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.

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.

Dry and lubricated sliding friction for rubber on concrete: the role of surface energies.

We study the influence of lubricant fluids (water-glycerol mixtures) on rubber sliding friction for two different rubber tread compounds on a concrete surface. We find that for the lubricated contacts the sliding friction below a critical velocity vc is similar to that of the dry contact, but for v > vc the friction drops fast with increasing sliding speed. We discuss the origin of this effect and show that it is not a "normal" mixed lubrication effect but depends on surface (or interfacial) energies.

Role of strain softening and viscoelastic memory for the rolling friction of two tire tread compounds.

Rolling friction is of great importance for many applications, such as tires and conveyor belts. We study the rolling friction for hard cylinders rolling on flat rubber sheets. The rolling friction depends on the number of rolling cycles, the rolling speed, and the temperature. We show that when the rubber is cooled down below the glass transition temperature, the deformations of the rubber surface are frozen-in, resulting in a non-flat rolling track where uphill and downhill rolling movements strongly affect the rolling force. The experimental data are analyzed using the Persson rolling friction theory; good agreement with the experiments is obtained when the non-linear (strain-softening) properties of the viscoelastic modulus are taken into account.

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.

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.

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.

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.

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.

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.

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.

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.

Interaction of Human Enterochromaffin Cells with Human Enteric Adenovirus 41 Leads to Serotonin Release and Subsequent Activation of Enteric Glia Cells.

Human adenovirus 41 (HAdV-41) causes acute gastroenteritis in young children. The main characteristics of HAdV-41 infection are diarrhea and vomiting. Nevertheless, the precise mechanism of HAdV-41-induced diarrhea is unknown, as a suitable small-animal model has not been described. In this study, we used the human midgut carcinoid cell line GOT1 to investigate the effect of HAdV-41 infection and the individual HAdV-41 capsid proteins on serotonin release by enterochromaffin cells and on enteric glia cell (EGC) activation. We first determined that HAdV-41 could infect the enterochromaffin cells. Immunofluorescence staining revealed that the cells expressed HAdV-41-specific coxsackievirus and adenovirus receptor (CAR); flow cytometry analysis supported these findings. HAdV-41 infection of the enterochromaffin cells induced serotonin secretion dose dependently. In contrast, control infection with HAdV-5 did not induce serotonin secretion in the cells. Confocal microscopy studies of enterochromaffin cells infected with HAdV-41 revealed decreased serotonin immunofluorescence compared to that in uninfected cells. Incubation of the enterochromaffin cells with purified HAdV-41 short fiber knob and hexon proteins increased the serotonin levels in the harvested cell supernatant significantly. HAdV-41 infection could also activate EGCs, as shown in the significantly altered expression of glia fibrillary acidic protein (GFAP) in EGCs incubated with HAdV-41. The EGCs were also activated by serotonin alone, as shown in the significantly increased GFAP staining intensity. Likewise, EGCs were activated by the cell supernatant of HAdV-41-infected enterochromaffin cells.IMPORTANCE The nonenveloped human adenovirus 41 causes diarrhea, vomiting, dehydration, and low-grade fever mainly in children under 2 years of age. Even though acute gastroenteritis is well described, how human adenovirus 41 causes diarrhea is unknown. In our study, we analyzed the effect of human adenovirus 41 infection on human enterochromaffin cells and found it stimulates serotonin secretion in the cells, which is involved in regulation of intestinal secretion and gut motility and can also activate enteric glia cells, which are found in close proximity to enterochromaffin cells in vivo This disruption of gut barrier homeostasis as maintained by these cells following human adenovirus 41 infection might be a mechanism in enteric adenovirus pathogenesis in humans and could indicate a possible serotonin-dependent cross talk between human adenovirus 41, enterochromaffin cells, and enteric glia cells.

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.

Electroadhesion with application to touchscreens.

There is growing interest in touchscreens displaying tactile feedback due to their tremendous potential in consumer electronics. In these systems, the friction between the user's fingerpad and the surface of the touchscreen is modulated to display tactile effects. One of the promising techniques used in this regard is electrostatic actuation. If, for example, an alternating voltage is applied to the conductive layer of a surface capacitive touchscreen, an attractive electrostatic force is generated between the finger and the surface, which results in an increase in frictional forces acting on the finger moving on the surface. By altering the amplitude, frequency, and waveform of this signal, a rich set of tactile effects can be generated on the touchscreen. Despite the ease of implementation and its powerful effect on our tactile sensation, the contact mechanics leading to an increase in friction due to electroadhesion has not been fully understood yet. In this paper, we present experimental results for how the friction between a finger and a touchscreen depends on the electrostatic attraction and the applied normal pressure. The dependency of the finger-touchscreen interaction on the applied voltage and on several other parameters is also investigated using a mean field theory based on multiscale contact mechanics. We present detailed theoretical analysis of how the area of real contact and the friction force depend on contact parameters, and show that it is possible to further augment the friction force, and hence the tactile feedback displayed to the user by carefully choosing those parameters.

Rolling friction of elastomers: role of strain softening.

