Identification of a new gregarine parasite associated with mass mortality events of freshwater pearl mussels (Margaritifera margaritifera) in Sweden.

Freshwater bivalves play key ecological roles in lakes and rivers, largely contributing to healthy ecosystems. The freshwater pearl mussel, Margaritifera margaritifera, is found in Europe and on the East coast of North America. Once common in oxygenated streams, M. margaritifera is rapidly declining and consequently assessed as a threatened species worldwide. Deterioration of water quality has been considered the main factor for the mass mortality events affecting this species. Yet, the role of parasitic infections has not been investigated. Here, we report the discovery of three novel protist lineages found in Swedish populations of M. margaritifera belonging to one of the terrestrial groups of gregarines (Eugregarinorida, Apicomplexa). These lineages are closely related-but clearly separated-from the tadpole parasite Nematopsis temporariae. In one lineage, which is specifically associated with mortality events of M. margaritifera, we found cysts containing single vermiform zoites in the gills and other organs of diseased individuals using microscopy and in situ hybridization. This represents the first report of a parasitic infection in M. margaritifera that may be linked to the decline of this mussel species. We propose a tentative life cycle with the distribution of different developmental stages and potential exit from the host into the environment.

Structural insights into the interaction between adenovirus C5 hexon and human lactoferrin.

Adenovirus (AdV) infection of the respiratory epithelium is common but poorly understood. Human AdV species C types, such as HAdV-C5, utilize the Coxsackie-adenovirus receptor (CAR) for attachment and subsequently integrins for entry. CAR and integrins are however located deep within the tight junctions in the mucosa where they would not be easily accessible. Recently, a model for CAR-independent AdV entry was proposed. In this model, human lactoferrin (hLF), an innate immune protein, aids the viral uptake into epithelial cells by mediating interactions between the major capsid protein, hexon, and yet unknown host cellular receptor(s). However, a detailed understanding of the molecular interactions driving this mechanism is lacking. Here, we present a new cryo-EM structure of HAdV-5C hexon at high resolution alongside a hybrid structure of HAdV-5C hexon complexed with human lactoferrin (hLF). These structures reveal the molecular determinants of the interaction between hLF and HAdV-C5 hexon. hLF engages hexon primarily via its N-terminal lactoferricin (Lfcin) region, interacting with hexon's hypervariable region 1 (HVR-1). Mutational analyses pinpoint critical Lfcin contacts and also identify additional regions within hLF that critically contribute to hexon binding. Our study sheds more light on the intricate mechanism by which HAdV-C5 utilizes soluble hLF/Lfcin for cellular entry. These findings hold promise for advancing gene therapy applications and inform vaccine development. IMPORTANCE: Our study delves into the structural aspects of adenovirus (AdV) infections, specifically HAdV-C5 in the respiratory epithelium. It uncovers the molecular details of a novel pathway where human lactoferrin (hLF) interacts with the major capsid protein, hexon, facilitating viral entry, and bypassing traditional receptors such as CAR and integrins. The study's cryo-EM structures reveal how hLF engages hexon, primarily through its N-terminal lactoferricin (Lfcin) region and hexon's hypervariable region 1 (HVR-1). Mutational analyses identify critical Lfcin contacts and other regions within hLF vital for hexon binding. This structural insight sheds light on HAdV-C5's mechanism of utilizing soluble hLF/Lfcin for cellular entry, holding promise for gene therapy and vaccine development advancements in adenovirus research.

Ice breakloose friction.

We discuss the origin of the breakloose (or static) friction force when an ice block is slid on a hard randomly rough substrate surface. If the substrate has roughness with small enough amplitude (of order a 1 nm or less), the breakloose force may be due to interfacial slip and is determined by the elastic energy per unit area, Uel/A0, stored at the interface after the block has been displaced a short distance from its original position. The theory assumes complete contact between the solids at the interface and that there is no elastic deformation energy at the interface in the original state before the application of the tangential force. The breakloose force depends on the surface roughness power spectrum of the substrate and is found to be in good agreement with experimental observations. We show that as the temperature decreases, there is a transition from interfacial sliding (mode II crack propagation, where the crack propagation energy GII = Uel/A0) to opening crack propagation (mode I crack propagation with GI the energy per unit area to break the ice-substrate bonds in the normal direction).

