Neutrophil Extracellular Traps Initiate Gallstone Formation.

The presence of gallstones (cholelithiasis) is a highly prevalent and severe disease and one of the leading causes of hospital admissions worldwide. Due to its substantial health impact, we investigated the biological mechanisms that lead to the formation and growth of gallstones. We show that gallstone assembly essentially requires neutrophil extracellular traps (NETs). We found consistent evidence for the presence of NETs in human and murine gallstones and describe an immune-mediated process requiring activation of the innate immune system for the formation and growth of gallstones. Targeting NET formation via inhibition of peptidyl arginine deiminase type 4 or abrogation of reactive oxygen species (ROS) production, as well as damping of neutrophils by metoprolol, effectively inhibit gallstone formation in vivo. Our results show that after the physicochemical process of crystal formation, NETs foster their assembly into larger aggregates and finally gallstones. These insights provide a feasible therapeutic concept to prevent cholelithiasis in patients at risk.

CD19 CAR T-Cell Therapy in Autoimmune Disease - A Case Series with Follow-up.

BACKGROUND: Treatment for autoimmune diseases such as systemic lupus erythematosus (SLE), idiopathic inflammatory myositis, and systemic sclerosis often involves long-term immune suppression. Resetting aberrant autoimmunity in these diseases through deep depletion of B cells is a potential strategy for achieving sustained drug-free remission. METHODS: We evaluated 15 patients with severe SLE (8 patients), idiopathic inflammatory myositis (3 patients), or systemic sclerosis (4 patients) who received a single infusion of CD19 chimeric antigen receptor (CAR) T cells after preconditioning with fludarabine and cyclophosphamide. Efficacy up to 2 years after CAR T-cell infusion was assessed by means of Definition of Remission in SLE (DORIS) remission criteria, American College of Rheumatology-European League against Rheumatism (ACR-EULAR) major clinical response, and the score on the European Scleroderma Trials and Research Group (EUSTAR) activity index (with higher scores indicating greater disease activity), among others. Safety variables, including cytokine release syndrome and infections, were recorded. RESULTS: The median follow-up was 15 months (range, 4 to 29). The mean (±SD) duration of B-cell aplasia was 112±47 days. All the patients with SLE had DORIS remission, all the patients with idiopathic inflammatory myositis had an ACR-EULAR major clinical response, and all the patients with systemic sclerosis had a decrease in the score on the EUSTAR activity index. Immunosuppressive therapy was completely stopped in all the patients. Grade 1 cytokine release syndrome occurred in 10 patients. One patient each had grade 2 cytokine release syndrome, grade 1 immune effector cell-associated neurotoxicity syndrome, and pneumonia that resulted in hospitalization. CONCLUSIONS: In this case series, CD19 CAR T-cell transfer appeared to be feasible, safe, and efficacious in three different autoimmune diseases, providing rationale for further controlled clinical trials. (Funded by Deutsche Forschungsgemeinschaft and others.).

Sustained pacifier use is associated with smaller vocabulary sizes at 1 and 2 years of age: A cross-sectional study.

Pacifier use during childhood has been hypothesized to interfere with language processing, but, to date, there is limited evidence revealing detrimental effects of prolonged pacifier use on infant vocabulary learning. In the present study, parents of 12- and 24-month-old infants were recruited in Oslo (Norway). The sample included 1187 monolingual full-term born (without visual, auditory, or cognitive impairments) infants: 452 (230 girls; 222 boys) 12-month-olds and 735 (345 girls; 390 boys) 24-month-olds. Parents filled out an online Norwegian Communicative Development Inventory (CDI), which assesses the vocabulary in comprehension and production for 12-month-old infants and in production only for 24-month-old infants. CDI scores were transformed into age- and sex-adjusted percentiles using Norwegian norms. Additionally, parents retrospectively reported their child's daytime pacifier use, in hours, at 2-month intervals, from birth to the assessment date. Maternal education was used to control, in the analyses, for the socio-economic status. We found that greater pacifier use in an infant's lifespan was associated with lower vocabulary size. Pacifier use later in life was more negatively associated with vocabulary size than precocious use, and increased the odds of being a low language scorer. In sum, our study moves beyond the findings of momentary effects of experimentally induced "impairment" in articulators' movement on speech perception and suggests that, from 12 months of age, constraints on the infant's speech articulators (pacifier use) may be negatively associated with word comprehension and production. RESEARCH HIGHLIGHT: We examined the relationship between pacifier use and vocabulary sizes in production at 24 months of age and comprehension and production at 12 months of age. Lifespan Pacifier Use (LPU) was negatively correlated with vocabulary sizes in comprehension and production among 12-month-old infants and negatively correlated with production for 24-month-olds. Later pacifier use was found to be more negatively correlated with vocabulary size in infants, as compared to more precocious use. The amount of pacifier use in the 2 months prior to a child's second birthday was predictive of a higher prevalence of low vocabulary scores in 24-month-olds.

