Anti-Inflammatory Properties of Acerola (Malpighia emarginata) Leaf and Ripe Fruit Genotypes for Protection Against LPS-Induced Inflammation in Macrophage Cells and Their Selectivity to Cyclooxygenase-2 (COX-2) Activity

Conventionally, NSAID has been proposed to have inhibitory action against COX and therefore has traditionally been used for treatment of acute and chronic inflammation. This study aims at exploring putative anti-inflammation mechanism of acerola. Previous studies have illustrated that phytochemicals like alkaloids, terpenoids, flavonoids, curcumin and phenolics have COX inhibitory activities as well. However, a natural occurring selective inhibitor of COX-2 that can modulate inflammation and can overcome the limitations of drugs like aspirin is still a priority. Aspirin is known to form an irreversible and non-competitive binding to COX which proves to be a potent cardiovascular protective agent. On the other hand, irreversible binding has implications by initiating inhibition of blood platelet aggregation. Acerola has been earlier studied for its antioxidant, antimicrobial, anti-inflammatory, anticancer, antigenotoxic and antihyperglycemic properties. In the present study, anti-inflammatory properties of acerola have been established where different genotypes of acerola fruit and leaf fractions were studied for their biological properties. A comparative study using TLC, LC-MS and bioassays using macrophages is employed to identify which groups of phytochemicals are responsible for scavenging and inflammation inhibitory effect of acerola. Initially, phytochemicals were extracted using methanolic and methanolic/acetone/water solvents which isolated different groups of compounds in two fractions, including polyphenols and a mixture of polyphenols/terpenoids, respectively. The two fractions were explored to elucidate mode of action for different acerola genotypes. Results indicated that the methanolic fractions of acerola showed higher activity exhibited suppression of ROS and partial decrease of nitric oxide levels in LPS-stimulated RAW264.7 macrophage cell line. This fraction also demonstrated inhibition of enzyme expression of COX-1/2. Moreover, BRS-238, a ripe fruit genotypes of acerola had a selective action against COX-2 - confirming the hypothesis that acerola’s mode of anti-inflammatory action is through selective inhibition of COX-2

Anti-Inflammatory Properties of Acerola (Malpighia emarginata) Leaf and Ripe Fruit Genotypes for Protection Against LPS-Induced Inflammation in Macrophage Cells and Their Selectivity to Cyclooxygenase-2 (COX-2) Activity

Conventionally, NSAID has been proposed to have inhibitory action against COX and therefore has traditionally been used for treatment of acute and chronic inflammation. This study aims at exploring putative anti-inflammation mechanism of acerola. Previous studies have illustrated that phytochemicals like alkaloids, terpenoids, flavonoids, curcumin and phenolics have COX inhibitory activities as well. However, a natural occurring selective inhibitor of COX-2 that can modulate inflammation and can overcome the limitations of drugs like aspirin is still a priority. Aspirin is known to form an irreversible and non-competitive binding to COX which proves to be a potent cardiovascular protective agent. On the other hand, irreversible binding has implications by initiating inhibition of blood platelet aggregation. Acerola has been earlier studied for its antioxidant, antimicrobial, anti-inflammatory, anticancer, antigenotoxic and antihyperglycemic properties. In the present study, anti-inflammatory properties of acerola have been established where different genotypes of acerola fruit and leaf fractions were studied for their biological properties. A comparative study using TLC, LC-MS and bioassays using macrophages is employed to identify which groups of phytochemicals are responsible for scavenging and inflammation inhibitory effect of acerola. Initially, phytochemicals were extracted using methanolic and methanolic/acetone/water solvents which isolated different groups of compounds in two fractions, including polyphenols and a mixture of polyphenols/terpenoids, respectively. The two fractions were explored to elucidate mode of action for different acerola genotypes. Results indicated that the methanolic fractions of acerola showed higher activity exhibited suppression of ROS and partial decrease of nitric oxide levels in LPS-stimulated RAW264.7 macrophage cell line. This fraction also demonstrated inhibition of enzyme expression of COX-1/2. Moreover, BRS-238, a ripe fruit genotypes of acerola had a selective action against COX-2 - confirming the hypothesis that acerola’s mode of anti-inflammatory action is through selective inhibition of COX-2

