hardness (155.1 to 102.0 N/mm2) and wear resistance (-1394.9 to -1742.1 µm). Materials with higher amounts of UMA were also more likely to be influenced by water storage. Specimens with 75% and 100% UMA content were partly not analyzable due to soft consistency. Optical properties showed only minor influence from UMA content and aging. Differences in surface roughness (3.9 to 2.4 µm) and color stability were insignificant. Gloss was partly influenced by the UDMA:UMA ratio and water storage. Mean survival rates for cyclic pull-off and insertion testing ranged from 2537 to 23,857 cycles. A correlation between the amount of UMA and survival rates was observed. SIGNIFICANCE: The addition of up to 25% UMA showed promising results, complying with clinical standards and delivering acceptable results in the cyclic pull-off and insertion test. Further investigation on increments between 0 and 25% UMA could help to find an optimum.

Machine-driven simulation of removing luting agent remnants from implant surfaces: An investigator-independent assessment of cleaning protocols.

INTRODUCTION: To simulate removing luting agent remnants from crowns fixed onto implant-abutment analogs using a standardized machine-driven protocol including a scaler and air polishing or sonic. MATERIAL AND METHODS: A motor-driven device was constructed that controlled the rotational speed of the specimens, machining distance, contact pressure, and working time. A standardized layer of cement (Provicol, VOCO; Cuxhaven, G; Ketac Cem, 3MEspe; Seefeld, G; or Rely X Unicem, 3MEspe, Seefeld, G) was placed onto the finishing line of the crowns luted onto titanium-abutment analogs. The cement layer was scaled with a fresh titanium scaler maneuvered by the motor-driven device and treated with air polishing or sonic. Protocol 1: Scaling only for 20s, 40s, or 60s; n=20; protocol 2: 40s of scaling plus 20s of air polishing; protocol 3: 20s of scaling plus 40s of air polishing; protocol 4: 20s of scaling plus 40s of sonic; protocol 5: 40s of scaling plus 20s of sonic; protocols 2-5: n=10. Cement remnants were counted digitally as "percentage of remnants". STATISTICS: mean, standard deviation, Bonferroni post hoc tests; α=0.05. RESULTS: Ketac Cem was easily removed by scaling only and Provicol by scaling and air polishing, but the self-adhesive resin composite cement Rely X Unicem was not removable with the device. Only remnants of Provicol could be significantly reduced by further treatment after scaling (p<0.001). CONCLUSION: The presented motor-driven device enables reproducible investigations of various cleaning protocols and is thus useful to create an overview of cleaning protocols needed for the different types of cement.

Dynamic fatigue of 3D-printed splint materials.

OBJECTIVES: Printed splints may be an alternative treatment for functional disorders. In addition to the selection of materials, the influence of cleaning or polymerisation can affect the dynamic behaviour and fatigue limit of printed materials. MATERIAL AND METHODS: 96 discs (n = 6 per group, 16mmx2mm) were printed (P30+ DLP-printer, Straumann, CH; 100 µm layer) from splint materials (M1: Luxaprint OrthoPlus, DMG, G; M2: V-Print Splint, Voco, G). Specimens were either automatically cleaned (C1: Straumann P Wash, Straumann, CH) or manually cleaned (C2: Voco Pre-/Main-Clean protocol, Voco, G). Post polymerisation was performed with LED (P1: Cure, Straumann, CH) or Xenon light (P2: Otoflash N171, Ernst Hinrichs Dental, G). The flexural fatigue limit was determined under cyclic loading in terms of a staircase approach with a piston-on-3-ball-test according to ISO 6872 after 24 h or 60 days water storage (37 °C). Specimens were preloaded with 50N and dynamic force was applied for 105 loadings per step (f = 3Hz; steps 1: 50N-100N, 2: 50N-150N, 3: 50N-200N, 4: 50N-250N; F1000, Prematec, G; water at 37 °C). STATISTICS: Kaplan Maier Log Rank (Mantel-Cox) test, ANOVA, Pearson correlations, Levene-test (α = 0.05, SPSS 26.0, IBM, Armonk, NY, USA)). RESULTS: Mean survival cycles after 24 h of storage varied between 40388 (M1C2P2) and 195140 (M2C2P1) cycles and after 60 d decreased to 14022 (M1C2P2) and 173237 (M2C1P1). Kaplan Maier Log Rank test revealed significant differences between the material combinations. For M1 cleaning (Pearson: 0.346, p = 0.016) and for M2 polymerisation (Pearson: 0.616, p = 0.000) significantly influenced the number of loading cycles. Intermediate effects were found for material (p = 0.026), cleaning (p = 0.024) and polymerisation (p = 0.000) as well as the combination of material and polymerisation (p = 0.008). CONCLUSIONS: The results show that the number of possible loading cycles of additively manufactured splint specimens depends on the type of material, their cleaning and post-polymerisation. CLINICAL SIGNIFICANCE: Materials, cleaning and post-polymerisation of additive manufacturing processes should be matched to improve dynamic loading performance of splint materials.