Sliding friction on ice.

We study the friction when rectangular blocks made from rubber, polyethylene, and silica glass are sliding on ice surfaces at different temperatures ranging from -40 to 0 °C, and sliding speeds ranging from 3 µm/s to 1 cm s-1. We consider a winter tire rubber compound both in the form of a compact block and as a foam with ∼10% void volume. We find that both rubber compounds exhibit a similar friction on ice for all studied temperatures. As in a previous study at low temperatures and low sliding speeds, we propose that an important contribution to the friction force is due to slip between the ice surface and ice fragments attached to the rubber surface. At temperatures around 0 °C (or for high enough sliding speeds), a thin pre-melted water film will occur at the rubber-ice interface, and the contribution to the friction from shearing the area of real contact is small. In this case, the dominant contribution to the friction force is due to viscoelastic deformations of the rubber by the ice asperities. The sliding friction for polyethylene (PE) and silica glass (SG) blocks on ice differs strongly from that of rubber. The friction coefficient for PE is ∼0.04-0.15 and is relatively weakly velocity dependent except close to the ice melting temperature where the friction coefficient increases toward low sliding speeds. Silica glass exhibits a similarly low friction as PE for T > -10 °C but very large friction coefficients (of order unity) at low temperatures. For both PE and SG, unless the ice track is very smooth, the friction force depends on the position x along the sliding track. This is due to bumps on the ice surface, which are sheared off by the elastically stiff PE and SG blocks, resulting in a plowing-type of contribution to the friction force. This results in friction coefficients, which locally can be very large ∼1, and visual inspection of the ice surface after the sliding acts show ice wear particles (white powder) in regions where ice bumps occur. Similar effects can be expected for rubber blocks below the rubber glass transition temperature, and the rubber is in the (elastically stiff) glassy state.

Relationship between iron deficiency and expression of genes involved in iron metabolism in human myocardium and skeletal muscle.

BACKGROUND: Iron deficiency (ID) is associated with adverse prognosis in patients with heart failure. This study aims to investigate the relationship between ID and expression of genes involved in iron metabolism in human myocardium and skeletal muscle, focusing on Transferrin 1 receptor (TfR1), the main pathway of cellular iron uptake. METHODS: Patients undergoing elective CABG were assessed prior to surgery with echocardiography and serum iron parameters. Core needle biopsies were collected from the left and right ventricle (LV, RV), the right atrium and intercostal skeletal muscle (SM). Gene expression analyses were done by mRNA sequencing. RESULTS: Of 69 patients (median age 69 years, 91% men), 28% had ID. 26% had HFrEF, 25% had HFpEF physiology according to echocardiographic findings and NT-proBNP levels, and 49% had normal LV function. The expression of TfR1 was increased in patients with ID compared to patients without ID in ventricular tissue (p = 0.04) and in intercostal SM (p = 0.01). The increase in TfR1 expression in LV and RV was more pronounced when analysing patients with absolute ID (S-Ferritin<100 µg/L). Analysing the correlation between various iron parameters, S-Ferritin levels showed the strongest correlation with TfR1 expression. There was no correlation with NT-proBNP levels and no difference in TfR1 expression between different HF phenotypes. CONCLUSIONS: In patients undergoing elective CABG we found an association between ID and increased TfR1 expression in myocardium regardless of LV function, indicating physiologically upregulated TfR1 expression in the presence of ID to restore intracellular iron needs. CLINICAL TRIAL REGISTRATION: Clinicaltrials.govNCT03671122.

Characterization of a Novel Infectious Pancreatic Necrosis Virus (IPNV) from Genogroup 6 Identified in Sea Trout (Salmo trutta) from Lake Vänern, Sweden.

