Cancer Immunotherapy by Blocking Immune Checkpoints on Innate Lymphocytes.

Immune checkpoints refer to a plethora of inhibitory pathways of the immune system that play a crucial role in maintaining self-tolerance and in tuning the duration and amplitude of physiological immune responses to minimize collateral tissue damages. The breakdown of this delicate balance leads to pathological conditions, including cancer. Indeed, tumor cells can develop multiple mechanisms to escape from immune system defense, including the activation of immune checkpoint pathways. The development of monoclonal antibodies, targeting inhibitory immune checkpoints, has provided an immense breakthrough in cancer therapy. Immune checkpoint inhibitors (ICI), initially developed to reverse functional exhaustion in T cells, recently emerged as important actors in natural killer (NK)-cell-based immunotherapy. Moreover, the discovery that also helper innate lymphoid cells (ILCs) express inhibitory immune checkpoints, suggests that these molecules might be targeted on ILCs, to modulate their functions in the tumor microenvironment. Recently, other strategies to achieve immune checkpoint blockade have been developed, including miRNA exploiting systems. Herein, we provide an overview of the current knowledge on inhibitory immune checkpoints on NK cells and ILCs and we discuss how to target these innate lymphocytes by ICI in both solid tumors and hematological malignancies

Innate control of actin nucleation determines two distinct migration behaviours in dendritic cells

Dendritic cell (DC) migration in peripheral tissues serves two main functions: antigen sampling by immature DCs, and chemokine-guided migration towards lymphatic vessels (LVs) on maturation. These migratory events determine the efficiency of the adaptive immune response. Their regulation by the core cell locomotion machinery has not been determined. Here, we show that the migration of immature DCs depends on two main actin pools: a RhoA mDial-dependent actin pool located at their rear, which facilitates forward locomotion; and a Cdc42 Arp2/3-dependent actin pool present at their front, which limits migration but promotes antigen capture. Following TLR4 MyD88-induced maturation, Arp2/3-dependent actin enrichment at the cell front is markedly reduced. Consequently, mature DCs switch to a faster and more persistent mDial-dependent locomotion mode that facilitates chemotactic migration to LVs and lymph nodes. Thus, the differential use of actin-nucleating machineries optimizes the migration of immature and mature DCs according to their specific function

Assessment of Cardiac, Vascular, and Pulmonary Pathobiology In Vivo During Acute COVID-19.

Background Acute COVID-19-related myocardial, pulmonary, and vascular pathology and how these relate to each other remain unclear. To our knowledge, no studies have used complementary imaging techniques, including molecular imaging, to elucidate this. We used multimodality imaging and biochemical sampling in vivo to identify the pathobiology of acute COVID-19. Specifically, we investigated the presence of myocardial inflammation and its association with coronary artery disease, systemic vasculitis, and pneumonitis. Methods and Results Consecutive patients presenting with acute COVID-19 were prospectively recruited during hospital admission in this cross-sectional study. Imaging involved computed tomography coronary angiography (identified coronary disease), cardiac 2-deoxy-2-[fluorine-18]fluoro-D-glucose positron emission tomography/computed tomography (identified vascular, cardiac, and pulmonary inflammatory cell infiltration), and cardiac magnetic resonance (identified myocardial disease) alongside biomarker sampling. Of 33 patients (median age 51 years, 94% men), 24 (73%) had respiratory symptoms, with the remainder having nonspecific viral symptoms. A total of 9 patients (35%, n=9/25) had cardiac magnetic resonance-defined myocarditis. Of these patients, 53% (n=5/8) had myocardial inflammatory cell infiltration. A total of 2 patients (5%) had elevated troponin levels. Cardiac troponin concentrations were not significantly higher in patients with and without myocarditis (8.4 ng/L [interquartile range, IQR: 4.0-55.3] versus 3.5 ng/L [IQR: 2.5-5.5]; P=0.07) or myocardial cell infiltration (4.4 ng/L [IQR: 3.4-8.3] versus 3.5 ng/L [IQR: 2.8-7.2]; P=0.89). No patients had obstructive coronary artery disease or vasculitis. Pulmonary inflammation and consolidation (percentage of total lung volume) was 17% (IQR: 5%-31%) and 11% (IQR: 7%-18%), respectively. Neither were associated with the presence of myocarditis. Conclusions Myocarditis was present in a third patients with acute COVID-19, and the majority had inflammatory cell infiltration. Pneumonitis was ubiquitous, but this inflammation was not associated with myocarditis. The mechanism of cardiac pathology is nonischemic and not attributable to a vasculitic process. Registration URL: https://www.isrctn.com; Unique identifier: ISRCTN12154994

