Localized lung insult, such as pneumonia, can increase respiratory drive and effort. The interplay between the initial injury, the inflammatory reaction and increased drive and effort could damage other areas and progress to bilateral lung injury. Localized pulmonary inflammation was induced through instillation of 0.5 mg/kg lipopolysaccharide (LPS) in the left lower lobe of 18 pigs, while 5 animals served as sham controls. After 24 hours of spontaneous breathing, the severity of right lung injury was assessed by the validated histological score and correlated with potential physiological and biological determinants. Animals challenged with lobar inflammation developed bilateral lung injury, associated with increased respiratory drive and effort. Histological score of the right lung was characterized by wide inter-individual variability (median 11 [8-14], range 3-25). Right lung injury score was correlated with respiratory drive and effort; with respiratory rate and minute ventilation, but not with tidal volume; with peak inspiratory and driving transpulmonary pressures, and with EIT-derived lung strain; with lower sub-atmospheric alveolar pressure and with more negative end-expiratory transpulmonary pressure. Right lung injury score was also correlated with inflammatory plasma cytokines: higher SDF-1α and lower IL-1Ra, IL-5 and GM-CSF. An experimental model of localized lung inflammation allowed us to investigate the role of specific pathophysiological mechanisms for the development of injury in previously healthy lung regions.

Differentiating reactive lymphadenopathies in the context of autoimmune disease from Idiopathic Multicentric Castleman Disease (iMCD) poses a significant diagnostic challenge. Castleman-like histological features have been described in various autoimmune disorders, necessitating a strict and comprehensive integration of clinical and laboratory findings to reach the correct diagnosis. Although the Castleman Disease Collaborative Network (CDCN) consensus guidelines list several autoimmune conditions as exclusion criteria for an iMCD diagnosis, mixed connective tissue disease (MCTD) is not currently among them. We report the case of a  77-year-old woman presenting with fatigue, Raynaud's  phenomenon, sclerodactyly, mild generalized lymphadenopathy, in whom the lymph node biopsy revealed a Castleman-like histology. The absence of systemic inflammatory symptoms and the presence of high-titer anti-U1- RNP antibodies were, however, inconsistent with iMCD, favouring the diagnosis of a reactive Castleman-like lymphadenitis secondary to MCTD. This report highlights that Castleman-like lymphadenopathy can occur in MCTD, closely mimicking iMCD. Therefore, in patients with autoimmune diseases not explicitly listed among the CDCN exclusion criteria, comprehensive clinicopathological integration is essential to avoid misdiagnosis and potentially inappropriate antiIL-6-based therapy.

The onset of chronic lung allograft dysfunction (CLAD) represents the greatest long-term challenge in lung transplantation (LT). Here we aimed to identify early molecular signals of CLAD by analyzing the effects of bronchoalveolar lavage (BAL)-derived extracellular vesicles (EVs) on airway cells and validating these findings in patient lung tissues. In our BAL biobank, we identified 13 LT patients with a BAL sample at CLAD diagnosis and 13 patients with a stable graft function and a BAL sample obtained at least 12 months post LT (Ctrl). All patients were then followed for at least 18 months. EVs were isolated, immunophenotyped, and co-cultured with airway cells. The cells’ transcriptome and proteome were profiled. Selected targets were validated by immunohistochemistry. Logistic regression and survival analyses were performed for prediction of CLAD progression. During follow-up, 7 CLAD patients experienced allograft dysfunction aggravation, and one control developed CLAD. CLAD patients showed more EVs originating from epithelial cells and leukocytes than stable LT recipients. Exposure of airway cells to CLAD-EVs led to the up-regulation of p70S6K and canonical NF-κB signaling, altering their intracellular and extracellular proteome. Activation of NF-κB was also detected at the onset of CLAD in transbronchial biopsies and BAL cytology, and it persisted throughout the progression to end-stage CLAD. RelA overexpression was associated with poorer graft performance and worse outcomes. RelA-driven NF-κB activation is a key factor in the development of CLAD by promoting persistent inflammation. This pathway may be a promising therapeutic target to improve long-term graft survival after LT.