Butyrophilin 2a2 (Btn2a2) expression on thymic epithelial cells promotes central T cell tolerance and prevents autoimmune disease.

Butyrophilins are surface receptors belonging to the immunoglobulin superfamily. While several members of the butyrophilin family have been implicated in the development of unconventional T cells, butyrophilin 2a2 (Btn2a2) has been shown to inhibit conventional T cell activation. Here, we demonstrate that in steady state, the primary source of Btn2a2 are thymic epithelial cells (TEC). Absence of Btn2a2 alters thymic T cell maturation and bypasses central tolerance mechanisms. Furthermore, Btn2a2-/- mice develop spontaneous autoimmunity resembling human primary Sjögren's Syndrome (pSS), including formation of tertiary lymphoid structures (TLS) in target organs. Ligation of Btn2a2 on developing thymocytes is associated with reduced TCR signaling and CD5 levels, while absence of Btn2a2 results in increased TCR signaling and CD5 levels. These results define a novel role for Btn2a2 in promoting central tolerance by modulating TCR signaling strength and indicate a potential mechanism of pSS development.

Neutrophil extracellular traps drive epithelial-mesenchymal transition of human colon cancer.

Neutrophil extracellular traps (NETs) are extracellular structures, composed of nuclear DNA and various proteins released from neutrophils. Evidence is growing that NETs exert manifold functions in infection, immunity and cancer. Recently, NETs have been detected in colorectal cancer (CRC) tissues, but their association with disease progression and putative functional impact on tumourigenesis remained elusive. Using high-resolution stimulated emission depletion (STED) microscopy, we showed that citrullinated histone H3 (H3cit) is sufficient to specifically detect citrullinated NETs in colon cancer tissues. Among other evidence, this was supported by the close association of H3cit with de-condensed extracellular DNA, the hallmark of NETs. Extracellular DNA was reliably differentiated from nuclear condensed DNA by staining with an anti-DNA antibody, providing a novel and valuable tool to detect NETs in formalin-fixed paraffin-embedded tissues. Using these markers, the clinical association of NETs was investigated in a cohort of 85 patients with colon cancer. NETs were frequently detected (37/85, 44%) in colon cancer tissue sections and preferentially localised either only in the tumour centre or both in the tumour centre and the invasive front. Of note, citrullinated NETs were significantly associated with high histopathological tumour grades and lymph node metastasis. In vitro, purified NETs induced filopodia formation and cell motility in CRC cell lines. This was associated with increased expression of mesenchymal marker mRNAs (vimentin [VIM], fibronectin [FN1]) and epithelial-mesenchymal transition promoting transcription factors (ZEB1, Slug [SNAI2]), as well as decreased expression of the epithelial markers E-cadherin (CDH1) and epithelial cell adhesion molecule (EPCAM). These findings indicated that NETs activate an epithelial-mesenchymal transition-like process in CRC cells and may contribute to the metastatic progression of CRC. © 2021 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Neuropathic Pain and Positive Sensory Symptoms in Brachial Plexus Neuropathy: An Exploratory Study of Outcomes after Surgical Decompression and Proposal of a New Sensory Frequency of Symptoms Scale.

