Growth and life history of freshwater chelydrid turtles (Testudines: Cryptodira): A bone histological approach.

The current study examines the growth pattern and lifestyle habits of the freshwater snapping turtles Chelydra and Macrochelys based on limb bone histology. Femora, humeri, and tibiae of 25 individuals selected from a range of ontogenetic stages were assessed to determine inter-element and intraskeletal histological variation. Osteohistological assessment of multiple elements is consistent with overall moderate growth rates as revealed by the dominance of parallel-fibered bone. However, the growth was cyclical as shown by deposition of multiple lines of arrested growths in the compacta. It appears that the bone tissue of C. serpentina is more variable through ontogeny with intermittent higher growth rates. M. temminckii appears to grow more slowly than C. serpentina possessing compact and thick cortices in accordance with their larger size. Overall, vascularization decreases through ontogeny with humeri and femora being well-vascularized in both species. Contrarily, epipodials are poorly vascularized, though simple longitudinal and radial canals are present, suggesting differences in growth patterns when compared with associated diaphyseal sections. The tibiae were found to be the least remodeled of the limb bones and therefore better suited for skeletochronology for snapping turtles. Intra-elementally, femora and humeri preserved higher cortical vascularity ventrally, suggestive of faster relative growth. We hypothesize that the differential growth pattern in limb bones of snapping turtles may relate to differential functional constraints, where forelimbs are operational in swimming while the hindlimbs provide stability.

Ex vivo biomechanical evaluation of an adhered fiberglass and polymethyl methacrylate sole-hoof wall cast on stabilization of type III distal phalanx fractures under simulated physiologic midstance loads.

OBJECTIVE: To evaluate the effect of the application of a novel fiberglass-glue cast (FGC) on the fracture gap width in experimentally created type III distal phalanx fractures in cadaveric specimens under simulated physiologic loads. STUDY DESIGN: Ex vivo biomechanical laboratory study. ANIMALS: Nine unilateral adult equine cadaver forelimbs. METHODS: Type III distal phalanx fractures were created in forelimb specimens, which maintained distal components of the passive stay apparatus. The fracture gap was measured at 5%, 20%, 25%, 50%, 75%, and 95% of fracture length (palmar articular border to solar margin) using D65Pr-PaDiO radiographs. The limb was axially loaded (700, 3600, 4600, and 6700 N) before, during, and after removal of a woven fiberglass cloth and polymethyl methacrylate cast that encompassed the sole and distal portion of the hoof wall (FGC). Fracture gap widths were compared among loads and treatments using a mixed model ANOVA. RESULTS: On average, under simulated physiological midstance loads, the fracture gap width was 0.2 mm smaller after FGC application, with the greatest decrease (0.5 mm) near the articular surface. On average, it was 0.3 mm smaller than after FGC removal. Fracture gap width was 0.1 mm greater when midstance loads transitioned from standing load to walking, trotting, and gallop loads. The fracture gap width increased by 1.3 mm with increasing distance from the articular surface. CONCLUSION: The FGC reduced the fracture gap width and prevented the fracture gap widening that occurred after FGC removal. CLINICAL SIGNIFICANCE: The findings support consideration of FGC use in the treatment of horses with type III distal phalangeal fractures.

The nature of science: The fundamental role of natural history in ecology, evolution, conservation, and education.

There is a contemporary trend in many major research institutions to de-emphasize the importance of natural history education in favor of theoretical, laboratory, or simulation-based research programs. This may take the form of removing biodiversity and field courses from the curriculum and the sometimes subtle maligning of natural history research as a "lesser" branch of science. Additional threats include massive funding cuts to natural history museums and the maintenance of their collections, the extirpation of taxonomists across disciplines, and a critical under-appreciation of the role that natural history data (and other forms of observational data, including Indigenous knowledge) play in the scientific process. In this paper, we demonstrate that natural history knowledge is integral to any competitive science program through a comprehensive review of the ways in which they continue to shape modern theory and the public perception of science. We do so by reviewing how natural history research has guided the disciplines of ecology, evolution, and conservation and how natural history data are crucial for effective education programs and public policy. We underscore these insights with contemporary case studies, including: how understanding the dynamics of evolutionary radiation relies on natural history data; methods for extracting novel data from museum specimens; insights provided by multi-decade natural history programs; and how natural history is the most logical venue for creating an informed and scientifically literate society. We conclude with recommendations aimed at students, university faculty, and administrators for integrating and supporting natural history in their mandates. Fundamentally, we are all interested in understanding the natural world, but we can often fall into the habit of abstracting our research away from its natural contexts and complexities. Doing so risks losing sight of entire vistas of new questions and insights in favor of an over-emphasis on simulated or overly controlled studies.

