Aija Line

Effects of kaempferol and myricetin on inducible nitric oxide synthase expression and nitric oxide production in rats.

When administered as drugs or consumed as food components, polyphenolic compounds synthesized in plants interfere with intracellular signal transduction pathways, including pathways of nitric oxide synthase expression. However, effects of these compounds in vivo do not always correlate with nitric oxide synthase-inhibiting activities revealed in experiments with cultured cells. The initial goal of this work was to compare effects of flavonoids kaempferol and myricetin on inducible nitric oxide synthase mRNA and protein expression monitored by real-time RT-PCR and immunohistochemistry and to evaluate the impact of these effects on nitric oxide production in rat organs measured by means of electron paramagnetic resonance spectroscopy. Kaempferol and myricetin attenuated the lipopolysaccharide-induced outburst of inducible nitric oxide synthase gene expression; kaempferol also significantly decreased the lipopolysaccharide-induced outburst of inducible nitric oxide synthase protein expression in the liver. Myricetin decreased nitric oxide production in intact rat liver. Kaempferol did not decrease nitric oxide production neither in intact rats nor in the lipopolysaccharide-treated animals. Kaempferol even enhanced the lipopolysaccharide-induced increase of nitric oxide production in blood. Myricetin did not interfere with lipopolysaccharide effects. As both kaempferol and myricetin are known as inhibitors of inducible nitric oxide synthase expression, our results suggest that modifications of nitric oxide level in tissues by these compounds cannot be predicted from data about its effects on nitric oxide synthase expression or activity.

Biodistribution, Uptake and Effects Caused by Cancer-Derived Extracellular Vesicles

Extracellular vesicles (EVs) have recently emerged as important mediators of intercellular communication. They are released in the extracellular space by a variety of normal and cancerous cell types and have been found in all human body fluids. Cancer-derived EVs have been shown to carry lipids, proteins, mRNAs, non-coding and structural RNAs and even extra-chromosomal DNA, which can be taken up by recipient cells and trigger diverse physiological and pathological responses. An increasing body of evidence suggests that cancer-derived EVs mediate paracrine signalling between cancer cells. This leads to the increased invasiveness, proliferation rate and chemoresistance, as well as the acquisition of the cancer stem cell phenotype. This stimulates angiogenesis and the reprogramming of normal stromal cells into cancer-promoting cell types. Furthermore, cancer-derived EVs contribute to the formation of the pre-metastatic niche and modulation of anti-tumour immune response. However, as most of these data are obtained by in vitro studies, it is not entirely clear which of these effects are recapitulated in vivo. In the current review, we summarize studies that assess the tissue distribution, trafficking, clearance and uptake of cancer-derived EVs in vivo and discuss the impact they have, both locally and systemically.

Effects of lycopene, indole-3-carbinol, and luteolin on nitric oxide production and iNOS expression are organ-specific in rats.

