Il biofeedback preoperatorio migliora il recupero della continenza a seguito di prostatectomia radicale: una revisione sistematica con meta-analisi

BACKGROUND:  Postoperative urinary incontinence is the overall result of urethral sphincter incompetence and modifications in urethral length after radical prostatectomy. Findings for preoperative interventions targeted at preventing post-prostatectomy incontinence include preoperative pelvic floor muscle training (PFMT) and biodfeedback (BFB), which can be managed by nurses in many countries and have been used for decades to speed up continence recovery after surgery. AIM: to determine the effectiveness of preoperative biofeedback (BFB) for post-prostatectomy urinary incontinence compared to pelvic training without BFB, considering the variability between the results of the available studies.  METHODS: A systematic review and meta-analysis was conducted, analyzing the indications provided by the literature regarding preoperative biofeedback for preventing urinary incontinence after open radical prostatectomy, in terms of treatment regimens, timing for beginning the sessions, number of contraction and relaxation exercises, and scheduled work at home. Literature search on Pubmed, CINAHL, Cochrane Library, Web of Science, Scopus, EMBASE, and PEdro. RESULTS: Despite only three papers being suitable for metanalysis, our results support BFB over written instructions for continence recovery after both 3 and 6 moths from surgery. Implementing progressive programs with many different muscular exercises and including relaxation are the main recommendations. CONCLUSIONS: Preoperative biofeedback leads to improved urinary continence after 3 and 6 months from radical prostatectomy. Future studies should focus on the characteristics and number of pelvic muscle contractions required during biofeedback in order to maximize effectiveness.BACKGROUND: L'incontinenza urinaria postoperatoria è il risultato complessivo dell'incompetenza dello sfintere uretrale e delle modifiche della lunghezza dell'uretra dopo la prostatectomia radicale. I risultati degli interventi preoperatori mirati a prevenire l'incontinenza post-prostatectomia includono l'allenamento preoperatorio dei muscoli del pavimento pelvico (PFMT) e il biodfeedback (BFB), che possono essere gestiti dal personale infermieristico in molti Paesi. OBIETTIVO: determinare l'efficacia del biofeedback preoperatorio (BFB) per l'incontinenza urinaria post-prostatectomia rispetto al training pelvico senza BFB, considerando la variabilità dei risultati degli studi disponibili.  METODI: è stata condotta una revisione sistematica con meta-analisi, analizzando le indicazioni fornite dalla letteratura sul biofeedback preoperatorio per la prevenzione dell'incontinenza urinaria in termini di regimi di trattamento, tempi di inizio delle sessioni, numero di esercizi di contrazione e rilassamento e lavoro programmato a casa. La ricerca della letteratura è stata effettuata su Pubmed, CINAHL, Cochrane Library, Web of Science, Scopus, EMBASE e PEdro. RISULTATI: nonostante solo tre articoli siano stati adatti alla metanalisi, i nostri risultati supportano il BFB rispetto alle istruzioni scritte per il recupero della continenza dopo 3 e 6 mesi dall'intervento. L'implementazione di programmi progressivi con molti esercizi muscolari diversi e l'inclusione del rilassamento sono le principali raccomandazioni. CONCLUSIONI: Il biofeedback preoperatorio porta a un miglioramento della continenza urinaria dopo 3 e 6 mesi dalla prostatectomia radicale. Gli studi futuri dovrebbero concentrarsi sulle caratteristiche e sul numero di contrazioni muscolari pelviche richieste durante il biofeedback per massimizzare l'efficacia.Il biofeedback preoperatorio migliora il recupero della continenza dopo la prostatectomia aperta: una revisione sistematica e una meta-analis

Mitochondrial dysfunction induced by a SH2 domain-Targeting STAT3 inhibitor leads to metabolic synthetic lethality in cancer cells

