CREB activation drives acinar to ductal reprogramming and promote pancreatic cancer progression in animal models of alcoholic chronic pancreatitis.

BACKGROUND AND AIMS: In vivo induction of alcoholic chronic pancreatitis (ACP) causes significant acinar damage, increased fibroinflammatory response, and heightened activation of cyclic response element binding protein 1 (CREB) when compared with alcohol (A) or chronic pancreatitis (CP) mediated pancreatic damage. However, the study elucidating the cooperative interaction between CREB and the oncogenic Kras G12D/+ (Kras*) in promoting pancreatic cancer progression with ACP remains unexplored. METHODS: Experimental ACP induction was established in multiple mouse models, followed by euthanization of the animals at various time intervals during the recovery periods. Tumor latency was determined in these mice cohorts. Here, we established CREB deletion (Creb fl/fl ) in Ptf1a CreERTM/+ ;LSL-Kras G12D+/-(KC) genetic mouse models (KCC-/-). Western blot, phosphokinase array, and qPCR were used to analyze the pancreata of Ptf1a CreERTM+/-, KC and KCC -/- mice. The pancreata of ACP-induced KC mice were subjected to single-cell RNA sequencing (scRNAseq). Further studies involved conducting lineage tracing and acinar cell explant cultures. RESULTS: ACP induction in KC mice had detrimental effects on the pancreatic damage repair mechanism. The persistent existence of acinar cell-derived ductal lesions demonstrated a prolonged state of hyperactivated CREB. Persistent CREB activation leads to acinar cell reprogramming and increased pro-fibrotic inflammation in KC mice. Acinar-specific Creb ablation reduced advanced PanINs lesions, hindered tumor progression, and restored acinar cell function in ACP-induced mouse models. CONCLUSIONS: Our findings demonstrate that CREB cooperates with Kras* to perpetuate an irreversible ADM and PanIN formation. Moreover, CREB sustains oncogenic activity to promote the progression of premalignant lesions toward cancer in the presence of ACP.

Characterization and Hydrolysis Studies of a Prodrug Obtained as Ester Conjugate of Geraniol and Ferulic Acid by Enzymatic Way.

Ferulic acid (Fer) and geraniol (Ger) are natural compounds whose antioxidant and anti-inflammatory activity confer beneficial properties, such as antibacterial, anticancer, and neuroprotective effects. However, the short half-lives of these compounds impair their therapeutic activities after conventional administration. We propose, therefore, a new prodrug (Fer-Ger) obtained by a bio-catalyzed ester conjugation of Fer and Ger to enhance the loading of solid lipid microparticles (SLMs) designed as Fer-Ger delivery and targeting systems. SLMs were obtained by hot emulsion techniques without organic solvents. HPLC-UV analysis evidenced that Fer-Ger is hydrolyzed in human or rat whole blood and rat liver homogenates, with half-lives of 193.64 ± 20.93, 20.15 ± 0.75, and 3.94 ± 0.33 min, respectively, but not in rat brain homogenates. Studies on neuronal-differentiated mouse neuroblastoma N2a cells incubated with the reactive oxygen species (ROS) inductor H2O2 evidenced the Fer-Ger ability to prevent oxidative injury, despite the fact that it appears ROS-promoting. The amounts of Fer-Ger encapsulated in tristearin SLMs, obtained in the absence or presence of glucose, were 1.5 ± 0.1%, allowing the control of the prodrug release (glucose absence) or to sensibly enhance its water dissolution rate (glucose presence). These new "green" carriers can potentially prolong the beneficial effects of Fer and Ger or induce neuroprotection as nasal formulations.

