Search bioRxiv⌕ Search

Biology subjects

Hughes, K.

Publications and source records attributed to Hughes, K..

At least 19 recordsLinked to original sources

Macrophage calcium signaling dynamics revealed using genetically-encoded sensors

Macrophages (M{Phi}s) inhabit all mammalian organs, adapting to and influencing the tissues in which they reside. These versatile cells rapidly respond to tissue-specific signals, clean up debris and serve as the first line of defense against pathogens. Their ability to act quickly relates to their abundance and specific arrangement in tissues, as well as the systems they possess for detecting, processing and integrating emergent signals. Ca2+ is a fast second messenger that has been linked to many of the core functions of M{Phi}s. However, spatiotemporal features of physiologically-relevant Ca2+ signals in M{Phi}s remain largely uncharacterized. Using mouse models expressing genetic biosensors and fluorescently tagged channels, we visualize M{Phi} Ca2+ signal dynamics and heterogeneity, uncover a remarkable degree of mechanosensitivity in these cells, and characterize the physiological consequences of genetic ablation of Piezo1 channels via analysis of knockout models across their lifetime. Our in-depth investigation of M{Phi} Ca2+ signaling dynamics has broad relevance for the field of M{Phi} biology and the tissues that these cells support.

immunology↗

Structure of the proton-powered secretion motor at the heart of the bacterial flagellum

Bacterial type III secretion systems export proteins across the inner membrane using the proton-motive force (PMF), but how proton flow is coupled to opening of the export channel is unknownXX,XX. Here we present single-particle cryo-EM structures at 2.5-4.2 [A] resolution of the intact flagellar Export Apparatus from Salmonella enterica, obtained using optimized extraction conditions of the endogenous assembly that retain the complete transmembrane complex. The transmembrane domain of FlhA forms a nonameric funnel-shaped basket beneath the Export Gate, with each subunit harboring a buried pathway of conserved hydrophilic residues spanning the hydrophobic core of the membrane. FlhB is resolved for the first time within the intact gate, revealing helices that thread through the FlhA channel, completely sealing it at rest. A symmetry-free reconstruction captures one FlhA protomer hinged outward at the water-filled cavity, breaking the rotational symmetry. Together, these structures define the architecture of the proton-transducing element of the type III secretion system and suggest a rotary gating mechanism, with parallels to the F0 motor of ATP synthase, in which PMF-driven conformational cycling of FlhA drives regulated opening of the export channel.

microbiology↗

Selective conservation of symbiont cell-surface glycans across generations in a vertically transmitting coral

Coral resilience under climate change depends on the stability of coral-Symbiodiniaceae symbioses. While vertically transmitting corals inherit symbionts directly from parental colonies, the extent to which symbiont cellular traits are conserved across life stages remains unclear. Here, we examined cell-surface glycan profiles of Symbiodiniaceae in parental colonies and eggs of the coral Montipora capitata. Glycan signatures were structured by symbiont genus and differed between life stages, with mannose/glucose- and galactose-containing glycoproteins as primary drivers of variation. Despite life-stage differences, parent-offspring comparisons revealed significant conservation of glycan profiles, indicating intergenerational transmission of symbiont cellular traits that differed between Cladocopium and Durusdinium and were driven by distinct glycan classes. These results suggest that vertical transmission preserves key recognition-relevant glycans while allowing flexibility in other symbionts surface traits, providing a mechanistic basis for symbiosis stability.

cell biology↗

Structures and broad-spectrum growth-inhibiting activity of formomarinobactin, formylated marinobactin analogues from the Pseudomonas lutea clade