In addition to its canonical role in nuclear transcription, signal transducer and activator of transcription 3 (STAT3) is emerging as an important regulator of mitochondrial function. Here, we demonstrate that a novel inhibitor that binds with high affinity to the STAT3 SH2 domain triggers a complex cascade of events initiated by interference with mitochondrial STAT3 (mSTAT3). The mSTAT3\u2013drug interaction leads to mitochondrial dysfunction, accumulation of proteotoxic STAT3 aggregates, and cell death. The cytotoxic effects depend directly on the drug\u2019s ability to interfere with mSTAT3 and mitochondrial function, as demonstrated by site-directed mutagenesis and use of STAT3 knockout and mitochondria-depleted cells. Importantly, the lethal consequences of mSTAT3 inhibition are enhanced by glucose starvation and by increased reliance of cancer cells and tumor-initiating cells on mitochondria, resulting in potent activity in cell cultures and tumor xenografts in mice. These findings can be exploited for eliciting synthetic lethality in metabolically stressed cancer cells using highaffinity STAT3 inhibitors. Thus, this study provides insights on the role of mSTAT3 in cancer cells and a conceptual framework for developing more effective cancer therapies

Mitochondrial dysfunction induced by a SH2 domain-targeting STAT3 inhibitor leads to metabolic synthetic lethality in cancer cells

In addition to its canonical role in nuclear transcription, signal transducer and activator of transcription 3 (STAT3) is emerging as an important regulator of mitochondrial function. Here, we demonstrate that a novel inhibitor that binds with high affinity to the STAT3 SH2 domain triggers a complex cascade of events initiated by interference with mitochondrial STAT3 (mSTAT3). The mSTAT3–drug interaction leads to mitochondrial dysfunction, accumulation of proteotoxic STAT3 aggregates, and cell death. The cytotoxic effects depend directly on the drug’s ability to interfere with mSTAT3 and mitochondrial function, as demonstrated by site-directed mutagenesis and use of STAT3 knockout and mitochondria- depleted cells. Importantly, the lethal consequences of mSTAT3 inhibition are enhanced by glucose starvation and by increased reliance of cancer cells and tumor-initiating cells on mitochondria, resulting in potent activity in cell cultures and tumor xenografts in mice. These findings can be exploited for eliciting synthetic lethality in metabolically stressed cancer cells using high-affinity STAT3 inhibitors. Thus, this study provides insights on the role of mSTAT3 in cancer cells and a conceptual framework for developing more effective cancer therapies

Basic Digital Image Acquisition, Design, Processing, Analysis, Management, and Presentation

A guide for understanding the basic concepts of light microscopy involved in the acquisition, analysis, and unbiased presentation of bioimages. The chapter will help the reader to understand the simplest techniques used for image acquisition, read the metadata, and be aware of the limits deriving from the use of those imaging techniques. Applying the protocols contained in this chapter, it is possible to visualize and render image data sets using the most common software tools and apply basic image analysis workflows to get data out of images. Finally, the reader is introduced to how to present images and data analysis results in an unbiased way. D’Antuono, R. (2022). Basic Digital Image Acquisition, Design, Processing, Analysis, Management, and Presentation. In: Nechyporuk-Zloy, V. (eds) Principles of Light Microscopy: From Basic to Advanced . Springer, Cham. https://doi.org/10.1007/978-3-031-04477-9_4  https://link.springer.com/chapter/10.1007/978-3-031-04477-9_4</p

Towards super‐resolved terahertz microscopy for cellular imaging

Abstract: Biomedical imaging includes the use of a variety of techniques to study organs and tissues. Some of the possible imaging modalities are more spread at clinical level (CT, MRI, PET), while others, such as light and electron microscopy are preferred in life sciences research. The choice of the imaging modalities can be based on the capability to study functional aspects of an organism, the delivered radiation dose to the patient, and the achievable resolution. In the last few decades, spectroscopists and imaging scientists have been interested in the use of terahertz (THz) frequencies (30 μm to 3 mm wavelength) due to the low photon energy associated (E∼1 meV, not causing breaking of the molecular bonds but still interacting with some vibrational modes) and the high penetration depth that is achievable. THz has been already adopted in security, quality control and material sciences. However, the adoption of THz frequencies for biological and clinical imaging means to face, as a major limitation, the very scarce resolution associated with the use of such long wavelengths. To address this aspect and reconcile the benefit of minimal harmfulness for bioimaging with the achievable resolving power, many attempts have been made. This review summarises the state‐of‐the‐art of THz imaging applications aimed at achieving super‐resolution, describing how practical aspects of optics and quasi‐optics may be treated to efficaciously implement the use of THz as a new low‐dose and versatile modality in biomedical imaging and clinical research

Multilayered regulation of autophagy by the Atg1 kinase orchestrates spatial and temporal control of autophagosome formation