BACKGROUND: There are no articles that aim to evaluate the specific role of surgical decompression on the recovery of pain and positive sensory symptoms (PSS) in patients with brachial plexus neuropathy (BPN), as well as the relationship between pain and frequency of sensory manifestations. METHODS: A prospective before and after study was performed, considering the pain intensity through the visual analogue scale (VAS), and the frequency of PSS through a proposed new scale: Sensory Frequency of Symptoms Scale (SFSS). To compare the patients before and after the intervention, a paired T-test, a Wilcoxon signed-rank test, and Cohen's D test were made, coupled with a Spearman analysis in order to establish the relationship between pain and PSS. RESULTS: Sixteen patients were included in the study, the clinical evaluation showed changes in pain according with VAS, going from a mean preoperative state of 8.19 to 1.31 after surgery, showing significant changes (84%, p < 0.00006, Δ = 2.776). Within the PSS, a significant decrease was observed in paresthesias (74%, p < 0.0001, Δ = 1.645), dysesthesias (80%, p < 0.002, Δ = 1.453), and allodynia (70%, p = 0.031, Δ = 0.635). Conversely, the preoperative correlation analysis between pain and dysesthesias/allodynia showed a low and non-significant relationship (R < 0.4, p > 0.05). CONCLUSIONS: Surgical decompression is an effective technique for the relief of pain and sensory manifestations in adult patients with BPN of compressive origin. No relationship was observed between pain and dysesthesias/allodynia. Therefore, during clinical evaluation, they should be considered as independent manifestations, highlighting the need to validate new scales.

On-Line Evaluation and Monitoring of Security Features of an RO-Based PUF/TRNG for IoT Devices.

The proliferation of devices for the Internet of Things (IoT) and their implication in many activities of our lives have led to a considerable increase in concern about the security of these devices, posing a double challenge for designers and developers of products. On the one hand, the design of new security primitives, suitable for resource-limited devices, can facilitate the inclusion of mechanisms and protocols to ensure the integrity and privacy of the data exchanged over the Internet. On the other hand, the development of techniques and tools to evaluate the quality of the proposed solutions as a step prior to their deployment, as well as to monitor their behavior once in operation against possible changes in operating conditions arising naturally or as a consequence of a stress situation forced by an attacker. To address these challenges, this paper first describes the design of a security primitive that plays an important role as a component of a hardware-based root of trust, as it can act as a source of entropy for True Random Number Generation (TRNG) or as a Physical Unclonable Function (PUF) to facilitate the generation of identifiers linked to the device on which it is implemented. The work also illustrates different software components that allow carrying out a self-assessment strategy to characterize and validate the performance of this primitive in its dual functionality, as well as to monitor possible changes in security levels that may occur during operation as a result of device aging and variations in power supply or operating temperature. The designed PUF/TRNG is provided as a configurable IP module, which takes advantage of the internal architecture of the Xilinx Series-7 and Zynq-7000 programmable devices and incorporates an AXI4-based standard interface to facilitate its interaction with soft- and hard-core processing systems. Several test systems that contain different instances of the IP have been implemented and subjected to an exhaustive set of on-line tests to obtain the metrics that determine its quality in terms of uniqueness, reliability, and entropy characteristics. The results obtained prove that the proposed module is a suitable candidate for various security applications. As an example, an implementation that uses less than 5% of the resources of a low-cost programmable device is capable of obfuscating and recovering 512-bit cryptographic keys with virtually zero error rate.

Anti-DNA-IgM Favors the Detection of NET-Associated Extracellular DNA.

During inflammatory responses, neutrophils enter the sites of attack where they execute various defense mechanisms. They (I) phagocytose microorganisms, (II) degranulate to release cytokines, (III) recruit various immune cells by cell-type specific chemokines, (IV) secrete anti-microbials including lactoferrin, lysozyme, defensins and reactive oxygen species, and (V) release DNA as neutrophil extracellular traps (NETs). The latter originates from mitochondria as well as from decondensed nuclei. This is easily detected in cultured cells by staining of DNA with specific dyes. However, in tissues sections the very high fluorescence signals emitted from the condensed nuclear DNA hamper the detection of the widespread, extranuclear DNA of the NETs. In contrast, when we employ anti-DNA-IgM antibodies, they are unable to penetrate deep into the tightly packed DNA of the nucleus, and we observe a robust signal for the extended DNA patches of the NETs. To validate anti-DNA-IgM, we additionally stained the sections for the NET-markers histone H2B, myeloperoxidase, citrullinated histone H3, and neutrophil elastase. Altogether, we have described a fast one-step procedure for the detection of NETs in tissue sections, which provides new perspectives to characterize neutrophil-associated immune reactions in disease.

