Interferon-γ primes macrophages for pathogen ligand-induced killing via a caspase-8 and mitochondrial cell death pathway.

Cell death plays an important role during pathogen infections. Here, we report that interferon-γ (IFNγ) sensitizes macrophages to Toll-like receptor (TLR)-induced death that requires macrophage-intrinsic death ligands and caspase-8 enzymatic activity, which trigger the mitochondrial apoptotic effectors, BAX and BAK. The pro-apoptotic caspase-8 substrate BID was dispensable for BAX and BAK activation. Instead, caspase-8 reduced pro-survival BCL-2 transcription and increased inducible nitric oxide synthase (iNOS), thus facilitating BAX and BAK signaling. IFNγ-primed, TLR-induced macrophage killing required iNOS, which licensed apoptotic caspase-8 activity and reduced the BAX and BAK inhibitors, A1 and MCL-1. The deletion of iNOS or caspase-8 limited SARS-CoV-2-induced disease in mice, while caspase-8 caused lethality independent of iNOS in a model of hemophagocytic lymphohistiocytosis. These findings reveal that iNOS selectively licenses programmed cell death, which may explain how nitric oxide impacts disease severity in SARS-CoV-2 infection and other iNOS-associated inflammatory conditions.

Caspase-8-driven apoptotic and pyroptotic crosstalk causes cell death and IL-1ß release in X-linked inhibitor of apoptosis (XIAP) deficiency.

Genetic lesions in X-linked inhibitor of apoptosis (XIAP) pre-dispose humans to cell death-associated inflammatory diseases, although the underlying mechanisms remain unclear. Here, we report that two patients with XIAP deficiency-associated inflammatory bowel disease display increased inflammatory IL-1ß maturation as well as cell death-associated caspase-8 and Gasdermin D (GSDMD) processing in diseased tissue, which is reduced upon patient treatment. Loss of XIAP leads to caspase-8-driven cell death and bioactive IL-1ß release that is only abrogated by combined deletion of the apoptotic and pyroptotic cell death machinery. Namely, extrinsic apoptotic caspase-8 promotes pyroptotic GSDMD processing that kills macrophages lacking both inflammasome and apoptosis signalling components (caspase-1, -3, -7, -11 and BID), while caspase-8 can still cause cell death in the absence of both GSDMD and GSDME when caspase-3 and caspase-7 are present. Neither caspase-3 and caspase-7-mediated activation of the pannexin-1 channel, or GSDMD loss, prevented NLRP3 inflammasome assembly and consequent caspase-1 and IL-1ß maturation downstream of XIAP inhibition and caspase-8 activation, even though the pannexin-1 channel was required for NLRP3 triggering upon mitochondrial apoptosis. These findings uncouple the mechanisms of cell death and NLRP3 activation resulting from extrinsic and intrinsic apoptosis signalling, reveal how XIAP loss can co-opt dual cell death programs, and uncover strategies for targeting the cell death and inflammatory pathways that result from XIAP deficiency.

The Pyroptotic Cell Death Effector Gasdermin D Is Activated by Gout-Associated Uric Acid Crystals but Is Dispensable for Cell Death and IL-1ß Release.

The pyroptotic cell death effector gasdermin D (GSDMD) is required for murine models of hereditary inflammasome-driven, IL-1ß-dependent, autoinflammatory disease, making it an attractive therapeutic target. However, the importance of GSDMD for more common conditions mediated by pathological IL-1ß activation, such as gout, remain unclear. In this study, we address whether GSDMD and the recently described GSDMD inhibitor necrosulfonamide (NSA) contribute to monosodium urate (MSU) crystal-induced cell death, IL-1ß release, and autoinflammation. We demonstrate that MSU crystals, the etiological agent of gout, rapidly activate GSDMD in murine macrophages. Despite this, the genetic deletion of GSDMD or the other lytic effector implicated in MSU crystal killing, mixed lineage kinase domain-like (MLKL), did not prevent MSU crystal-induced cell death. Consequently, GSDMD or MLKL loss did not hinder MSU crystal-mediated release of bioactive IL-1ß. Consistent with in vitro findings, IL-1ß induction and autoinflammation in MSU crystal-induced peritonitis was not reduced in GSDMD-deficient mice. Moreover, we show that the reported GSDMD inhibitor, NSA, blocks inflammasome priming and caspase-1 activation, thereby preventing pyroptosis independent of GSDMD targeting. The inhibition of cathepsins, widely implicated in particle-induced macrophage killing, also failed to prevent MSU crystal-mediated cell death. These findings 1) demonstrate that not all IL-1ß-driven autoinflammatory conditions will benefit from the therapeutic targeting of GSDMD, 2) document a unique mechanism of MSU crystal-induced macrophage cell death not rescued by pan-cathepsin inhibition, and 3) show that NSA inhibits inflammasomes upstream of GSDMD to prevent pyroptotic cell death and IL-1ß release.

