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EGFR-targeted therapies are efficacious, but toxicity is common and can be severe. Urokinase type plasminogen activator receptor (uPAR)-targeted drugs are only emerging, so neither their efficacy nor toxicity is fully established. Recombinant eBAT was created by combining cytokines EGF and uPA on the same single-chain molecule with truncated Pseudomonas toxin. Its purpose was to simultaneously target tumors and their vasculature in the tumor microenvironment. In prior studies on mice and dogs, the drug proved efficacious. Here, we report the safety of eBAT in normal wildtype, uPAR knockout, and immunoreplete and immunodeficient tumor-bearing mice, as well as in dogs with spontaneous sarcoma that more closely mirror human cancer onset. In immunocompetent mice, tumor-bearing mice, uPAR knockout mice, and mice receiving species-optimized eBAT, toxicities were mild and self-limiting. Likewise, in dogs with life-threatening sarcoma given dosages found to be biologically active, eBAT was well tolerated. In mice receiving higher doses, eBAT was associated with dose-dependent evidence of liver injury, including portal biliary hyperplasia, oval cell proliferation, lymphoplasmacytic inflammation, periportal hepatocellular microvesicular change, hemorrhage, necrosis, and apoptosis. The results support continuing the clinical development of eBAT as a therapeutic agent for individuals with sarcoma and other cancers.

Hemangiosarcoma and angiosarcoma are soft-tissue sarcomas of blood vessel-forming cells in dogs and humans, respectively. These vasoformative sarcomas are aggressive and highly metastatic, with disorganized, irregular blood-filled vascular spaces. Our objective was to define molecular programs which support the niche that enables progression of canine hemangiosarcoma and human angiosarcoma. Dog-in-mouse hemangiosarcoma xenografts recapitulated the vasoformative and highly angiogenic morphology and molecular characteristics of primary tumors. Blood vessels in the tumors were complex and disorganized, and they were lined by both donor and host cells. In a series of xenografts, we observed that the transplanted hemangiosarcoma cells created exuberant myeloid hyperplasia and gave rise to lymphoproliferative tumors of mouse origin. Our functional analyses indicate that hemangiosarcoma cells generate a microenvironment that supports expansion and differentiation of hematopoietic progenitor populations. Furthermore, gene expression profiling data revealed hemangiosarcoma cells expressed a repertoire of hematopoietic cytokines capable of regulating the surrounding stromal cells. We conclude that canine hemangiosarcomas, and possibly human angiosarcomas, maintain molecular properties that provide hematopoietic support and facilitate stromal reactions, suggesting their potential involvement in promoting the growth of hematopoietic tumors., Significance: We demonstrate that hemangiosarcomas regulate molecular programs supporting hematopoietic expansion and differentiation, providing insights into their potential roles in creating a permissive stromal-immune environment for tumor progression., (© 2024 The Authors; Published by the American Association for Cancer Research.)

Objective: To determine if dogs with neoplasia produce more coated platelets, a subpopulation of activated platelets generated by dual stimulation with thrombin and convulxin, a glycoprotein VI agonist, than healthy control dogs., Animals: Client-owned dogs diagnosed with lymphoma (n = 19) or solid tumors (14) and healthy control dogs (14)., Procedures: Platelets were stimulated ex vivo with thrombin and convulxin. Flow cytometry was used to quantify the percentage of coated platelets based on high levels of surface fibrinogen. To compare the percentage of coated platelets between the three groups, an ANOVA was performed followed by pairwise 95% confidence intervals (CI) adjusted for multiple comparisons using Tukey's method., Results: We observed a greater mean percentage of coated platelets in dogs with solid tumors, compared with healthy control dogs, by 10.9 percentage points (95% CI: -1.0, 22.8), and a mean percentage of coated platelets in dogs with lymphoma that was less than healthy control dogs by 0.3 percentage points (95% CI: -11.4, 10.8)., Clinical Relevance: This study provides the first data-based evidence that dogs with solid tumors may have a greater mean coated platelet percentage when compared with healthy control dogs, although there is overlap between groups. Further studies are needed investigating coated platelets in specific subsets of neoplasia and investigating additional mechanisms of hypercoagulability in dogs with neoplasia.