The Molecular Role of Polyamines in Age-Related Diseases: An Update.

Polyamines (Pas) are short molecules that exhibit two or three amine groups that are positively charged at a physiological pH. These small molecules are present in high concentrations in a wide variety of organisms and tissues, suggesting that they play an important role in cellular physiology. Polyamines include spermine, spermidine, and putrescine, which play important roles in age-related diseases that have not been completely elucidated. Aging is a natural process, defined as the time-related deterioration of the physiological functions; it is considered a risk factor for degenerative diseases such as cardiovascular, neurodegenerative, and musculoskeletal diseases; arthritis; and even cancer. In this review, we provide a new perspective on the participation of Pas in the cellular and molecular processes related to age-related diseases, focusing our attention on important degenerative diseases such as Alzheimerߣs disease, Parkinsonߣs disease, osteoarthritis, sarcopenia, and osteoporosis. This new perspective leads us to propose that Pas function as novel biomarkers for age-related diseases, with the main purpose of achieving new molecular alternatives for healthier aging.

Matched linac stereotactic radiotherapy: An assessment of delivery similarity and distributive patient-specific quality assurance feasibility.

Matching multiple linacs to common baseline data allows patients to be treated, and patient-specific quality assurance (PSQA) to be completed on any linac. Stereotactic body radiotherapy (SBRT) requires higher levels of accuracy and quality assurance than routine radiotherapy. The achieved linac matching must therefore be evaluated before distributive treatment or PSQA models can be implemented safely. This investigation aimed to propose metrics for defining linacs to be matched for SBRT deliveries, assess 12 linacs against these criteria, and determine if a distributive PSQA model could be implemented by reviewing the rates of false PSQA results. Ten SBRT spine plans were delivered by 12 matched Elekta linacs and measured using one of seven SRS MapCHECK devices. For gamma criteria of (3%, 2 mm), 96.9% of equivalent location detectors, showed a range of gamma ≤ 1.0 and 99.9% showed a standard deviation of ≤ 0.5. For criteria of (3%,1 mm) and (2%,1 mm), these ranges decreased to 92.1% and 80.2% while the standard deviations decreased to 99.3% and 95.7%, respectively. The dose differences showed that 43.6%, 82.7%, and 91.4% of detectors had a dose range of ≤ 3.0%, ≤ 5.0%, and ≤ 6.0%, respectively. Standard deviations of dose differences were 1.5%, 2.5%, and 3.0% for 94.1%, 98.3%, and 99.5% of detectors, respectively. For the fleet of linacs, distributive PSQA yielded false results for 0.0%, 17.7%, and 33.0% of plans, equivalent to 1.2%, 3.5%, and 9.4% of detectors when using gamma criteria of (3%,2 mm), (3%,1 mm), or (2%,1 mm), respectively. These linacs could be considered matched for SBRT treatments and implement a distributive PSQA model when gamma analysis was completed with a criterion of (3%, 2 mm). For stricter criterion of (3%,1 mm) or (2%,1 mm), they did not meet the proposed metrics.

Standing-Wave Feeding for High-Gain Linear Dielectric Resonator Antenna (DRA) Array.

A novel feeding method for linear DRA arrays is presented, illuminating the use of the power divider, transitions, and launchers, and keeping uniform excitation to array elements. This results in a high-gain DRA array with low losses with a design that is simple, compact and inexpensive. The proposed feeding method is based on exciting standing waves using discrete metallic patches in a simple design procedure. Two arrays with two and four DRA elements are presented as a proof of concept, which provide high gains of 12 and 15dBi, respectively, which are close to the theoretical limit based on array theory. The radiation efficiency for both arrays is about 93%, which is equal to the array element efficiency, confirming that the feeding method does not add losses as in the case of standard methods. To facilitate the fabrication process, the entire array structure is 3D-printed, which significantly decreases the complexity of fabrication and alignment. Compared to state-of-the-art feeding techniques, the proposed method provides higher gain and higher efficiency with a smaller electrical size.