Cell-specific nanoengineering strategy disrupts tolerogenic signaling from myeloid-derived suppressor cells to invigorate antitumor immunity in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by intratumoral abundance of neutrophilic/polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) which inhibit T-cell function through JAK2/STAT3-regulated arginase activity. To overcome limitations of systemic inhibition of PMN-MDSCs in cancer-bearing patients-i.e., neutropenia and compensatory myelopoietic adaptations-we develop a nanoengineering strategy to target cell-specific signaling exclusively in PMN-MDSCs without provoking neutropenia. We conjugate a chemically modified small-molecule inhibitor of MDSC-surface receptor CXCR2 (AZD5069) with polyethylene glycol (PEG) and chemically graft AZD5069-PEG constructs onto amphiphilic polysaccharide derivatives to engineer CXCR2-homing nanoparticles (CXCR2-NP). Cy5.5 dye-loaded CXCR2-NP showed near-exclusive uptake in PMN-MDSCs compared with PDAC tumor-cells, cancer-associated fibroblasts, and macrophages. Encapsulation of JAK2/STAT3i Ruxolitinib (CXCR2-NP Ruxo ) resulted in more durable attenuation in STAT3-regulated arginase activity from PMN-MDSCs and induction of cytolytic T-cell activity vs. free Ruxolitinib in-vitro and in-vivo . Cell-specific delivery of payloads via CXCR2-homing immunonanoparticles represents a novel strategy to disrupt MDSC-mediated immunosuppression and invigorate antitumor immunity in PDAC.

EEG connectivity in functional brain networks supporting visuomotor integration processes in dominant and non-dominant hand movements.

Objective.This study explores the changes in the organization of functional brain networks induced by performing a visuomotor integration task, as revealed by noninvasive spontaneous electroencephalographic traces (EEG).Approach.EEG data were acquired during the execution of the Nine Hole Peg Test (NHPT) with the dominant and non-dominant hands in a group of 44 right-handed volunteers. Both spectral analysis and phase-based connectivity analysis were performed in the theta (ϑ), mu (µ) and beta (ß) bands. Graph Theoretical Analysis (GTA) was also performed to investigate the topological reorganization induced by motor task execution.Main results.Spectral analysis revealed an increase of frontoparietal ϑ power and a spatially diffused reduction ofµand ß contribution, regardless of the hand used. GTA showed a significant increase in network integration induced by movement performed with the dominant limb compared to baseline in the ϑ band. Theµand ß bands were associated with a reduction in network integration during the NHPT. In theµrhythm, this result was more evident for the right-hand movement, while in the ß band, results did not show dependence on the laterality. Finally, correlation analysis highlighted an association between frequency-specific topology measures and task performance for both hands.Significance.Our results show that functional brain networks reorganize during visually guided movements in a frequency-dependent manner, differently depending on the hand used (dominant/non dominant).

Assessing cardiovascular stress based on heart rate variability in female shift workers: a multiscale-multifractal analysis approach.

Introduction: Sleep-wake cycle disruption caused by shift work may lead to cardiovascular stress, which is observed as an alteration in the behavior of heart rate variability (HRV). In particular, HRV exhibits complex patterns over different time scales that help to understand the regulatory mechanisms of the autonomic nervous system, and changes in the fractality of HRV may be associated with pathological conditions, including cardiovascular disease, diabetes, or even psychological stress. The main purpose of this study is to evaluate the multifractal-multiscale structure of HRV during sleep in healthy shift and non-shift workers to identify conditions of cardiovascular stress that may be associated with shift work. Methods: The whole-sleep HRV signal was analyzed from female participants: eleven healthy shift workers and seven non-shift workers. The HRV signal was decomposed into intrinsic mode functions (IMFs) using the empirical mode decomposition method, and then the IMFs were analyzed using the multiscale-multifractal detrended fluctuation analysis (MMF-DFA) method. The MMF-DFA was applied to estimate the self-similarity coefficients, α(q, τ), considering moment orders (q) between -5 and +5 and scales (τ) between 8 and 2,048 s. Additionally, to describe the multifractality at each τ in a simple way, a multifractal index, MFI(τ), was computed. Results: Compared to non-shift workers, shift workers presented an increase in the scaling exponent, α(q, τ), at short scales (τ < 64 s) with q < 0 in the high-frequency component (IMF1, 0.15-0.4 Hz) and low-frequency components (IMF2-IMF3, 0.04-0.15 Hz), and with q> 0 in the very low frequencies (IMF4, < 0.04 Hz). In addition, at large scales (τ> 1,024 s), a decrease in α(q, τ) was observed in IMF3, suggesting an alteration in the multifractal dynamic. MFI(τ) showed an increase at small scales and a decrease at large scales in IMFs of shift workers. Conclusion: This study helps to recognize the multifractality of HRV during sleep, beyond simply looking at indices based on means and variances. This analysis helps to identify that shift workers show alterations in fractal properties, mainly on short scales. These findings suggest a disturbance in the autonomic nervous system induced by the cardiovascular stress of shift work.