In November 2016, infectious pancreatic necrosis virus (IPNV) was isolated from a broodstock female of landlocked sea trout (Salmo trutta) in Lake Vänern in Sweden. VP2 gene sequencing placed the IPNV isolate in genogroup 6, for which pathogenicity is largely unknown. Lake Vänern hosts landlocked sea trout and salmon populations that are endangered, and thus the introduction of new pathogens poses a major threat. In this study we characterized the novel isolate by conducting an infection trial on three salmonid species present in Lake Vänern, whole genome sequencing of the isolate, and prevalence studies in the wild sea trout and salmon in Lake Vänern. During the infection trial, the pathogenicity of the Swedish isolate was compared to that of a pathogenic genogroup 5 isolate. Dead or moribund fish were collected, pooled, and analyzed by cell culture to identify infected individuals. In the trial, the Swedish isolate was detected in fewer sample pools in all three species compared to the genogroup 5 isolate. In addition, the prevalence studies showed a low prevalence (0.2-0.5%) of the virus in the feral salmonids in Lake Vänern. Together the data suggest that the novel Swedish IPNV genogroup 6 isolate is only mildly pathogenic to salmonids.

Serine Protease Inhibitors Restrict Host Susceptibility to SARS-CoV-2 Infections.

The coronavirus disease 2019, COVID-19, is a complex disease with a wide range of symptoms from asymptomatic infections to severe acute respiratory syndrome with lethal outcome. Individual factors such as age, sex, and comorbidities increase the risk for severe infections, but other aspects, such as genetic variations, are also likely to affect the susceptibility to SARS-CoV-2 infection and disease severity. Here, we used a human 3D lung cell model based on primary cells derived from multiple donors to identity host factors that regulate SARS-CoV-2 infection. With a transcriptomics-based approach, we found that less susceptible donors show a higher expression level of serine protease inhibitors SERPINA1, SERPINE1, and SERPINE2, identifying variation in cellular serpin levels as restricting host factors for SARS-CoV-2 infection. We pinpoint their antiviral mechanism of action to inhibition of the cellular serine protease, TMPRSS2, thereby preventing cleavage of the viral spike protein and TMPRSS2-mediated entry into the target cells. By means of single-cell RNA sequencing, we further locate the expression of the individual serpins to basal, ciliated, club, and goblet cells. Our results add to the importance of genetic variations as determinants for SARS-CoV-2 susceptibility and suggest that genetic deficiencies of cellular serpins might represent risk factors for severe COVID-19. Our study further highlights TMPRSS2 as a promising target for antiviral intervention and opens the door for the usage of locally administered serpins as a treatment against COVID-19. IMPORTANCE Identification of host factors affecting individual SARS-CoV-2 susceptibility will provide a better understanding of the large variations in disease severity and will identify potential factors that can be used, or targeted, in antiviral drug development. With the use of an advanced lung cell model established from several human donors, we identified cellular protease inhibitors, serpins, as host factors that restrict SARS-CoV-2 infection. The antiviral mechanism was found to be mediated by the inhibition of a serine protease, TMPRSS2, which results in a blockage of viral entry into target cells. Potential treatments with these serpins would not only reduce the overall viral burden in the patients, but also block the infection at an early time point, reducing the risk for the hyperactive immune response common in patients with severe COVID-19.

Risk factors for complicated Mohs surgery in the South Sweden Mohs Cohort.

BACKGROUND: Mohs micrographic surgery (MMS) is a precise, tissue-sparing surgical technique that offers superior cure rates compared to traditional surgical excision. However, the degree of difficulty of MMS depends on many variables, and consequently, the number of surgical stages required for each case is quite unpredictable. OBJECTIVES: To identify risk factors for complicated MMS, defined as MMS requiring ≥3 stages. METHODS: In a cohort study design, data were prospectively collected from 612 patients that underwent MMS for basal cell carcinoma (BCC) at the Department of Dermatology, Skåne University Hospital, Lund, between 2009 and 2020. Univariate and multivariate logistic regression were used to estimate the risk of MMS requiring ≥3 stages. Due to the risk of multicollinearity between recurrent or incompletely excised BCC and previous treatments, a partially and a fully adjusted multivariate logistic regression model were constructed. RESULTS: In fully adjusted multivariate analyses, age (odds ratio (OR) 1.02; confidence interval (CI) 95% 1.00-1.04), previous cryotherapy (OR 2.3; CI 95% 1.1-4.8), and >1 previous surgery (OR 3.4; CI 95% 1.5-7.7) were significantly associated with risk of complicated MMS. Recurrent BCC was associated with the risk of complicated MMS in partially adjusted multivariate analyses, but not in the fully adjusted analyses. In this highly selected cohort, histopathological subtype, and tumour localization were not associated with the risk of complicated MMS. CONCLUSIONS: Older age and tumours previously treated with cryotherapy or multiple prior surgeries increased the risk of MMS requiring ≥3 stages. Whether recurrent BCC is an independent risk factor for complicated MMS needs further evaluation. Knowledge of these risk factors may ameliorate the planning of Mohs surgeries.