An Integrated Millimeter-Wave Satellite Radiometer Working at Room-Temperature with High Photon Conversion Efficiency.

In this work, the design of an integrated 183GHz radiometer frontend for earth observation applications on satellites is presented. By means of the efficient electro-optic modulation of a laser pump with the observed millimeter-wave signal followed by the detection of the generated optical sideband, a room-temperature low-noise receiver frontend alternative to conventional Low Noise Amplifiers (LNAs) or Schottky mixers is proposed. Efficient millimeter-wave to 1550 nm upconversion is realized via a nonlinear optical process in a triply resonant high-Q Lithium Niobate (LN) Whispering Gallery Mode (WGM) resonator. By engineering a micromachined millimeter-wave cavity that maximizes the overlap with the optical modes while guaranteeing phase matching, the system has a predicted normalized photon-conversion efficiency ≈10-1 per mW pump power, surpassing the state-of-the-art by around three orders of magnitude at millimeter-wave frequencies. A piezo-driven millimeter-wave tuning mechanism is designed to compensate for the fabrication and assembly tolerances and reduces the complexity of the manufacturing process.

A Pleomorphic Puzzle: Heterogeneous Pulmonary Vascular Occlusions in Patients with COVID-19.

Vascular occlusions in patients with coronavirus diseases 2019 (COVID-19) have been frequently reported in severe outcomes mainly due to a dysregulation of neutrophils mediating neutrophil extracellular trap (NET) formation. Lung specimens from patients with COVID-19 have previously shown a dynamic morphology, categorized into three types of pleomorphic occurrence based on histological findings in this study. These vascular occlusions in lung specimens were also detected using native endogenous fluorescence or NEF in a label-free method. The three types of vascular occlusions exhibit morphology of DNA rich neutrophil elastase (NE) poor (type I), NE rich DNA poor (type II), and DNA and NE rich (type III) cohort of eleven patients with six males and five females. Age and gender have been presented in this study as influencing variables linking the occurrence of several occlusions with pleomorphic contents within a patient specimen and amongst them. This study reports the categorization of pleomorphic occlusions in patients with COVID-19 and the detection of these occlusions in a label-free method utilizing NEF.

Synthesis of Benzo[4,5]thiazolo[2,3-c][1,2,4]triazole Derivatives via C-H Bond Functionalization of Disulfide Intermediates.

Many nitrogen- and sulfur-containing heterocyclic compounds exhibit biological activity. Among these heterocycles are benzo[4,5]thiazolo[2,3-c][1,2,4]triazoles for which two main synthetic approaches exist. Here we report a new synthetic protocol that allows the preparation of these tricyclic compounds via the oxidation of a mercaptophenyl moiety to its corresponding disulfide. Subsequent C-H bond functionalization is thought to enable an intramolecular ring closure, thus forming the desired benzo[4,5]thiazolo[2,3-c][1,2,4]triazole. This method combines a high functional group tolerance with short reaction times and good to excellent yields.

Annexin A5 regulates surface αvß5 integrin for retinal clearance phagocytosis.

Diurnal clearance phagocytosis by the retinal pigment epithelium (RPE) is a conserved efferocytosis process whose binding step is mediated by αvß5 integrin receptors. Two related annexins, A5 (ANXA5) and A6 (ANXA6), share an αvß5 integrin-binding motif. Here, we report that ANXA5, but not ANXA6, regulates the binding capacity for spent photoreceptor outer segment fragments or apoptotic cells by fibroblasts and RPE. Similar to αvß5-deficient RPE, ANXA5-/- RPE in vivo lacks the diurnal burst of phagocytosis that follows photoreceptor shedding in wild-type retina. Increasing ANXA5 in cells lacking αvß5 or increasing αvß5 in cells lacking ANXA5 does not affect particle binding. Association of cytosolic ANXA5 and αvß5 integrin in RPE in culture and in vivo further supports their functional interdependence. Silencing ANXA5 is sufficient to reduce levels of αvß5 receptors at the apical phagocytic surface of RPE cells. The effect of ANXA5 on surface αvß5 and on particle binding requires the C-terminal ANXA5 annexin repeat but not its unique N-terminus. These results identify a novel role for ANXA5 specifically in the recognition and binding step of clearance phagocytosis, which is essential to retinal physiology.This article has an associated First Person interview with the first author of the paper.