Cancer is among the most common causes of death for dogs (and cats) and humans in the developed world, even though it is uncommon in wildlife and other domestic animals. We provide a rationale for this observation based on recent advances in our understanding of the evolutionary basis of cancer. Over the course of evolutionary time, species have acquired and fine-tuned adaptive cancer protective mechanisms that are intrinsically related to their energy demands, reproductive strategies, and expected lifespan. These cancer protective mechanisms are general across species and/or specific to each species and their niche, and they do not seem to be limited in diversity. The evolutionarily acquired cancer-free longevity that defines a species' life history can explain why the relative cancer risk, rate, and incidence are largely similar across most species in the animal kingdom despite differences in body size and life expectancy. The molecular, cellular, and metabolic events that promote malignant transformation and cancerous growth can overcome these adaptive, species-specific protective mechanisms in a small proportion of individuals, while independently, some individuals in the population might achieve exceptional longevity. In dogs and humans, recent dramatic alterations in healthcare and social structures have allowed increasing numbers of individuals in both species to far exceed their species-adapted longevities (by 2-4 times) without allowing the time necessary for compensatory natural selection. In other words, the cancer protective mechanisms that restrain risk at comparable levels to other species for their adapted lifespan are incapable of providing cancer protection over this recent, drastic and widespread increase in longevity., Competing Interests: Conflict of Interest JFM is an inventor on U.S. patent 7910315, “Early Detection of Hemangiosarcoma and Angiosarcoma,” assigned to the Regents of the University of Colorado. JFM is an inventor on U.S. patent application, “Reduction of EGFR Therapeutic Toxicity,” filed on behalf of the Regents of the University of Minnesota. JFM and TAD are inventors on U.S. patent application, “Artificial Intelligence for Early Cancer Detection,” filed on behalf of the Regents of the University of Minnesota.

Angiosarcomas are soft-tissue sarcomas that form malignant vascular tissues. Angiosarcomas are very rare, and due to their aggressive behavior and high metastatic propensity, they have poor clinical outcomes. Hemangiosarcomas commonly occur in domestic dogs, and share pathological and clinical features with human angiosarcomas. Typical pathognomonic features of this tumor are irregular vascular channels that are filled with blood and are lined by a mixture of malignant and nonmalignant endothelial cells. The current gold standard is the histological diagnosis of angiosarcoma; however, microscopic evaluation may be complicated, particularly when tumor cells are undetectable due to the presence of excessive amounts of nontumor cells or when tissue specimens have insufficient tumor content. In this study, we implemented machine learning applications from next-generation transcriptomic data of canine hemangiosarcoma tumor samples (n = 76) and nonmalignant tissues (n = 10) to evaluate their training performance for diagnostic utility. The 10-fold cross-validation test and multiple feature selection methods were applied. We found that extra trees and random forest learning models were the best classifiers for hemangiosarcoma in our testing datasets. We also identified novel gene signatures using the mutual information and Monte Carlo feature selection method. The extra trees model revealed high classification accuracy for hemangiosarcoma in validation sets. We demonstrate that high-throughput sequencing data of canine hemangiosarcoma are trainable for machine learning applications. Furthermore, our approach enables us to identify novel gene signatures as reliable determinants of hemangiosarcoma, providing significant insights into the development of potential applications for this vascular malignancy., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Sporadic angiosarcomas are aggressive vascular sarcomas whose rarity and genomic complexity present significant obstacles in deciphering the pathogenic significance of individual genetic alterations. Numerous fusion genes have been identified across multiple types of cancers, but their existence and significance remain unclear in sporadic angiosarcomas. In this study, we leveraged RNA-sequencing data from 13 human angiosarcomas and 76 spontaneous canine hemangiosarcomas to identify fusion genes associated with spontaneous vascular malignancies. Ten novel protein-coding fusion genes, including TEX2-PECAM1 and ATP8A2-FLT1 , were identified in seven of the 13 human tumors, with two tumors showing mutations of TP53 . HRAS and NRAS mutations were found in angiosarcomas without fusions or TP53 mutations. We found 15 novel protein-coding fusion genes including MYO16-PTK2, GABRA3-FLT1 , and AKT3-XPNPEP1 in 11 of the 76 canine hemangiosarcomas; these fusion genes were seen exclusively in tumors of the angiogenic molecular subtype that contained recurrent mutations in TP53, PIK3CA, PIK3R1 , and NRAS . In particular, fusion genes and mutations of TP53 cooccurred in tumors with higher frequency than expected by random chance, and they enriched gene signatures predicting activation of angiogenic pathways. Comparative transcriptomic analysis of human angiosarcomas and canine hemangiosarcomas identified shared molecular signatures associated with activation of PI3K/AKT/mTOR pathways. Our data suggest that genome instability induced by TP53 mutations might create a predisposition for fusion events that may contribute to tumor progression by promoting selection and/or enhancing fitness through activation of convergent angiogenic pathways in this vascular malignancy. IMPLICATIONS: This study shows that, while drive events of malignant vasoformative tumors of humans and dogs include diverse mutations and stochastic rearrangements that create novel fusion genes, convergent transcriptional programs govern the highly conserved morphologic organization and biological behavior of these tumors in both species., (©2021 American Association for Cancer Research.)