Objective Both primary optic nerve sheath meningiomas (pONSMs) and secondary optic nerve sheath meningiomas (sONSMs) pose clinical challenges because standard treatments such as surgical debulking and radiation therapy can further damage the optic nerve, producing permanent visual loss. The molecular pathology of primary skull base meningiomas is becoming clearer. However, by comparison, pONSMs and sONSMs have not been studied adequately with contemporary high-throughput molecular genetic techniques, which is the primary aim of this study. This is a crucial issue because these tumors may harbor distinct genetic alterations that render them susceptible to targeted therapy, allowing for vision preservation or even visual improvement.Methods A total of 18 optic nerve sheath meningiomas, of which 11 were pONSMs and 7 were sONSMs, were obtained from 3 different institutions and underwent next-generation sequencing.Results We found that pONSMs and sONSMs harbor gene variants previously identified in other meningiomas but also distinct alterations in genes implicated in cell signaling, transcriptional regulation, and DNA damage repair.Conclusions These findings expand our understanding of a relatively understudied specific meningioma with unique therapeutic challenges.

Introduction Neurofibromatosis type 1 (NF1), also known as von Recklinghausen disease, is an autosomal dominant neurocutaneous disorder caused by pathogenic variants in the NF1 gene. This gene encodes neurofibromin, a tumor suppressor involved in negative regulation of the RAS/mTOR signaling pathway. While cutaneous neurofibromas and gliomas are common manifestations, glioneuronal tumors are rare in NF1. Case presentation We report the case of a 33-year-old man with genetically confirmed NF1, who presented with café-au-lait macules, cutaneous neurofibromas, and bilateral sensorineural hearing loss. Follow-up brain MRI revealed two contrast-enhancing lesions: one at the apex of the left petrous ridge, and another intraparenchymal mass in the left frontobasal region involving the ipsilateral olfactory nerve. The frontobasal lesion showed increased FDG uptake on PET imaging. The patient underwent microsurgical resection of this lesion. Intraoperatively, the mass was found to infiltrate the left olfactory bulb. Histopathological analysis confirmed a WHO grade I ganglioglioma, with no evidence of malignant transformation. Conclusion To our knowledge, this represents the first reported case of an olfactory nerve ganglioglioma in a patient with NF1. This finding suggests a possible novel association between NF1 and neuronal tumors affecting cranial nerve I. Further studies are warranted to explore the role of NF1 in glioneuronal tumorigenesis.

BACKGROUND:O6-Methylguanine-DNA methyltransferase (MGMT) promoter percentage of methylation in gliomas has been proved to be the most important predictive factor in temozolomide (TMZ) response. Nevertheless, an agreement about the cut-off to discriminate between a "methylated" and "unmethylated" status has not been reached yet. Many reports have analyzed the correlation between methylated status cut-off and survival, but they lacked sample homogeneity. Our aim was to calculate a clinical significant cut-off considering a homogenous group of patients. METHODS:We retrospectively analyzed 96 patients who underwent a complete removal of glioblastoma in our Institution. All the patients underwent to radiation therapy plus concomitant TMZ and twelve cycles of adjuvant TMZ as described by Stupp. Receiver operating characteristic (ROC) curve analysis was performed and 21% was determined as the optimal cut-off. RESULTS:The median OS was significantly higher in methylated patients compared to unmethylated ones (median 48 months vs. 22 months respectively 95% CI 30-42 vs. 15-19, P<0.001). No difference was observed for PFS. The multivariate analysis with Cox regression model identified MGMT methylation status as an independent predictive factor for OS (P<0.001). CONCLUSIONS:We confirmed the prognostic role of MGMT methylation status even in a highly selective group of patients with the best outcome. We calculated a cut-off of 21% to be highly predictable of survival.

RECQL4 plays an important role in maintaining the integrity of the genome and regulating DNA replication. However, the role of RECQL4 in CNS tumors remains unknown. Sequencing data were reviewed and immunohistochemistry was performed on a variety of glial and nerve sheath tumors. Functional studies were performed in glioma (U251) and malignant peripheral nerve sheath tumors (MPNSTs) (NF90-8, ST88-14) cell lines following RECQL4 knockdown and treatment with ATR-inhibitors. Across 1580 CNS tumors, RECQL4 gene variants were identified in 71 cases (4.5%), with 21 (29.6%) of probable pathogenic significance. RECQL4 expression differed significantly across glioma subgroups (P = 0.012). Low-grade gliomas (diffuse: median H-score 57.5; circumscribed: median 130) showed lower expression than high-grade gliomas (median 145, P < 0.05). Neurofibromas displayed higher RECQL4 expression (median 160) compared with MPNSTs (median 97.5, P < 0.001). Among MPNSTs, NF1-associated cases (n = 24, median 95) expressed significantly less RECQL4 than sporadic cases (n = 8, median 162.5, P < 0.001). RECQL4 knockdown in glioma and MPNST cell lines resulted in increased apoptosis and susceptibility to ATR-inhibitors. Our findings show that RECQL4 expression has divergent patterns across tumor types and that targeting RECQL4 may dampen tumor survival and enhance susceptibility to ATR inhibitor therapy in CNS tumors.