Theropod dinosaur facial reconstruction and the importance of soft tissues in paleobiology.

Large theropod dinosaurs are often reconstructed with their marginal dentition exposed because of the enormous size of their teeth and their phylogenetic association to crocodylians. We tested this hypothesis using a multiproxy approach. Regressions of skull length and tooth size for a range of theropods and extant varanid lizards confirm that complete coverage of theropod dinosaur teeth with extraoral tissues (gingiva and labial scales) is both plausible and consistent with patterns observed in living ziphodont amniotes. Analyses of dental histology from crocodylians and theropod dinosaurs, including Tyrannosaurus rex, further indicate that the most likely condition was complete coverage of the marginal dentition with extraoral tissue when the mouth was closed. This changes our perceptions about the appearance and oral configuration of these iconic predators and has broad implications for our interpretations of other terrestrial animals with large teeth.

Large-bodied ornithomimosaurs inhabited Appalachia during the Late Cretaceous of North America.

Reconstructing the evolution, diversity, and paleobiogeography of North America's Late Cretaceous dinosaur assemblages require spatiotemporally contiguous data; however, there remains a spatial and temporal disparity in dinosaur data on the continent. The rarity of vertebrate-bearing sedimentary deposits representing Turonian-Santonian ecosystems, and the relatively sparse record of dinosaurs from the eastern portion of the continent, present persistent challenges for studies of North American dinosaur evolution. Here we describe an assemblage of ornithomimosaurian materials from the Santonian Eutaw Formation of Mississippi. Morphological data coupled with osteohistological growth markers suggest the presence of two taxa of different body sizes, including one of the largest ornithomimosaurians known worldwide. The regression predicts a femoral circumference and a body mass of the Eutaw individuals similar to or greater than that of large-bodied ornithomimosaurs, Beishanlong grandis, and Gallimimus bullatus. The paleoosteohistology of MMNS VP-6332 demonstrates that the individual was at least ten years of age (similar to B. grandis [~375 kg, 13-14 years old at death]). Additional pedal elements share some intriguing features with ornithomimosaurs, yet suggest a larger-body size closer to Deinocheirus mirificus. The presence of a large-bodied ornithomimosaur in this region during this time is consistent with the relatively recent discoveries of early-diverging, large-bodied ornithomimosaurs from mid-Cretaceous strata of Laurasia (Arkansaurus fridayi and B. grandis). The smaller Eutaw taxon is represented by a tibia preserving seven growth cycles, with osteohistological indicators of decreasing growth, yet belongs to an individual approaching somatic maturity, suggesting the co-existence of medium- and large-bodied ornithomimosaur taxa during the Late Cretaceous Santonian of North America. The Eutaw ornithomimosaur materials provide key information on the diversity and distribution of North American ornithomimosaurs and Appalachian dinosaurs and fit with broader evidence of multiple cohabiting species of ornithomimosaurian dinosaurs in Late Cretaceous ecosystems of Laurasia.

New giant carnivorous dinosaur reveals convergent evolutionary trends in theropod arm reduction.

Giant carnivorous dinosaurs such as Tyrannosaurus rex and abelisaurids are characterized by highly reduced forelimbs that stand in contrast to their huge dimensions, massive skulls, and obligate bipedalism.1,2 Another group that follows this pattern, yet is still poorly known, is the Carcharodontosauridae: dominant predators that inhabited most continents during the Early Cretaceous3-5 and reached their largest sizes in Aptian-Cenomanian times.6-10 Despite many discoveries over the last three decades, aspects of their anatomy, especially with regard to the skull, forearm, and feet, remain poorly known. Here we report a new carcharodontosaurid, Meraxes gigas, gen. et sp. nov., based on a specimen recovered from the Upper Cretaceous Huincul Formation of northern Patagonia, Argentina. Phylogenetic analysis places Meraxes among derived Carcharodontosauridae, in a clade with other massive South American species. Meraxes preserves novel anatomical information for derived carcharodontosaurids, including an almost complete forelimb that provides evidence for convergent allometric trends in forelimb reduction among three lineages of large-bodied, megapredatory non-avian theropods, including a remarkable degree of parallelism between the latest-diverging tyrannosaurids and carcharodontosaurids. This trend, coupled with a likely lower bound on forelimb reduction, hypothesized to be about 0.4 forelimb/femur length, combined to produce this short-armed pattern in theropods. The almost complete cranium of Meraxes permits new estimates of skull length in Giganotosaurus, which is among the longest for theropods. Meraxes also provides further evidence that carchardontosaurids reached peak diversity shortly before their extinction with high rates of trait evolution in facial ornamentation possibly linked to a social signaling role.

