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Abstract Inspired by biology, great progress has been made in creating artificial molecular motors. However, the dream of harnessing proteins – the building blocks selected by nature – to design autonomous motors has so far remained elusive. Here we report the synthesis and characterization of the Lawnmower, an autonomous, protein-based artificial molecular motor comprised of a spherical hub decorated with proteases. Its “burnt-bridge” motion is directed by cleavage of a peptide lawn, promoting motion towards unvisited substrate. We find that Lawnmowers exhibit directional motion with average speeds of up to 80 nm/s, comparable to biological motors. By selectively patterning the peptide lawn on microfabricated tracks, we furthermore show that the Lawnmower is capable of track-guided motion. Our work opens an avenue towards nanotechnology applications of artificial protein motors.

Summary: The axon initial segment (AIS) is located at the proximal axon demarcating the boundary between axonal and somatodendritic compartments. The AIS facilitates the generation of action potentials and maintenance of neuronal polarity. In this study, we show that the location of AIS assembly, as marked by Ankyrin G, corresponds to the nodal plane of the lowest-order harmonic of the Laplace-Beltrami operator solved over the neuronal shape. This correlation establishes a coupling between location of AIS assembly and neuronal cell morphology. We validate this correlation for neurons with atypical morphology and neurons containing multiple AnkG clusters on distinct neurites, where the nodal plane selects the appropriate axon showing enriched Tau. Based on our findings, we propose that Turing patterning systems are candidates for dynamically governing AIS location. Overall, this study highlights the importance of neuronal cell morphology in determining the precise localization of the AIS within the proximal axon.

Abstract Cryptophyte algae have a unique phycobiliprotein light-harvesting antenna that fills a spectral gap in chlorophyll absorption from photosystems. However, it is unclear how the antenna transfers energy efficiently to these photosystems. We show that the cryptophyte Hemiselmis andersenii expresses an energetically complex antenna comprising three distinct spectrotypes of phycobiliprotein, each composed of two αβ protomers but with different quaternary structures arising from a diverse α subunit family. We report crystal structures of the major phycobiliprotein from each spectrotype. Two-thirds of the antenna consists of open quaternary form phycobiliproteins acting as primary photon acceptors. These are supplemented by a newly discovered open-braced form (~15%), where an insertion in the α subunit produces ~10 nm absorbance red-shift. The final components (~15%) are closed forms with a long wavelength spectral feature due to substitution of a single chromophore. This chromophore is present on only one β subunit where asymmetry is dictated by the corresponding α subunit. This chromophore creates spectral overlap with chlorophyll, thus bridging the energetic gap between the phycobiliprotein antenna and the photosystems. We propose that the macromolecular organization of the cryptophyte antenna consists of bulk open and open-braced forms that transfer excitations to photosystems via this bridging closed form phycobiliprotein.

Cryptophytes acquired plastids from red algae but replaced the light-harvesting phycobilisome with a unique cryptophyte antenna. Here via analysis of phycobilisome cryo-EM structures, Rathbone et al. propose that the α subunit of the cryptophyte antenna originated from phycobilisome linker proteins

The authors examined the research and activism of 10th graders (ages 15-16) involved in a youth participatory action research project in a course in qualitative research at their small public school in New York City, New York. The authors, a classroom researcher and a high school teacher, looked closely at how the youth researchers used photography, collage, and videography to transform their school hallways into a space for critical conversations about race and gender. The authors examined how the hallways became a civic space shaped by collective youth resistance, multimodal counterstories, and negotiated civic engagement.

Cecal volvulus is an uncommon cause of acute intestinal obstruction accounting for around 10% of intestinal volvuli. There are three main variants of cecal volvuli including the axial, loop, and bascule types. Diagnosis is confirmed via a computed tomography scan and surgery is the mainstay treatment due to a high risk of morbidity and mortality. Here we report a rare presentation of cecal volvulus in a COVID-19 positive patient that was complicated by an anastomotic leak.