Transmembrane TNF-TNFR2 signaling as a critical immunoregulatory node in pancreatic cancer.

Pancreatic cancer is characterized by extreme therapeutic resistance. In pancreatic cancers harboring high-risk genomes, we describe that cancer cell-neutrophil signaling circuitry provokes neutrophil-derived transmembrane (tm)TNF-TNFR2 interactions that dictate inflammatory polarization in cancer-associated fibroblasts and T-cell dysfunction - two hallmarks of therapeutic resistance. Targeting tmTNF-TNFR2 signaling may sensitize pancreatic cancer to chemo±immunotherapy.

Early sleep after action observation plus motor imagery improves gait and balance abilities in older adults.

Action observation plus motor imagery (AOMI) is a rehabilitative approach to improve gait and balance performance. However, limited benefits have been reported in older adults. Early sleep after motor practice represents a strategy to enhance the consolidation of trained skills. Here, we investigated the effects of AOMI followed by early sleep on gait and balance performance in older adults. Forty-five older adults (mean age: 70.4 ± 5.2 years) were randomized into three groups performing a 3-week training. Specifically, AOMI-sleep and AOMI-control groups underwent observation and motor imagery of gait and balance tasks between 8:00 and 10:00 p.m. or between 8:00 and 10:00 a.m. respectively, whereas Control group observed landscape video-clips. Participants were assessed for gait performance, static and dynamic balance and fear of falling before and after training and at 1-month follow-up. The results revealed that early sleep after AOMI training sessions improved gait and balance abilities in older adults compared to AOMI-control and Control groups. Furthermore, these benefits were retained at 1-month after the training end. These findings suggested that early sleep after AOMI may represent a safe and easy-applicable intervention to minimize the functional decay in older adults.

Tumor Cell-Intrinsic p38 MAPK Signaling Promotes IL1α-Mediated Stromal Inflammation and Therapeutic Resistance in Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a KRAS-driven inflammatory program and a desmoplastic stroma, which contribute to the profoundly chemoresistant phenotype. The tumor stroma contains an abundance of cancer-associated fibroblasts (CAF), which engage in extensive paracrine cross-talk with tumor cells to perpetuate protumorigenic inflammation. IL1α, a pleiotropic, tumor cell-derived cytokine, plays a critical role in shaping the stromal landscape. To provide insights into the molecular mechanisms regulating IL1A expression in PDAC, we performed transcriptional profiling of The Cancer Genome Atlas datasets and pharmacologic screening in PDAC cells and identified p38α MAPK as a key positive regulator of IL1A expression. Both genetic and pharmacologic inhibition of p38 MAPK significantly diminished IL1α production in vitro. Chromatin- and coimmunoprecipitation analyses revealed that p38 MAPK coordinates the transcription factors Sp1 and the p65 subunit of NFκB to drive IL1A overexpression. Single-cell RNA sequencing of a highly desmoplastic murine PDAC model, Ptf1aCre/+; LSL-KrasG12D/+; Tgfbr2flox/flox (PKT), confirmed that p38 MAPK inhibition significantly decreases tumor cell-derived Il1a and attenuates the inflammatory CAF phenotype in a paracrine IL1α-dependent manner. Furthermore, p38 MAPK inhibition favorably modulated intratumoral immunosuppressive myeloid populations and augmented chemotherapeutic efficacy to substantially reduce tumor burden and improve overall survival in PKT mice. These findings illustrate a cellular mechanism of tumor cell-intrinsic p38-p65/Sp1-IL1α signaling that is responsible for sustaining stromal inflammation and CAF activation, offering an attractive therapeutic approach to enhance chemosensitivity in PDAC. SIGNIFICANCE: Inhibition of p38 MAPK suppresses tumor cell-derived IL1α and attenuates the inflammatory stroma and immunosuppressive tumor microenvironment to overcome chemotherapeutic resistance in pancreatic cancer.

