Adenoid ameloblastoma harbors beta-catenin mutations.

Adenoid ameloblastoma is a very rare benign epithelial odontogenic tumor characterized microscopically by epithelium resembling conventional ameloblastoma, with additional duct-like structures, epithelial whorls, and cribriform architecture. Dentinoid deposits, clusters of clear cells, and ghost-cell keratinization may also be present. These tumors do not harbor BRAF or KRAS mutations and their molecular basis appears distinct from conventional ameloblastoma but remains unknown. We assessed CTNNB1 (beta-catenin) exon 3 mutations in a cohort of 11 samples of adenoid ameloblastomas from 9 patients. Two of the 9 patients were female and 7 male and in 7/9 patients the tumors occurred in the maxilla. Tumors of 4 of these 9 patients harbored CTNNB1 mutations, specifically p.Ser33Cys, p.Gly34Arg, and p.Ser37Phe. Notably, for one patient 3 samples were analyzed including the primary tumour and two consecutive recurrences, and results were positive for the mutation in all three tumors. Therefore, 6/11 samples tested positive for the mutation. In the 6 mutation-positive samples, ghost cells were present in only 2/6, indicating beta-catenin mutations are not always revealed by ghost cell formation. Dentinoid matrix deposition was observed in 5/6 mutation-positive samples and clear cells in all 6 cases. None of the cases harbored either BRAF or KRAS mutations. Beta-catenin immunoexpression was assessed in the samples of 8 patients. Except for one wild-type case, all cases showed focal nuclear expression irrespective of the mutational status. Together with the absence of BRAF mutation, the detection of beta-catenin mutation in adenoid ameloblastomas supports its classification as a separate entity, and not as a subtype of ameloblastoma. The presence of this mutation may help in the diagnosis of challenging cases.

Cytosolic Delivery of Small Protein Scaffolds Enables Efficient Inhibition of Ras and Myc.

The ability to deliver small protein scaffolds intracellularly could enable the targeting and inhibition of many therapeutic targets that are not currently amenable to inhibition with small-molecule drugs. Here, we report the engineering of small protein scaffolds with anionic polypeptides (ApPs) to promote electrostatic interactions with positively charged nonviral lipid-based delivery systems. Proteins fused with ApPs are either complexed with off-the-shelf cationic lipids or encapsulated within ionizable lipid nanoparticles for highly efficient cytosolic delivery (up to 90%). The delivery of protein inhibitors is used to inhibit two common proto-oncogenes, Ras and Myc, in two cancer cell lines. This report demonstrates the feasibility of combining minimally engineered small protein scaffolds with tractable nanocarriers to inhibit intracellular proteins that are generally considered "undruggable" with current small molecule drugs and biologics.

Synchronous Chemoradiation Nanovesicles by X-Ray Triggered Cascade of Drug Release.

The approach of concurrent-to-synchronous chemoradiation has now been advanced by well-designed nanovesicles that permit X-ray irradiation-triggered instant drug release. The nanovesicles consist of Au nanoparticles tethered with irradiation labile linoleic acid hydroperoxide (LAHP) molecules and oxidation-responsive poly(propylene sulfide)-poly(ethylene glycol) (PPS-PEG) polymers, where DOX were loaded in the inner core of the vesicles (Au-LAHP-vDOX). Upon irradiation, the in situ formation of hydroxyl radicals from LAHP molecules triggers the internal oxidation of PPS from being hydrophobic to hydrophilic, leading to degradation of the vesicles and burst release of cargo drugs. In this manner, synchronous chemoradiation showed impressive anticancer efficacy both in vitro and in a subcutaneous mouse tumor model by one-dose injection and one-time irradiation.

Lipid-mediated intracellular delivery of recombinant bioPROTACs for the rapid degradation of undruggable proteins.

Recently, targeted degradation has emerged as a powerful therapeutic modality. Relying on "event-driven" pharmacology, proteolysis targeting chimeras (PROTACs) can degrade targets and are superior to conventional inhibitors against undruggable proteins. Unfortunately, PROTAC discovery is limited by warhead scarcity and laborious optimization campaigns. To address these shortcomings, analogous protein-based heterobifunctional degraders, known as bioPROTACs, have been developed. Compared to small-molecule PROTACs, bioPROTACs have higher success rates and are subject to fewer design constraints. However, the membrane impermeability of proteins severely restricts bioPROTAC deployment as a generalized therapeutic modality. Here, we present an engineered bioPROTAC template able to complex with cationic and ionizable lipids via electrostatic interactions for cytosolic delivery. When delivered by biocompatible lipid nanoparticles, these modified bioPROTACs can rapidly degrade intracellular proteins, exhibiting near-complete elimination (up to 95% clearance) of targets within hours of treatment. Our bioPROTAC format can degrade proteins localized to various subcellular compartments including the mitochondria, nucleus, cytosol, and membrane. Moreover, substrate specificity can be easily reprogrammed, allowing modular design and targeting of clinically-relevant proteins such as Ras, Jnk, and Erk. In summary, this work introduces an inexpensive, flexible, and scalable platform for efficient intracellular degradation of proteins that may elude chemical inhibition.

Lipid Nanoparticle Delivery of Small Proteins for Potent In Vivo RAS Inhibition.