BAF45b Is Required for Efficient Zika Virus Infection of HAP1 Cells.

The 2016 Zika virus (ZIKV) epidemic illustrates the impact of flaviviruses as emerging human pathogens. For unknown reasons, ZIKV replicates more efficiently in neural progenitor cells (NPCs) than in postmitotic neurons. Here, we identified host factors used by ZIKV using the NCI-60 library of cell lines and COMPARE analysis, and cross-analyzed this library with two other libraries of host factors with importance for ZIKV infection. We identified BAF45b, a subunit of the BAF (Brg1/Brm-associated factors) protein complexes that regulate differentiation of NPCs to post-mitotic neurons. ZIKV (and other flaviviruses) infected HAP1 cells deficient in expression of BAF45b and other BAF subunits less efficiently than wildtype (WT) HAP1 cells. We concluded that subunits of the BAF complex are important for infection of ZIKV and other flavivirus. Given their function in cell and tissue differentiation, such regulators may be important determinants of tropism and pathogenesis of arthropod-borne flaviviruses.

Side-leakage of face mask.

Face masks are used to trap particles (or fluid drops) in a porous material (filter) in order to avoid or reduce the transfer of particles between the human lungs (or mouth and nose) and the external environment. The air exchange between the lungs and the environment is assumed to occur through the face mask filter. However, if the resistance to air flow through the filter is high some air (and accompanied particles) will leak through the filter-skin interface. In this paper I will present a model study of the side-leakage problem.

Heparan Sulfate Is a Cellular Receptor for Enteric Human Adenoviruses.

Human adenovirus (HAdV)-F40 and -F41 are leading causes of diarrhea and diarrhea-associated mortality in children under the age of five, but the mechanisms by which they infect host cells are poorly understood. HAdVs initiate infection through interactions between the knob domain of the fiber capsid protein and host cell receptors. Unlike most other HAdVs, HAdV-F40 and -F41 possess two different fiber proteins-a long fiber and a short fiber. Whereas the long fiber binds to the Coxsackievirus and adenovirus receptor (CAR), no binding partners have been identified for the short fiber. In this study, we identified heparan sulfate (HS) as an interaction partner for the short fiber of enteric HAdVs. We demonstrate that exposure to acidic pH, which mimics the environment of the stomach, inactivates the interaction of enteric adenovirus with CAR. However, the short fiber:HS interaction is resistant to and even enhanced by acidic pH, which allows attachment to host cells. Our results suggest a switch in receptor usage of enteric HAdVs after exposure to acidic pH and add to the understanding of the function of the short fibers. These results may also be useful for antiviral drug development and the utilization of enteric HAdVs for clinical applications such as vaccine development.

A simple model for viscoelastic crack propagation.

 When a crack propagates in a viscoelastic solid, energy dissipation can occur very far from the crack tip where the stress field may be very different from the [Formula: see text] singular form expected close to the crack tip. Most theories of crack propagation focus on the near crack tip region. Remarkable, here I show that a simple theory which does not account for the nature of the stress field in the near crack tip region results in a crack propagation energy in semiquantitative agreement with a theory based on the stress field in the near crack tip region. I consider both opening and closing crack propagation and show that for closing crack propagation in viscoelastic solids, some energy dissipation processes must occur in the crack tip process zone. The theory is illustrated by new experimental results for the adhesive interaction between a silica glass ball and a silicone rubber surface.