Dimeric ferulic acid conjugate as a prodrug for brain targeting after nasal administration of loaded solid lipid microparticles.

OBJECTIVE: Ferulic acid (Fer) displays antioxidant/anti-inflammatory properties useful against neurodegenerative diseases. To increase Fer uptake and its central nervous system residence time, a dimeric prodrug, optimizing the Fer loading on nasally administrable solid lipid microparticles (SLMs), was developed. METHODS: The prodrug was synthesized as Fer dimeric conjugate methylated on the carboxylic moiety. Prodrug antioxidant/anti-inflammatory properties and ability to release Fer in physiologic environments were evaluated. Tristearin or stearic acid SLMs were obtained by hot emulsion technique. In vivo pharmacokinetics were quantified by HPLC. RESULTS: The prodrug was able to release Fer in physiologic environments (whole blood and brain homogenates) and induce in vitro antioxidant/anti-inflammatory effects. Its half-life in rats was 18.0 ± 1.9 min. Stearic acid SLMs, exhibiting the highest prodrug loading and dissolution rate, were selected for nasal administration to rats (1 mg/kg dose), allowing to obtain high prodrug bioavailability and prolonged residence in the cerebrospinal fluid, showing AUC (Area Under Concentration) values (108.5 ± 3.9 µg∙mL-1∙min) up to 30 times over those of Fer free drug, after its intravenous/nasal administration (3.3 ± 0.3/5.16 ± 0.20 µg∙mL-1∙min, respectively) at the same dose. Chitosan presence further improved the prodrug brain uptake. CONCLUSIONS: Nasal administration of prodrug-loaded SLMs can be proposed as a noninvasive approach for neurodegenerative disease therapy.

Energy imparted and ionisation yield at the nanometre scale: results for extended beams.

The metrological problem of interpreting ionisation-based micro- and nanodosimetric measurements in terms of quantities proportional to energy imparted becomes particularly relevant when the sensitive volume (SV) size is in the nanometre range. At these scales, a constant W-value cannot be assumed, and the stochastics of the energy transfer per single collision could play a more important role. This problem was recently analysed by our group by means of track-structure Monte Carlo simulations with the Geant4-DNA code, finding a strong correlation between the energy imparted and ionisation yield also for SV diameters of 1 nm. As the previous study was limited to primary beams of radius zero crossing the sensitive sphere along its diameter, it is the aim of the present work to extend the analysis to beams with a radius larger than the dimensions of the SV, to better assess the role played by secondary electrons.

First microdosimetric measurements with a tissue-equivalent proportional counter at the MedAustron ion-beam therapy facility.

The aim of this work is to present the first microdosimetric spectra measured with a miniaturised tissue-equivalent proportional counter in the clinical environment of the MedAustron ion-beam therapy facility. These spectra were gathered with a 62.4-MeV proton beam and have been compared with microdosimetric spectra measured in the 62-MeV clinical proton beam of the CATANA beam line. Monte Carlo simulations were performed using the Geant4 toolkit GATE and a fully commissioned clinical beam line model. Finally, similarities and discrepancies of the measured data to simulations based on a simple and complex detector geometry are discussed.

