Laboratory for Biomedical Neurosciences

The role of tumor-associated neutrophils (TANs) infiltrating colorectal cancers (CRCs) is still debated. Here, we unravel that TAN recruitment and functions are modulated by intratumoral microbiota and only defined bacterial species can unleash neutrophil cytotoxic potential. Fusobacterium nucleatum promotes the production of neutrophil-recruiting chemokines by tumor cells and enhances neutrophil migration more efficiently than Bacteroides fragilis. Importantly, Fusobacterium nucleatum, but not Bacteroides fragilis, triggers neutrophils to release cytotoxic proteins showing tumoricidal activity in vitro and in xenograft models. Mechanistically, these effects are elicited upon Fusobacterium nucleatum binding to sialic-acid-binding immunoglobulin-like lectin (Siglec)-14 expressed by neutrophils but are impaired upon Siglec-14 blockade or loss-of-function polymorphisms. Supporting these findings, in human CRCs, elevated Fusobacterium nucleatum loads and high TAN densities correlate with improved prognosis, whereas lack of Siglec-14 expression is associated with reduced patient survival. Our findings identify microbiota composition and host genetic background as critical determinants of neutrophil functional profiles, offering insights into neutrophil-targeted therapeutic strategies in CRC.

Traumatic and overuse injuries of bones and tendons of the wrist represent a significant proportion of the musculoskeletal disorders encountered in both athletic and occupational settings. Conventional radiography remains the first-line imaging modality for osseous trauma, but it can miss up to a third of scaphoid and hamate fractures. Computed tomography is superior for characterizing occult or nondisplaced fractures. Magnetic resonance imaging provides important information on occult fractures, overuse injuries of osseous structures, and tendinous abnormalities.A comprehensive understanding of wrist injuries is essential to ensure accurate and timely diagnosis, in turn facilitating the selection of appropriate patient-specific treatment strategies. By minimizing diagnostic delays, clinicians can reduce the risk of adverse sequelae such as chronic instability, degenerative joint changes, and functional impairment. The implementation of optimized diagnostic and therapeutic pathways not only lessens the long-term burden of disability for patients, but it also plays a crucial role in enabling a safe and efficient return to both occupational duties and sporting activities.

Chronic stress is a risk factor for neuropsychiatric disorders, making the ability to adapt to repeated stress a crucial determinant of mental health. It remains unclear whether repeated stress exposure leads to habituation or new adaptive responses. Here, we investigated how the transcriptomic response to acute restraint stress changes - in the mouse ventralhippocampus - with repeated exposure. Using bulk RNA-seq across multiple timepoints, we observed widespread transcriptional habituation: stress-induced gene expression was blunted without the emergence of new response profiles or changes in baseline gene expression. Temporally resolved single-nucleus multi-omic profiling (RNA and chromatin accessibility) confirmed that the transcriptional habituation occurs across cell types, revealing cell type-specific patterns. We identify gene clusters whose expression habituates with repeated exposure, which appear primarily related to two distinct mechanisms: an early blunting of cAMP-associated genes chiefly linked to a reduced number of activated cells, and a shortened transcriptional response of corticosterone-associated genes, independent of the number of activated cells. Our study provides a comprehensive resource, accessible as an interactive app, that characterizes the dynamic transcriptomic response to stress in the mouse hippocampus and how it habituates with repeated exposure to stress.

Abstract BACKGROUND In glioblastoma (GBM) patients two factors are relevant for prediction of survival: MGMT promoter hypermethylation, which improves the patient’s response to temozolomide (TMZ), and the extent of resection (EOR), with a better overall survival (OS) described in patients subjected to gross total resection (GTR). The aim of our study is to analyze the level of concordance between the results obtained with the standard non-quantitative MGMT analysis methods compared to the quantitative real-time method EpiDirect® and to verify whether the different levels (frequencies) of MGMT promoter hypermethylation, assessed by EpiDirect®, can better predict the response to GBM treatment. MATERIAL AND METHODS MGMT promoter methylation in GBM IDH1 wt cases diagnosed from 2004 to 2022 in two neurosurgical centers (EOC and Insubria University Hospital, Italy) and treated by temozolomide (TMZ) was evaluated for diagnosis by sodium bisulfite pretreatment and methylation-specific PCR (MSP) and then we tested the samples by the new real-time EpiDirect® (Pentabase ApS, Denmark). Cohen index and Kaplan-Meier curves were evaluated for statistics. RESULTS We enrolled 119 GBM cases treated by standard adjuvant partial brain radiation and TMZ based chemotherapy for which MGMT promoter methylation data by MSP or by PCR-pyrosequencing were available for diagnostic purposes. With the common standard methods 41/119 (34.5%) cases were methylated in the MGMT promoter (M) and 72/119 (60.5%) were unmethylated (UM). In addition, 6/119 cases (5.05%) were not evaluable. By EpiDirect® all the cases provided reproducible results, MGMT M was detected in 45/119 cases (37.8%) and UM in 74/119 (62.2%). Comparing Epidirect® to the commonly used methodologies, nine evaluable cases (5 M and 4 UM cases), corresponding to 8% of the entire cohort, showed discrepant methylation results. Despite this, the level of concordance between methodologies was high (K Cohen index corresponding to 0.829). The comparisons of OS, progression-free survival and time-to-progression in the five categories corresponding to different MGMT promoter methylation levels identified by Epidirect® resulted in not statistical significant results considering the whole cohort and then the different kind of surgery (biopsy, partial resection or GTR) (p›0.05). CONCLUSION EpiDirect® is a robust methodology enabling a faster, more sensible and easier evaluation of cases even in presence of a highly damaged DNA. In addition, it shares a high level of superimposition with the most diffused technologies currently used for assessing the MGMT promoter methylation for diagnostic purposes. The EpiDirect® ability to quantify the percentage of MGMT promoter methylation is another advantage of this methodology however we found that it has not clinical relevance on survival.

