bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Functional Nanopore Screen: A Versatile High-throughput Assay to Study and Engineer Protein Nanopores inEscherichia coli
Authors: Weber Wadim , Roeder Markus , Probanowski Tobias , Yang Jie , Abujubara Helal , Koeppl Heinz , Tietze Alesia , Stein Viktor 
Published: April 2021

AbstractNanopores comprise a versatile class of membrane proteins that carry out a range of key physiological functions and are increasingly developed for different biotechnological applications. Yet, a capacity to study and engineer protein nanopores by combinatorial means has so far been hampered by a lack of suitable assays that combine sufficient experimental resolution with throughput. Addressing this technological gap, the Functional Nanopore (FuN) screen now provides a quantitative and dynamic read-out of nanopore assembly and function inE. coli. The assay is based on genetically-encoded fluorescent protein sensors that resolve the nanopore-dependent influx of Ca2+across the inner membrane ofE. coli. Illustrating its versatile capacity, the FuN screen is first applied to dissect the molecular features that underlie the assembly and stability of nanopores formed by the S2168 holin. In a subsequent step, nanopores are engineered by recombining the transmembrane module of S2168 with different ring-shaped oligomeric protein structures that feature defined hexa-, hepta- and octameric geometries. Library screening highlights substantial plasticity in the ability of the S2168 transmembrane module to oligomerize in alternative geometries while the functional properties of the resultant nanopores can be fine-tuned through the identity of the connecting linkers. Overall, the FuN screen is anticipated to facilitate both fundamental studies and complex nanopore engineering endeavors with many potential applications in biomedicine, biotechnology and synthetic biology.
A shuttle-vector system allows heterologous gene expression in the thermophilic methanogenMethanothermobacter thermautotrophicusΔH
Authors: Fink Christian , Beblawy Sebastian , Enkerlin Andreas M. , Mühling Lucas , Angenent Largus T. , Molitor Bastian 
Published: April 2021

AbstractThermophilicMethanothermobacterspp. are used as model microbes to study the physiology and biochemistry of the conversion of hydrogen and carbon dioxide into methane (i.e., hydrogenotrophic methanogenesis), because of their short doubling times and robust growth with high growth yields. Yet, a genetic system for these model microbes was missing despite intense work for four decades. Here, we report the establishment of tools for genetic modification ofM. thermautotrophicus. We developed the modularMethanothermobactervector system, which provided shuttle-vector plasmids (pMVS) with exchangeable selectable markers and replicons for bothEscherichia coliandM. thermautotrophicus. ForM. thermautotrophicus, a thermostable neomycin-resistance cassette served as the selectable marker for positive selection with neomycin, and the cryptic plasmid pME2001 fromMethanothermobacter marburgensisserved as the replicon. The pMVS-plasmid DNA was transferred fromE. coliintoM. thermautotrophicus viainterdomain conjugation. After the successful validation of DNA transfer and positive selection inM. thermautotrophicus, we demonstrated heterologous gene expression of a thermostable β-galactosidase-encoding gene (bgaB) fromGeobacillus stearothermophilusunder the expression control of four distinct synthetic and native promoters. In quantitativein-vitroenzyme activity assays, we found significantly different β-galactosidase activity with these distinct promoters. With a formate dehydrogenase operon-encoding shuttle vector, we allowed growth ofM. thermautotrophicuson formate as the sole growth substrate, while this was not possible for the empty vector control. These genetic tools provide the basis to investigate hypotheses from four decades of research on the physiology and biochemistry ofMethanothermobacterspp. on a genetic level.Significance StatementThe world economies are facing permanently increasing energy demands. At the same time, carbon emissions from fossil sources need to be circumvented to minimize harmful effects from climate change. The power-to-gas platform is utilized to store renewable electric power and decarbonize the natural gas grid. The microbeMethanothermobacter thermautotrophicusis already applied as the industrial biocatalyst for the biological methanation step in large-scale power-to-gas processes. To improve the biocatalyst in a targeted fashion, genetic engineering is required. With our shuttle-vector system for heterologous gene expression inM. thermautotrophicus, we set the cornerstone to engineer the microbe for optimized methane production, but also for production of high-value platform chemicals in power-to-x processes.
Wolbachia cifBinduces cytoplasmic incompatibility in the malaria mosquito
Authors: Adams Kelsey L. , Abernathy Daniel G. , Willett Bailey C. , Selland Emily K. , Itoe Maurice A. , Catteruccia Flaminia 
Published: April 2021