Pseudomonas graminis LMG 21661T, an environmental strain of the P. lutea clade, produces the siderophore formomarinobactin, a novel marinobactin-like siderophore. Mass spectrometry revealed that formomarinobactin shares the same six-residue peptide backbone as marinobactin but contains formylated rather than acetylated N-hydroxyornithines. Alike marinobactins, formomarinobactins are produced as a suite of siderophores with a conserved hexapeptide core but varying lipid tail lengths (C10-C14), shorter than the C12 to C18 characteristic of marinobactins. Both the biosynthesis and cognate receptor genes of the formomarinobactin system in P. graminis are iron regulated but unaffected by zinc or nickel underscoring their role in iron homeostasis. Genome mining combined with mass analyses demonstrated that formomarinobactin production is a conserved trait across the P. lutea clade, with one exception which appears to represent an intraspecific cheater that has lost siderophore production. Beyond the producing strains themselves, we identified a widespread distribution of putative formomarinobactin receptors among diverse Pseudomonas species, revealing a substantial capacity within the P. fluorescens super clade to pirate formomarinobactin as an iron source. Putative receptors were also found in genera outside the Pseudomonas genus. Growth stimulation assays confirmed functional formomarinobactin uptake in several Pseudomonas spp. and a Phytopseudomonas strain, with genetic validation in Pseudomonas rhodesiae. Importantly, formomarinobactin production confers more than a nutritional advantage. Members of the P. lutea clade producing formomarinobactin display pronounced growth inhibiting activity against a broad spectrum of clinical and environmental Gram-positive and with lower efficacy against Gram-negative bacteria. Purified formomarinobactin was able to inhibit growth under iron-limiting conditions and to a lesser extent in iron-rich conditions, highlighting a dual role for this molecule in both iron acquisition and microbial growth inhibition.

microbiology↗

Avasopasem Manganese acts as both a Radioprotector and a Radiomitigator of Radiation-Induced Acute or Late Effects.

The pentaazamacrocyclic superoxide dismutase mimetic, Avasopasem Manganese (AVA), has been shown in clinical trials to reduce the severity and duration of acute oral mucositis (OM) and acute esophagitis in patients treated for head and neck and lung cancers, respectively, by radiotherapy using conventional fractionation protocols. Here, the radioprotective effects of AVA were tested in normal tissues after high dose per fraction radiation exposures to determine whether: radioprotective effects of AVA were still present after doses like those used with stereotactic ablative radiotherapy (SAbR); AVA protected against late normal tissue responses; and, whether AVA could act as a radiomitigator of adverse normal tissue events. With AVA, residual DNA lesions and micronuclei were reduced in HBEC3 KT but increased in H1299 cells 24h post-irradiation. Furthermore, radiation-induced mutations and chromosome aberrations were reduced in WTK-1 lymphoblast cells. The radioprotective effects of AVA at high dose per fraction were then tested against both acute and late normal tissue effects. When provided prior to radiation, AVA reduced the extent of epithelial cell layer degradation of mouse tongue irradiated with a single dose of 17 Gy and reduced radiation recall when a second dose of radiation of 12 or 17 Gy was given two weeks following the initial 17 Gy dose. In addition, when provided after radiation, there was a modest but significant reduction in adverse epithelial layer response. Radiation-induced lung fibrosis, determined at 24 weeks post-irradiation, was also reduced when AVA was delivered 1 hour prior to irradiation after a single dose of 54 Gy. When AVA was provided 24h after 54 Gy and given daily (Monday-Friday) for increasing numbers of weeks, fibrosis was progressively reduced as the length of AVA treatment increased. However, AVAs effect on fibrosis decreased as the time between irradiation and post-irradiation AVA application increased. These studies confirm the efficacy of AVA as a radioprotector and mitigator of both radiation-induced acute and late effects after high dose per fraction exposures while not protecting tumor cells to radiation exposure.

cancer biology↗

A novel chimeric coronavirus spike vaccine combining SARS-CoV-2 RBD and scaffold domains from HKU-1 elicits potent neutralising antibody responses