We investigated research processes applied in recent publications in the "European Journal of Engineering Education" (EJEE), exploring how papers link to theoretical work and how research processes have been designed and reported. We analysed all 155 papers published in EJEE in 2009, 2010 and 2013, classifying the papers using a taxonomy of research processes in engineering education research (EER) (Malmi et al. 2012). The majority of the papers presented either empirical work (59%) or were case reports (27%). Our main findings are as follows: (1) EJEE papers build moderately on a wide selection of theoretical work; (2) a great majority of papers have a clear research strategy, but data analysis methods are mostly simple descriptive statistics or simple/undocumented qualitative research methods; and (3) there are significant shortcomings in reporting research questions, methodology and limitations of studies. Our findings are consistent with and extend analyses of EER papers in other publishing venues; they help to build a clearer picture of the research currently published in EJEE and allow us to make recommendations for consideration by the editorial team of the journal. Our employed procedure also provides a framework that can be applied to monitor future global evolution of this and other EER journals.

Introduction Osteoporosis is a bone disease that is both preventable and treatable. It usually becomes evident when a fragility fracture occurs. Unfortunately, most studies show that only a small percentage of individuals at increased risk of fracture are assessed and treated, even following a fragility fracture. Objective The aim of this study was to determine whether patients suffering from a low-energy hip fractures in the Maltese Islands are given osteoporosis treatment. Method All patients older than 50 years presenting to the acute care hospitals in Malta and Gozo with a fragility hip fracture during December 1, 2015 and November 30, 2016 were included. Data on mortality, other fragility fractures, prescription of calcium, vitamin D, and antiresorptive therapy were collected. Results Calcium with vitamin D supplements were prescribed to 40% of patients; however, only 2.64% of patients were given pharmacological therapy. Following a hip fracture, the mortality rate was 18.5% at 1 year and 26.21% at 2 years. Apart from a high mortality rate, 28.19% of individuals sustained another fragility fracture before or after the hip fracture. Conclusion There should be increased osteoporosis awareness in Malta and a national bone mineral density screening program should be set up. An active role of the orthogeriatrics team in the management and treatment of osteoporosis following a fragility fracture might improve treatment rate and decrease refracture and mortality rates.

Young people are constantly targeted by online advertisements. This systematic literature review aims to identify the principal factors that influence young people's attitude towards online advertising. It seeks to understand if the influence of online advertising is dependent on gender and age differences in young people. The methodology involves the systematic review of empirical studies published from 1994 to 2020 and identifies the factors that impact young people's attitude towards online advertising. The review commences with a scoping study and follows the PRISMA structure, which includes identifying studies, screening and evaluation, the analysis and synthesis of studies, and the presentation of the final studies. This work reveals two main observations from a thematic content analysis of the appraised studies. The first relates to the principal factors influencing young people's attitude towards advertising. These factors comprise informativeness, entertainment, irritation, credibility, personalisation, and interactivity. In general, studies show that when informativeness, entertainment, credibility and interactivity are present, the more positive the attitude towards advertising is, while irritation fosters an unfavourable attitude. Studies provide a dichotomous stand on personalisation. The second relates to young people's age and gender. Our review notes that earlier studies lack focus on audiences predominantly between 10 and 15 years. Keywords: Attitude towards online advertising, advertising value, young people JEL Classifications: M31, M37 DOI: https://doi.org/10.32479/irmm.11398

The impact of drip irrigation on structural stability of soil aggregates was studied in soils of an apple (Malus domestica Borkh.) orchard, developed on marl. The field study was carried out in a sloping (20%) terrain in the north-eastern Slovenia at three slope positions (upslope, mid-slope and downslope), involving a comparison of irrigated versus non-irrigated situations after 6 years of drip irrigation practice. Structural stability was studied in three soil layers (0-5, 5-15 and 15-30 cm) at the end of the irrigation season (in September). In the same samples, soil organic carbon, total carbonates and soil moisture contents were determined. Drip irrigation significantly reduced structural stability and soil organic carbon in the surface soil layer (0-5 cm), while total carbonates increased. Based on the whole set of data, structural stability was strongly positively correlated with total carbonates and negatively correlated with soil organic carbon. This means that the effect of higher level of organic matter mineralisation on structural stability, due to irrigation, is counterbalanced by the increase of total carbonates content in the fine textured calcareous soils. Thus, a negative effect of irrigation on soil organic carbon had less destructive consequences on structural stability than expected.