Immunomodulatory Pathways and Metabolism

Energy metabolism plays a vital role in normal physiology, adaptive responses and host defense mechanisms. Research throughout the last decade has shown evidence that immune pathways communicate with metabolic pathways to alter the metabolic status in response to physiological or pathological signals. In this thesis, I will explore how immunomodulatory molecules affect metabolic homeostasis and conversely, how metabolic sensing pathways modulate immune responses. The first part my work utilizes an immunomodulatory sugar motif to determine mechanisms by which immune cells influence metabolism. Specifically, I show in chapter 2 that lacto-N-fucopentaose III (LNFPIII), a motif used by pathogens to attenuate inflammation, is capable of improving systemic insulin sensitivity by increasing Il-10 production in macrophages and dendritic cells and subsequently improving white adipose tissue insulin sensitivity. Chapter 3 will address the observation that this same glycan is capable of directly activating Fxra in hepatocytes. This direct effect manifests as a reduction in high-fat-diet-induced hepatic triglyceride accumulation and improvement in liver function. Lastly, in chapter 4, I will discuss the role of metabolic regulators in the macrophage and how this affects the ability of the macrophage to kill bacteria. Specifically, I will show that lipid sensing nuclear receptors, such as Ppard and Pparg, are critical regulators of phagosomal function and bacterial killing. Macrophage-specific deletion of these receptors prevents efficient killing of Streptococcus pneumoniae, the causative bacterium in many cases of respiratory pneumonia. Ligand activation improves survival, suggesting a potential therapeutic role for Ppar activation during infection. Taken together, all the data suggest a critical role for the evolutionary interaction between metabolic and immune pathways. These interactions may be important when developing new therapeutics for complex metabolic and immunological dysfunctions

Immunomodulatory glycan LNFPIII alleviates hepatosteatosis and insulin resistance through direct and indirect control of metabolic pathways

Parasitic worms express host-like glycans to attenuate the immune response of human hosts. The therapeutic potential of this immunomodulatory mechanism in controlling metabolic dysfunction associated with chronic inflammation remains unexplored. We demonstrate here that administration of Lacto-N-fucopentaose III (LNFPIII), a LewisX containing immunomodulatory glycan found in human milk and on parasitic helminths, improves glucose tolerance and insulin sensitivity in diet-induced obese mice. This effect is mediated partly through increased Il-10 production by LNFPIII activated macrophages and dendritic cells, which reduces white adipose tissue inflammation and sensitizes the insulin response of adipocytes. Concurrently, LNFPIII treatment up-regulates nuclear receptor Fxr-α (or Nr1h4) to suppress lipogenesis in the liver, conferring protection against hepatosteatosis. At the signaling level, the extracellular signal-regulated kinase (Erk)-Ap1 pathway appears to mediate the effects of LNFPIII on both inflammatory and metabolic pathways. Our results suggest that LNFPIII may provide novel therapeutic approaches to treat metabolic diseases

Antibiotic-Induced Changes to the Host Metabolic Environment Inhibit Drug Efficacy and Alter Immune Function

Bactericidal antibiotics alter microbial metabolism as part of their lethality and can damage mitochondria in mammalian cells. In addition, antibiotic susceptibility is sensitive to extracellular metabolites, but it remains unknown whether metabolites present at an infection site can affect either treatment efficacy or immune function. Here, we quantify local metabolic changes in the host microenvironment following antibiotic treatment for a peritoneal Escherichia coli infection. Antibiotic treatment elicits microbiome-independent changes in local metabolites, but not those distal to the infection site, by acting directly on host cells. The metabolites induced during treatment, such as AMP, reduce antibiotic efficacy and enhance phagocytic killing. Moreover, antibiotic treatment impairs immune function by inhibiting respiratory activity in immune cells. Collectively, these results highlight the immunomodulatory potential of antibiotics and reveal the local metabolic microenvironment to be an important determinant of infection resolution

A diurnal serum lipid integrates hepatic lipogenesis and peripheral fatty acid utilization

Food intake increases the activity of hepatic de novo lipogenesis, which mediates the conversion of glucose to fats for storage or utilization. In mice, this program follows a circadian rhythm that peaks with nocturnal feeding1,2 and is repressed by Rev-erbα/β and an HDAC3-containing complex3–5 during the day. The transcriptional activators controlling rhythmic lipid synthesis in the dark cycle remain poorly defined. Disturbances in hepatic lipogenesis are also associated with systemic metabolic phenotypes6–8, suggesting that lipogenesis in the liver communicates with peripheral tissues to control energy substrate homeostasis. Here we identify a PPARδ-dependent de novo lipogenic pathway in the liver that modulates fat utilization by muscle via a circulating lipid. The nuclear receptor PPARδ controls diurnal expression of lipogenic genes in the dark/feeding cycle. Liver-specific PPARδ activation increases, while hepatocyte-Ppard deletion reduces, muscle fatty acid (FA) uptake. Unbiased metabolite profiling identifies PC(18:0/18:1), or 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), as a serum lipid regulated by diurnal hepatic PPARδ activity. PC(18:0/18:1) reduces postprandial lipid levels and increases FA utilization through muscle PPARα. High fat feeding diminishes rhythmic production of PC(18:0/18:1), whereas PC(18:0/18:1) administration in db/db mice improves metabolic homeostasis. These findings reveal an integrated regulatory circuit coupling lipid synthesis in the liver to energy utilization in muscle by coordinating the activity of two closely related nuclear receptors. These data implicate alterations in diurnal hepatic PPARδ-PC(18:0/18:1) signaling in metabolic disorders including obesity