Introduction: There is a growing trend of multidrug-resistant organisms (MDRO). The goal of this study was to characterize MDRO at a single center from ophthalmic cultures to better understand how treatments were tailored and to assess effect on visual acuity., Materials and Methods: The MDRO data were collected by the Multidrug-Resistant Organism Repository and Surveillance Network from the Brooke Army Medical Center clinical laboratory. Both patient- and isolate-specific data were collected and qualitatively analyzed. Primary outcome measures were organism and type of resistance, anatomic location of isolate, initial and final antibiotic choice, and visual acuity., Results: Thirty-one bacterial culture samples were analyzed from 29 patients. Twenty-two (72%) were Gram-positive and all were methicillin-resistant Staphylococcus aureus (MRSA). Nine (29%) were Gram-negative and of these five were Pseudomonas spp. Fourteen (45%) isolates were cultured from the cornea, nine (29%) from the lid, four (13%) from the conjunctiva, and four (13%) from other locations. The majority (66.6%) required adjustment of initial antibiotics following ocular culture results. Sixteen adult patients had recorded initial and final visual acuities. Fifteen of those 16 patients had stable or improved visual acuities following treatment of the infection, but five patients had a final visual acuity less than 20/200., Conclusion: This study demonstrated a high frequency of corneal MDRO infections and specifically MRSA and Pseudomonas spp. isolates. Antibiotic treatments frequently required adjustment. Further prospective study of visual outcomes from ophthalmic MDRO cultures is needed., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2020. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

Angiosarcoma is a highly aggressive cancer of blood vessel-forming cells with few effective treatment options and high patient mortality. It is both rare and heterogenous, making large, well-powered genomic studies nearly impossible. Dogs commonly suffer from a similar cancer, called hemangiosarcoma, with breeds like the golden retriever carrying heritable genetic factors that put them at high risk. If the clinical similarity of canine hemangiosarcoma and human angiosarcoma reflects shared genomic etiology, dogs could be a critically needed model for advancing angiosarcoma research. We assessed the genomic landscape of canine hemangiosarcoma via whole-exome sequencing (47 golden retriever hemangiosarcomas) and RNA sequencing (74 hemangiosarcomas from multiple breeds). Somatic coding mutations occurred most frequently in the tumor suppressor TP53 (59.6% of cases) as well as two genes in the PI3K pathway: the oncogene PIK3CA (29.8%) and its regulatory subunit PIK3R1 (8.5%). The predominant mutational signature was the age-associated deamination of cytosine to thymine. As reported in human angiosarcoma, CDKN2A/B was recurrently deleted and VEGFA, KDR , and KIT recurrently gained. We compared the canine data to human data recently released by The Angiosarcoma Project, and found many of the same genes and pathways significantly enriched for somatic mutations, particularly in breast and visceral angiosarcomas. Canine hemangiosarcoma closely models the genomic landscape of human angiosarcoma of the breast and viscera, and is a powerful tool for investigating the pathogenesis of this devastating disease. IMPLICATIONS: We characterize the genomic landscape of canine hemangiosarcoma and demonstrate its similarity to human angiosarcoma., (©2019 American Association for Cancer Research.)

Angiosperms and their pollinators are adapted in a close co-evolution. For both the plants and pollinators, the functioning of the visual signaling system is highly relevant for survival. As the frequency range of visual perception in many insects extends into the ultraviolet (UV) region, UV-patterns of plants play an important role in the flower-pollinator interaction. It is well known that many flowers contain UV-absorbing pigments in their petal cells, which are localized in vacuoles. However, the contribution of the petal surface microarchitecture to UV-reflection remains uncertain. The correlation between the surface structure and its reflective properties is also relevant for biomimetic applications, for example, in the field of photovoltaics. Based on previous work, we selected three model species with distinct UV-patterns to explore the possible contribution of the surface architecture to the UV-signaling. Using a replication technique, we transferred the petal surface structure onto a transparent polymer. Upon illumination with UV-light, we observed structural-based patterns in the replicas that were surprisingly comparable to those of the original petals. For the first time, this experiment has shown that the parameters of the surface structure lead to an enhancement in the amount of absorbed UV-radiation. Spectrophotometric measurements revealed up to 50% less reflection in the UV-absorbing regions than in the UV-reflecting areas. A comparative characterization of the micromorphology of the UV-reflecting and UV-absorbing areas showed that, in principle, a hierarchical surface structure results in more absorption. Therefore, the results of our experiments demonstrate the structural-based amplification of UV-reflection and provide a starting point for the design of bioinspired antireflective and respectively strongly absorbing surfaces.