Purpose: The purpose of this paper was to enhance and complement teaching about resource system feedbacks and environmental modelling. Students were given an interactive exercise based on a research model (ForeseerTM), developed by an inter-disciplinary research team, that explores the interconnectivity of water, energy and land resources. Two groups of students were involved, one of undergraduates and the other of graduates. Design/methodology/approach: The Foreseer model represents physical flows of the three resources (water, energy and land) using an interactive visual interface. The exercise was set up by giving students short instructions about how to use the tool to create four scenarios, and an online questionnaire was used to capture their understanding and their ability to extract information from the model. Findings: The exercise proved to be a helpful way to connect research and teaching in higher education, to the benefit of both. For students, it was an interactive and engaging way to learn about these complex sustainability issues. At the same time, it provided tangible feedback to researchers working on the model about the clarity of its user interface and its pedagogic value. Originality/value: This exercise represents a novel use of a resource model as a teaching tool in the study of the water, energy and land nexus, and is relevant to sustainability educators as an example of a model-centred learning approach on this topic.

We generated draft genome sequences for two cold-adapted Archaea, Methanogenium frigidum and Methanococcoides burtonii, to identify genotypic characteristics that distinguish them from Archaea with a higher optimal growth temperature (OGT). Comparative genomics revealed trends in amino acid and tRNA composition, and structural features of proteins. Proteins from the cold-adapted Archaea are characterized by a higher content of non-charged polar amino acids, particularly Gln and Thr and a lower content of hydrophobic amino acids, particularly Leu. Sequence data from nine methanogen genomes (OGT 15-98oC) was used to generate 1 111 modeled protein structures. Analysis of the models from the cold-adapted Archaea showed a strong tendency in the solvent accessible area for more Gln, Thr an hydrophobic residues and fewer charged residues. A cold shock domain (CSD) protein (CspA homolog) was identified in M. frigidum, two hypothetical proteins with CSD-folds in M. burtonii, and a unique winged helix DNA-binding domain protein in M. burtonii. This suggests that these types of nucleic acid binding proteins have a critical role in cold-adapted Archaea. Structural analysis of tRNA sequences from the Archaea indicated that GC content is the major factor influencing tRNA stability in hyperthermophiles, but not in the psychrophiles, mesophiles or moderate thermophiles. Below an OGT of 60oC, the GC content in tRNA was largely unchanged, indicating that any requirement for flexibility of tRNA in psychrophiles is mediated by other means. This is the first time that comparisons have been performed with genome data from Archaea spanning the growth temperature extremes from psychrophiles to hyperthermophiles.

The aim of this research was to evaluate the physical attributes of Cambisol in apple (Malus domestica Borkh. cv. Gala) orchard under different weed management systems in the state of Santa Catarina (SC), Brazil. The treatments were as follows: control (no weed management), chemical desiccation and mowing. We took samples of undisturbed soil at depths of 0-5, 5-10, 10-15 and 15-20 cm. We evaluated hydraulic conductivity (HC); bulk density (BD); soil penetration resistance (Pr); total porosity (TP), macropores (MaP), micropores (MiP); soil aggregation indices such as arithmetic and geometric mean diameter of air-dried and water-stable aggregates (AMDad, AMDws, GMDad, GMDws, respectively); aggregate stability index (ASIAMD and ASIGMD) and volumetric water content (VWC). We also carried out principal component analysis (PCA) with the data. The mowing or desiccation treatments increased BD at the 15-20 cm depth compared to control. The mowing treatment had the highest ASIGMD while desiccation contributed to the lowest ASIGMD. The desiccation treatment decreased AMDws (0-10 cm), ASIAMD (0-5 cm) GMDws (5-10 cm) and reduced HC by 26% and 22%, respectively, compared to the mowing and control treatments. Through PCA we were able to separate the three treatments. The mowing treatment was correlated with HC, VWC at pressure of 600 kPa and ASIGMD; desiccation was correlated with BD and Pr; and control with ASIAMD, MaP and TP.