Comparison of biological weighting functions to estimate the microdosimetric RBE in BNCT.

In the framework of the MUNES project, a prototype accelerator-based thermal neutron source was developed and installed at the Legnaro National Laboratories of INFN. The microdosimetric characterization of this radiation field was performed with a Tissue-Equivalent Proportional Counter with interchangeable cathode walls, either doped with 100 ppm of 10B or without boron doping. A suitable subtraction procedure allowed to discriminate the gamma, neutron and BNC dose components (Selva et al., 2022, Appl. Radiat. Isot. 182, 110144). The measured microdosimetric spectra can be weighted with a biological weighting function to estimate the Relative Biological Effectiveness of the radiation field, for the purpose of intercomparison between different thermal neutron sources. This work compares, therefore, the biological doses resulting from four different weighting functions applied to the same initial microdosimetric spectrum, discussing strengths and limitations of each of them.

Monopolar tDCS might affect brainstem reflexes: A computational and neurophysiological study.

OBJECTIVE: To assess whether monopolar multi-electrode transcranial direct current stimulation (tDCS) montages might selectively affect deep brain structures through computational predictions and neurophysiological assessment. METHODS: Electric field distribution in deep brain structures (i.e., thalamus and midbrain) were estimated through computational models simulating tDCS with two monopolar and two monopolar multi-electrode montages. Monopolar multi-electrode tDCS was then applied to healthy subject, and effects on pontine and medullary circuitries was evaluated studying changes in blink reflex (BR) and masseter inhibitory reflex (MIR). RESULTS: Computational results suggest that tDCS with monopolar multi-electrode montages might induce electric field intensities in deep brain structure comparable to those in grey matter, while neurophysiological results disclosed that BR and MIR were selectively modulated by tDCS only when cathode was placed over the right deltoid. CONCLUSIONS: Multi-electrode tDCS (anodes over motor cortices, cathode over right deltoid) could induce significant electric fields in the thalamus and midbrain, and selectively affect brainstem neural circuits. SIGNIFICANCE: Multi-electrode tDCS (anodes over motor cortices, cathode over right deltoid) might be further explored to affect brainstem activity, also in the context of non-invasive deep brain stimulation.

Sentinel Lymph Node Biopsy vs No Axillary Surgery in Patients With Small Breast Cancer and Negative Results on Ultrasonography of Axillary Lymph Nodes: The SOUND Randomized Clinical Trial.