Quantification of cooling effects on basic tissue measurements and exposed cross-sectional brain area of cadaver heads from market pigs.

The objective of this project was to determine the impact of cooling on the soft tissue thickness, cranial thickness, and cross-sectional brain area of cadaver heads from market pigs. Documenting the effect of cooling on tissue dimensions of swine heads is valuable and important for future investigations of physical stunning and euthanasia methods that use cadaver heads. Scalded and dehaired cadaver heads with intact jowls were sourced from market pigs stunned with CO2 gas. After transport to the data collection location, a penetrating captive bolt (PCB) shot (Jarvis Model PAS-Type P 0.25R Caliber Captive Bolt Pistol with Medium Rod Assembly and Blue Powder Cartridges) was applied in the frontal position. Following PCB application, each head (n = 36) underwent an UNCHILLED treatment followed by CHILLED treatment. The UNCHILLED treatment involved images collected immediately after splitting each head along the bolt path, and the CHILLED treatment involved images of the same heads after storage in a walk-in cooler for 24 h at 2 to 4°C. All measurements for each treatment were collected from images of the heads on the plane of the bolt path immediately prior to and immediately after the refrigeration treatment. Measurements were performed by two observers. Across all measurements, mean interobserver coefficient of variation was 11.3 ± 0.6%. The soft tissue caudal to the bolt path was different (P = 0.0120) between treatments (CHILLED: 6.4 ± 0.2 mm; UNCHILLED: 7.2 ± 0.2 mm). The soft tissue thickness rostral to the bolt path was different (P = 0.0378) between treatments (CHILLED: 5.5 ± 0.2 mm; UNCHILLED: 6.1 ± 0.2 mm). Cranial thickness caudal to the bolt path was not different (P = 0.8659; CHILLED: 18.1 ± 0.6 mm; UNCHILLED: 18.3 ± 0.6 mm), nor was there a significant difference (P = 0.2593) in cranial thickness rostral to the bolt path between treatments (CHILLED: 16.2 ± 0.6 mm; UNCHILLED: 15.2 ± 0.6 mm). Cross-sectional brain area did not differ (P = 0.0737; CHILLED: 3633.4 ± 44.1 mm; UNCHILLED: 3519.9 ± 44.1 mm). A correction factor of 1.12 was determined from this study for cases where estimation of UNCHILLED soft tissue thickness from CHILLED soft tissue thickness is necessary.

How Young German General Practitioners View and Use Complementary and Alternative Medicine: A Qualitative Study.

BACKGROUND: Qualitative and quantitative surveys show that many German general practitioners (GPs) use complementary and alternative medicine (CAM) in their daily work. However, participants in such studies were mostly experienced GPs. OBJECTIVE: The aim of this study was to investigate systematically how young GPs view CAM and to what extent and how they use CAM treatments. METHODS: Thirteen young physicians in specialist training for general practice as well as GPs who had completed their specialist training within the last 2 years participated in problem-oriented interviews. Interviews were evaluated by qualitative content analysis. RESULTS: The attitudes of participants towards CAM in general and towards specific CAM modalities varied strongly, but doubts regarding a firm evidence base and specific effects over placebo were expressed frequently. Nevertheless, participants were generally open to herbal medicines and all seemed to use them more or less regularly in clinical practice. Other CAM treatments seemed to be used infrequently. Participants used CAM often for minor illnesses; largely as a first low-intensity therapeutic trial to avoid potentially harmful conventional treatments; and to comply with patient preferences. CONCLUSIONS: Compared to more experienced GPs, the participants in our study tended to be more critical towards CAM. Apart from herbal medicines, CAM treatments were used rarely.