Maternal and perinatal outcomes related to COVID-19 and pregnancy: An overview of systematic reviews.

INTRODUCTION: Evidence about coronavirus disease 2019 (COVID-19) and pregnancy has rapidly increased since December 2019, making it difficult to make rigorous evidence-based decisions. The objective of this overview of systematic reviews is to conduct a comprehensive analysis of the current evidence on prognosis of COVID-19 in pregnant women. MATERIAL AND METHODS: We used the Living OVerview of Evidence (L·OVE) platform for COVID-19, which continually retrieves studies from 46 data sources (including PubMed/MEDLINE, Embase, other electronic databases, clinical trials registries, and preprint repositories, among other sources relevant to COVID-19), mapping them into PICO (population, intervention, control, and outcomes) questions. The search covered the period from the inception date of each database to 13 September 2020. We included systematic reviews assessing outcomes of pregnant women with COVID-19 and/or their newborns. Two authors independently screened the titles and abstracts, assessed full texts to select the studies that met the inclusion criteria, extracted data, and appraised the risk of bias of each included systematic review. We measured the overlap of primary studies included among the selected systematic reviews by building a matrix of evidence, calculating the corrected covered area, and assessing the level of overlap for every pair of systematic reviews. RESULTS: Our search yielded 1132 references. 52 systematic reviews met inclusion criteria and were included in this overview. Only one review had a low risk of bias, three had an unclear risk of bias, and 48 had a high risk of bias. Most of the included reviews were highly overlapped among each other. In the included reviews, rates of maternal death varied from 0% to 11.1%, admission to intensive care from 2.1% to 28.5%, preterm deliveries before 37 weeks from 14.3% to 61.2%, and cesarean delivery from 48.3% to 100%. Regarding neonatal outcomes, neonatal death varied from 0% to 11.7% and the estimated infection status of the newborn varied between 0% and 11.5%. CONCLUSIONS: Only one of 52 systematic reviews had a low risk of bias. Results were heterogeneous and the overlap of primary studies was frequently very high between pairs of systematic reviews. High-quality evidence syntheses of comparative studies are needed to guide future clinical decisions.

Consistency of small-field dosimetry, on and off axis, in beam-matched linacs used for stereotactic radiosurgery.

PURPOSE: Stereotactic radiosurgery (SRS) can be delivered with a standard linear accelerator (linac). At institutions having more than one linac, beam matching is common practice. In the literature, there are indications that machine central axis (CAX) matching for broad fields does not guarantee matching of small fields with side ≤2 cm. There is no indication on how matching for broad fields on axis translates to matching small fields off axis. These are of interest to multitarget single-isocenter (MTSI) SRS planning and the present work addresses that gap in the literature. METHODS: We used 6 MV flattening filter free (FFF) beams from four Elekta VersaHD® linacs equipped with an Agility™ multileaf collimator (MLC). The linacs were strictly matched for broad fields on CAX. We compared output factors (OPFs) and effective field size, measured concurrently using a novel 2D solid-state dosimeter "Duo" with a spatial resolution of 0.2 mm, in square and rectangular static fields with sides from 0.5 to 2 cm, either on axis or away from it by 5 to 15 cm. RESULTS: Among the four linacs, OPF for fields ≥1 × 1 cm2 ranged 1.3% on CAX, whereas off axis a maximum range of 1.9% was observed at 15 cm. A larger variability in OPF was noted for the 0.5 × 0.5 cm2 field, with a range of 5.9% on CAX, which improved to a maximum of 2.3% moving off axis. Two linacs showed greater consistency with a range of 1.4% on CAX and 2.2% at 15 cm off axis. Between linacs, the effective field size varied by <0.04 cm in most cases, both on and off axis. Tighter matching was observed for linacs with a similar focal spot position. CONCLUSIONS: Verification of small-field consistency for matched linacs used for SRS is an important task for dosimetric validation. A significant benefit of concurrent measurement of field size and OPF allowed for a comprehensive assessment using a novel diode array. Our study showed the four linacs, strictly matched for broad fields on CAX, were still matched down to a field size of 1 x 1 cm2 on and off axis.