The receptor binding domain (RBD) of the SARS-CoV-2 spike is the major target for neutralising antibodies elicited by current vaccines. Using small domains such as the RBD as vaccine immunogens, however, may constrain the availability of CD4 T follicular helper (TFH) cells and impact immunogenicity. We engineered a novel chimeric trimeric RBD (CTR) glycoprotein, replacing the RBD of human coronavirus HKU-1 spike with SARS-CoV-2 RBD of either ancestral (WT) or Omicron BA.2 strains. This strategy maintains a native trimeric conformation of the RBD, while providing additional sources of CD4 T cell help via the HKU-1 spike scaffold. In C57BL/6 mice, CTR-BA.2 prime-boost vaccination elicited high anti-BA.2-RBD IgG and neutralising titres, matching responses in animals immunised with native SARS-CoV-2 spike proteins. GC B cells elicited by CTR-BA.2 were predominantly WT+/BA.2+ cross- reactive, and TFH cells predominantly recognised HKU-1 epitopes, demonstrating scaffold-directed T cell help. Macaques prime-boost immunised with CTR-WT similarly elicited high anti-RBD IgG, anti-spike IgG and neutralising responses, comparable to native spike-vaccinated animals. In draining lymph nodes of CTR-WT vaccinated macaques, RBD-specific GC B cells were present at elevated levels. In contrast to the murine studies, lymph node-draining TFH responses in macaques were broadly elicited against RBD, NTD/S2 or HKU-1-derived peptides. Although native SARS- CoV-2 spike was also highly immunogenic in animal models, our findings establish the chimeric glycoprotein design as a strategy to overcome the poor immunogenicity of the SARS-CoV-2 RBD by engaging CD4 TFH cells, while maintaining the ability to elicit protective neutralising responses. One sentence summaryA chimeric glycoprotein design preserves SARS-CoV-2 RBD antigenic conformation enabling elicitation of neutralising responses, while allowing recruitment of HKU-1 scaffold-directed CD4 helper responses to support the humoral response.

immunology↗

Proactive Coral Reef Restoration Using Thermally Tolerant Corals in Hawaii

Effective conservation of degraded ecosystems requires mitigation of the original cause of decline, but this step can be difficult in the context of global climate change. On coral reefs, persistent environmental stress which causes coral bleaching may be addressed by using coral restoration stock which is naturally more resilient, often termed "proactive restoration" in terrestrial management. To explore the feasibility and consequences of this approach, we outplanted 391 colonies of 7 species of reef-building coral designated as thermally tolerant or thermally sensitive during stress testing and monitored them for 2 years using photogrammetry to evaluate tradeoffs and return-on-effort. We found no growth, complexity or effort tradeoffs when using thermally tolerant corals, but tolerant corals had lower survivorship during our monitoring period, driven primarily by one genus. These data illustrate nuanced tradeoffs and consequences to proactive reef restoration and suggest that the potential benefits of this approach may only be fully realized during future coral bleaching events.

ecology↗

Greening rates are sensitive to methodology and biology; comment to Sustained greening of the Antarctic Peninsula observed from satellites

Roland et al.1 claim to provide evidence for a greening trend throughout the Antarctic Peninsula region over the last four decades, based on satellite remote sensing data. However, the early period vegetation cover estimates do not match with the likely extent of vegetated areas in this region at that time, raising doubts about the magnitude of any greening trend. Furthermore, growth rates of mosses and higher plants in Antarctica are insufficient to explain the 14-fold green cover expansion claimed, and neither have such changes been observed at long-term monitoring sites or from field warming studies. The reported satellite time series analyses of the presented trend seems biased by satellite image availability, lack of consistency in the areas covered by imagery and processing pitfalls. Antarctic terrestrial ecosystems are indeed predicted to become greener with climate change, but at much slower rates than reported by Roland et al.1.

ecology↗

MYH11 rare variant augments aortic growth and induces cardiac hypertrophy and heart failure with pressure overload