AbstractWolbachiainfections are a fascinating example of reproductive parasitism with strong potential to combat vector-borne diseases, due to their combined ability to spread in insect populations and block pathogen replication. Though theWolbachiafactors mediating the notable reproductive manipulation cytoplasmic incompatibility (CI) have now been identified as prophage WO genescifAandcifB, the relative role of these genes is still intensely debated, with different models claiming that CI requires either both factors orcifBalone. Here we investigated whethercifAandcifBare sufficient to induce conditional sterility in the major malaria vectorAnopheles gambiae, a species that appears to have limited susceptibility to invasion byWolbachia. We report that CI can be fully recapitulated in these mosquitoes, and thatcifBis sufficient to cause this reproductive manipulation.cifB-induced sterility is fully rescued by high levels ofcifAexpression in females. Surprisingly, however, whencifAis highly expressed in males alongsidecifB, the CI phenotype is attenuated.cifBstrongly impairs fertility also when expressed in the female germline, again mitigated bycifA. These data support a system wherebycifBandcifAmust be fine-tuned to exercise CI and rescue, respectively, possibly explaining the limited success ofWolbachiaat invadingAnopheles. Our findings pave the way towards facilitatingWolbachiainfections in anopheline vectors, for use in malaria control strategies.
Multifactorial seroprofiling dissects the contribution of pre-existing human coronaviruses responses to SARS-CoV-2 immunity
Authors: Abela Irene A. , Pasin Chloé , Schwarzmüller Magdalena , Epp Selina , Sickmann Michèle E. , Schanz Merle M. , Rusert Peter , Weber Jacqueline , Schmutz Stefan , Audigé Annette , Maliqi Liridona , Hunziker Annika , Hesselman Maria C. , Niklaus Cyrille R. , Gottschalk Jochen , Schindler Eméry , Wepf Alexander , Karrer Urs , Wolfensberger Aline , Rampini Silvana K. , Meyer Sauteur Patrick M. , Berger Christoph , Huber Michael , Böni Jürg , Braun Dominique L. , Marconato Maddalena , Manz Markus G. , Frey Beat M. , Günthard Huldrych F. , Kouyos Roger D. , Trkola Alexandra 
Published: April 2021

AbstractDetermination of SARS-CoV-2 antibody responses in the context of pre-existing immunity to circulating human coronavirus (HCoV) is critical to understanding protective immunity. Here we perform a multifactorial analysis of SARS-CoV-2 and HCoV antibody responses in pre-pandemic (N=825) and SARS-CoV-2-infected donors (N=389) using a custom-designed multiplex ABCORA assay. ABCORA seroprofiling, when combined with computational modeling, enables accurate definition of SARS-CoV-2 seroconversion and prediction of neutralization activity, and reveals intriguing interrelations with HCoV immunity. Specifically, higher HCoV antibody levels in SARS-CoV-2-negative donors suggest that preexisting HCoV immunity may provide protection against SARS-CoV-2 acquisition. In those infected, higher HCoV activity is associated with elevated SARS-CoV-2 responses, indicating cross-stimulation. Most importantly, HCoV immunity may impact disease severity, as patients with high HCoV reactivity are less likely to require hospitalization. Collectively, this evidence points to HCoV immunity promoting the rapid development of SARS-CoV-2-specific immunity, underscoring the importance of exploring cross-protective responses for comprehensive coronavirus prevention.
Clinical evaluation of the SD Biosensor saliva antigen rapid test with symptomatic and asymptomatic, non-hospitalized patients
Authors: Igloi Zsofia , Velzing Jans , Huisman Robin , Geurtsvankessel Corine , Comvalius Anoushka , van Beek Janko , Ensing Roel , Boelsums Timo , Koopmans Marion , Molenkamp Richard 
Published: April 2021