Mutated RAS proteins are potent oncogenic drivers and have long been considered "undruggable". While RAS-targeting therapies have recently shown promise, there remains a clinical need for RAS inhibitors with more diverse targets. Small proteins represent a potential new therapeutic option, including K27, a designed ankyrin repeat protein (DARPin) engineered to inhibit RAS. However, K27 functions intracellularly and is incapable of entering the cytosol on its own, currently limiting its utility. To overcome this barrier, we have engineered a lipid nanoparticle (LNP) platform for potent delivery of functional K27-D30─a charge-modified version of the protein─intracellularly in vitro and in vivo. This system efficiently encapsulates charge-modified proteins, facilitates delivery in up to 90% of cells in vitro, and maintains potency after at least 45 days of storage. In vivo, these LNPs deliver K27-D30 to the cytosol of cancerous cells in the liver, inhibiting RAS-driven growth and ultimately reducing tumor load in an HTVI-induced mouse model of hepatocellular carcinoma. This work shows that K27 holds promise as a new cancer therapeutic when delivered using this LNP platform. Furthermore, this technology has the potential to broaden the use of LNPs to include new cargo types─beyond RNA─for diverse therapeutic applications.

Treatment of humeral condylar fractures and humeral intracondylar fissures in cats with patellar fracture and dental anomaly syndrome.

OBJECTIVES: The aim of this study is to describe the treatment and outcome of humeral condylar fractures and humeral intracondylar fissures in cats with patellar fracture and dental anomaly syndrome (PADS) and to provide advice on how to manage these cases in practice. METHODS: Data were collated on cats with PADS that were reported to have sustained humeral fractures or had fractures or fissures of the humerus identified on radiographs. The details of the fractures were recorded in addition to any treatment and outcome information. RESULTS: Of the 207 cases reported with PADS, 18 cats (8.7%) were found to have humeral condylar fractures, none of which was known to have resulted from significant trauma. Where treatment occurred, it involved the placement of transcondylar positional or lag screws. In some cases additional implants, including supracondylar bone plates and screws or Kirschner wires (K-wires), were used. Follow-up data revealed that only two cats were euthanased owing to the presence of the humeral fractures, with at least eight achieving some degree of recovery of function. CONCLUSIONS AND RELEVANCE: These humeral fractures all have the characteristics of stress insufficiency fractures, being simple isolated fractures that are short oblique, with increased radio-density at the fracture line and occurring following minimal or no trauma. Humeral intracondylar fissures were identified in two cats and it is possible that some of the other fractures may have occurred secondary to pre-existing fissures. To our knowledge, no prior reports exist of fissures in cats that do not meet the criteria for PADS. Surgical repair primarily consisted of the placement of transcondylar lag or positional screws with, in some cases, adjunct implants such as bone plates and screws or K-wires. Though there were insufficient data to determine the prognosis for these fractures in the long term, unlike patellar fractures, many of these fractures will heal if treated appropriately.

Metallic implant-associated lymphoma: a distinct subgroup of large B-cell lymphoma related to pyothorax-associated lymphoma?

Primary non-Hodgkin lymphoma arising at the site of metallic implant is very rare, and the possible carcinogenic effects of the metallic components and wear particles of the implant have not been answered despite many years of investigation. We report a case of large B-cell lymphoma occurring in a 78-year-old man who had a knee prosthesis implant for more than 30 years. The lymphoma was of microscopic size and found incidentally in the wear debris removed at surgical revision of the loosened prosthesis. The lymphoma expressed CD20, showed clonal rearrangements of immunoglobulin gene, and harbored Epstein-Barr virus (EBV). This case, together with previously reported cases, suggests that metallic implant-associated lymphoma is a distinctive subgroup of large B-cell lymphoma that shares many similarities with pyothorax-associated lymphoma and osteomyelitis-associated lymphoma, in that the lymphoma is an EBV-associated large B-cell lymphoma arising in a setting of chronic inflammation or irritation in a confined body space.

Remote and local control of stimuli responsive materials for therapeutic applications.

Materials offering the ability to change their characteristics in response to presented stimuli have demonstrated application in the biomedical arena, allowing control over drug delivery, protein adsorption and cell attachment to materials. Many of these smart systems are reversible, giving rise to finer control over material properties and biological interaction, useful for various therapeutic treatment strategies. Many smart materials intended for biological interaction are based around pH or thermo-responsive materials, although the use of magnetic materials, particularly in neural regeneration, has increased over the past decade. This review draws together a background of literature describing the design principles and mechanisms of smart materials. Discussion centres on recent literature regarding pH-, thermo-, magnetic and dual responsive materials, and their current applications for the treatment of neural tissue.

Diagnostic cytological features of polyacrylamide gel injection augmentation mammoplasty.

AIMS: To study the cytological features of breast lesions associated with hydrophilic polyacrylamide gel (PAAG) injection augmentation mammoplasty and to determine the specific diagnostic features. METHODS: Fine needle aspiration (FNA) cytology smears from 14 patients who presented with breast lump and a clinical history of PAAG injection were reviewed. The staining properties of the PAAG material in Papanicolaou (Pap), Haematoxylin and Eosin (H&E) and Diff-Quik stains, as well as the cellular background, were studied. Cell blocks were also studied with H&E, periodic acid-Schiff (PAS), periodic acid-Schiff with diastase digestion (PASD) and mucicarmine (MC) stains. RESULTS: PAAG was stained consistently pinkish with a homogenous smooth texture on H&E (100%), polychromasia on Pap (100%) and magenta violet with bubbly vacuoles on Diff-Quik (100%). Half (7) of the cases showed sharp borders. The presence of foreign body type giant cells with histiocytes (5 cases, 36%) and scanty clusters of benign ductal cells (4 cases, 29%) were seen in the background of a minority of the cases. PAAG was represented by blocks of homogenous grey-purplish acellular material with a sharp border on cell block sections, which were negative for PAS, PASD and MC. CONCLUSION: PAAG shows a consistent staining pattern in various common cytological staining preparations. Blocks of acellular gelatinous material, stained homogenous pinkish on H&E, polychromasia on Pap and magenta violet with bubbly vacuoles on Diff-Quik, are the reliable cytological features of PAAG.