Antigen-based vaccination and prevention of type 1 diabetes.

Insulin-dependent or type 1 diabetes (T1D) is a paradigm for prevention of autoimmune disease: Pancreatic ß-cell autoantigens are defined, at-risk individuals can be identified before the onset of symptoms, and autoimmune diabetes is preventable in rodent models. Intervention in asymptomatic individuals before or after the onset of subclinical islet autoimmunity places a premium on safety, a requirement met only by lifestyle-dietary approaches or autoantigen-based vaccination to induce protective immune tolerance. Insulin is the key driver of autoimmune ß-cell destruction in the nonobese diabetic (NOD) mouse model of T1D and is an early autoimmune target in children at risk for T1D. In the NOD mouse, mucosal administration of insulin induces regulatory T cells that protect against diabetes. The promise of autoantigen-specific vaccination in humans has yet to be realized, but recent trials of oral and nasal insulin vaccination in at-risk humans provide grounds for cautious optimism.

Study the charging process of moving quantum batteries inside cavity.

In quantum mechanics, quantum batteries are devices that can store energy by utilizing the principles of quantum mechanics. While quantum batteries has been investigated largely theoretical, recent research indicates that it may be possible to implement such a device using existing technologies. The environment plays an important role in the charging of quantum batteries. If a strong coupling exists between the environment and the battery, then battery can be charged properly. It has also been demonstrated that quantum battery can be charged even in weak coupling regime just by choosing a suitable initial state for battery and charger. In this study, we investigate the charging process of open quantum batteries mediated by a common dissipative environment. We will consider a wireless-like charging scenario, where there is no external power and direct interaction between charger and battery. Moreover, we consider the case in which the battery and charger move inside the environment with a particular speed. Our results demonstrate that the movement of the quantum battery inside the environment has a negative effect on the performance of the quantum batteries during the charging process. It is also shown that the non-Markovian environment has a positive effect on improving battery performance.

Curcumin-loaded porous particles functionalized with pH-responsive cell-penetrating peptide for colorectal cancer targeted drug delivery.

The anticancer properties of curcumin have been broadly examined in several shapes, such as nanoparticles and nanocomposite structures. Despite its benefits, curcumin also has some disadvantages, including rapid metabolism, poor absorption, and rapid systemic excretion. Therefore, numerous strategies have been used to increase curcumin's bioavailability. One of these approaches is the use of porous particles like aerogels as drug carriers. Aerogels are special due to their peculiar physical structure. They have a high specific surface area, a significant amount of porosity, and a solid composition, which make them a good choice for drug delivery systems. In the present study, a pH-sensitive aerogel was constructed and evaluated for targeted drug delivery of curcumin to colon cancer. To control the release of curcumin, trehalose was used as a coating agent, and PLP (poly(l-lysine isophthalamide)) was used as a targeted drug delivery agent. PLP is a pseudo-peptidic polymer that increases the cell permeability. In order to investigate and compare the synthesized aerogel before and after loading curcumin and coating with trehalose, physicochemical characterization analyses were performed. Finally, the efficacy of the final formulation was evaluated on HT29 colon cells using the cell bioavailability test. The results indicated the successful synthesis of the aerogel with porous structure with solitary cavities. The trehalose coating performed well, preventing drug release at lower pH but allowing the drug to be released at its intended site. The designed curcumin-loaded porous particles functionalized with PLP showed significant efficacy due to increasing penetration of curcumin into cells, and has potential for use as a new drug carrier with dual effectivity in cancer therapy.