AbstractPerformance of saliva antigen rapid test was evaluated in non-hospitalized patients, with or without symptoms. Overall sensitivity was 66.1% compared to RT-PCR in saliva. Using cycle threshold <30 cutoff or virus culture as reference, sensitivity increased to 88.6% or 96.7% respectively. Specificity was 99.6%.
Clinical profile of cutaneous adverse events of immune checkpoint inhibitors in a single tertiary center
Authors: Park Ji‐Hye , Yoon Dokyoung , Lee Jeeyun , Oh Se Jin , Kim Hyun Je , Lee Jong Hee , Lee Dong‐Youn 
Published: April 2021

AbstractProgrammed death 1 (PD‐1)/programmed death ligand 1 (PD‐L1) inhibitors have demonstrated their efficacy in the treatment of various malignancies. Despite their benefits, their immunomodulatory activities can cause unpredictable cutaneous adverse events (CAE). This study aimed to identify characteristics of CAE in patients treated with PD‐1/PD‐L1 inhibitors through the medical records, photographs, and pathology reports. Fifty CAE occurred in 47 (2.75%) of 1711 patients treated with PD‐1/PD‐L1 inhibitors. Pruritic, psoriasiform, urticarial, and acneiform eruptions were the four most common types. Melanoma patients showed CAE more frequently than other malignancies. Acneiform eruption occurred more often at ages under 60 years. Urticarial eruption appeared earlier, while keratoacanthoma appeared later after immunotherapy. The overall survival times were not significantly different between the two groups with and without CAE by Kaplan–Meier analysis (p = 0.055). Studies on CAE may provide more information to understand these drugs and to help manage the patients.
Improving performance of species distribution model in mountainous areas with complex topography
Authors: Liao Chi‐Cheng , Chen Yi‐Huey 
Published: April 2021

AbstractThis study aims to improve performance of species distribution model in mountainous areas with complex topography. Random Forest algorithm was used to evaluate distribution ranges ofTrochodendron aralioidesSiebold & Zucc. in northern Taiwan to identify the determinants of the plant distributions and to assess the effects of data collections on the spatial bias of model predictions. Gridded climate dataset with spatial resolution of 50 × 50 m2for the study area was generated from daily data of 18 meteorological stations and environmental variables were interpolated by inverse distance weighted method and altitudinal adjusted by linear regression model using empirical lapse rates varied among slope inclinations. Model performances were much better when model was run with larger sizes of training datasets generated by randomized resample of presence/absence data records. Surprisingly, distribution ranges of the plant were drastically affected by marginal individuals that is likely to have reached its geographical border as well as climatic limits. Wind speed in winter and temperature differences between warmest and coldest months contributed noticeably to the model predictions. In contrast, index of cloud occurrence contributed little to the model predictions, despiteT. aralioideswas a characteristic species of cloud zone. The interpolated bioclimatic predictors had precisely reflected the habitat differentiation among topographical positions and accurately captured habitat requirements of plant species at fine scale. Our results presented practical and applicable example of novel interpolation method to generate gridded climate dataset that is useful to improve performance of model predictions in mountainous areas with complex topography.
Increased variation in sex organ positions can increase reciprocity and pollination success in heterostylous plant populations
Authors: Ganguly Shatarupa , Shreenidhi PM , Barua Deepak 
Published: April 2021

AbstractThe reciprocal position of sexual organs in complementary floral morphs is central to our understanding of heterostyly. Reciprocity indices are used to quantify the spatial match between complementary sex organs, but previous indices fail to appropriately account for intra‐population variation in sex organ positions. In this study, we examine how an increase in intra‐population variation in sex organ heights affects reciprocity and consequently reproductive success. We formulated a reciprocity index that incorporates this variation and asked if estimates of reciprocity can predict reproductive success in naturally occurring heterostylous populations. We developed a reciprocity index that assumed pollen transfer success equalled one for a perfectly matched stigma–anther pair, and decreased to zero with increasing mismatch. We examined the relationship between intra‐population variation in organ position and reciprocity, compared previously proposed indices using simulated populations and empirical data from natural populations, and tested the ability of the indices to predict reproductive success. We observed that when differences between mean complementary sex‐organ heights are small, increasing intra‐population variation in heights resulted in a decrease in reciprocity. However, when this difference is larger, reciprocity increased, reached a peak, and then decreased with increasing variation. Previous indices failed to capture this behavior. Seed set was positively related to reciprocity for our index. These results challenge the current understanding that increasing variation in sex‐organ heights will always decrease reciprocity in heterostylous populations. This may help explain why heterostylous systems exhibit and tolerate high amounts of intra‐population variation in sex organ heights.
Nanoliposomes for encapsulation and calcium delivery of egg white peptide–calcium complex
Authors: Zhang Penglin , Bao Zhijie , Jiang Pengfei , Zhang Simin , Zhang Xiumin , Lin Songyi , Sun Na 
Published: April 2021