Introduction The Littre hernia is a rare complication of Meckel's diverticulum. Meckel's diverticulum is vestigial remnant of the omphalomesenteric duct occurring in approximately 2% of the general population with an estimated 4 to 16% risk of complications. Usual sites of the Littre hernia include inguinal (50%), umbilical (20%), and femoral (20%). We report a case of an acquired transthoracic Littre's hernia occurring through the left part of the diaphragm triggered by a history of traumatic rib fractures associated with alcohol abuse. Case Report A 71-year-old man presented with 4-day history of worsening shortness of breath, colicky lower abdominal pain, and inability to open bowels despite passing flatus, without nausea or vomiting. His past medical history was remarkable for multiple traumatic rib fractures caused by falls which were associated with excessive alcohol consumption. A noncontrast computed tomography (CT) scan of the abdomen and pelvis showed distended jejunal loops containing air/fluid levels likely resulting from herniated jejunum between the left chest wall and left diaphragm. An urgent laparotomy was performed which revealed small bowel and omentum herniating through a small defect in the left posterior hemidiaphragm. The contents of the sac were reduced and a Meckel's diverticulum was found inside the sac, characteristic of Littre's hernia. The diaphragmatic defect was closed and the Meckel diverticulum stapled and excised. Discussion Herniation of Meckel's diverticulum through the diaphragm most commonly occurs in the pediatric population. Acquired transthoracic Littre's hernia is rare and may arise following thoracobdominal trauma caused by surgery, motor vehicle accidents, and falls from height. Left-diaphragmatic tears are characteristically more clinically apparent and symptomatic than the right since the liver often has a protective effect on the right part of the diaphragm. Herniation of abdominal contents in the chest cavity causes respiratory distress and requires urgent surgical correction. Diagnosis is often delayed since diaphragmatic hernia tends to present very late after the initial trauma, subjecting the patient to possible life-threatening complications. While it is easier to reduce the herniated contents and repair the diaphragm via a thoracic approach, laparotomy is often preferred in cases of acute trauma associated with intra-abdominal injuries. Repair of Littre's hernia then consists of resection of the diverticulum and herniorraphy. Conclusion Internal Littre's hernia is usually of congenital origin. This is the first case of a transthoracic Littre's hernia caused by traumatic rib fractures. Hence, it is of utter importance that a clinician is aware of such uncommon pathology.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Intracellular chloride channel protein 1 (CLIC1) participates in inflammatory processes by regulating macrophage phagosomal functions such as pH and proteolysis. Here, we sought to determine if CLIC1 can regulate adaptive immunity by actions on dendritic cells (DCs), the key professional antigen presenting cells. To do this, we first generated bone marrow-derived DCs (BMDCs) from germline CLIC1 gene-deleted (CLIC1−/−) and wild-type (CLIC1+/+) mice, then studied them in vitro and in vivo. We found phagocytosis triggered cytoplasmic CLIC1 translocation to the phagosomal membrane where it regulated phagosomal pH and proteolysis. Phagosomes from CLIC1−/− BMDCs displayed impaired acidification and proteolysis, which could be reproduced if CLIC1+/+, but not CLIC1−/− cells, were treated with IAA94, a CLIC family ion channel blocker. CLIC1−/− BMDC displayed reduced in vitro antigen processing and presentation of full-length myelin oligodendrocyte glycoprotein (MOG) and reduced MOG-induced experimental autoimmune encephalomyelitis. These data suggest that CLIC1 regulates DC phagosomal pH to ensure optimal processing of antigen for presentation to antigen-specific T-cells. Further, they indicate that CLIC1 is a novel therapeutic target to help reduce the adaptive immune response in autoimmune diseases.

Many important cellular processes require the accurate positioning of subcellular structures. Underpinning many of these are protein systems that spontaneously generate spatiotemporal patterns. In some cases, these systems can be described by non-linear reaction-diffusion equations, however, a full description of such equations is rarely available. A well-studied patterning system is the Min protein system that underpins the positioning of the FtsZ contractile ring during cell division in Escherichia coli. Using a coordinate-free linear stability analysis, the reaction terms can be separated from the geometry of a cell. The reaction terms produce a dispersion relation that can be used to predict patterning on any cell shape and size. Applying linear stability analysis to an accurate mathematical model of the Min system shows that while it correctly predicts the onset of patterning, the dispersion relation fails to predict oscillations and quantitative mode transitions. However, we show that data from full solutions of the Min model can be used to generate a heuristic dispersion relation. We show that this heuristic dispersion relation can be used to approximate the Min protein patterning obtained by full simulations of the non-linear reaction-diffusion equations. Moreover, it also predicts Min patterning obtained from experiments where the shapes of E. coli cells have been deformed into rectangles or arbitrary shapes. Using this procedure, it should be possible to generate heuristic dispersion relations from protein patterning data or simulations for any patterning process and subsequently use these to predict patterning for arbitrary cell shapes.