Effects of Lycopene, Indole-3-Carbinol, and Luteolin on Nitric Oxide Production and iNOS Expression are Organ-Specific in Rats Natural compounds are known to modify NO content in tissues; however, the biological activity of polyphenol-rich food often does not correspond to the effects of individual polyphenols on NO synthase activity. The aim of this study was to see how natural compounds luteolin, indole-3-carbinol, and lycopene modify NO production in rat tissues and change the expression of the iNOS gene and protein. Indole-3-carbinol produced multiple effects on the NO level; it significantly decreased NO concentration in blood, lungs, and skeletal muscles and increased it in the liver. Indole-3-carbinol enhanced lipopolyssaccharide (LPS)-induced NO production in all rat organs. It decreased iNOS gene expression in the brain cortex of animals that did not receive LPS and up-regulated it in the LPS-treated animals. Lycopene increased the iNOS gene transcription rate in the brain cortex of LPS-treated animals. Luteolin did not modify NO production in any organ of LPS-untreated rats, nor did it affect gene expression in the liver. In the brain it slightly decreased iNOS gene expression. Luteolin decreased NO production in the blood of LPS-treated animals and the number of iNOS-positive cells in these animals. Our results suggest that changes in tissue NO levels caused by natural compounds cannot be predicted from their effect on NOS expression or activity obtained in model systems. This stresses the importance of direct measurements of NO and NOS expression in animal tissues. Djelovanje likopena, indol-3-karbinola i luteolina na stvaranje dušikova monoksida i ekspresiju iNOS-gena u štakora ovisi o organu Zna se da prirodni spojevi utječu na sadržaj NO u pojedinome tkivu. Ipak, biološka aktivnost hrane bogate polifenolima često ne odgovara djelovanju pojedinih polifenola na aktivnost NO-sintaze. Cilj je ovoga ispitivanja bio utvrditi kako prirodni spojevi luteolin, indol-3-karbinol i likopen utječu na stvaranje NO u tkivu štakora te kako utječu na ekspresiju gena i razine inducibilne sintaze dušikova monoksida (iNOS). Indol-3-karbinol višestruko je utjecao na razinu NO; značajno je snizio njegovu koncentraciju u krvi, plućima i skeletnim mišićima, ali ju je povisio u jetri. Također je povećao njegovo stvaranje u svim organima štakora koji su primili lipopolisaharide (LPS). U kori mozga životinja koje nisu primile LPS doveo je do smanjenja ekspresije iNOS-gena, a u onih koje su primile LPS do njegova povećanja. Likopen je doveo do ubrzane transkripcije iNOS-gena u kori mozga životinja koje su primile LPS, ali nije utjecao na stvaranje NO u životinja koje nisu primile LPS, odnosno na ekspresiju gena u jetri. U mozgu životinja koje nisu primile LPS doveo je do laganoga smanjenja ekspresije iNOS-gena. Luteolin je povećao stvaranje NO u krvi životinja koje su primile LPS te njihov broj iNOS-pozitivnih stanica. Naši rezultati upućuju na to da se promjene razina NO uzrokovane prirodnim spojevima ne daju predvidjeti na temelju ekspresije gena iNOS odnosno aktivnosti toga enzima dobivene na modelu. Stoga je veoma važno izravno mjeriti razine NO i ekspresiju NO-sintaze u tkivu životinja.

Proteome Analysis of Colorectal Cancer Cell Line SW480 Released Extracellular Vesicles

The detection and profiling of disease-specific extracellular vesicles (EVs) from body fluids has been challenging research area during recent years. However, the question – can EVs surface proteins be exploited as a credible tool for early cancer diagnosis – is still not answered. Objective of the current study was to find out whether hypoxia induces differences in protein profiles of EVs released from hypoxic human colorectal cancer cells SW480 (EVHyp) and EVs released from these cells grown in normoxic conditions – EVNorm. Obtained results show differences in EVs surface protein samples. Some protein fragments were found only or mostly in EVHyp surface protein samples. Finding of one or two such EVHyp protein fragments allows us to suggest that deciphered EVHyp surface proteins might be indices of hypoxia-induced proteome changes and might serve as a hint to find a cancer specific protein.

Abstract B85: Tertiary lymphoid structures in chemotherapy-treated and untreated lung squamous cell carcinoma patients

Tertiary lymphoid structures (TLS) form in chronically inflamed tissues and contain organized compartments of T cells, B cells and follicular dendritic cells (FDCs). TLS were first noticed in autoimmune diseases where they support ectopic germinal center (GC) reactions and facilitate the activation and infiltration of immune cells. In cancer B cell and mature DC markers have been used for TLS detection because of their frequent localisation in TLS, and show a significant association with improved survival in several tumor types. Together these studies have driven the interest in TLS as a potential site of anti-tumor immune response activation and/or facilitation of lymphocyte infiltration. Here we aimed to characterise TLS as a microanatomical structure excluding non-organised lymphocytic infiltrates and to determine their prognostic value and association with clinico-pathologic parameters in non-small cell lung cancer patients. We characterised TLS density and presence of GCs in tumor center and periphery using HE December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B85.