Hyalinizing clear cell carcinoma (HCCC) is a rare tumor. Most lesions arise in the salivary gland; however, other primary sites include the nasal and oral cavities, and rarely lung. Lesions are characterized by tumor lobules with pale or clear cytoplasm, arranged in nests or cords, surrounded by a mixed hyalinized and fibrocellular stroma. EWSR1-ATF1 gene fusion is often reported in these lesions. Typically, HCCC bears a good prognosis. We report a unique case of primary lung HCCC (PLHCCC) with cutaneous metastasis and aggressive clinical course. To the best of our knowledge, cutaneous metastasis by PLHCCC has not been previously observed and may signal an aggressive clinical course with poor prognosis.

Mycoplasma hyopneumoniae (M. hyopneumoniae), the etiological agent of porcine enzootic pneumonia, is one of the most prevalent and economically significant respiratory pathogens in the swine industry. Vaccination constitutes a primary strategy for controlling M. hyopneumoniae infections. Live-attenuated vaccines confer protection against wild-type M. hyopneumoniae through competitive exclusion of early lung colonization and induce multifaceted immune responses, including humoral, mucosal, and cellular immunity, thereby exhibiting superior protective efficacy. However, the lack of methods for differentiating live-attenuated vaccine immunization from wild-type infection has hindered the widespread application of live-attenuated vaccines. In this study, we developed a quantitative real-time polymerase chain reaction (qPCR) method based on the ribonuclease H2 (RNase H2) cleavage principle, which emits specific fluorescent signals exclusively when the probe is fully complementary to the template of the live-attenuated vaccine strains. This qPCR method exhibited high specificity and sensitivity for the M. hyopneumoniae live-attenuated vaccines (168-L and RM48), with a lower detection limit of 10 copies/µL. Using this qPCR method, the bacterial shedding dynamics of the 168-L vaccine strain in piglets were effectively monitored, with the first detection in the nasal cavity at 3 days postvaccination (dpv), and a peak detection rate of 100% between 10 and 17 dpv. This established assay can specifically monitor the respiratory colonization and shedding of the vaccine strain after M. hyopneumoniae live-attenuated vaccine immunization, thereby providing crucial technical support for the clinical evaluation of immunization efficacy.

Limited understanding of the biological processes that govern metastatic dissemination hinders its prevention and treatment[1]. Here, using 501 longitudinally collected primary and metastatic tumour samples from 24 patients with non-small cell lung cancer (NSCLC) enrolled in the TRACERx lung study and PEACE autopsy programme, we infer tumour evolution from diagnosis to death. With DNA-sequencing data encompassing 70% of the metastases that were radiologically detected before death and paired multi-region sampled primary tumours, we show that the genomes of metastases diverge markedly from those of their ancestral primary tumour, with additional driver alterations and genome doubling events occurring after metastatic dissemination. In 62.5% of patients, multiple primary tumour subclones disseminated, each founding a distinct metastasis. These metastases served as sources of onward spread: more than half of the metastases sampled were seeded by other metastases. The duration that metastases existed in situ influenced their likelihood of seeding further metastases. Most metastatic migrations started and ended in the same anatomical cavity. The few subclones that exited the thorax to seed metastases disseminated widely and were enriched for somatic copy-number alterations, suggesting that chromosomal instability may facilitate extrathoracic spread. This spatial and temporal evolutionary analysis sheds light on the extent of metastatic diversity and seeding in advanced NSCLC-which tends to be underestimated in single metastasis biopsies-and identifies genomic and clinical mediators of metastatic progression.

An amoebic liver abscess (ALA) caused by Entamoeba histolytica remains a significant cause of morbidity and mortality in tropical regions. Although hepatic rupture into adjacent thoracic structures is a recognised but rare complication, it can be fatal if undiagnosed or untreated. Hepatic abscesses can rupture and spread the infection to the thoracic cavity, resulting in the formation of a hepato-thoracic fistula, rarely. We report an autopsy case of a middle-aged female who died suddenly following nonspecific systemic symptoms. Postmortem examination revealed a large ALA that had ruptured through the diaphragm, leading to massive pleural empyema. This autopsy case report underscores the importance of early clinical suspicion, imaging, and surgical management to prevent such catastrophic outcomes.