Mutations of the SPG4 (SPAST) gene encoding for spastin protein are the main causes of hereditary spastic paraplegia. Spastin binds to microtubules and severs them through the enzymatic activity of its AAA domain. Several missense mutations located in this domain lead to stable, nonsevering spastins that decorate a subset of microtubules, suggesting a possible negative gain-of-function mechanism for these mutants. Of the two main isoforms of spastin, only mutations of the long isoform, M1, are supposed to be involved in the onset of the pathology, leaving the role of the ubiquitously expressed shorter one, M87, not fully investigated and understood. Here, we show that two isoforms of spastin harboring the same missense mutation bind and bundle different subsets of microtubules in HeLa cells, and likely stabilize them by increasing the level of acetylated tubulin. However, only mutated M1 has the ability to interact with wild-type M1, and decorates a subset of perinuclear microtubules associated with the endoplasmic reticulum that display higher resistance to microtubule depolymerization and increased intracellular ionic strength, compared with those decorated by mutated M87. We further show that only mutated M1 decorates microtubules of proximal axons and dendrites, and strongly impairs axonal transport in cortical neurons through a mechanism likely independent of the microtubule-severing activity of this protein.

The Y-box binding protein 1 (YB-1) is an RNA/DNA-binding protein regulating gene expression in the cytoplasm and the nucleus. Although mostly cytoplasmic, YB-1 accumulates in the nucleus under stress conditions. Its nuclear localization is associated with aggressiveness and multidrug resistance of cancer cells, which makes the understanding of the regulatory mechanisms of YB-1 subcellular distribution essential. Here, we report that inhibition of RNA polymerase II (RNAPII) activity results in the nuclear accumulation of YB-1 accompanied by its phosphorylation at Ser102. The inhibition of kinase activity reduces YB-1 phosphorylation and its accumulation in the nucleus. The presence of RNA in the nucleus is shown to be required for the nuclear retention of YB-1. Thus, the subcellular localization of YB-1 depends on its post-translational modifications (PTMs) and intracellular RNA distribution.

Decorrente dos sistemas de manejo empregados no solo, como o sistema de preparo convencional (SPC) versus o sistema de plantio direto de hortaliças (SPDH), modificações nos atributos edáficos ocorrem; por exemplo, nos índices de agregação do solo e seu teor de carbono orgânico total (COT). Objetivaram-se quantificar os teores de COT e avaliar os índices de agregação do solo e a distribuição dos agregados por classes de diâmetro sob cultivo de cebola em SPDH e SPC, comparados a uma área de mata adjacente em Ituporanga, SC. Os tratamentos constituíram-se da semeadura de plantas de cobertura, solteiras e consorciadas, em SPDH: vegetação espontânea (VE); 100 % aveia; 100 % centeio; 100 % nabo-forrageiro; consórcio de nabo-forrageiro (14 %) e centeio (86 %); e consórcio de nabo-forrageiro (14 %) e aveia (86 %). Adicionalmente, foram avaliadas uma área de cultivo de cebola em SPC por ±37 anos e uma área de mata (floresta secundária; ±30 anos), ambas adjacentes ao experimento. Em setembro de 2013, cinco anos após a implantação dos tratamentos com plantas de cobertura, foram coletadas amostras indeformadas do solo nas camadas de 0-5, 5-10 e 10-20 cm e separados os agregados para avaliar a estabilidade via úmida. Nos agregados, foi quantificado o COT; após a separação em classes de diâmetro (8,00 mm>Ø≥0,105 mm), calcularam-se o diâmetro médio ponderado (DMP) e o geométrico (DMG) dos agregados; a distribuição deles em macroagregados (Ø≥2,0 mm), mesoagregados (2,0>Ø≥0,25 mm) e microagregados (Ø

