Inflammasome independent leukotriene-b4 production drives crystalline silica induced sterile inflammation.

Silicosis is an irreversible lung inflammatory disease caused by chronic exposure to crystalline silica (CS) and is one of the most prevalent occupational diseases worldwide. Lipid chemoattractant Leukotriene B4 (LTB4) plays an important role in neutrophilic inflammation that drives silicosis and promotes lung cancer. Previous studies in our laboratory have demonstrated that CS-induced neutrophilic inflammation and lung tumor burden in K-rasLA1 mice is abrogated in the LTB4-receptor deficient mice. Another pathway whose importance is well studied in the progression of silicosis is the Nalp3 inflammasome pathway. Studies have shown inflammasome- dependent IL-1b to be important for the development of CS-induced pulmonary fibrosis. In this study, we examined the cellular mechanisms involved in CS-induced inflammatory pathways. We demonstrate that phagocytosis of CS particles is essential for the production of LTB4 and IL-1b in macrophages, mast cells and neutrophils. CS uptake induced rapid formation of lipid bodies in the cytoplasm independent of inflammasome activation. The appearance of these lipid bodies correlated with LTB4 production in mouse bone marrow-derived macrophages. LTB4 synthesis enzymes 5-LO, FLAP and LTA4H, co-localized within the lipid bodies suggesting that they are not merely storage vesicles but the sites of CS-induced LTB4 production. We coined the term “lipidosome” to define the functional unit of LTB4 synthesis in these lipid bodies. Our studies with bafilomycin-A1 and NLRP3 deficient mice, confirmed that LTB4 synthesis in the lipidosome is independent of inflammasome activation. siRNA knockdown and confocal microscopy studies revealed that lipidosome is closely associated with phagosome and their formation appeared to be seamlessly linked to the phagosome maturation pathway. GTPases Rab5c, Rab40c along with JNK1 are essential for lipidosome formation and LTB4 production. Additionally, activation of JNK pathway is necessary for both LTB4 and IL-1b production. BI-78D3, a JNK inhibitor, completely abrogated CS-induced neutrophilic inflammation in an in-vivo air pouch model. In conclusion, these results highlight an inflammasome independent and JNK activation dependent lipidosome pathway as a major regulator of LTB4 synthesis and CS-induced sterile inflammation

Genetically Engineered T-Cells for Malignant Glioma: Overcoming the Barriers to Effective Immunotherapy.

Malignant gliomas carry a dismal prognosis. Conventional treatment using chemo- and radiotherapy has limited efficacy with adverse events. Therapy with genetically engineered T-cells, such as chimeric antigen receptor (CAR) T-cells, may represent a promising approach to improve patient outcomes owing to their potential ability to attack highly infiltrative tumors in a tumor-specific manner and possible persistence of the adaptive immune response. However, the unique anatomical features of the brain and susceptibility of this organ to irreversible tissue damage have made immunotherapy especially challenging in the setting of glioma. With safety concerns in mind, multiple teams have initiated clinical trials using CAR T-cells in glioma patients. The valuable lessons learnt from those trials highlight critical areas for further improvement: tackling the issues of the antigen presentation and T-cell homing in the brain, immunosuppression in the glioma microenvironment, antigen heterogeneity and off-tumor toxicity, and the adaptation of existing clinical therapies to reflect the intricacies of immune response in the brain. This review summarizes the up-to-date clinical outcomes of CAR T-cell clinical trials in glioma patients and examines the most pressing hurdles limiting the efficacy of these therapies. Furthermore, this review uses these hurdles as a framework upon which to evaluate cutting-edge pre-clinical strategies aiming to overcome those barriers

Genetically Engineered T-Cells for Malignant Glioma: Overcoming the Barriers to Effective Immunotherapy

Malignant gliomas carry a dismal prognosis. Conventional treatment using chemo- and radiotherapy has limited efficacy with adverse events. Therapy with genetically engineered T-cells, such as chimeric antigen receptor (CAR) T-cells, may represent a promising approach to improve patient outcomes owing to their potential ability to attack highly infiltrative tumors in a tumor-specific manner and possible persistence of the adaptive immune response. However, the unique anatomical features of the brain and susceptibility of this organ to irreversible tissue damage have made immunotherapy especially challenging in the setting of glioma. With safety concerns in mind, multiple teams have initiated clinical trials using CAR T-cells in glioma patients. The valuable lessons learnt from those trials highlight critical areas for further improvement: tackling the issues of the antigen presentation and T-cell homing in the brain, immunosuppression in the glioma microenvironment, antigen heterogeneity and off-tumor toxicity, and the adaptation of existing clinical therapies to reflect the intricacies of immune response in the brain. This review summarizes the up-to-date clinical outcomes of CAR T-cell clinical trials in glioma patients and examines the most pressing hurdles limiting the efficacy of these therapies. Furthermore, this review uses these hurdles as a framework upon which to evaluate cutting-edge pre-clinical strategies aiming to overcome those barriers