[How do young general practitioners experience the transition to general practice? A qualitative study]. / Wie erleben junge Allgemeinärzt*innen den Übergang in die hausärztliche Praxis? Eine qualitative Studie.

BACKGROUND: Although the role of general practice has been strengthened in recent years, undergraduate teaching at medical schools and the clinical phase of specialist training remain dominated by specialized care of seriously ill people in hospitals. It is to be assumed that young doctors' views on medical care are strongly shaped by this clinical focus. OBJECTIVE: To investigate how young general practitioners (GPs) perceive transition from medical school and hospital work to general practice. METHODS: In a qualitative study, a total of 13 physicians in specialist training for general practice as well as general practitioners who had completed their specialist examination up to two years ago participated in problem-oriented interviews. The interviews were analyzed using content analysis. RESULTS: The significant differences between hospital-based and primary care practice initially came as quite a shock to the study participants. Key differences and challenges compared to working in a hospital included: 1) the totally different type of patients or complaints they faced; 2) learning that in many situations one can and should bide one's time ("wait-and-see" approach); 3) ruling out avoidable dangerous developments in patients reliably and coping with the corresponding residual risk; 4) the discovery that sometimes it makes sense not to make a diagnosis; 5) that the doctor-patient relationship should be more cooperative in general practice; and 6) that GPs are often under pressure to act although there is no clear need for taking action or no clear treatment option from a medical and scientific point of view. CONCLUSIONS: Our findings confirm that young doctors' initial views on medical care are strongly shaped by the clinical focus of medical schools and hospital work. Working in general practice is perceived as being very different from working in a hospital.

Cardiorespiratory fitness, sleep, and physiological responses to stress in women.

Epidemiological studies suggest that cardiorespiratory fitness (CRfitness) is associated with reduced risk of depression and anxiety in women, however, the mechanisms by which CRfitness may be protective against the development of these disorders are less clear. Because sleep problems are associated with both a higher risk for mental illness and altered physiological responses to stress, this study investigated whether sleep quality might influence the relationship between CRfitness and physiological stress responses in women. Thirty healthy women (18-45 y) who were medication-free, with regular menstrual cycles completed: (1) enrolment visit [including the assessment of CRfitness via maximal oxygen consumption during exercise]; (2) one-week sleep monitoring period including subjective (daily sleep diaries) and objective (wrist actigraphy) sleep measures; and (3) psychosocial stressor protocol (the Trier Social Stress Test; TSST) for the collection of heart rate (HR), blood pressure (BP), and salivary cortisol stress responses. Higher CRfitness was associated with reduced wake after sleep onset (WASO) duration (r = -.38, p = 0.04), higher self-reported sleep quality (higher scores reflect poorer sleep quality; r = -.37, p = 0.05), and lower HR (r = -.43, p = 0.02) during the stressor. Higher sleep quality was associated with a lower HR during the stressor (r = .44, p = 0.01). Increased WASO duration and WASO number were associated with blunted cortisol output during the stressor (r = -.44, p = 0.02, and r = -.46, p = 0.02, respectively). Results suggest that, in women, CRfitness may be protective against the deleterious effects of stress via improved sleep quality.

Design of the Enzyme-Carrier Interface to Overcome the O2 and NADH Mass Transfer Limitations of an Immobilized Flavin Oxidase.

Understanding how the immobilization of enzymes on solid carriers affects their performance is paramount for the design of highly efficient heterogeneous biocatalysts. An efficient supply of substrates onto the solid phase is one of the main challenges to maximize the activity of the immobilized enzymes. Herein, we apply advanced single-particle analysis to decipher the optimal design of an immobilized NADH oxidase (NOX) whose activity depends both on O2 and NADH concentrations. Carrier physicochemical properties and its functionality along with the enzyme distribution across the carrier were implemented as design variables to study the effects of the intraparticle concentration of substrates (O2 and NADH) on the activity. Intraparticle O2-sensing analysis revealed the superior performance of the enzyme immobilized at the outer surface in terms of effective supply of O2. Furthermore, the co-immobilization of NADH and NOX within the tuned surface of porous microbeads increases the effective concentration of NADH in the surroundings of the enzyme. As a result, the optimal spatial organization of NOX and its confinement with NADH allow a 100% recovery of the activity of the soluble enzyme upon the immobilization process. By engineering these variables, we increase the NADH oxidation activity of the heterogeneous biocatalyst by up to 650% compared to NOX immobilized under suboptimal conditions. In conclusion, this work highlights the rational design and engineering of the enzyme-carrier interface to maximize the efficiency of heterogeneous biocatalysts.