Pancreatic cancer is expected to become the second leading cause of death by 2030 in Western countries. There is a need to pinpoint high-risk populations since extensive screening would be economically impractical. Methods: This study, conducted on liquid biopsies of patients affected by pancreatic ductal adenocarcinoma (PDAC), sequenced, by NGS, the main genes involved in pancreatic carcinogenesis. Results: The study was discontinued due to a low recruitment rate. NGS analysis, conducted on included patients, revealed the TP53 variant rs1042522 in 30 out of 35 patients, with a cytosine (C) replaced by a guanine (G), hence inserting an Arginine in the final protein instead of a Proline. The presence of the rs1042522 variant confers an odds ratio of 6.11 for PaC and an OR of 20 for homozygosity G/G when comparing our cohort of PaC patients to a healthy population from the 1000GenomeProject. Conclusion: These findings could identify a very-high-risk population deserving of being screened for PDAC, even though a wider validation of rs1042522 as a risk factor is needed. Impact: These preliminary data may open the way for identification of a population more prone to developing pancreatic cancer.

Background: Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a mature T-cell lymphoma linked to textured breast implants. A leading hypothesis suggests that chronic inflammation, combined with immunological and genetic factors, drives its pathogenesis. Two previous studies investigating bacterial biofilms on breast implant capsules have produced conflicting results, particularly regarding the enrichment of Ralstonia spp. Methods: We analyzed the microbiota profiles in seroma samples from 10 BIA-ALCL patients and 12 patients with non-neoplastic effusion, subclassified into acute-, mixed-, and chronic-type based on cellular composition. We used two metagenomic approaches: 16S rRNA gene sequencing and Nanopore sequencing with the "What's in My Pot?" (WIMP) taxonomic classifier. Our analyses included alpha and beta diversity metrics, as well as comparisons of Gram status and oxygen requirements. Results: Both sequencing methods identified Staphylococcaceae, Propionibacteriaceae, and Bradyrhizobiaceae as the most prevalent bacterial families in both BIA-ALCL and benign seroma samples. Notably, the Burkholderiaceae family was more abundant in some of the benign seromas according to the 16S rRNA sequencing, but Ralstonia spp. were not detected. BIA-ALCL showed higher richness (based on Nanopore data) and higher evenness (based on 16S rRNA data) compared to acute-type seromas, indicating a more homogenous representation of the different taxa identified. BIA-ALCL seromas did not cluster together based on Nanopore data, but they did form a distinct cluster with 16S rRNA data. This cluster was differentiated from the other two clusters by a relatively balanced presence of multiple families without overt dominance. We observed no significant differences in Gram staining between BIA-ALCL and benign samples using either method. However, non-aerobic bacterial families were enriched in BIA-ALCL cases only when analyzed with the Nanopore pipeline. Conclusions: Overall, our findings did not identify a distinctive microbial signature specifically associated with BIA-ALCL.

The exclusion of one lung from ventilation or pulmonary artery perfusion triggers pathological mechanisms that can lead to lung injury. Although the final effect is similar for both insults, the underlying mechanisms may differ. Primary aim of this study was to compare severity of lung injury between non-ventilated (NVLI) and non-perfused (NPLI) lung injury. Secondary aims were to compare physiologic, imaging and proteomic signatures of NVLI vs NPLI. Sedated and paralyzed healthy female piglets (weight = 36 ± 5 kg) were mechanically ventilated for 24 h after left pulmonary artery ligation (NPLI group, n = 11) or exclusion from ventilation of the left lung (NVLI group, n = 10). Physiological data including electrical impedance tomography imaging of regional ventilation and perfusion were collected. Histological scoring was performed blindly as well as proteomic analysis of broncho-alveolar lavage (BAL) fluids and lung tissue samples at the end of the experiment. The left lung of both groups received similarly low fraction (< 20

A man in his 60s, under dual immunosuppression following bilateral lung transplantation (BiLuTx), developed bacteraemia and septic shock by Pseudomonas aeruginosa with ecthyma gangrenosum. Symptoms emerged acutely a few weeks after he had received high-dose intravenous steroid treatment for acute cellular graft rejection (ACR). The suspected route of entry of the pathogen was a minor accidental skin injury. Despite the severity of his condition, he successfully recovered after an intensive 26-day course of intravenous antibiotic therapy.