Comparison of the effects of canola oil versus sunflower oil on the biochemical markers of bone metabolism in osteoporosis.

BACKGROUND: Recently, omega-3 fatty acids are in the center of attention for their potent anti-inflammatory effects. Osteoporosis as a chronic senile disease is associated with inflammation, and the role of inflammatory mediators has been demonstrated in the recent years. The beneficial effects of n-3 fatty acids on bone were proven in many animal studies, while to date, no conclusive data is available in human. The aim of this study was to evaluate the impact of n-3 fatty acids on bone biomarkers in osteoporotic women. MATERIAL AND METHODS: Forty osteoporotic post-menopausal women were recruited in the study and randomized in receiving either 40 g canola oil or the same amount sunflower oil per day as their dietary oil for 3 months. Serum levels of osteocalcin, bone alkaline phosphatase (BALP), N telo peptide collagen (NTX) and 25- hydroxy vitamin D3 were measured at baseline and at the end of the third month in both groups. RESULTS: In the canola oil group, BALP and NTX were increased after 3 months while Osteocalcin decreased in both groups slightly; however,none of these changes were significant. In both groups, serum vitamin D3 was increased significantly; however, this change between groups was not significant. CONCLUSION: Canola oil did not affect bone formation and resorption significantly after 3 months consumption. Further investigations with longer follow up are recommended.

Ubiquitylation of RIPK3 beyond-the-RHIM can limit RIPK3 activity and cell death.

Pathogen recognition and TNF receptors signal via receptor interacting serine/threonine kinase-3 (RIPK3) to cause cell death, including MLKL-mediated necroptosis and caspase-8-dependent apoptosis. However, the post-translational control of RIPK3 is not fully understood. Using mass-spectrometry, we identified that RIPK3 is ubiquitylated on K469. The expression of mutant RIPK3 K469R demonstrated that RIPK3 ubiquitylation can limit both RIPK3-mediated apoptosis and necroptosis. The enhanced cell death of overexpressed RIPK3 K469R and activated endogenous RIPK3 correlated with an overall increase in RIPK3 ubiquitylation. Ripk3 K469R/K469R mice challenged with Salmonella displayed enhanced bacterial loads and reduced serum IFNγ. However, Ripk3 K469R/K469R macrophages and dermal fibroblasts were not sensitized to RIPK3-mediated apoptotic or necroptotic signaling suggesting that, in these cells, there is functional redundancy with alternate RIPK3 ubiquitin-modified sites. Consistent with this idea, the mutation of other ubiquitylated RIPK3 residues also increased RIPK3 hyper-ubiquitylation and cell death. Therefore, the targeted ubiquitylation of RIPK3 may act as either a brake or accelerator of RIPK3-dependent killing.

An appropriate test for diagnosis of gestational diabetes mellitus.

Early diagnosis of GDM is necessary to reduce maternal and fetal morbidity and mortality. As all approaches to diagnosis of GDM are costly and difficult, we meant to find an appropriate and simple way to perform this test. One-thousand six-hundred and fifty-three pregnant women were screened for GDM at 24-28 weeks of gestation. Initial screening was done by a glucose challenge test with 50 g glucose. If the 1-h blood glucose level exceeded 130 mg/dl, then a 3-h oral glucose tolerance test (OGTT) with 100 g glucose was performed, and diagnosis was established according to ADA criteria. For determining which plasma glucose level is the best test for diagnosis of GDM, we used receiver operative characteristic cures (ROC) by plotting sensitivity versus one minus specificity at different times of plasma glucose levels in OGTT. In 732 pregnant women with a positive GCT, a 2-h plasma glucose level above 150 mg/dl was the most powerful test for detecting GDM, which revealed a sensitivity and specificity of 0.84 (0.81-0.86) and 0.94 (CI: 0.92-0.96), respectively. The results of this study suggest that 2-h 100 g OGTT could be an appropriate approach to diagnose GDM, which is cost-effective and could reduce laboratory workload.