Autophagy is a conserved intracellular degradation pathway exerting various cytoprotective and homeostatic functions by using de novo double-membrane vesicle (autophagosome) formation to target a wide range of cytoplasmic material for vacuolar/lysosomal degradation. The Atg1 kinase is one of its key regulators, coordinating a complex signaling program to orchestrate autophagosome formation. Combining in vitro reconstitution and cell-based approaches, we demonstrate that Atg1 is activated by lipidated Atg8 (Atg8-PE), stimulating substrate phosphorylation along the growing autophagosomal membrane. Atg1-dependent phosphorylation of Atg13 triggers Atg1 complex dissociation, enabling rapid turnover of Atg1 complex subunits at the pre-autophagosomal structure (PAS). Moreover, Atg1 recruitment by Atg8-PE self-regulates Atg8-PE levels in the growing autophagosomal membrane by phosphorylating and thus inhibiting the Atg8-specific E2 and E3. Our work uncovers the molecular basis for positive and negative feedback imposed by Atg1 and how opposing phosphorylation and dephosphorylation events underlie the spatiotemporal regulation of autophagy

Macrophage Death following Influenza Vaccination Initiates the Inflammatory Response that Promotes Dendritic Cell Function in the Draining Lymph Node

The mechanism by which inflammation influences the adaptive response to vaccines is not fully understood. Here, we examine the role of lymph node macrophages (LNMs) in the induction of the cytokine storm triggered by inactivated influenza virus vaccine. Following vaccination, LNMs undergo inflammasome-independent necrosis-like death that is reliant on MyD88 and Toll-like receptor 7 (TLR7) expression and releases pre-stored interleukin-1α (IL-1α). Furthermore, activated medullary macrophages produce interferon-β (IFN-β) that induces the autocrine secretion of IL-1α. We also found that macrophage depletion promotes lymph node-resident dendritic cell (LNDC) relocation and affects the capacity of CD11b+ LNDCs to capture virus and express co-stimulatory molecules. Inhibition of the IL-1α-induced inflammatory cascade reduced B cell responses, while co-administration of recombinant IL-1α increased the humoral response. Stimulation of the IL-1α inflammatory pathway might therefore represent a strategy to enhance antigen presentation by LNDCs and improve the humoral response against influenza vaccines.ISSN:2666-3864ISSN:2211-124

Multicenter Comparison of 22C3 PharmDx (Agilent) and SP263 (Ventana) Assays to Test PD-L1 Expression for NSCLC Patients to Be Treated with Immune Checkpoint Inhibitors

Introduction Among the several agents targeting the programmed cell death 1 (PD-1) pathway, pembrolizumab is currently the only one approved for the treatment of patients with NSCLC in association with a companion diagnostic assay, the anti–PD-L1 immunohistochemical (IHC) 22C3 PharmDx (Agilent Technologies, Santa Clara, CA) using the Dako Autostainer (Dako, Carpinteria, CA). However, the Dako platform is not present in each pathology department, and this technical limitation is a major problem for the diffusion of the PD-L1 IHC predictive test for pembrolizumab. Methods The Italian Society of Anatomic Pathology and Cytopathology and the Italian Association of Medical Oncology in an independent, multicenter study compared the in vitro diagnostics PD-L1 IHC 22C3 pharmDx test (Agilent) on the Dako Autostainer and the in vitro diagnostics Ventana PD-L1 (SP263) test on the Ventana BenchMark platform (Ventana Medical Systems, Tucson, AZ). Using serial sections from tissue microarrays, 100 lung adenocarcinomas were locally stained and scored in four centers with the same antibody batches. Results A high analytical correlation (more than 90% at the lower 95% confidence interval [CI] value) between PD-L1 expression levels obtained with the 22C3 and SP263 assays was observed. At the proposed clinically relevant cutoffs (≥50% and ≥1%), the overall concordances between 22C3 and SP263 data were 0.99 (95% CI: 0.96–1) and 0.80 (95% CI: 0.68–0.91), respectively. The lower agreement between data obtained with the 22C3 and SP263 clones at the cutoff of 1% or higher was mainly related to the lower (about 80%) interrater agreement at this cutoff with each clone. Conclusions These results indicate a high correlation between PD-L1 IHC expression data obtained with the Agilent PD-L1 IHC 22C3 pharmDx and the Ventana PD-L1 (SP263) tests in NSCLC and suggest that the two assays could be utilized interchangeably as an aid to select patients for first-line and second-line treatment with pembrolizumab and potentially with other anti–PD-1/PD-L1 checkpoint inhibitors