An overview on microfluidic systems for nucleic acids extraction from human raw samples

Nucleic acid (NA) extraction is a basic step for genetic analysis, from scientific research to diagnostic and forensic applications. It aims at preparing samples for its application with biomolecular technologies such as isothermal and non\u2010isothermal amplification, hybridization, electrophoresis, Sanger sequencing and next\u2010generation sequencing. Multiple steps are involved in NA collection from raw samples, including cell separation from the rest of the specimen, cell lysis, NA isolation and release. Typically, this process needs molecular biology facilities, specialized instrumentation and labor\u2010intensive operations. Microfluidic devices have been developed to analyze NA samples with high efficacy and sensitivity. In this context, the integration within the chip of the sample preparation phase is crucial to leverage the promise of portable, fast, user-friendly and economic point\u2010of\u2010care solutions. This review presents an overview of existing lab\u2010on-a\u2010chip (LOC) solutions designed to provide automated NA extraction from human raw biological fluids, such as whole blood, excreta (urine and feces), saliva. It mainly focuses on LOC implementation aspects, aiming to describe a detailed panorama of strategies implemented for different human raw sample preparations

Measurement of torque generated during intracanal instrumentation in vivo

Aim: To compare two different clinical techniques when using endodontic rotary instruments by monitoring the torque in vivo. Methodology: Ten single-rooted maxillary anterior teeth were prepared by an endodontist using size 25,.06 taper TF rotary instruments (KerrEndo, Orange, CA, USA). All instruments were rotated at 500 rpm with maximum torque set at 2.5 N cm using an endodontic motor (Kavo, Biberach, Germany), which automatically recorded the torque values every 1/10 of a second. Instruments were used with two modi operandi: (i) Inward action: short-length intermittent progression with slight apical pressure aiming to reach the working length (also known as pecking motion), followed by (ii) Outward action: lateral brushing action selectively directed to address the canal circumference whilst removing instruments in the coronal direction. The mean, maximum and accumulated torque values were recorded and analysed statistically (t-test and Wilcoxon test) with a significance level set at 5%. None of the recorded values exceeded the selected torque limit. Results: A significant difference between inward and outward actions was found regarding the evaluated parameters (P < 0.05). The outward action (brushing) was significantly safer (requiring less torque) compared to the inward action (pecking). Conclusions: Torque measurements during in vivo instrumentation provided useful information regarding the techniques evaluated. Although the torque limit was not reached by any of the two actions, the technique impacted significantly on the torque applied to the instrument

Operative torque analysis to evaluate cutting efficiency of two nickel-titanium rotary instruments for glide path: an in vitro comparison

Aim and objective: The aim and objective of this study was to evaluate and compare the cutting efficacy of two different nickel titanium rotary instruments by a novel methodology: the operative torque (torque and time needed to progress toward the apex). Materials and methods: Ten fresh extracted upper first premolars with two canals were instrumented with a KaVo (Biberach, Germany) and a KaVo 1:1 handpiece at 300 rpm with maximum torque set at 2 N. One canal was instrumented with ProGlider NiTi rotary instruments (Dentsply Sirona Endodontics, Ballaigues, Switzerland), with tip size of 16.02, and the other one with EgdeGlidePath rotary instrument (EdgeEndo, Albuquerque, New Mexico). Mean instrumentation time, mean torque values, and maximum torque values were evaluated for each instrument. The significance level was set at p <0.05. Results: EdgeGlidePath instruments reached the working length in significantly less time with a significantly smaller amount of torque when compared to ProGlider (p >0.05). No instruments exhibited flute deformation or underwent intracanal failure. Conclusions: Operative torque is related to the capability to cut dentin and progress toward the apex: the smaller the torque values, the higher the cutting ability (and safety). Operative torque is also dependent on debris removal and irrigation techniques. Nevertheless, both operative torque and instrumentation time are clinically relevant parameters for evaluating instruments' performance (i.e., cutting ability). Clinical significance: Operative torque during endodontic instrumentation helps understanding the overall performance in terms of both cutting efficiency and safety