Fibrillary aggregation of alpha-synuclein in Lewy body inclusions and nigrostriatal dopaminergic neuron degeneration define Parkinson's disease neuropathology. Mutations in GBA1, encoding glucocerebrosidase, are the most frequent genetic risk factor for Parkinson's disease. However, the lack of reliable experimental models able to reproduce key neuropathological signatures has hampered clarification of the link between mutant glucocerebrosidase and Parkinson's disease pathology.Here, we describe an innovative protocol for the generation of human induced pluripotent stem cell-derived midbrain organoids containing dopaminergic neurons with nigral identity that reproduce characteristics of advanced maturation. When applied to patients with GBA1-related Parkinson's disease, this method enabled the differentiation of midbrain organoids recapitulating dopaminergic neuron loss and fundamental features of Lewy pathology observed in human brains, including the generation of alpha-synuclein fibrillary aggregates with seeding activity that also propagate pathology in healthy control organoids. Concurrently, we found that the retention of mutant glucocerebrosidase in the endoplasmic reticulum and increased levels of its substrate, glucosylceramide, are determinants of alpha-synuclein aggregation into Lewy body-like inclusions, and the reduction of glucocerebrosidase activity accelerated alpha-synuclein pathology by promoting fibrillary alpha-synuclein deposition.Finally, we demonstrated the efficacy of ambroxol and GZ667161 (two modulators of the glucocerebrosidase pathway in clinical development for the treatment of GBA1-related Parkinson's disease) in reducing alpha-synuclein pathology in this model, supporting the use of midbrain organoids as a relevant preclinical platform for investigational drug screening. Frattini et al. present a new human iPSC-derived midbrain organoid model for the study of Parkinson's disease. The midbrain organoids contain mature dopaminergic neurons with nigrostriatal-like characteristics, and reproduce key signatures of Lewy pathology observed in the human brain.

Current therapy for advanced adrenocortical carcinoma (ACC) is represented by EDP-M (etoposide, doxorubicin, cisplatin + mitotane), but its efficacy is limited and new approaches are needed. Our previous in vitro findings showed that AZD1775, an inhibitor of the G2/M checkpoint gatekeeper Wee1, reduces proliferation and increases apoptosis in ACC cell models. The compensatory upregulation of Myt1, a Wee1-redundant kinase, has been involved in the onset of AZD1775 resistance in other tumour types. Aim of this study was to investigate in vitro and in vivo the effects of AZD1775 alone or combined with EDP-M, and to explore the onset of molecular mechanisms of resistance. In vitro experiments in human ACC cell line NCI-H295R demonstrated that coincubation of AZD1775 and EDP-M exerted synergistic effects in reducing cell viability and proliferation and additive effects in inhibiting cortisol secretion. In NCI-H295R xenografts in nude mice, AZD1775 demonstrated an antitumor efficacy comparable to EDP-M, without synergistic effects. Myt1 upregulation was observed after Wee1 silencing or inhibition by AZD1775 in NCI-H295R and patient-derived ACC cells, but not in mice tumours after 22-days treatment with AZD1775 and/or EDP-M. Interestingly, Myt1 increase after AZD1775 treatment in primary ACC cells was reverted by EDP-M cotreatment. Overall, our data in in vitro and in vivo preclinical ACC models support AZD1775 as a promising ACC therapeutic option, and its combination with EDP-M as a useful strategy to enhance drug efficacy, reduce cortisol secretion, prevent drug resistance and minimize side effects by reducing the therapeutic dosage.