Smooth muscle cell-specific myosin heavy chain, encoded by MYH11, is selectively expressed in smooth muscle cells (SMCs). Pathogenic variants in MYH11 predispose to a number of disorders, including heritable thoracic aortic disease associated with patent ductus arteriosus, visceral myopathy, and megacystis-microcolon-intestinal hypoperistalsis syndrome. Rare variants of uncertain significance occur throughout the gene, including MYH11 p.Glu1892Asp, and we sought to determine if this variant causes thoracic aortic disease in mice. Genomic editing was used to generate Myh11E1892D/E1892D mice. Wild-type (WT) and mutant mice underwent cardiovascular phenotyping and with transverse aortic constriction (TAC). Myh11E1892D/E1892D and WT mice displayed similar growth, blood pressure, root and ascending aortic diameters, and cardiac function up to 13 months of age, along with similar contraction and relaxation on myographic testing. TAC induced hypertension similarly in Myh11E1892D/E1892D and WT mice, but mutant mice showed augmented ascending aortic enlargement and increased elastic fragmentation on histology. Unexpectedly, male Myh11E1892D/E1892D mice two weeks post-TAC had decreased ejection fraction, stroke volume, fractional shortening, and cardiac output compared to similarly treated male WT mice. Importantly, left ventricular mass increased significantly due to primarily posterior wall thickening, and cardiac histology confirmed cardiomyocyte hypertrophy and increased collagen deposition in the myocardium and surrounding arteries. These results further highlight the clinical heterogeneity associated with MYH11 rare variants. Given that MYH11 is selectively expressed in SMCs, these results implicate a role of vascular SMCs in the heart contributing to cardiac hypertrophy and failure with pressure overload. Author SummaryIn this study, we explore the impact of a specific genetic variant, MYH11 p.Glu1892Asp, on the heart and blood vessels in mice. The MYH11 gene is crucial for smooth muscle cells, which are found in the walls of blood vessels and play an important role in various vascular diseases. We created mice with this genetic variant to see if it would lead to thoracic aortic disease, a condition affecting the main artery from the heart. We found that mice with the variant were similar to normal mice in many aspects, such as growth, blood pressure, and heart function, for up to 13 months. However, when we induced high blood pressure in the mice, the mutant mice showed more significant enlargement of the aorta and damage to the elastic fibers in the aortic walls. Interestingly, male mutant mice also developed heart problems, such as reduced heart pumping ability and increased heart muscle thickness, after the high blood pressure challenge. This was accompanied by signs of heart muscle cell enlargement and increased tissue stiffness. These findings suggest that this rare MYH11 variant can contribute to a range of heart and vascular issues, particularly under conditions of pressure overload, and highlight the importance of smooth muscle cells in the development of these problems.

genetics↗

Horizontal transfer of nuclear DNA in transmissible cancer

Although somatic cell genomes are usually entirely clonally inherited, nuclear DNA exchange between cells of an organism can occur sporadically by cell fusion, phagocytosis or other mechanisms1-3. This phenomenon has long been noted in the context of cancer, where it could be envisaged that DNA horizontal transfer plays a functional role in disease evolution4-13. However, an understanding of the frequency and significance of this process in naturally occurring tumours is lacking. The host-tumour genetic discordance of transmissible cancers, malignant clones which pass between animals as allogeneic grafts, provides an opportunity to investigate this. We screened for host-to-tumour horizontal transfer of nuclear DNA in 174 tumours from three transmissible cancers affecting dogs and Tasmanian devils, and detected a single instance in the canine transmissible venereal tumour (CTVT). This involved introduction of a 15-megabase dicentric genetic element, composed of 11 rearranged fragments of six chromosomes, to a CTVT sublineage occurring in Asia around 2,000 years ago. The element forms the short arm of a small submetacentric chromosome, and derives from a dog with ancestry associated with the ancient Middle East. The introduced DNA fragment is transcriptionally active and has adopted the expression profile of CTVT. Its 143 genes do not, however, confer any obvious advantage to its spatially restricted CTVT sublineage. Our findings indicate that nuclear DNA horizontal transfer, although likely a rare event in tumour evolution, provides a viable mechanism for the acquisition of genetic material in naturally occurring cancer genomes.