Effect of pan retinal photocoagulation on the serum levels of vascular endothelial growth factor in diabetic patients.

This study tests the hypothesis that subjects with proliferative diabetic retinopathy (PDR) have a detectable rise in levels of serum vascular endothelial growth factor (VEGF), which is an important regulator of angiogenesis. Our investigation aims to evaluate plasma VEGF changes after pan-retinal photocoagulation (PRP) in diabetic patients. Twenty-nine type two diabetic patients (17 male, 12 female: mean age 53.13±12.22 years) with PDR secondary to diabetes were studied. Blood samples were obtained before and at 2 months after the last PRP session. Serum VEGF levels were measured by ELISA. After PRP, the mean serum VEGF decreased, but this reduction was not remarkable (88.68±71.09 vs. 77.01±60.33 ng/ml) (P=0.18). There was a statistically significant difference in serum VEGF changes between patients who had regressed PDR with patients who had progressed PDR (-25.98±47.37 vs. 56.44±31.7 ng/ml) (P=0.003). Our results showed a significant reduction in levels of serum VEGF in the patients who had successful laser treatment. Our findings suggest that serum VEGF levels could be used for monitoring diabetic retinopathy outcome.

Necroptotic movers and shakers: cell types, inflammatory drivers and diseases.

The necroptotic cell death pathway has received significant attention for its ability to trigger inflammatory responses and its potential involvement in related conditions. Recent insights into the essential membrane damaging necroptotic pseudokinase effector, Mixed lineage kinase domain like (MLKL), have revealed a number of diverse MLKL functions that contribute to the inflammatory nature of necroptosis. Here we review distinct MLKL signalling roles and document the immunogenic molecules released by necroptosis. We discuss specific in vivo MLKL-driven responses, the activation of inflammasome complexes and innate lymphoid cells, which have been documented to drive disease. Finally, we list necroptotic competent cell types and their involvement in MLKL-driven cell death-associated and inflammatory-associated conditions.

The ubiquitylation of IL-1ß limits its cleavage by caspase-1 and targets it for proteasomal degradation.

Interleukin-1ß (IL-1ß) is activated by inflammasome-associated caspase-1 in rare autoinflammatory conditions and in a variety of other inflammatory diseases. Therefore, IL-1ß activity must be fine-tuned to enable anti-microbial responses whilst limiting collateral damage. Here, we show that precursor IL-1ß is rapidly turned over by the proteasome and this correlates with its decoration by K11-linked, K63-linked and K48-linked ubiquitin chains. The ubiquitylation of IL-1ß is not just a degradation signal triggered by inflammasome priming and activating stimuli, but also limits IL-1ß cleavage by caspase-1. IL-1ß K133 is modified by ubiquitin and forms a salt bridge with IL-1ß D129. Loss of IL-1ß K133 ubiquitylation, or disruption of the K133:D129 electrostatic interaction, stabilizes IL-1ß. Accordingly, Il1bK133R/K133R mice have increased levels of precursor IL-1ß upon inflammasome priming and increased production of bioactive IL-1ß, both in vitro and in response to LPS injection. These findings identify mechanisms that can limit IL-1ß activity and safeguard against damaging inflammation.

Inflammasomes and Cell Death: Common Pathways in Microparticle Diseases.

The accumulation of cellular and environmental microparticles has been linked to many diseases associated with tissue inflammation. These particulate-driven diseases include joint, lung, kidney, cardiovascular, and neurodegenerative disorders. Recently a conserved proinflammatory inflammasome signaling pathway elicited by such microparticles has become apparent. Here, we review disease-promoting microparticles and the mechanisms by which they trigger activation of the inflammasome complexes responsible for generating bioactive interleukin-1ß (IL-1ß) and inducing cell death. We highlight how microparticle-induced inflammasome and cell death responses diverge from canonical inflammasome activators, and discuss the preclinical and clinical targeting of inflammasomes to treat microparticle-driven diseases.