AbstractNanoliposomes and crude liposomes loaded with egg white peptide–calcium complex (EWP–Ca) were fabricated by thin‐film dispersion with or without dynamic high‐pressure microfluidization. Their physiochemical properties, in vitro stability, and calcium release profiles were investigated in this study. Results showed that the EWP–Ca‐loaded nanoliposomes exhibited spherical structures with a lower particle size and polydispersity index as well as a higher thermal stability as compared to the corresponding crude liposomes. Further investigations revealed that EWP–Ca was embedded into the liposomes mainly through hydrogen bonding and present in an amorphous form within the liposomes. Additionally, the EWP–Ca‐loaded nanoliposomes effectively slowed the release of calcium in gastric digestion, allowing more soluble calcium to enter the intestinal tract; in the subsequent intestinal digestion, the EWP–Ca‐loaded nanoliposomes were more electrically and physically stable than the crude liposomes. Therefore, the EWP–Ca‐loaded nanoliposomes could be used as a favorable dietary calcium delivery system to promote calcium bioavailability.Practical ApplicationNanoliposomes were fabricated in this study to encapsulate the egg white peptide–calcium complex (EWP–Ca) for calcium delivery. The EWP–Ca‐loaded nanoliposomes effectively slowed the release of calcium in gastric digestion, allowing more soluble calcium to enter the intestinal tract, and were more electrically and physically stable in the subsequent intestinal digestion. Therefore, the EWP–Ca‐loaded nanoliposomes may be incorporated in calcium‐fortified food to enhance calcium delivery for maintaining bone health.
General anaesthesia in end‐of‐life care: extending the indications for anaesthesia beyond surgery
Authors: Takla A. , Savulescu J. , Wilkinson D. J. C. , Pandit J. J. 
Published: April 2021

SummaryIn this article, we describe an extension of general anaesthesia – beyond facilitating surgery – to the relief of suffering during dying. Some refractory symptoms at the end of life (pain, delirium, distress, dyspnoea) might be managed by analgesia, but in high doses, adverse effects (e.g. respiratory depression) can hasten death. Sedation may be needed for agitation or distress and can be administered as continuous deep sedation (also referred to as terminal or palliative sedation) generally using benzodiazepines. However, for some patients these interventions are not enough, and others may express a clear desire to be completely unconscious as they die. We summarise the historical background of an established practice that we refer to as ‘general anaesthesia in end‐of‐life care’. We discuss its contexts and some ethical and legal issues that it raises, arguing that these are largely similar issues to those already raised by continuous deep sedation. To be a valid option, general anaesthesia in end‐of‐life care will require a clear multidisciplinary framework and consensus practice guidelines. We see these as an impending development for which the specialty should prepare. General anaesthesia in end‐of‐life care raises an important debate about the possible role of anaesthesia in the relief of suffering beyond the context of surgical/diagnostic interventions.
The Energetic Brain Club in Life and Death
Authors: Hames Aaron 
Published: April 2021

SummaryJapan’s aging society leaves many of its eldest residents to encounter loneliness and social isolation. However, the elderly in Japan and elsewhere are rarely passive vessels to fate and often use available tools to meet their needs. Exploring loneliness and its remedy, this essay presents an ethnographic account of a single Energetic Brain Club meeting in the Tokyo sprawl. While, on paper, the club is aimed at preventing dementia, cognitive declines frequently remain a peripheral topic. Elderly members partake in hospitality rituals and engage in long, meandering conversations about family, memories, struggles, and politics. In the process, they find commiseration, connection, and a salve for loneliness.
Measuring optic nerve head perfusion to monitor glaucoma: a study on structure–function relationships using laser speckle flowgraphy
Authors: Calzetti Giacomo , Mursch‐Edlmayr Anna Sophie , Bata Ahmed M. , Ungaro Nicola , Mora Paolo , Chua Jacqueline , Schmidl Doreen , Bolz Matthias , Garhöfer Gerhard , Gandolfi Stefano , Schmetterer Leopold , Wong Damon 
Published: April 2021