cancer biology↗

Mineral Phase Changes During Intervertebral Disc Degeneration

Intervertebral disc disease is a common cause of pain and neurological deficits and is known to be associated with degeneration and calcification. Here we analysed samples of herniated disc material and compared it to material taken from non-herniated discs following surgical treatment in dogs. Our clinical approach to these cases allows collection of samples providing a unique opportunity for a case-controlled study such as this, an opportunity which is not available to the human neurosurgeon. We analysed all samples using Fourier transform infrared (FTIR) spectroscopy, as well as a proportion with X-ray diffraction (XRD) and transmission electron microscopy (TEM). FTIR spectra of the majority of herniated samples were consistent with the presence of crystalline hydroxyapatite, whereas most of the non-herniated discs showed spectra consistent with amorphous phosphate material. XRD analysis and TEM confirmed these findings and identified the amorphous material as amorphous calcium phosphate nanoparticle clusters of [~] 20 nm diameter and the crystalline hydroxyapatite material as needles up to 100 nm in length. The differences between the herniated and non-herniated discs indicate that the degenerative process involves a conversion of amorphous calcium phosphate into crystalline hydroxyapatite which precedes and may predispose the disc to herniate.

neuroscience↗

Single cell RNA sequencing reveals immunomodulatory mechanism of stem cell factor and granulocyte colony-stimulating factor treatment in the brains of aged APP/PS1 mice

Alzheimers disease (AD) leads to progressive neurodegeneration and dementia. AD primarily affects older adults with neuropathological changes including amyloid-beta (A{beta}) deposition, neuroinflammation, and neurodegeneration. We have previously demonstrated that systemic treatment with combined stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) (SCF+G-CSF), reduces A{beta} load, increases A{beta} uptake by activated microglia and macrophages, reduces neuroinflammation, and restores dendrites and synapses in the brains of aged APPswe/PS1dE9 (APP/PS1) mice. However, the mechanisms underlying SCF+G-CSF-enhanced brain repair in aged APP/PS1 mice remain unclear. This study used a transcriptomic approach to identify potential mechanisms by which SCF+G-CSF treatment modulates microglia and peripheral myeloid cells to mitigate AD pathology in the aged brain. After injections of SCF+G-CSF for 5 consecutive days, single-cell RNA sequencing was performed on CD11b+ cells isolated from the brains of 28-month-old APP/PS1 mice. The vast majority of cell clusters aligned with transcriptional profiles of microglia in various activation states. However, SCF+G-CSF treatment dramatically increased a cell population showing upregulation of marker genes related to peripheral myeloid cells. Flow cytometry data also revealed an SCF+G-CSF-induced increase of cerebral CD45high/CD11b+ active phagocytes. SCF+G-CSF treatment robustly increased the transcription of genes implicated in immune cell activation, including gene sets that regulate inflammatory processes and cell migration. Expression of S100a8 and S100a9 were robustly enhanced following SCF+G-CSF treatment in all CD11b+ cell clusters. Moreover, the topmost genes differentially expressed with SCF+G-CSF treatment were largely upregulated in S100a8/9-positive cells, suggesting a well-conserved transcriptional profile related to SCF+G-CSF treatment in resident and peripherally derived CD11b+ immune cells. This S100a8/9-associated transcriptional profile contained notable genes related to pro-inflammatory and anti-inflammatory responses, neuroprotection, and A{beta} plaque inhibition or clearance. Altogether, this study reveals immunomodulatory effects of SCF+G-CSF treatment in the aged brain with AD pathology, which will guide future studies to further uncover the therapeutic mechanisms.

neuroscience↗

An ovine model for investigation of the microenvironment of the male mammary gland