Chronic stress-induced changes in REM sleep on theta oscillations in the rat hippocampus and amygdala

The present study investigated the effect of Chronic Immobilization Stress (CIS) on theta oscillations in the hippocampus and amygdala during Rapid Eye Movement (REM) sleep. Adult male Wistar rats were subjected to 2 h of CIS daily for 10 days. Polysomnographic recordings with Electroencephalogram (EEG) from hippocampus (CA3 and CA1 subregion) and lateral nucleus of amygdala (LA) were carried out after termination of CIS protocol on the 7th, 14th and 21st day. The results showed a bimodal distribution on the total REM sleep duration in CIS rats: group of rats exhibited increased REM sleep duration considered as a stress-enhanced REM (SER) and rats with reduced REM sleep as stress-reduced REM sleep (SRR) group. The bimodal distribution in REM sleep was continued to exhibit even after 21 days of termination of stress, showing increased REM sleep in SER and reversible REM sleep in SRR rats. In addition to changes in sleep, increased REM sleep in SER rats was associated with attenuated theta activity in the hippocampus and amygdala, while the SRR rats did not show attenuated theta activities during the stress recovery period. Thus, the study demonstrates the dependence of synchronized amygdalo-hippocampal theta activity with the CIS-induced enhanced REM sleep duration. This raises the possibility that CIS-induced manifestations of the anxiety may be associated with synchronized theta oscillations in the hippocampus and amygdala

Considerations when treating high-grade pediatric glioma patients with immunotherapy

IntroductionChildren with high-grade gliomas (pHGGs) represent a clinical population in substantial need of new therapeutic options given the inefficacy and toxicity of current standard-of-care modalities. Although immunotherapy has emerged as a promising modality, it has yet to elicit a significant survival benefit for pHGG patients. While preclinical studies address a variety of underlying challenges, translational clinical trial design and management also need to reflect the most updated progress and lessons from the field.Areas coveredThe authors will focus our discussion on the design of clinical trials, the management of potential toxicities, immune monitoring, and novel biomarkers. Clinical trial design should integrate appropriate patient populations, novel, and preclinically optimized trial design, and logical treatment combinations, particularly those which synergize with standard of care modalities. However, there are caveats due to the nature of immunotherapy trials, such as patient selection bias, evidenced by the frequent exclusion of patients on high-dose corticosteroids. Robust immune-modulating effects of modern immunotherapy can have toxicities. As such, it is important to understand and manage these, especially in pHGG patients.Expert opinionAdequate integration of these considerations should allow us to effectively gain insights on biological activity, safety, and biomarkers associated with benefits for patients

Nonintrusive TCP Connection Admission Control for Bandwidth Management of an Internet Access Link

We describe our approach to monitoring and managing the bandwidth of an Internet edge link with a view towards certain quality of service objectives for the services it carries. Such a link could be, for example, a campus's Internet access link, or a small ISP's backbone access link. We use SNMP polls and packet snooping to obtain traffic statistics, and TCP admission control for bandwidth management. Our implementation is completely nonintrusive: we use Ethernet packet capture in the promiscuous mode for traffic analysis, and use IP masquerading for blocking new TCP connections. This approach has been implemented by us in a software system for traffic management. We first justify our approach with a simple analytical model. We give an overview of our software implementation, and discuss some implementation issues. Then we provide measurement results that show the effectiveness of the techniques. Keywords: Internet bandwidth management; admission control; quality of service; TCP perfo..

Crystalline silica-induced leukotrieneB4-dependent inflammation promotes lung tumor growth

Chronic exposure to crystalline silica (CS) causes silicosis, an irreversible lung inflammatory disease that may eventually lead to lung cancer. In this study, we demonstrate that in K-rasLA1 mice, CS exposure markedly enhances the lung tumor burden and genetic deletion of leukotriene B4 receptor1 (BLT1−/−) attenuates this increase. Pulmonary neutrophilic inflammation induced by CS is significantly reduced in BLT1−/−K-rasLA1 mice. CS exposure induces LTB4 production by mast cells and macrophages independent of inflammasome activation. In an air pouch model, CS-induced neutrophil recruitment is dependent on LTB4 production by mast cells and BLT1 expression on neutrophils. In an implantable lung tumor model, CS exposure results in rapid tumor growth and decrease survival that is attenuated in the absence of BLT1. These results suggest that LTB4/BLT1 axis sets the pace of CS-induced sterile inflammation that promotes lung cancer progression. This knowledge will facilitate development of immunotherapeutic strategies to fight silicosis and lung cancer