Hierarchically structured flower leaves (petals) of many plants are superhydrophobic, but water droplets do not roll-off when the surfaces are tilted. On such surfaces water droplets are in the "Cassie impregnating wetting state", which is also known as the "petal effect". By analyzing the petal surfaces of different species, we discovered interesting new wetting characteristics of the surface of the flower of the wild pansy (Viola tricolor). This surface is superhydrophobic with a static contact angle of 169° and very low hysteresis, i.e., the petal effect does not exist and water droplets roll-off as from a lotus (Nelumbo nucifera) leaf. However, the surface of the wild pansy petal does not possess the wax crystals of the lotus leaf. Its petals exhibit high cone-shaped cells (average size 40 µm) with a high aspect ratio (2.1) and a very fine cuticular folding (width 260 nm) on top. The applied water droplets are in the Cassie-Baxter wetting state and roll-off at inclination angles below 5°. Fabricated hydrophobic polymer replicas of the wild pansy were prepared in an easy two-step moulding process and possess the same wetting characteristics as the original flowers. In this work we present a technical surface with a new superhydrophobic, low adhesive surface design, which combines the hierarchical structuring of petals with a wetting behavior similar to that of the lotus leaf.

The contact area between liquids and solid surfaces plays the crucial role in the wetting and self-cleaning properties of surfaces. In this study, we have developed a cryo-preparation method to visualize the contact area between liquids and superhydrophobic biological surfaces by scanning electron microscopy. Aqueous liquids that do not crystallize during freezing, such as glycerol and phosphoric acid, were used. First, the samples in contact with the liquid droplets were cooled with liquid nitrogen. After this, the droplets were separated and the contact areas on the frozen droplets were visualized by scanning electron microscopy. The contact areas of droplets on various biological and artificial surfaces with microstructure, nanostructure, and hierarchical structures are shown in detail. It could be shown that spaces between nanostructures were not penetrated by the droplet, which rested only on top of the structures. Measurements of the contact areas showed the largest reduction in the solid-liquid contact area on hierarchically structured leaf surfaces. On these surfaces, the droplets are in the "Cassie state" at both levels of surface structuring. On plant surfaces, the varying height of the epidermal cells and the surface relief caused considerable variations in the contact between droplet and surface. The examples demonstrate that this new approach provides detailed insights into the wetting behavior of surfaces in the Cassie state with partial contact with the liquid.

Plant surfaces are characterized by a high diversity of structures which determine their optical properties, such as shiny, gleaming, silky, matt or iridescent. Replicas with different optical properties have been generated by using plant surfaces as templates and an improved replica technique. The technique allows the replication of complex surface structures with overhangs, cavities, and fragile or soft structures in a fast and cost-efficient way. Structures from some millimetres to some nanometres can be replicated. The transfer of complex architectures with different optical properties from plant surfaces onto technical surfaces implies a great potential for the development of new biomimetic surfaces with new optical properties.

Biological surfaces are multifunctional interfaces between the organisms and their environment. Properties such as the wettability and adhesion of particles are linked to the micro- and nanostructures of their surfaces. In this study, we used plant and artificial surfaces covered with wax crystals to develop a low-cost replication technique with high resolution. The technique is applicable for fragile surface structures, as demonstrated for three-dimensional wax crystals, and is fast to prevent shrinking of the biological material by water loss during the molding process. Thermal evaporation of octacosan-1-ol has been used to create microstructured surfaces with small platelets as templates for molding. Epoxy resin as filling material provided the smallest deviations from the original surface structures and can be used for replication of nanostructures as small as 4.5 nm. Contact angle measurements of leaves and their replicas show that this technique can be used to develop biomimetic surfaces with similar wettability as in the plant surfaces.

The use of the preperitoneal approach in the treatment of recurrent inguinal hernias is gradually increasing in the Netherlands. This may be concluded from an inquiry conducted among the General Surgery teaching hospitals. The technique, indications and early results are described. Fifty-six recurrent inguinal hernias in 48 patients were treated by this approach. When indicated prosthetic material was used. The follow-up after an average of 19.5 (4-36) months revealed 5 (8.9%) unilateral recurrences.