The specific biology of the male breast remains relatively unexplored in spite of the increasing global prevalence of male breast cancer. Delineation of the microenvironment of the male breast is restricted by the low availability of human samples and a lack of characterisation of appropriate animal models. Unlike the mouse, the male ovine gland persists postnatally. We suggest that the male ovine mammary gland constitutes a promising adjunctive model for the male human breast. In this study we evaluate the male ovine mammary gland microenvironment, comparing intact and neutered males. Assessment of the glandular histo-anatomy highlights the resemblance of the male gland to that of the neonatal female sheep and confirms the presence of rudimentary terminal duct lobular units. Irrespective of neutered status, cell proliferation in epithelial and stromal compartments is low in males, and cell proliferation in epithelial cells and in the intralobular stroma is significantly lower than in pubertal female sheep. Between 42% and 72% of the luminal mammary epithelial cells in the male gland express the androgen receptor, and expression is significantly reduced by neutering. Luminal epithelial cells within the intact and neutered male gland also express oestrogen receptor alpha, but minimal progesterone receptor expression is observed. The distrubution of mammary leukocytes within the ducts and stroma is similar in the female mammary gland of sheep and other species. Both macrophages and T lymphocytes are intercalated in the epithelial bilayer and are more abundant in the intralobular stroma than the interlobular stroma, suggesting that they may have a protective immunological function within the vestigial glandular tissue of the male sheep. Mast cells are also observed within the stroma. These cells cluster nearer the glandular tissue and are frequently located adjacent to blood vessels. The abundance of mast cells is significantly higher in intact males compared to neutered males, suggesting that hormone signalling may impact mast cell recruitment. In this study, we demonstrate the utility of the male ovine mammary gland as a model for furthering our knowledge of postnatal male mammary biology.

pathology↗

In the murine and bovine maternal mammary gland signal transducer and activator of transcription 3 is activated in clusters of epithelial cells around the day of birth

Signal Transducers and Activators of Transcription (STATs) regulate mammary gland development. Here we investigate the expression of pSTAT3 in the murine and bovine mammary gland around the day of birth. We identify polarisation of mammary alveoli towards either a low- or high-proportion of pSTAT3 positive alveolar epithelial cells. We present localised colocation analysis applicable to other mammary studies where identification, quantification and interrogation of significant, spatially congregated events is required. We demonstrate that pSTAT3-positive events are multifocally clustered in a non-random and statistically significant fashion. This finding represents a new facet of mammary STAT3 biology meriting further functional investigation.

cell biology↗

RNA structure and multiple weak interactions balance the interplay between RNA binding and phase separation of SARS-CoV-2 nucleocapsid

The nucleocapsid (N) protein of SARS-CoV-2 binds viral RNA, condensing it inside the virion, and phase separating with RNA to form liquid-liquid condensates. There is little consensus on what differentiates sequence-independent N-RNA interactions in the virion or in liquid droplets from those with specific genomic RNA motifs necessary for viral function inside infected cells. To identify the RNA structures and the N domains responsible for specific interactions and phase separation, we use the first 1000nt of viral RNA and short RNA segments designed as models for single-stranded and paired RNA. Binding affinities estimated from fluorescence anisotropy of these RNAs to the two folded domains of N (the NTD and CTD) and comparison to full-length N demonstrate that the NTD binds preferentially to single-stranded RNA, and while it is the primary RNA binding site, it is not essential to phase separation. Nuclear magnetic resonance spectroscopy identifies two RNA binding sites on the NTD: a previously characterized site and an additional although weaker RNA-binding face that becomes prominent when binding to the primary site is weak, such as with dsRNA or a binding-impaired mutant. Phase separation assays of nucleocapsid domains with different RNA structures support a model where multiple weak interactions, such as with the CTD or the NTDs secondary face promote phase separation, while strong, specific interactions do not. These studies indicate that both strong and multivalent weak N-RNA interactions underlie the multifunctional abilities of N. SignificanceThe nucleocapsid protein of the SARS-CoV-2 coronavirus binds to viral RNA, both to protect and condense it inside the viral particle and to facilitate viral transcription inside infected host cells. Evidence suggests that variations in RNA structure impact how and where it binds to the nucleocapsid, but these differences are not well understood at a structural level. Using nuclear magnetic resonance spectroscopy, we examine the interactions between each folded domain of the nucleocapsid and different RNA structures. Binding affinities and NMR chemical shift profiles demonstrate that binding between the N-terminal domain and single stranded RNA is driven by strong interactions at a specific site, while multiple weak nonspecific interactions at newly discovered sites lead to phase separation and RNA condensation.