We study the temperature and velocity dependency of rolling friction. Steel and PMMA cylinders are rolled on sheets of nitrile butadiene rubber (NBR), with and without filler, and fluoroelastomer (FKM) with filler. Measurements of the rolling friction are performed for temperatures between -40 °C and 20 °C, and for velocities between 5 µm s-1 and 0.5 cm s-1. For the unfilled NBR, a smooth rolling friction master curve is obtained using the bulk viscoelastic frequency-temperature shift factor aT. For the filled rubber compounds, a small deviation from the bulk viscoelastic shift factor is observed at low temperatures. The experimental data are analyzed using an analytical theory of rolling friction. For the filled compounds, good agreement with theory is obtained when strain softening is included, which increases the rolling friction by a factor ∼2 for the filled FKM and ∼3 for the filled NBR compounds. For the unfilled NBR, the maximum of the rolling friction occurs at higher sliding speeds than predicted by the theory. We discuss the role of the adhesive (crack-opening) contribution to the rolling friction, and the role of frozen-in elastic deformations as the rubber is cooled down below the rubber glass transition temperature.

Surface topography and water contact angle of sandblasted and thermally annealed glass surfaces.

Surface roughness has a huge influence on most tribology properties. Sandblasting is a standard way to produce surface roughness in a controlled and reproducible way. Sometimes the sandblasted surfaces are annealed to reduce the roughness and reduce the sharpness of the roughness. We study the nature of the surface roughness of sandblasted silica glass surfaces and how it is modified by annealing at different temperatures. The surface roughness decreases with increasing annealing temperature due to viscous flow of the glass driven by the surface tension. However, the skewness and kurtosis remain nearly unchanged. Optical pictures of the annealed glass surfaces exhibit cell-like structures (cell diameter ≈20-40 µm), which we interpret as micro-cracks. The concentration of micro-cracks increases with increasing annealing temperature. The micro-cracks result in a (advancing) water contact angle which decreases with increasing annealing temperature, which is opposite to what is expected from the theory if no micro-cracks would occur.

Human Adenovirus Type 37 Uses αVß1 and α3ß1 Integrins for Infection of Human Corneal Cells.

Epidemic keratoconjunctivitis (EKC) is a severe, contagious ocular disease that affects 20 to 40 million individuals worldwide every year. EKC is mainly caused by six types of human adenovirus (HAdV): HAdV-8, -19, -37, -53, -54, and -56. Of these, HAdV-8, -19, and -37 use sialic acid-containing glycans as cellular receptors. αVß3, αVß5, and a few additional integrins facilitate entry and endosomal release of other HAdVs. With the exception of a few biochemical analyses indicating that HAdV-37 can interact physically with αVß5, little is known about the integrins used by EKC-causing HAdVs. Here, we investigated the overall integrin expression on human corneal cells and found expression of α2, α3, α6, αV, ß1, and ß4 subunits in human corneal in situ epithelium and/or in a human corneal epithelial (HCE) cell line but no or less accessible expression of α4, α5, ß3, or ß5. We also identified the integrins used by HAdV-37 through a series of binding and infection competition experiments and different biochemical approaches. Together, our data suggest that HAdV-37 uses αVß1 and α3ß1 integrins for infection of human corneal epithelial cells. Furthermore, to confirm the relevance of these integrins in the HAdV-37 life cycle, we developed a corneal multilayer tissue system and found that HAdV-37 infection correlated well with the patterns of αV, α3, and ß1 integrin expression. These results provide further insight into the tropism and pathogenesis of EKC-causing HAdVs and may be of importance for future development of new antiviral drugs.IMPORTANCE Keratitis is a hallmark of EKC, which is caused by six HAdV types (HAdV-8, -19, -37, -53, -54, and -56). HAdV-37 and some other HAdV types interact with integrin αVß5 in order to enter nonocular human cells. In this study, we found that αVß5 is not expressed on human corneal epithelial cells, thus proposing other host factors mediate corneal infection. Here, we first characterized integrin expression patterns on corneal tissue and corneal cells. Among the integrins identified, competition binding and infection experiments and biochemical assays pointed out αVß1 and α3ß1 to be of importance for HAdV-37 infection of corneal tissue. In the absence of a good animal model for EKC-causing HAdVs, we also developed an in vitro system with multilayer HCE cells and confirmed the relevance of the suggested integrins during HAdV-37 infection.

Contact mechanics for polydimethylsiloxane: from liquid to solid.

Adhesion between a glass ball and a polydimethylsiloxane (PDMS) sample is dependent on the PDMS cross-link density, and the transformation of the material from the uncrosslinked liquid state to the fully crosslinked solid state is investigated in this study. The physical picture reflected a gradual transition from capillary forces driven contact mechanics to the classical Johnson-Kendall-Roberts (JKR)-type contact mechanics. PDMS was produced by mixing the base fluid and a cross-linker at a ratio of 10 : 1 and allowed to slowly cross-link at room temperature with simultaneous measurement of the ball-PDMS interaction force. The PDMS sample was in the liquid state during the first ≈16 hours, and in this case the ball-PDMS interaction was purely adhesive, i.e., no repulsive interaction was observed. Later at the PDMS gel-point the cross-linked PDMS clusters percolate, converting the fluid into a soft (fluid-filled) poroelastic solid. In the transition period, PDMS appears similar to pressure-sensitive adhesives. There we observe so-called "stringing" and permanent deformation of the material impacted by the ball. At room temperature, it takes more than ∼100 hours for PDMS to fully cross-link that can be confirmed by the comparison with the earlier-studied reference PDMS produced at elevated temperatures.