Importance: Sentinel lymph node biopsy (SLNB) is the standard of care for axillary node staging of patients with early breast cancer (BC), but its necessity can be questioned since surgery for examination of axillary nodes is not performed with curative intent. Objective: To determine whether the omission of axillary surgery is noninferior to SLNB in patients with small BC and a negative result on preoperative axillary lymph node ultrasonography. Design, Setting, and Participants: The SOUND (Sentinel Node vs Observation After Axillary Ultra-Sound) trial was a prospective noninferiority phase 3 randomized clinical trial conducted in Italy, Switzerland, Spain, and Chile. A total of 1463 women of any age with BC up to 2 cm and a negative preoperative axillary ultrasonography result were enrolled and randomized between February 6, 2012, and June 30, 2017. Of those, 1405 were included in the intention-to-treat analysis. Data were analyzed from October 10, 2022, to January 13, 2023. Intervention: Eligible patients were randomized on a 1:1 ratio to receive SLNB (SLNB group) or no axillary surgery (no axillary surgery group). Main Outcomes and Measures: The primary end point of the study was distant disease-free survival (DDFS) at 5 years, analyzed as intention to treat. Secondary end points were the cumulative incidence of distant recurrences, the cumulative incidence of axillary recurrences, DFS, overall survival (OS), and the adjuvant treatment recommendations. Results: Among 1405 women (median [IQR] age, 60 [52-68] years) included in the intention-to-treat analysis, 708 were randomized to the SLNB group, and 697 were randomized to the no axillary surgery group. Overall, the median (IQR) tumor size was 1.1 (0.8-1.5) cm, and 1234 patients (87.8%) had estrogen receptor-positive ERBB2 (formerly HER2 or HER2/neu), nonoverexpressing BC. In the SLNB group, 97 patients (13.7%) had positive axillary nodes. The median (IQR) follow-up for disease assessment was 5.7 (5.0-6.8) years in the SLNB group and 5.7 (5.0-6.6) years in the no axillary surgery group. Five-year distant DDFS was 97.7% in the SLNB group and 98.0% in the no axillary surgery group (log-rank P = .67; hazard ratio, 0.84; 90% CI, 0.45-1.54; noninferiority P = .02). A total of 12 (1.7%) locoregional relapses, 13 (1.8%) distant metastases, and 21 (3.0%) deaths were observed in the SLNB group, and 11 (1.6%) locoregional relapses, 14 (2.0%) distant metastases, and 18 (2.6%) deaths were observed in the no axillary surgery group. Conclusions and Relevance: In this randomized clinical trial, omission of axillary surgery was noninferior to SLNB in patients with small BC and a negative result on ultrasonography of the axillary lymph nodes. These results suggest that patients with these features can be safely spared any axillary surgery whenever the lack of pathological information does not affect the postoperative treatment plan. Trial Registration: ClinicalTrials.gov Identifier: NCT02167490.

Entropy and fractal analysis of brain-related neurophysiological signals in Alzheimer's and Parkinson's disease.

Brain-related neuronal recordings, such as local field potential, electroencephalogram and magnetoencephalogram, offer the opportunity to study the complexity of the human brain at different spatial and temporal scales. The complex properties of neuronal signals are intrinsically related to the concept of 'scale-free' behavior and irregular dynamic, which cannot be fully described through standard linear methods, but can be measured by nonlinear indexes. A remarkable application of these analysis methods on electrophysiological recordings is the deep comprehension of the pathophysiology of neurodegenerative diseases, that has been shown to be associated to changes in brain activity complexity. In particular, a decrease of global complexity has been associated to Alzheimer's disease, while a local increase of brain signals complexity characterizes Parkinson's disease. Despite the recent proliferation of studies using fractal and entropy-based analysis, the application of these techniques is still far from clinical practice, due to the lack of an agreement about their correct estimation and a conclusive and shared interpretation. Along with the aim of helping towards the realization of a multidisciplinary audience to approach nonlinear methods based on the concepts of fractality and irregularity, this survey describes the implementation and proper employment of the mostly known and applied indexes in the context of Alzheimer's and Parkinson's diseases.

The Role of Female Reproductive Hormones in the Association between Migraine and Breast Cancer: An Unanswered Question.

Accumulating epidemiological studies have investigated a possible interconnection between migraine (Mi) and breast cancer (BC) because of the strong link between these diseases and female reproductive hormones. This review aims to consolidate findings from epidemiological studies and explore biologically plausible hypothetical mechanisms related to hormonal pathways. Current evidence suggests a protective role of Mi in BC development, particularly in case-control studies but not in cohort ones. The inconsistency among studies may be due to several reasons, including diagnostic criteria for Mi and the age gap between the development of these two diseases. Furthermore, recent research has challenged the concept of a net beneficial effect of Mi on BC, suggesting a more complex relationship between the two conditions. Many polymorphisms/mutations in hormone-related pathways are involved in at least one of the two conditions. The most promising evidence has emerged for a specific alteration in the estrogen receptor 1 gene (rs2228480). However, the possible specific mutation or polymorphism involved in this association has not yet been identified. Further studies with robust methodologies are needed to validate the protective role of Mi in BC and fully elucidate the precise nature of this causal relationship.