biophysics↗

Modifications to the SR-Rich Region of the SARS-CoV-2 Nucleocapsid Regulate Self-Association and Attenuate RNA Interactions

The nucleocapsid protein (N) of SARS-CoV-2 is essential for virus replication, genome packaging, and maturation. N is comprised of two folded domains that are separated by a highly conserved, disordered, Ser/Arg-rich linker, and flanked by disordered tails. Using NMR spectroscopy and analytical ultracentrifugation we identify an alpha-helical region in the linker that undergoes concentration dependent self-association. NMR and gel shift assays show that the linker binds viral RNA but this binding is dampened by both phosphorylation and a naturally occurring mutation, whereas in contrast, RNA binding to the full-length protein is not affected. Interestingly, phase separation with RNA is significantly reduced upon phosphorylation but enhanced with the mutation. We attribute these differences to changes in the linker helix self-association which dissociates upon phosphorylation but forms more stable higher order oligomers in the variant. These data provide a structural mechanism for how the linker region contributes to protein-protein interactions, RNA-protein interactions, liquid-liquid phase separation and N protein regulation.

biophysics↗

AI-powered pan-species computational pathology: bridging clinic and wildlife care

Cancers occur across species. Understanding what is consistent and varies across species can provide new insights into cancer initiation and evolution, with significant implications for animal welfare and wildlife conservation. We built the pan-species cancer digital pathology atlas (PANCAD) and conducted the first pan-species study of computational comparative pathology using a supervised convolutional neural network algorithm trained on human samples. The artificial intelligence algorithm achieves high accuracy in measuring immune response through single-cell classification for two transmissible cancers (canine transmissible venereal tumour, 0.94; Tasmanian devil facial tumour disease, 0.88). Furthermore, in 18 other vertebrate species (mammalia=11, reptilia=4, aves=2, and amphibia=1), accuracy (0.57-0.94) was influenced by cell morphological similarity preserved across different taxonomic groups, tumour sites, and variations in the immune compartment. A new metric, named morphospace overlap, was developed to guide veterinary pathologists towards rational deployment of this technology on new samples. This study provides the foundation and guidelines for transferring artificial intelligence technologies to veterinary pathology based on a new understanding of morphological conservation, which could vastly accelerate new developments in veterinary medicine and comparative oncology.

cancer biology↗

Developing mammary terminal duct lobular units have a dynamic mucosal and stromal immune microenvironment

The human breast and ovine mammary gland undergo a striking degree of postnatal development, leading to formation of terminal duct lobular units (TDLUs). In this study we interrogated aspects of sheep TDLU growth to increase understanding of ovine mammogenesis and as a model for the study of breast development. Mammary epithelial proliferation is significantly higher in lambs less than two months old than in peri-pubertal animals. Ki67 expression is polarized to the leading edge of the developing TDLUs. Intraepithelial ductal macrophages exhibit striking periodicity and significantly increased density in lambs approaching puberty. Stromal macrophages are more abundant centrally than peripherally. The developing ovine mammary gland is infiltrated by intraepithelial and stromal T lymphocytes that are significantly more numerous in older lambs. In the stroma, hotspots of Ki67 expression colocalize with large aggregates of lymphocytes and macrophages. Multifocally these aggregates exhibit distinct organization consistent with tertiary lymphoid structures. The lamb mammary gland thus exhibits a dynamic mucosal and stromal immune microenvironment and, as such, constitutes a valuable model system that provides new insights into postnatal breast development. Summary statementDevelopment of terminal duct lobular units in the sheep mammary gland involves distinct growth phases and macrophage and lymphocyte fluxes. Tertiary lymphoid structures are present subjacent to the mucosal epithelium.

developmental biology↗