Multicenter Comparison of 22C3 PharmDx (Agilent) and SP263 (Ventana) Assays to Test PD-L1 Expression for NSCLC Patients to Be Treated with Immune Checkpoint Inhibitors

INTRODUCTION: Among the several agents targeting the programmed cell death 1 (PD-1) pathway, pembrolizumab is currently the only one approved for the treatment of patients with NSCLC in association with a companion diagnostic assay, the anti-PD-L1 immunohistochemical (IHC) 22C3 PharmDx (Agilent Technologies, Santa Clara, CA) using the Dako Autostainer (Dako, Carpinteria, CA). However, the Dako platform is not present in each pathology department, and this technical limitation is a major problem for the diffusion of the PD-L1 IHC predictive test for pembrolizumab. METHODS: The Italian Society of Anatomic Pathology and Cytopathology and the Italian Association of Medical Oncology in an independent, multicenter study compared the in vitro diagnostics PD-L1 IHC 22C3 pharmDx test (Agilent) on the Dako Autostainer and the in vitro diagnostics Ventana PD-L1 (SP263) test on the Ventana BenchMark platform (Ventana Medical Systems, Tucson, AZ). Using serial sections from tissue microarrays, 100 lung adenocarcinomas were locally stained and scored in four centers with the same antibody batches. RESULTS: A high analytical correlation (more than 90% at the lower 95% confidence interval [CI] value) between PD-L1 expression levels obtained with the 22C3 and SP263 assays was observed. At the proposed clinically relevant cutoffs (≥50% and ≥1%), the overall concordances between 22C3 and SP263 data were 0.99 (95% CI: 0.96-1) and 0.80 (95% CI: 0.68-0.91), respectively. The lower agreement between data obtained with the 22C3 and SP263 clones at the cutoff of 1% or higher was mainly related to the lower (about 80%) interrater agreement at this cutoff with each clone. CONCLUSIONS: These results indicate a high correlation between PD-L1 IHC expression data obtained with the Agilent PD-L1 IHC 22C3 pharmDx and the Ventana PD-L1 (SP263) tests in NSCLC and suggest that the two assays could be utilized interchangeably as an aid to select patients for first-line and second-line treatment with pembrolizumab and potentially with other anti-PD-1/PD-L1 checkpoint inhibitors

IFN-γ-dependent interactions between tissue-intrinsic γδ T cells and tissue-infiltrating CD8 T cells limit allergic contact dermatitis

BACKGROUND: Elicitation of allergic contact dermatitis (ACD), an inflammatory type 4 hypersensitivity disease, induces skin infiltration by polyclonal effector CD8 αβ T cells and precursors of tissue-resident memory T (TRM) cells. Because TRM have long-term potential to contribute to body-surface immunoprotection and immunopathology, their local regulation needs a fuller understanding.OBJECTIVE: We sought to investigate how TRM-cell maturation might be influenced by innate-like T cells pre-existing within many epithelia.METHODS: This study examined CD8+ TRM-cell maturation following hapten-induced ACD in wild-type mice and in strains harboring altered compartments of dendritic intraepidermal γδ T cells (DETCs), a prototypic tissue-intrinsic, innate-like T-cell compartment that reportedly regulates ACD, but by no elucidated mechanism.RESULTS: In addition to eliciting CD8 TRM, ACD induced DETC activation and an intimate coregulatory association of the 2 cell types. This depended on DETC sensing IFN-γ produced by CD8 cells and involved programmed death-ligand 1 (PD-L1). Thus, in mice lacking DETC or lacking IFN-γ receptor solely on γδ cells, ACD-elicited CD8 T cells showed enhanced proliferative and effector potentials and reduced motility, collectively associated with exaggerated ACD pathology. Comparable dysregulation was elicited by PD-L1 blockade in vitro, and IFN-γ-regulated PD-L1 expression was a trait of human skin-homing and intraepithelial γδ T cells.CONCLUSIONS: The size and quality of the tissue-infiltrating CD8 T-cell response during ACD can be profoundly regulated by local innate-like T cells responding to IFN-γ and involving PD-L1. Thus, interindividual and tissue-specific variations in tissue-intrinsic lymphocytes may influence responses to allergens and other challenges and may underpin inflammatory pathologies such as those repeatedly observed in γδ T-cell-deficient settings.</p