Comment on "The influence of juvenile dinosaurs on community structure and diversity".

Schroeder et al. (Reports, 26 February 2021, p. 941) reported a size gap among predatory dinosaur species. We argue that the supporting dataset is skewed toward Late Cretaceous North America and that the gap was likely absent during other intervals in most geographic regions. We urge broader consideration of this hypothesis, with quantitative evaluation of preservational and dataset biases.

Growth variability, dimensional scaling, and the interpretation of osteohistological growth data.

Osteohistological data are commonly used to study the life history of extant and extinct tetrapods. While recent advances have permitted detailed reconstructions of growth patterns, physiology and other features using these data, they are most commonly used in assessments of ontogenetic stage and relative growth in extinct animals. These methods have seen widespread adoption in recent years, rapidly becoming a common component of the taxonomic description of new fossil taxa, but are often applied without close consideration of the sources of variation present or the dimensional scaling relationships that exist among different osteohistological measurements. Here, we use a combination of theoretical models and empirical data from a range of extant and extinct tetrapods to review sources of variability in common osteohistological measurements, their dimensional scaling relationships and the resulting interpretations that can be made from those data. In particular, we provide recommendations on the usage and interpretation of growth mark spacing/zonal thickness data, when these are likely to be unreliable, and under what conditions they can provide useful inferences for studies of growth and life history.

Medial malleolus fragmentation following talocalcaneal arthrodesis by a dorsomedial approach in a horse.

A 16-year-old, Quarter Horse mare was presented for a 3/5 right hind lameness associated with osteoarthritis of the talocalcaneal joint (TCLJ). Positron emission tomography (PET) and computed tomography (CT) demonstrated marked increased uptake of 18F-sodium fluoride and bone remodeling at the medial facet of the TCLJ, respectively. Under general anesthesia 2 cortical screws (4.5 and 5.5 mm) were placed in neutral fashion via an arthrotomy from dorsomedial to plantaromedial through the medial facet of the TCLJ followed by copious lavage of the tarsocrural joint. Eight weeks after surgery, observable effusion of the tarsocrural joint was present and lameness had worsened. Radiographic examination revealed a fragmented medial malleolus of the tibia, likely secondary to repetitive trauma of the screw heads during tarsal flexion. Repeated CT showed partial fusion of the TCLJ. Both screws were removed and the tarsocrural joint was thoroughly lavaged arthroscopically. At a 20-month recheck the lameness had not improved, and ultrasound examination revealed severe thickening of the TCLJ capsule. Recheck examination 48 mo after surgery showed complete fusion of the TCLJ and resolution of the lameness. Key clinical message: Diagnosis of osteoarthritis of the TCLJ is challenging. Management by arthrodesis using a dorsomedial approach can result in fragmentation of the medial malleolus, with secondary synovitis and capsulitis of the tarsocrural joint.