Post-tuberculosis (TB) lung disease (PTLD) affects approximately 50% of persons with pulmonary TB. We recently discovered whole blood transcriptional signatures associated with PTLD. We examined whether a minimum gene signature predicts PTLD as a clinically useful biomarker.

The portfolio of next generation COVID-19 vaccines would benefit from candidates that induce durable systemic and mucosal immune responses that would lessen person-to-person transmission. We constructed an intranasal (IN) replication-competent adenovirus type 4 recombinant platform to express SARS-CoV-2 Spike variants (Ad4-S) and assessed immunogenicity and efficacy in the Syrian hamster model. Although both IN Ad4-S and intramuscular (IM) vaccines (Ad26.CoV2.S and mRNA-1273) induced serum binding antibodies, only Ad4-S induced a robust mucosal response in the nasal cavity. IN Ad4-S vaccination induced serum neutralizing titers equivalent to or greater than IM vaccination but more durable up to 6 months. Upon challenge, IN immunization also resulted in less weight loss, greater breadth and durability of restriction of viral replication, and less lung pathology than IM immunization up to 268 days after immunization. These data support the potential of the IN Ad4 vaccine platform to reduce transmission of SARS-CoV-2 and other respiratory viruses with pandemic potential.

The tumor suppressor p53 is the most frequently mutated protein in tumors and a target for drug development. More than 2000 cancer-associated p53 missense mutations have been reported, most of them located in the DNA-binding domain (DBD). Due to the low intrinsic thermostability of the latter, they often lead to unfolding at physiological temperature. Stabilizing the DBD with small molecules has been shown to be effective in reactivating the cavity-creating cancer mutant Y220C. Unfortunately, the majority of p53 mutants seem to lack druggable binding pockets for small molecules. Here we show that a designed ankyrin repeat protein (DARPin) that binds to the p53 DBD stabilizes temperature-sensitive (TS) p53 cancer mutants, thereby compensating for mutation-induced loss of stability. We determined high-resolution crystal structures of multiple DARPin-mutant p53 complexes, providing mechanistic insights into this mode of stabilization. Reporter gene assays across a comprehensive panel of cancer-associated mutants revealed reactivation of the majority of TS mutants, whereas DNA-contact mutants and those with local misfolding of the DNA-binding surface remained inactive, as expected. We demonstrate that this reactivation induces the transcription of canonical p53 target genes and elicits antiproliferative effects in cancer cell lines. A combination of this DARPin with an mRNA/lipid nanoparticle-based transfection approach may have the potential to reactivate most TS p53 mutants and resensitize cancer cells to chemotherapy.

Head and neck mucosal melanoma (HNMM) arises in the nasal and oral cavities and has the propensity to metastasize to local and distant body sites. HNMM is also notable for its resistance to available therapeutics. The rarity of this disease makes it difficult to conduct large-scale clinical studies to develop standard treatment protocols. In contrast to cutaneous melanoma, c-Kit-dependent pathways are well studied in HNNMM and provide a potential therapeutic target. We identified and isolated genetically distinct subpopulations with stem cell characteristics in HNMM samples bearing Kit wild-type and mutations. Functional analysis of these subpopulations reveals that, in addition to expressing the stem cell marker proteins CD20, CD117, CD133, and CD166, these subpopulations are characterized by self-renewal potential, migratory capacity, and resistance to Kit inhibitors such as Imatinib. Immunofluorescence staining and inhibition experiments demonstrate that the maintenance and resistance of HHMM subpopulations to Kit inhibitors is mediated by the Kit signal to the PI3K signaling pathway. The KIT signal to the PI3K signaling pathway does not result exclusively from a KIT mutation localized to Exon 17, but can also be triggered by mutations localized to Exons 11 and 13. In the present study, we identify and characterize an HNMM subpopulation with stemness properties in patients with c-Kit wild-type and mutation, and demonstrate for the first time the mechanisms by which the CD117[+]/CD133[+] HNMM subpopulations survive and confer resistance to the specific inhibitor of c-Kit mutation.