Objective:This nationwide survey aimed to assess current diagnostic practices, adherence to international guidelines, and challenges encountered by Italian pathologists in the diagnosis of diffuse pleural mesothelioma (PM). Methods:A structured questionnaire with 38 items was distributed electronically via Google Forms to Italian pathologists involved in mesothelioma diagnosis. Questions covered demographics, biopsy practices, pathology report, immunohistochemistry, molecular diagnostics, educational needs, and barriers to collaborative research. Results:Participants represented diverse experience levels and institutional affiliations, primarily academic medical centers. Significant variability was found in tissue sampling and biobanking practices. Major diagnostic challenges included identifying sarcomatoid/desmoplastic patterns and inadequate adipose tissue in biopsies. Most pathologists managed inconclusive cases via multidisciplinary discussions and molecular analyses (BAP1, MTAP). Barriers identified included inadequate digital pathology infrastructure and limited standardized protocols for tissue collection. Participants strongly favored enhanced molecular resources, standardized histopathological protocols, and national collaborative initiatives. Conclusions:Improved diagnostic accuracy requires targeted training, standardized protocols, enhanced molecular diagnostic capabilities, and structured national collaborations.

Background: Vitiligo is featured by the manifestation of white maculae and primarily results from inflammatory/immune-selective aggression to melanocytes. The trigger mechanism leading to the activation of resident immune cells in the skin still lacks a molecular description. There is growing evidence linking altered mitochondrial metabolism to vitiligo, suggesting that an underlying metabolic defect may enable a direct activation of the immune system. Recent evidence demonstrated the association of vitiligo with disorders related to systemic metabolism, including insulin resistance (IR) and lipid disarrangements. However, IR, defined as a cellular defect in the insulin-mediated control of glucose metabolism, and its possible role in vitiligo pathogenesis has not been proven yet. Methods: In this study, we compared the Ins/IGF-1 intracellular signaling of dermal and epidermal cells isolated from non-lesional vitiligo skin to that belonging to cells obtained from healthy donors. Results: We demonstrated that due to the intensified glucose uptake, S6, and insulin receptor substrate 1 (IRS1) chronic phosphorylation, their inducibilities were downsized, a condition that coincides with the definition of insulin resistance at the cellular level. Correspondingly, the mitogenic and metabolic activities normally provoked by Ins/IGF-1 exposure resulted in significantly compromised vitiligo cells (p ≤ 0.05). Besides all the vitiligo-derived skin cells manifesting an energetic disequilibrium consisting of a low ATP, catabolic processes activation, and chronic oxidative stress, the functional consequences of this state appear amplified in the keratinocyte lineage. Conclusion: The presented data argue for insulin and IGF-1 resistance collocating dysfunctional glucose metabolism in the mechanisms of vitiligo pathogenesis. In vitiligo keratinocytes, the intrinsic impairment of intracellular metabolic activities, particularly when associated with stimulation with Ins/IGF-1, converges into an aberrant pro-inflammatory phenotype that may initiate immune cell recruitment.

Human herpesvirus 8 (HHV8)-associated diseases include Kaposi sarcoma (KS), multicentric Castleman disease (MCD), germinotropic lymphoproliferative disorder (GLPD), Kaposi sarcoma inflammatory cytokine syndrome (KICS), HHV8-positive diffuse large B-cell lymphoma (HHV8+ DLBCL), primary effusion lymphoma (PEL), and extra-cavitary PEL (ECPEL). We report the case of a human immunodeficiency virus (HIV)-negative male treated for cutaneous KS, who developed generalized lymphadenopathy, hepatosplenomegaly, pleural and abdominal effusions, renal insufficiency, and pancytopenia. The excised lymph node showed features of concomitant involvement by micro-KS and MCD, with aggregates of HHV8+, Epstein Barr virus (EBV)-negative, IgM+, and lambda+ plasmablasts reminiscent of microlymphoma. Molecular investigations revealed a somatically hypermutated (SHM) monoclonal rearrangement of the immunoglobulin heavy chain (IGH), accounting for 4% of the B-cell population of the lymph node. Mutational analyses identified a pathogenic variant of KMT2D and variants of unknown significance in KMT2D, FOXO1, ARID1A, and KMT2A. The patient died shortly after surgery. The histological features (HHV8+, EBV−, IgM+, Lambda+, MCD+), integrated with the molecular findings (monoclonal IGH, SHM+, KMT2D mutated), supported the diagnosis of a monoclonal HHV8+ microlymphoma, with features intermediate between an incipient HHV8+ DLBCL and an EBV-negative ECPEL highlighting the challenges in the accurate classification of HHV8-driven lymphoid proliferations.