Fragmentation de la malléole médiale suite à une arthrodèse talo-calcanéenne par voie dorsomédiale chez un cheval. Une jument Quarter Horse âgée de 16 ans a été présentée pour une boiterie postérieure droite de 3/5 associée à une arthrose de l'articulation talo-calcanéenne (TCLJ). La tomographie par émission de positrons (TEP) et la tomodensitométrie (CT) ont démontré une augmentation marquée de l'absorption du fluorure de sodium-18F et un remodelage osseux significatif au niveau de la facette médiale du TCLJ, respectivement. Sous anesthésie générale, deux vis corticales (4,5 et 5,5 mm) ont été placées de façon neutre via une arthrotomie dorsomédiale à plantaro-médiale à travers la face médiale du TCLJ suivie d'un lavage abondant de l'articulation tarsocrurale. Huit semaines après la chirurgie, un épanchement significatif de l'articulation tarso-crurale était présent et la boiterie s'était aggravée. L'examen radiographique a révélé une malléole médiale du tibia fragmentée, probablement secondaire à un traumatisme répétitif des têtes de vis lors de la flexion du tarse. La tomodensitométrie répétée a montré une fusion partielle du TCLJ. Les deux vis ont été retirées et l'articulation tarso-crurale a été soigneusement lavée par arthroscopie. Lors d'un nouveau contrôle après 20 mois, la boiterie ne s'était pas améliorée, et l'échographie a révélé un épaississement sévère de la capsule TCLJ. Un nouvel examen 48 mois après la chirurgie a montré une fusion complète du TCLJ et une résolution de la boiterie.Message clinique clé :Le diagnostic de l'arthrose du TCLJ est difficile. La prise en charge par arthrodèse par voie dorso-médiale peut entraîner une fragmentation de la malléole médiale, avec synovite secondaire et capsulite de l'articulation tarso-crurale.(Traduit par Dr Serge Messier).

Osteohistological analyses reveal diverse strategies of theropod dinosaur body-size evolution.

The independent evolution of gigantism among dinosaurs has been a topic of long-standing interest, but it remains unclear if gigantic theropods, the largest bipeds in the fossil record, all achieved massive sizes in the same manner, or through different strategies. We perform multi-element histological analyses on a phylogenetically broad dataset sampled from eight theropod families, with a focus on gigantic tyrannosaurids and carcharodontosaurids, to reconstruct the growth strategies of these lineages and test if particular bones consistently preserve the most complete growth record. We find that in skeletally mature gigantic theropods, weight-bearing bones consistently preserve extensive growth records, whereas non-weight-bearing bones are remodelled and less useful for growth reconstruction, contrary to the pattern observed in smaller theropods and some other dinosaur clades. We find a heterochronic pattern of growth fitting an acceleration model in tyrannosaurids, with allosauroid carcharodontosaurids better fitting a model of hypermorphosis. These divergent growth patterns appear phylogenetically constrained, representing extreme versions of the growth patterns present in smaller coelurosaurs and allosauroids, respectively. This provides the first evidence of a lack of strong mechanistic or physiological constraints on size evolution in the largest bipeds in the fossil record and evidence of one of the longest-living individual dinosaurs ever documented.

Muscle tears as a primary cause of lameness in horses: 14 cases (2009-2016).

This study describes clinical and ultrasonographic findings and outcomes of horses with lameness associated with muscle tears. Records of horses diagnosed with muscle tears were retrospectively evaluated. Horses in which one or more muscle tears were confirmed ultrasonographically and lameness was associated with the tear were included in the study (n = 14). Eight horses had tearing of a single muscle, 6 horses had 2 or more muscles involved. Twelve of 14 horses survived to discharge and were alive at follow-up (8 to 90 months). Recovery time ranged from 6 to 52 weeks (mean: 19.8 weeks); 6 to 16 weeks for single muscle injuries and 12 to 52 weeks for multiple muscle injuries. Of the 12 long-term survivors 3 had chronic lameness/stiffness preventing return to their previous activity level and 9 returned to equal or greater level of activity. No statistically significant difference existed between outcomes and location of injury or number of muscles injured. Key clinical message: This study indicates that conservative management of muscle tears can result in favorable long-term outcomes.