Sequencing of 231 forensic genetic markers using the MiSeq FGx™ forensic genomics system - an evaluation of the assay and software.

The MiSeq FGx™ Forensic Genomics System types 231 genetic markers in one multiplex polymerase chain reaction (PCR) assay. The markers include core forensic short tandem repeats (STRs) as well as identity, ancestry and phenotype informative short nucleotide polymorphisms (SNPs). In this work, the MiSeq FGx™ Forensic Genomics System was evaluated by analysing reproducibility, sensitivity, mixture identification and forensic phenotyping capabilities of the assay. Furthermore, the genotype calling of the ForenSeq™ Universal Analysis Software was verified by analysing fastq.gz files from the MiSeq FGx™ platform using the softwares STRinNGS and GATK. Overall, the performance of the MiSeq FGx™ Forensic Genomics System was high. However, locus and allele drop-outs were relatively frequent at six loci (two STRs and four human identification SNPs) due to low read depth or skewed heterozygote balances, and the stutter ratios were larger than those observed with conventional STR genotyping methods. The risk of locus and allele drop-outs increased dramatically when the amount of DNA in the first PCR was lower than 250 pg. Two-person 50:1 mixtures were identified as mixtures, whereas 100:1 and 1 000:1 mixtures were not. Y-chromosomal short tandem repeats (Y-STRs) alleles were detected in the 100:1 and 1 000:1 female/male mixtures. The ForenSeq™ Universal Analysis Software provided the data analyst with useful alerts that simplified the analysis of the large number of markers. Many of the alerts were due to user-defined, locus-specific criteria. The results shown here indicated that the default settings should be altered for some loci. Also, recommended changes to the assay and software are discussed.

Pediatric material properties: a review of human child and animal surrogates.

Because pediatric tissue is difficult for researchers to obtain, the biomechanical responses of adult humans have been studied much more extensively than those of children. Piglets, chimpanzees, and other animals have been used as child surrogates, but the tissue properties and responses to impact forces obtained from these animals may not directly correlate with the human child, and this correlation is not well understood. Consequently, only a handful of human pediatric tissue properties are known. Child anthropomorphic test devices employed in automotive safety have been developed largely by scaling data obtained from adult human cadaveric tests, where various scaling methods have been used to account for differences in geometry, material properties, or a combination of these two parameters. Similar scaling techniques have also been implemented to develop injury assessment reference values for child anthropomorphic test devices. Nevertheless, these scaling techniques have not yet proven to be accurate, in part because of the lack of pediatric data. In this review, the properties of pediatric human and animal surrogate tissue that have been mechanically tested are evaluated. It was found that most of the pediatric tissue that has previously been tested pertains to the head, neck, cervical spine, and extremities. It is evident that some body regions, such as the head and neck, have been tested to some extent since injuries to these regions are critical from an injury perspective. On the other hand, there is limited pediatric data available for the thorax, abdomen, thoracic and lumbar spines and fetal-related tissue. This review presents the pediatric data available in the literature and highlights the body regions where further testing is needed.

Finite element model development of a child pelvis with optimization-based material identification.

A finite element (FE) model of a 10-years-old child pelvis was developed and validated against experimental data from lateral impacts of pediatric pelves. The pelvic bone geometry was reconstructed from a set of computed tomography images, and a hexahedral mesh was generated using a new octree-based hexahedral meshing technique. Lateral impacts to the greater trochanter and iliac wing of the seated pelvis were simulated. Sensitivity analysis was conducted to identify material parameters that substantially affected the model response. An optimization-based material identification method was developed to obtain the most favorable material property set by minimizing differences in biomechanical responses between experimental and simulation results. This study represents a pilot effort in the development and validation of age-dependent musculoskeletal FE models for children, which may ultimately serve to evaluate injury mechanisms and means of protection for the pediatric population.