Dendritic Cells Pulsed with Cytokine-Adjuvanted Tumor Membrane Vesicles Inhibit Tumor Growth in HER2-Positive and Triple Negative Breast Cancer Models.

Dendritic cells (DCs) are the most effective antigen presenting cells for the development of T cell responses. The only FDA approved DC-based immunotherapy to date is Sipuleucel-T, which utilizes a fusion protein to stimulate DCs ex vivo with GM-CSF and simultaneously deliver the antigen PAP for prostate cancer. This approach is restricted by the breadth of immunity elicited to a single antigen, and to cancers that have a defined tumor associated antigen. Other multi-antigen approaches have been restricted by poor efficacy of vaccine adjuvants. We have developed a vaccine platform that consists of autologous DCs pulsed with cytokine-adjuvanted tumor membrane vesicles (TMVs) made from tumor tissue, that encapsulate the antigenic landscape of individual tumors. Here we test the efficacy of DCs pulsed with TMVs incorporated with glycolipid-anchored immunostimulatory molecules (GPI-ISMs) in HER2-positive and triple negative breast cancer murine models. Pulsing of DCs with TMVs containing GPI-ISMs results in superior uptake of vesicles, DC activation and cytokine production. Adaptive transfer of TMV-pulsed DCs to tumor bearing mice results in the inhibition of tumor growth, reduction in lung metastasis, and an increase in immune cell infiltration into the tumors. These observations suggest that DCs pulsed with TMVs containing GPI-GM-CSF and GPI-IL-12 can be further developed to be used as a personalized immunotherapy platform for cancer treatment.

Connection between Periodontitis-Induced Low-Grade Endotoxemia and Systemic Diseases: Neutrophils as Protagonists and Targets.

Periodontitis is considered a promoter of many systemic diseases, but the signaling pathways of this interconnection remain elusive. Recently, it became evident that certain microbial challenges promote a heightened response of myeloid cell populations to subsequent infections either with the same or other pathogens. This phenomenon involves changes in the cell epigenetic and transcription, and is referred to as ''trained immunity''. It acts via modulation of hematopoietic stem and progenitor cells (HSPCs). A main modulation driver is the sustained, persistent low-level transmission of lipopolysaccharide from the periodontal pocket into the peripheral blood. Subsequently, the neutrophil phenotype changes and neutrophils become hyper-responsive and prone to boosted formation of neutrophil extracellular traps (NET). Cytotoxic neutrophil proteases and histones are responsible for ulcer formations on the pocket epithelium, which foster bacteremia and endoxemia. The latter promote systemic low-grade inflammation (SLGI), a precondition for many systemic diseases and some of them, e.g., atherosclerosis, diabetes etc., can be triggered by SLGI alone. Either reverting the polarized neutrophils back to the homeostatic state or attenuation of neutrophil hyper-responsiveness in periodontitis might be an approach to diminish or even to prevent systemic diseases.

Missing in action-The meaning of cell death in tissue damage and inflammation.

Billions of cells die every day in higher organisms as part of the normal process of tissue homeostasis. During special conditions like in development, acute infections, mechanical injuries, and immunity, cell death is a common denominator and it exerts profound effects in the outcome of these scenarios. To prevent the accumulation of aged, superfluous, infected, damaged and dead cells, professional phagocytes act in a rapid and efficient manner to clear the battle field and avoid spread of the destruction. Neutrophils are the most abundant effector immune cells that extravasate into tissues and can turn injured tissues into gory battle fields. In peace times, neutrophils tend to patrol tissues without provoking inflammatory reactions. We discuss in this review actual and forgotten knowledge about the meaning of cell death during homeostatic processes and drive the attention to the importance of the action of neutrophils during patrolling and for the maintenance or recovery of the homeostatic state once the organism gets attacked or injured, respectively. In this fashion, we disclose several disease conditions that arise as collateral damage of physiological responses to death.