Déchirures musculaires comme cause primaire de boiterie chez des chevaux: 14 cas (2009­2016). La présente étude décrit les trouvailles cliniques et échographiques ainsi que le devenir de chevaux avec une boiterie associée à des déchirures musculaires. Les dossiers de chevaux diagnostiqués avec une déchirure musculaire furent rétrospectivement évalués. Les chevaux chez qui une ou plusieurs déchirures musculaires furent confirmées par échographie et dont la boiterie était associée avec la déchirure furent inclus dans l'étude (n = 14). Huit chevaux avaient une déchirure de seulement un muscle, 6 chevaux avaient 2 muscles ou plus d'impliqués. Douze des 14 chevaux ont survécu jusqu'au congé et étaient vivants lors du suivi (8 à 90 mois). Le temps de guérison varia de 6 à 52 semaines (moyenne: 19,8 semaines); 6 à 16 semaines pour une blessure à seulement un muscle et 12 à 52 semaines pour des blessures musculaires multiples. Parmi les 12 survivants à long-terme, trois avaient une boiterie chronique/raideur empêchant un retour à leur niveau d'activité antérieur et neuf retournèrent à un niveau d'activité égal ou plus grand. Aucune différence statistiquement significative n'existait entre le devenir des chevaux et la localisation de la blessure ou le nombre de muscles blessés.Message clinique clé:Cette étude démontre qu'une gestion conservatrice des déchirures musculaires peut résulter en une issu favorable à long terme.(Traduit par Dr Serge Messier).

Total Lipopolysaccharide from the Human Gut Microbiome Silences Toll-Like Receptor Signaling.

Cohabitation of microbial communities with the host enables the formation of a symbiotic relationship that maintains homeostasis in the gut and beyond. One prevailing model suggests that this relationship relies on the capacity of host cells and tissues to remain tolerant to the strong immune stimulation generated by the microbiota such as the activation of Toll-like receptor 4 (TLR4) pathways by lipopolysaccharide (LPS). Indeed, gut microbial LPS is thought to be one of the most potent activators of innate immune signaling and an important mediator of the microbiome's influence on host physiology. In this study, we performed computational and experimental analyses of healthy human fecal samples to examine the TLR4 signaling capacity of the gut microbiota. These analyses revealed that an immunoinhibitory activity of LPS, conserved across the members of the order Bacteroidales and derived from an underacylated structural feature, silences TLR4 signaling for the entire consortium of organisms inhabiting the human gut. Comparative analysis of metagenomic data from the Human Microbiome Project and healthy-donor samples indicates that immune silencing via LPS is a microbe-intrinsic feature in all healthy adults. These findings challenge the current belief that robust TLR4 signaling is a feature of the microbiome and demonstrate that microbiome-derived LPS has the ability to facilitate host tolerance of gut microbes. These findings have broad implications for how we model host-microbe interactions and for our understanding of microbiome-linked disease. IMPORTANCE While the ability for humans to host a complex microbial ecosystem is an essential property of life, the mechanisms allowing for immune tolerance of such a large microbial load are not completely understood and are currently the focus of intense research. This study shows that an important proinflammatory pathway that is commonly triggered by pathogenic bacteria upon interaction with the host is, in fact, actively repressed by the bacteria of the gut microbiome, supporting the idea that beneficial microbes themselves contribute to the immune tolerance in support of homeostasis. These findings are important for two reasons. First, many currently assume that proinflammatory signaling by lipopolysaccharide is a fundamental feature of the gut flora. This assumption influences greatly how host-microbiome interactions are theoretically modeled but also how they are experimentally studied, by using robust TLR signaling conditions to simulate commensals. Second, elucidation of the mechanisms that support host-microbe tolerance is key to the development of therapeutics for both intestinal and systemic inflammatory disorders.

Correlations Between Life-Detection Techniques and Implications for Sampling Site Selection in Planetary Analog Missions.

We conducted an analog sampling expedition under simulated mission constraints to areas dominated by basaltic tephra of the Eldfell and Fimmvörðuháls lava fields (Iceland). Sites were selected to be "homogeneous" at a coarse remote sensing resolution (10-100 m) in apparent color, morphology, moisture, and grain size, with best-effort realism in numbers of locations and replicates. Three different biomarker assays (counting of nucleic-acid-stained cells via fluorescent microscopy, a luciferin/luciferase assay for adenosine triphosphate, and quantitative polymerase chain reaction (qPCR) to detect DNA associated with bacteria, archaea, and fungi) were characterized at four nested spatial scales (1 m, 10 m, 100 m, and >1 km) by using five common metrics for sample site representativeness (sample mean variance, group F tests, pairwise t tests, and the distribution-free rank sum H and u tests). Correlations between all assays were characterized with Spearman's rank test. The bioluminescence assay showed the most variance across the sites, followed by qPCR for bacterial and archaeal DNA; these results could not be considered representative at the finest resolution tested (1 m). Cell concentration and fungal DNA also had significant local variation, but they were homogeneous over scales of >1 km. These results show that the selection of life detection assays and the number, distribution, and location of sampling sites in a low biomass environment with limited a priori characterization can yield both contrasting and complementary results, and that their interdependence must be given due consideration to maximize science return in future biomarker sampling expeditions. Key Words: Astrobiology-Biodiversity-Microbiology-Iceland-Planetary exploration-Mars mission simulation-Biomarker. Astrobiology 17, 1009-1021.