Maternal serum adiponectin concentration in gestational diabetes.

AIM: Adiponectin is an insulin sensitizing protein. Because gestational diabetes mellitus is associated with insulin resistance, we compared serum adiponectin levels in women with gestational diabetes mellitus and healthy pregnant women. STUDY DESIGN: Twenty-nine women with gestational diabetes and 26 women with impaired glucose tolerance were compared with 27 normal pregnant women in control group. Controls were matched for gestational age, age and body mass index (BMI) before pregnancy with two other groups. At 28 weeks of gestation serum concentration of adiponectin, insulin and insulin resistance (calculated by the homeostasis model assessment) were measured in three groups. MAIN FINDINGS: The serum adiponectin level in gestational diabetes (6379.31 + or - 1934.90 ng/ml), was significantly lower than the impaired glucose tolerance test (7384.61 + or - 1626.70 ng/ml) and control groups (7962.96 + or - 2667.20 ng/ml),(p = 0.02). Serum level of insulin and HOMA index in gestational diabetes were higher than the normal group (p > 0.05). In patients with gestational diabetes, there was a significant correlation between serum adiponectin level and BMI before pregnancy (r = -0.531, p = 0.013). Also, the correlation between maternal serum adiponectin levels and neonatal birth weight was not significant (r = -0.07, p value = 0.73). CONCLUSION: Our data show that serum adiponectin level was significantly lower in gestational diabetes in comparison with healthy pregnant women.

A nutritional status survey of older adults in long-term care in the Yazd province of Iran.

It is important to assess the nutritional status of older adults because of its role in ensuring health and quality of life and its association with functional status. The purpose of this study was to evaluate the nutritional status of an older adult population living in long-term care institutions in the Yazd province of Iran. Fifty elderly subjects were randomly selected from each of two long-term care institutions in Yazd. A 3-day food intake survey was conducted using the direct weighing method and anthropometric measurements for calculating body mass index (BMI) were also collected. Of the participants, 54% of women and 41% of men had a BMI less than 19.9 kg/m(2). The mean intakes of energy, protein, vitamins A and C, riboflavin, and niacin as well as the minerals calcium, phosphorus, and iron, were significantly less than Dietary Reference Intakes (DRIs) for both genders. Thiamin intake was more than adequate in both women and men. In our study, the majority of elderly subjects displayed a poor reported nutritional intake according to the DRIs. Our findings support the development of national nutrition plans for older adults living in long-term care institutions as an important necessity.

Prevalence of dry eye syndrome and diabetic retinopathy in type 2 diabetic patients.

BACKGROUND: This study was performed to assess the prevalence of dry eye syndrome and diabetic retinopathy (DR) in type 2 diabetic patients and their contributing factors. METHODS: 199 type 2 diabetic patients referred to Yazd Diabetes Research Center were consecutively selected. All Subjects were assessed by questionnaire about other diseases and drugs. Dry eye syndrome was assessed with Tear break up time tests and Schirmer. All the subjects underwent indirect ophthalmoscopy and retinal color photography. DR was graded according to early Treatment Diabetic Retinopathy (ETDRS) criteria. RESULTS: Of 199 subjects, 108 patients (54.3%) suffer from dry eye syndrome. Although dry eye syndrome was more common in older and female patients, this association was not significant. But there was significantly association between dry eye syndrome and duration of diabetes (P = 0.01). Dry eye syndrome was more frequent in diabetic patients with DR (P = 0.02). DR was found in 140 patients (70.35%), which included 34 patients (17.1%) with mild non proliferative DR (NPDR), 34 patients (17.1%) with moderate NPDR, 22 patients (11.1%) with severe NPDR and 25 patients (25.1%) with proliferative DR (PDR). There were significant relation between age, sex and duration of diabetes and DR. CONCLUSION: In this study the prevalence of dry eye syndrome was 54.3%. Diabetes and dry eyes appear to have a common association. Further studies need to be undertaken to establish an etiologic relationship. However, examination for dry eye should be an integral part of the assessment of diabetic eye disease.