Human species D adenovirus hexon capsid protein mediates cell entry through a direct interaction with CD46.

Human adenovirus species D (HAdV-D) types are currently being explored as vaccine vectors for coronavirus disease 2019 (COVID-19) and other severe infectious diseases. The efficacy of such vector-based vaccines depends on functional interactions with receptors on host cells. Adenoviruses of different species are assumed to enter host cells mainly by interactions between the knob domain of the protruding fiber capsid protein and cellular receptors. Using a cell-based receptor-screening assay, we identified CD46 as a receptor for HAdV-D56. The function of CD46 was validated in infection experiments using cells lacking and overexpressing CD46, and by competition infection experiments using soluble CD46. Remarkably, unlike HAdV-B types that engage CD46 through interactions with the knob domain of the fiber protein, HAdV-D types infect host cells through a direct interaction between CD46 and the hexon protein. Soluble hexon proteins (but not fiber knob) inhibited HAdV-D56 infection, and surface plasmon analyses demonstrated that CD46 binds to HAdV-D hexon (but not fiber knob) proteins. Cryoelectron microscopy analysis of the HAdV-D56 virion-CD46 complex confirmed the interaction and showed that CD46 binds to the central cavity of hexon trimers. Finally, soluble CD46 inhibited infection by 16 out of 17 investigated HAdV-D types, suggesting that CD46 is an important receptor for a large group of adenoviruses. In conclusion, this study identifies a noncanonical entry mechanism used by human adenoviruses, which adds to the knowledge of adenovirus biology and can also be useful for development of adenovirus-based vaccine vectors.

Adhesion paradox: Why adhesion is usually not observed for macroscopic solids.

The adhesion paradox refers to the observation that for most solid objects no adhesion can be detected when they are separated from a state of molecular contact. The adhesion paradox results from surface roughness, and we present experimental and theoretical results that show that adhesion in most cases is "killed" by the longest-wavelength roughness. In addition, adhesion experiments between a human finger and a clean glass plate were carried out, and for a dry finger no macroscopic adhesion occurred. We suggest that the observed decrease in the contact area with increasing shear force results from nonadhesive finger-glass contact mechanics, involving large deformations of complex layered material.

Cylinder-flat-surface contact mechanics during sliding.

Using molecular dynamics we study the dependency of the contact mechanics on the sliding speed when an elastic block (cylinder) with a cos(q_{0}x) surface height profile is sliding in adhesive contact on a rigid flat substrate. The atoms on the block interact with the substrate atoms by Lennard-Jones potentials, and we consider both commensurate and (nearly) incommensurate contacts. For the incommensurate system the friction force fluctuates between positive and negative values, with an amplitude proportional to the sliding speed, but with the average close to zero. For the commensurate system the (time-averaged) friction force is much larger and nearly velocity independent. For both types of systems the width of the contact region is velocity independent even when, for the commensurate case, the frictional shear stress increases from zero (before sliding) to ≈0.1MPa during sliding. This frictional shear stress, and the elastic modulus used, are typical for polydimethylsiloxane rubber sliding on a glass surface, and we conclude that the reduction in the contact area observed in some experiments when increasing the tangential force must be due to effects not included in our model study, such as viscoelasticity or elastic nonlinearity.

Lactoferrin-Hexon Interactions Mediate CAR-Independent Adenovirus Infection of Human Respiratory Cells.