Palaeoenvironmental drivers of vertebrate community composition in the Belly River Group (Campanian) of Alberta, Canada, with implications for dinosaur biogeography.

BACKGROUND: The Belly River Group of southern Alberta is one of the best-sampled Late Cretaceous terrestrial faunal assemblages in the world. This system provides a high-resolution biostratigraphic record of terrestrial vertebrate diversity and faunal turnover, and it has considerable potential to be a model system for testing hypotheses of dinosaur palaeoecological dynamics, including important aspects of palaeoecommunity structure, trophic interactions, and responses to environmental change. Vertebrate fossil microsites (assemblages of small bones and teeth concentrated together over a relatively short time and thought to be representative of community composition) offer an unparalleled dataset to better test these hypotheses by ameliorating problems of sample size, geography, and chronostratigraphic control that hamper other palaeoecological analyses. Here, we assembled a comprehensive relative abundance dataset of microsites sampled from the entire Belly River Group and performed a series of analyses to test the influence of environmental factors on site and taxon clustering, and assess the stability of faunal assemblages both temporally and spatially. We also test the long-held idea that populations of large dinosaur taxa were particularly sensitive to small-scale environmental gradients, such as the paralic (coastal) to alluvial (inland) regimes present within the time-equivalent depositional basin of the upper Oldman and lower Dinosaur Park Formations. RESULTS: Palaeoenvironment (i.e. reconstructed environmental conditions, related to relative amount of alluvial, fluvial, and coastal influence in associated sedimentary strata) was found to be strongly associated with clustering of sites by relative-abundance faunal assemblages, particularly in relation to changes in faunal assemblage composition and marine-terrestrial environmental transitions. Palaeogeography/palaeolandscape were moderately associated to site relative abundance assemblage clustering, with depositional setting and time (i.e. vertical position within stratigraphic unit) more weakly associated. Interestingly, while vertebrate relative abundance assemblages as a whole were strongly correlated with these marine-terrestrial transitions, the dinosaur fauna does not appear to be particularly sensitive to them. CONCLUSIONS: This analysis confirms that depositional setting (i.e. the sediment type/sorting and associated characteristics) has little effect on faunal assemblage composition, in contrast to the effect of changes in the broader palaeoenvironment (e.g. upper vs. lower coastal plain, etc.), with marine-terrestrial transitions driving temporal faunal dynamics within the Belly River Group. The similarity of the dinosaur faunal assemblages between the time-equivalent portions of the Dinosaur Park Formation and Oldman Formation suggests that either these palaeoenvironments are more similar than characterized in the literature, or that the dinosaurs are less sensitive to variation in palaeoenvironment than has often been suggested. A lack of sensitivity to subtle environmental gradients casts doubt on these forces acting as a driver of putative endemism of dinosaur populations in the Late Cretaceous of North America.

Variation in Microbiome LPS Immunogenicity Contributes to Autoimmunity in Humans.

According to the hygiene hypothesis, the increasing incidence of autoimmune diseases in western countries may be explained by changes in early microbial exposure, leading to altered immune maturation. We followed gut microbiome development from birth until age three in 222 infants in Northern Europe, where early-onset autoimmune diseases are common in Finland and Estonia but are less prevalent in Russia. We found that Bacteroides species are lowly abundant in Russians but dominate in Finnish and Estonian infants. Therefore, their lipopolysaccharide (LPS) exposures arose primarily from Bacteroides rather than from Escherichia coli, which is a potent innate immune activator. We show that Bacteroides LPS is structurally distinct from E. coli LPS and inhibits innate immune signaling and endotoxin tolerance; furthermore, unlike LPS from E. coli, B. dorei LPS does not decrease incidence of autoimmune diabetes in non-obese diabetic mice. Early colonization by immunologically silencing microbiota may thus preclude aspects of immune education.