People with spinal cord injury (SCI) may suffer from anejaculation due to functional obstructive azoospermia (OA). Testicular sperm extraction (TESE) may successfully overcome this problem, even if the optimal timing is controversial. The primary aim of this study was to report our experience with TESE in SCI, focusing on the effect of time since SCI event on the TESE outcomes. We included all consecutive people with SCI and functional OA undergoing TESE between January 2011 and December 2021 in four Italian tertiary referral centers. We recorded TESE sample parameters, sperm retrieval rate (SRR) and intracytoplasmic sperm injection (ICSI) outcomes. Logistic regression analysis was performed to assess whether time since SCI was significantly associated with these outcomes. The time since SCI was considered in three different ways: 1) continuously; 2) ≤9 years vs >9 years; 3) ≤5 years, >5 and ≤10 years, >10 years. We included 32 patients with tetraplegia and 75 with paraplegia, undergoing 107 TESE procedures. The median age at surgery and time since SCI were 33 years (IQR 29-38) and 9 years (IQR 3-14), respectively. The SRR was 81.3%. Thirty-three out of 87 patients underwent ICSI, achieving pregnancy in 63.6% after one cycle. The final live birth rate was 90.5%. Logistic regression analyses outlined that the SRR was not affected by considered variables, including time since SCI, considered both continuously and categorically. Our SRR did not prove to be negatively affected by all considered variables, especially by the time since SCI. Clinicians should not deter SCI patients with functional OA from undergoing TESE after a long time since SCI.

A 40-year-old woman presented with a 3-week history of nonradiating left-sided cervical pain, followed by a 3-day history of left upper limb weakness (affecting shoulder abduction, external rotation, and elbow flexion).Electromyography revealed deltoid muscle denervation, indicating possible subacute C5 radiculopathy.Weakness with cervical pain raised concern for a cervical spine abnormality, prompting an unremarkable initial MRI.However, neck pain persisted, and eco-color Doppler revealed decreased flow in the left vertebral artery, suggesting vertebral artery dissection (VAD).Dedicated T1 fat-saturated (Figures 1A and2A) and gradient-echo (Figure 1B) sequences showed a mural hematoma compressing the C5 nerve root.MR angiography confirmed a nonocclusive VAD at the extradural segment (Figure 2B).The patient showed clinical and radiologic improvement after 6 months of single antiplatelet therapy (Figure 2, C andD).VAD-related radiculopathy is rare but should be considered in unexplained causes of acute neck pain.Possible mechanisms include compression from a mural hematoma or impaired vasa nervorum perfusion, with the C5 nerve root being especially vulnerable. 1,2 Axial T1 fat-saturated (fs) sequence at the level of C4-C5 (A) shows eccentric crescent-shaped hyperintense subacute intramural hematoma of the left vertebral artery (VA) extradural segment (white arrowhead), consistent with vertebral artery dissection, compressing the C5 nerve root (yellow arrow) on axial gradient-echo T2-weighted sequence (B).

IntroductionMild Cognitive Impairment (MCI) is a transitional stage between normal aging and dementia, frequently associated with subtle deficits in social cognition. Empathy, a core component of social cognition, encompasses both affective and cognitive dimensions and may be compromised even in prodromal phases of neurodegenerative conditions. Despite its clinical relevance, empathy in MCI remains underexplored, and standardized assessment tools are seldom used in routine diagnostics. Theory of mind is another important aspect of social cognition, and, as with empathy, it is unclear how it is affected at the MCI stage. This study aimed to investigate empathy and theory of mind abilities in individuals with MCI, excluding cases attributable to prodromal frontotemporal dementia, using two validated instruments: the Interpersonal reactivity index (IRI) and the Story-based empathy task (SET).MethodsWe conducted a case–control study involving 23 individuals with MCI and 25 cognitively healthy controls. All participants completed the IRI, while a subsample (19 MCI patients) also underwent the SET. The clinical group included both amnestic and non-amnestic MCI subtypes, with heterogeneous etiologies. Group comparisons were performed on IRI subscales and SET indices to assess both self-reported empathy traits and performance-based socio-cognitive abilities.ResultsThe MCI group exhibited significantly lower scores in the IRI subscales of Empathic Concern and Perspective Taking compared to controls, indicating concurrent affective and cognitive empathy impairments. While no significant differences emerged in individual SET subcomponents, the MCI group showed a significantly lower global SET score, suggesting reduced integrative socio-cognitive performance. These findings should be interpreted with caution given the limited sample size and clinical heterogeneity, which make this a preliminary study.ConclusionThese findings provide preliminary evidence of early empathy-related and theory of mind alterations in MCI, supporting the inclusion of social cognition assessments in standard neuropsychological protocols. The combined use of self-report and task-based instruments may enhance early identification of socio-emotional dysfunctions and inform personalized clinical interventions. The preliminary nature of this study is mainly due to the small sample size and the heterogeneous clinical profiles, which limit generalizability but highlight the need for replication in larger cohorts.