Virus entry into host cells is a complex process that is largely regulated by access to specific cellular receptors. Human adenoviruses (HAdVs) and many other viruses use cell adhesion molecules such as the coxsackievirus and adenovirus receptor (CAR) for attachment to and entry into target cells. These molecules are rarely expressed on the apical side of polarized epithelial cells, which raises the question of how adenoviruses-and other viruses that engage cell adhesion molecules-enter polarized cells from the apical side to initiate infection. We have previously shown that species C HAdVs utilize lactoferrin-a common innate immune component secreted to respiratory mucosa-for infection via unknown mechanisms. Using a series of biochemical, cellular, and molecular biology approaches, we mapped this effect to the proteolytically cleavable, positively charged, N-terminal 49 residues of human lactoferrin (hLF) known as human lactoferricin (hLfcin). Lactoferricin (Lfcin) binds to the hexon protein on the viral capsid and anchors the virus to an unknown receptor structure of target cells, resulting in infection. These findings suggest that HAdVs use distinct cell entry mechanisms at different stages of infection. To initiate infection, entry is likely to occur at the apical side of polarized epithelial cells, largely by means of hLF and hLfcin bridging HAdV capsids via hexons to as-yet-unknown receptors; when infection is established, progeny virions released from the basolateral side enter neighboring cells by means of hLF/hLfcin and CAR in parallel.IMPORTANCE Many viruses enter target cells using cell adhesion molecules as receptors. Paradoxically, these molecules are abundant on the lateral and basolateral side of intact, polarized, epithelial target cells, but absent on the apical side that must be penetrated by incoming viruses to initiate infection. Our study provides a model whereby viruses use different mechanisms to infect polarized epithelial cells depending on which side of the cell-apical or lateral/basolateral-is attacked. This study may also be useful to understand the biology of other viruses that use cell adhesion molecules as receptors.

Interfacial fluid flow for systems with anisotropic roughness.

I discuss fluid flow at the interface between solids with anisotropic roughness. I show that the Bruggeman effective medium theory and the critical junction theory give nearly the same results for the fluid flow conductivity. This shows that, in most cases, the surface roughness observed at high magnification is irrelevant for fluid flow problems such as the leakage of static seals, and fluid squeeze-out. The effective medium theory predicts that the fluid flow conductivity vanishes at the relative contact area A/A0 = 0.5 independent of the anisotropy. However, the effective medium theory does not solve the elastic contact mechanics problem but is based on a purely geometric argument. Thus, for anisotropic roughness the contact area may percolate at different values of A/A0 depending on the direction. We discuss how this may be taken into account in the effective medium and critical junction theories.

Electric field effect in heat transfer in 2D devices.

We calculate heat transfer between a 2D sheet (e.g. graphene) and a dielectric in presence of a gate voltage. The gate potential induces surface charge densities on the sheet and dielectric, which results in electric field, which is coupled to the surface displacements and, as a consequence, resulting an additional contributions to the radiative heat transfer. The electrostatic and van der Waals interactions between the surface displacement result in the phonon heat transfer, which we calculate taking into account the nonlocality of these interactions. Numerical calculations are presented for heat transfer between graphene and a SiO2 substrate.

Lubricated sliding friction: Role of interfacial fluid slip and surface roughness.

We derive approximate mean field equations for the fluid flow between elastic solids with randomly rough surfaces including interfacial fluid slip and shear thinning. We present numerical results for the fluid flow and friction factors for realistic systems, in particular, we consider the case of an elastic cylinder with random surface roughness in relative sliding contact with a flat rigid (low-energy) counter-surface. We present experimental data for the sliding friction between rubber stoppers and glass barrels lubricated with baked-on silicone oil. We find that the frictional shear stress acting in the rubber asperity contact regions is nearly velocity independent for velocities in the 10-1000µm/s range, and very small [Formula: see text] MPa, while for bare glass in silicone oil [Formula: see text] is much larger and velocity dependent.

Physics of suction cups.

We have developed a theory of air leakage at interfaces between two elastic solids with application to suction cups in contact with randomly rough surfaces. We present an equation for the airflow in narrow constrictions which interpolates between the diffusive and ballistic (Knudsen) air-flow limits. To test the theory, we performed experiments using two different suction cups, made from soft polyvinylchloride (PVC), in contact with sandblasted polymethylmethacrylate (PMMA) plates. We found that the measured time to detach (lifetime) of the suction cups was in good agreement with theory, except for surfaces with a root-mean-square (rms) roughness below ≈1 µm, where diffusion of plasticizer from the PVC to the PMMA surface caused blockage of critical constrictions. The suction cup volume, stiffness, and elastic modulus have a huge influence on the air leakage and hence the failure time of the cups. Based on our research we propose an improved biomimetic design of suction cups that could show improved failure times with varying degrees of roughness under dry and wet environments.