Helicobacter pylori Resists the Antimicrobial Activity of Calprotectin via Lipid A Modification and Associated Biofilm Formation.

UNLABELLED: Helicobacter pylori is one of several pathogens that persist within the host despite a robust immune response. H. pylori elicits a proinflammatory response from host epithelia, resulting in the recruitment of immune cells which manifests as gastritis. Relatively little is known about how H. pylori survives antimicrobials, including calprotectin (CP), which is present during the inflammatory response. The data presented here suggest that one way H. pylori survives the nutrient sequestration by CP is through alteration of its outer membrane. CP-treated H. pylori demonstrates increased bacterial fitness in response to further coculture with CP. Moreover, CP-treated H. pylori cultures form biofilms and demonstrate decreased cell surface hydrophobicity. In response to CP, the H. pylori Lpx lipid A biosynthetic enzymes are not fully functional. The lipid A molecules observed in H. pylori cultures treated with CP indicate that the LpxF, LpxL, and LpxR enzyme functions are perturbed. Transcriptional analysis of lpxF, lpxL, and lpxR indicates that metal restriction by CP does not control this pathway through transcriptional regulation. Analyses of H. pylori lpx mutants reveal that loss of LpxF and LpxL results in increased fitness, similar to what is observed in the presence of CP; moreover, these mutants have significantly increased biofilm formation and reduced cell surface hydrophobicity. Taken together, these results demonstrate a novel mechanism of H. pylori resistance to the antimicrobial activity of CP via lipid A modification strategies and resulting biofilm formation. IMPORTANCE: Helicobacter pylori evades recognition of the host's immune system by modifying the lipid A component of lipopolysaccharide. These results demonstrate for the first time that the lipid A modification pathway is influenced by the host's nutritional immune response. H. pylori's exposure to the host Mn- and Zn-binding protein calprotectin perturbs the function of 3 enzymes involved in the lipid A modification pathway. Moreover, CP treatment of H. pylori, or mutants with an altered lipid A, exhibit increased bacterial fitness and increased biofilm formation. This suggests that H. pylori modifies its cell surface structure to survive under the stress imposed by the host immune response. These results provide new insights into the molecular mechanisms that influence the biofilm lifestyle and how endotoxin modification, which renders H. pylori resistant to cationic antimicrobial peptides, can be inactivated in response to sequestration of nutrient metals.

Mechanisms of Toll-like Receptor 4 Endocytosis Reveal a Common Immune-Evasion Strategy Used by Pathogenic and Commensal Bacteria.

Microbe-induced receptor trafficking has emerged as an essential means to promote innate immune signal transduction. Upon detection of bacterial lipopolysaccharides (LPS), CD14 induces an inflammatory endocytosis pathway that delivers Toll-like receptor 4 (TLR4) to endosomes. Although several regulators of CD14-dependent TLR4 endocytosis have been identified, the cargo-selection mechanism during this process remains unknown. We reveal that, in contrast to classic cytosolic interactions that promoted the endocytosis of transmembrane receptors, TLR4 was selected as cargo for inflammatory endocytosis entirely through extracellular interactions. Mechanistically, the extracellular protein MD-2 bound to and dimerized TLR4 in order to promote this endocytic event. Our analysis of LPS variants from human pathogens and gut commensals revealed a common mechanism by which bacteria prevent inflammatory endocytosis. We suggest that evasion of CD14-dependent endocytosis is an attribute that transcends the concept of pathogenesis and might be a fundamental feature of bacteria that inhabit eukaryotic hosts.

Should we screen heterosexuals for extra-genital chlamydial and gonococcal infections?

Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT) are two of the most prevalent bacterial sexually transmitted infections in the UK. Although the high burden of extra-genital infections with GC and CT in men who have sex with men has been well established, a significant number of extra-genital site infections with CT and GC could similarly be present in heterosexual women. For this reason we started to routinely offer extra-genital site testing for GC and CT in all patients attending our sexual health clinic who reported having had receptive anal sex and/or giving oral sex. This followed a review of current evidence by the clinical team and a change in local testing policy. This study not only confirmed a large reservoir of extra-genital infection amongst men who have sex with men, but also demonstrates that a comparable reservoir of extra-genital infection is present amongst heterosexual women. Our study adds to the mounting evidence that extra-genital site testing in heterosexual women should occur when oral or anal sexual activity is reported.