Harnessing single-cell genomics to improve the physiological fidelity of organoid-derived cell types

Background: Single-cell genomic methods now provide unprecedented resolution for characterizing the component cell types and states of tissues such as the epithelial subsets of the gastrointestinal tract. Nevertheless, functional studies of these subsets at scale require faithful in vitro models of identified in vivo biology. While intestinal organoids have been invaluable in providing mechanistic insights in vitro, the extent to which organoid-derived cell types recapitulate their in vivo counterparts remains formally untested, with no systematic approach for improving model fidelity. Results: Here, we present a generally applicable framework that utilizes massively parallel single-cell RNA-seq to compare cell types and states found in vivo to those of in vitro models such as organoids. Furthermore, we leverage identified discrepancies to improve model fidelity. Using the Paneth cell (PC), which supports the stem cell niche and produces the largest diversity of antimicrobials in the small intestine, as an exemplar, we uncover fundamental gene expression differences in lineage-defining genes between in vivo PCs and those of the current in vitro organoid model. With this information, we nominate a molecular intervention to rationally improve the physiological fidelity of our in vitro PCs. We then perform transcriptomic, cytometric, morphologic and proteomic characterization, and demonstrate functional (antimicrobial activity, niche support) improvements in PC physiology. Conclusions: Our systematic approach provides a simple workflow for identifying the limitations of in vitro models and enhancing their physiological fidelity. Using adult stem cell-derived PCs within intestinal organoids as a model system, we successfully benchmark organoid representation, relative to that in vivo, of a specialized cell type and use this comparison to generate a functionally improved in vitro PC population. We predict that the generation of rationally improved cellular models will facilitate mechanistic exploration of specific disease-associated genes in their respective cell types. Electronic supplementary material The online version of this article (10.1186/s12915-018-0527-2) contains supplementary material, which is available to authorized users

Elucidating the Anti-Inflammatory and Anti-Diabetic Properties of Urolithin Bioactive Metabolites from Pecan

. Pecans are a rich source of ellagitannins which are not absorbed by humans, rather they are catabolized into urolithins by human gut microbiota and enterohepatic circulation. Urolithin A and B have been studied for their antioxidant, antimicrobial, anti-inflammatory, anticancer, and antihyperglycemic properties. This project combined a holistic approach of traditional medicine with scientific evidences. A diseased state, with proper human diet, could potentially be reversed by a combination of bioactive compounds. Polyphenols mitigate insulin resistance, obesity, inflammation, cardiovascular diseases and many others. The challenge is poor systemic bioavailability of these secondary metabolites. However, biotransformation of polyphenols by intestinal microflora conserves their functions and makes them bioavailable. The aim of this investigation was to elucidate role of oxidative stresses in anti-inflammatory effect of urolithin A and B and understand their role in a diabetic model. Urolithin A and B exerted anti-inflammatory properties in inflamed colon Caco-2 and endothelial HUVEC cells by suppressing ROS and upregulating LXR⍺. ⍺. In presence of LPS and PA and TNF ⍺ urolithin A suppressed increased levels of intracellular ROS and downregulated the pathways such as inflammation and insulin signaling in cell models. Whereas urolithin B, in a ROS independent mechanism worked in HUVEC, Caco-2 and C2C12 cells. Urolithin A and B suppressed insulin resistance induced by palmitic acid in muscle C2C12 cells and AML 12 by upregulating protein expression of pIRS and pAKT. They also downregulated the glucose production in insulin resistant hepatic AML 12 cells. Thereby, this study successfully demonstrated the multifaceted nature of urolithin A and B to ameliorate inflammation and insulin resistance in in-vitro model, key events in the scenario of the metabolic syndrom

Boosting Bacterial Metabolism to Combat Antibiotic Resistance

The metabolic state of a bacterial cell influences its susceptibility to antibiotics. In this issue, Peng et al. (2015) show that resistant bacteria can be sensitized to antibiotic treatment through the addition of exogenous metabolites that stimulate central metabolic pathways and increase drug uptake

PSORIASIS - A Mini Review

Psoriasis is a chronic inflammatory squamous cell disease of papules that is characterized by multiple remissions and relapses. This has long been considered a serious violation of keratinization. The successful use of traditional immune suppressants and new immune modulators in the treatment of psoriasis has led to the belief that psoriasis is, in fact, a disorder of Th1 cell immunodeficiency. Recent developments have led to several new discoveries such as the role of Th17 cells and evidence of skin barrier dysfunction in psoriasis similar to atopic dermatitis. Psoriasis is an atypical, systemic autoimmune disease of unknown cause, mainly affecting the skin, spreading further to the nails and joints and causing arthritis. The treatment protocol should be based on the patient’s condition. People with mild psoriasis can be treated primarily with topical medications, while people with moderate to severe psoriasis can be treated with systemic medications