Effect of opium addiction on diabetes.

BACKGROUND: Type 2 diabetes is a common disorder that is recognized as a major health problem in Iran. Diabetes is a major cause of morbidity, mortality, and economic burden to the society. Some people believe that smoking opium can reduce serum glucose and lipids in diabetes mellitus. This study was designed to compare blood glucose and lipids in opium addicts with non-addicts among patients with type 2 diabetes. METHODS: In this case, control study subjects were chosen from type 2 diabetic patients. Twenty-three males with type 2 diabetes and addicted to opium were selected as the case group, and 46 patients with no addiction to opioid drugs were chosen as control group. Blood Sugar (BS), glycated hemoglobin (HbA1C), lipids and microalbumin in urine were measured in two groups. RESULTS: Our results showed that the mean FBS and 2-hour post prandial were significantly different between two groups. (P = .04). No significant difference was observed in HbA1C, triglyceride, total cholesterol, LDL, and HDL between the two groups (P > .05). CONCLUSIONS: Our finding showed that while opium might decrease blood glucose temporarily, it had no clear and long-lasting effects on blood glucose, as it had no significant effect on HbA1c.

CD52 inhibits Toll-like receptor activation of NF-κB and triggers apoptosis to suppress inflammation.

Soluble CD52 is a small glycoprotein that suppresses T-cell activation, but its effect on innate immune cell function is unknown. Here we demonstrate that soluble CD52 inhibits Toll-like receptor and tumor necrosis factor receptor signaling to limit activation of NF-κB and thereby suppress the production of inflammatory cytokines by macrophages, monocytes and dendritic cells. At higher concentrations, soluble CD52 depletes the short-lived pro-survival protein MCL-1, contributing to activation of the BH3-only proteins BAX and BAK to cause intrinsic apoptotic cell death. In vivo, administration of soluble CD52 suppresses lipopolysaccharide (LPS)-induced cytokine secretion and other features of endotoxic shock, whereas genetic deletion of CD52 exacerbates LPS responses. Thus, soluble CD52 exhibits broad immune suppressive effects that signify its potential as an immunotherapeutic agent.

New polyacetylenes from Echinophora cinerea (Boiss.) Hedge et Lamond.

Echinophora cinerea aerial parts are used in folk medicine to cure gastric diseases and as a food seasoning in cheese and yogurt. Besides several pharmacological effects have been assigned to Echinophora spp., there is no phytochemical investigation on this genus other than our previous publication on flavonoids. An acetone extract of E. cinerea afforded three new (1-3) polyacetylenes, one rare monoterpenoid glycoside as verbenone-5-O-ß-D-glycopyranoside (4) and one prenylated coumarin as osthol (5). The structures of all new compounds were elucidated using modern spectroscopic methods, including 2D NMR and mass analyses. The potency of the compounds to induce cell death was determined on SKNMC, PC3 and MCF-7 cell lines using MTT method in which compounds 1 and 2 showed moderate cytotoxic effects, especially against PC3 cells.

Quercetin-3-O-ß-D-glucopyranoside, a dietary flavonoid, protects PC12 cells from H2O2-induced cytotoxicity through inhibition of reactive oxygen species.

Since flavonols are antioxidant agents, they could in principle, beneficially affect neurodegenerative diseases where reactive oxygen species are involved. Quercetin derivatives are the most abundant dietary flavonoids, and we have investigated the capacity of quercetin-3-O-ß-d-glucopyranoside (Q3G) isolated from Echinophora cinerea to protect PC12 cells from H2O2-induced cytotoxicity. Direct cytotoxic effects of H2O2 on PC12 in presence and absence of Q3G were evaluated. H2O2 induced cytotoxicity in a concentration dependent manner (IC50=118 ± 5.09 µM, 24h). Pretreatment of cells with non-toxic concentrations of Q3G protected cells from H2O2-induced cytotoxicity, leading to a decrease in the generation of reactive oxygen species. These observations qualify Q3G as an interesting dietary compound worth further investigation as a cytoprotective agent.