The Min protein system creates a dynamic spatial pattern in Escherichia coli cells where the proteins MinD and MinE oscillate from pole to pole. MinD positions MinC, an inhibitor of FtsZ ring formation, contributing to the mid-cell localization of cell division. In this paper, Fourier analysis is used to decompose experimental and model MinD spatial distributions into time-dependent harmonic components. In both experiment and model, the second harmonic component is responsible for producing a mid-cell minimum in MinD concentration. The features of this harmonic are robust in both experiment and model. Fourier analysis reveals a close correspondence between the time-dependent behaviour of the harmonic components in the experimental data and model. Given this, each molecular species in the model was analysed individually. This analysis revealed that membrane-bound MinD dimer shows the mid-cell minimum with the highest contrast when averaged over time, carrying the strongest signal for positioning the cell division ring. This concurs with previous data showing that the MinD dimer binds to MinC inhibiting FtsZ ring formation. These results show that non-linear interactions of Min proteins are essential for producing the mid-cell positioning signal via the generation of second-order harmonic components in the time-dependent spatial protein distribution.

The merlin-ERM (ezrin, radixin, moesin) family of proteins plays a central role in linking the cellular membranes to the cortical actin cytoskeleton. Merlin regulates contact inhibition and is an integral part of cell–cell junctions, while ERM proteins, ezrin, radixin and moesin, assist in the formation and maintenance of specialized plasma membrane structures and membrane vesicle structures. These two protein families share a common evolutionary history, having arisen and separated via gene duplication near the origin of metazoa. During approximately 0.5 billion years of evolution, the merlin and ERM family proteins have maintained both sequence and structural conservation to an extraordinary level. Comparing crystal structures of merlin-ERM proteins and their complexes, a picture emerges of the merlin-ERM proteins acting as switchable interaction hubs, assembling protein complexes on cellular membranes and linking them to the actin cytoskeleton. Given the high level of structural conservation between the merlin and ERM family proteins we speculate that they may function together.

One challenge to synthetic biology is to design functional machines from natural building blocks, from individual amino acids up to larger motifs such as the coiled coil. Here we investigate a novel bipedal motor concept, the Bar-Hinge Motor (BHM), a peptide-based motor capable of executing directed motion via externally controlled conformational switching between a straight bar and a V -shaped hinged form. Incorporating ligand-regulated binding to a DNA track and periodic control of ligand supply makes the BHM an example of a ‘clocked walker’. Here, we employ a coarse-grained computational model for the BHM to assess the feasibility of a proposed experimental realization, with conformational switching regulated through the photoisomerization of peptide-bound azobenzene molecules. The results of numerical simulations using the model show that the incorporation of this conformational switch is necessary for the BHM to execute directional, rather than random, motion on a one-dimensional track. The power-stroke-driven directed motion is seen in the model even under conditions that underestimate the level of control we expect to be able to produce in the experimental realisation, demonstrating that this type of design should be an excellent vehicle for exploring the physics behind protein motion. By investigating its force-dependent dynamics, we show that the BHM is capable of directional motion against an applied load, even in the more relaxed conformational switching regimes. Thus, BHM appears to be an excellent candidate for a motor design incorporating a power stroke, enabling us to explore the ability of switchable coiled-coil designs to deliver power strokes within synthetic biology.

SUMMARY Mutations in SPG4, encoding the microtubule-severing protein spastin, are responsible for the most frequent form of hereditary spastic paraplegia (HSP), a heterogeneous group of genetic diseases characterized by degeneration of the corticospinal tracts. We previously reported that mice harboring a deletion in Spg4, generating a premature stop codon, develop progressive axonal degeneration characterized by focal axonal swellings associated with impaired axonal transport. To further characterize the molecular and cellular mechanisms underlying this mutant phenotype, we have assessed microtubule dynamics and axonal transport in primary cultures of cortical neurons from spastin-mutant mice. We show an early and marked impairment of microtubule dynamics all along the axons of spastin-deficient cortical neurons, which is likely to be responsible for the occurrence of axonal swellings and cargo stalling. Our analysis also reveals that a modulation of microtubule dynamics by microtubule-targeting drugs rescues the mutant phenotype of cortical neurons. Together, these results contribute to a better understanding of the pathogenesis of SPG4-linked HSP and ascertain the influence of microtubule-targeted drugs on the early axonal phenotype in a mouse model of the disease.