AbstractPurposeWe aimed to describe the global and localized correlations among visual field (VF) sensitivity, optic nerve head (ONH) perfusion measured by laser speckle flowgraphy (LSFG) and neural structure measured by optical coherence tomography (OCT) in open‐angle glaucoma (OAG) and to compare the floor effect for LSFG and OCT.MethodsCross‐sectional, multicenter study including one eye each from fifty OAG patients (mean age 69.3 years; average VF mean deviation, MD, −8.5 dB, range −25.17 to 0.85 dB) and fifty‐one controls. Patients underwent SITA standard 24‐2 automated perimetry and measurement of ONH perfusion, peripapillary retinal nerve fibre layer thickness (RNFLT) and macular ganglion cell‐inner plexiform layer thickness (GCIPLT). We tested the presence of a significant change (breakpoint) in the correlation slope with VF sensitivity to assess floor effect.ResultsThe correlation between the LSFG parameter Mean All (MA) of the global disc area and MD (r = 0.56, p < 0.001) did not show a breakpoint, in contrast to the correlations between MD and OCT global parameters, which showed breakpoints at −8.53 and −4.05 dB for RNFLT and GCIPLT, respectively. Global and localized correlations with VF sensitivity were stronger for LSFG compared to OCT. In particular, LSFG outperformed OCT in the correlation with the central VF sector (r = 0.50, p < 0.001 and r = 0.06, p = 0.67 for MA and RNFLT, respectively).ConclusionThe global and sectoral correlations with VF sensitivity and the favourable floor effect compared to OCT indicate LSFG as a promising tool to monitor progression particularly in late‐stage glaucoma. Further longitudinal studies are warranted.
Efficacy of bleb‐independent penetrating canaloplasty in primary angle‐closure glaucoma: one‐year results
Authors: Zhang Shaodan , Hu Cheng , Cheng Huanhuan , Gu Juan , Samuel Kwizera , Lin Haishuang , Deng Yuxuan , Xie Yanqian , Hu Jingjing , Le Rongrong , Xu Shuxia , Tham Clement C. , Liang Yuanbo 
Published: April 2021

AbstractPurposeTo report the efficacy and safety of bleb‐independent penetrating canaloplasty in the management of primary angle‐closure glaucoma (PACG).MethodsThis single‐centre prospective interventional case series enrolled 57 eyes from 53 PACG patients with medically uncontrolled intraocular pressure (IOP) and peripheral anterior synechiae of over 270°. Penetrating canaloplasty, mainly consisted of tensioning suture‐aided Schlemm’s canal dilation and a trabeculectomy, was performed to create a direct communication between the anterior chamber and the Schlemm’s canal. Postoperative IOP, number of glaucoma medications and procedure‐related complications were evaluated. Rate of success was defined as IOP ≤ 21, ≤18 and ≤15 mmHg, and a ≥30% IOP reduction without (complete) or with/without (qualified) IOP‐lowering medications.ResultsA total of 45 eyes had 360° catheterization successfully completed. The mean preoperative IOP was 33.9 ± 11.7 mmHg (range, 13–59.6 mmHg), on 3.2 ± 0.8 glaucoma medications (range 2–5), which was decreased to 15.4 ± 3.7 mmHg (range, 8.6–22.5) and 0.2 ± 0.6 (range, 0–3) medications at 6 months and 14.8 ± 3.5 mmHg (range, 9–24) and 0.1 ± 0.3 (range, 0–1) medications at 12 months postoperatively. Complete success at 12 months were achieved in 78.9% (95% CI: 0.65–0.93), 71.1% (0.56–0.86) and 50.0% (0.33–0.67) eyes at IOP ≤ 21, ≤18 and ≤15 mmHg, respectively. Transient IOP elevation (>30 mmHg, 26.7%) and hyphema (11.1%) were the most common early surgical complications.ConclusionPenetrating canaloplasty in PACG appeared to have good efficacy and safety profiles in this pilot study. Further studies are justified.