Cloudberry (Rubus chamaemorus) and its components as chemopreventive constituents in ApcMin mice and human colon adenocarcinoma cells

A diet rich in fruits and vegetables has been suggested to decrease the risk of colorectal cancer and the protective effects of these foods could probably be at least partly mediated by their polyphenolic compounds. The number of different polyphenols is huge and their effects on cells and tissues may differ. The promising results on chemopreventive effects of ellagitannin-rich pomegranate suggest that the other dietary sources of ellagitannins may also be effective in cancer prevention. The overall aim of the research reported in this thesis was to study the effects of cloudberry (Rubus chamaemorus), which is rich in ellagitannins, on all phases of intestinal tumour development. Specifically, the effects of cloudberry on the first two phases of carcinogenesis, initiation and promotion, were studied using the ApcMin mouse model. In addition, the effects of cloudberry extract on the indicators of the last phase of cancer development, progression and metastasis, and particularly cancer cell migration that is essential for this phase, were studied using scattering and wound healing assays in hepatocyte growth factor-induced HT29 and HCA7 human colon adenocarcinoma cell lines. A whole-cloudberry diet containing 10% (w/w) freeze-dried cloudberries significantly decreased the number and size of intestinal adenomas in ApcMin mice. In contrast, cloudberry seeds, cloudberry pulp, or pure ellagic acid, when incorporated into the diets in concentrations that corresponded to the whole-cloudberry diet, had no apparent effect. The effects of cloudberry on the development of adenomas were also compared with the effects of bilberry (Vaccinium myrtillus), which is rich in anthocyanins. Both berries decreased the number of adenomas, but their effects of adenoma size were different: thus, the adenoma size was decreased by cloudberry and increased by bilberry in comparison to the control diet. The opposite effects of the berries on tumour growth were associated with the changes in the gut microbiota, intestinal immunity, and the expression of energy metabolism-related genes. The activation of Met, which is also known as hepatocyte growth factor receptor induces cell migration and is considered to play an important role in tumour metastasis. Cloudberry extract inhibited cell migration by inhibiting the activation of Met in hepatocyte growth factor-induced human HT29 and HCA7 colon adenocarcinoma cells, and thus Met signalling and consequent activation of the phosphatidylinositol 3-kinase/AKT pathway. The activation of Met signalling in the tumours of cloudberry-fed ApcMin mice was also inhibited. The results of the research presented in this thesis suggest that consumption of cloudberry or cloudberry preparations may reduce the risk of colon cancer, slow down the growth of colon adenomas, and have therapeutic value in reducing cancer progression and metastasis. Long-term studies in human subjects are needed to confirm these results.Runsaasti kasvikunnan tuotteita sisältävä ruokavalio voi pienentää vaaraa sairastua paksusuolisyöpään. Tämä syövältä suojaava vaikutus perustuu todennäköisesti ainakin osittain näiden tuotteiden sisältämiin polyfenolisiin yhdisteisiin. Näiden yhdisteiden kirjo on laaja ja niillä on runsaasti erilaisia vaikutuksia soluissa ja kudoksissa. Lupaavat tulokset ellagitanniinien syöpää estävistä vaikutuksista tekevät runsaasti ellagitanniineja sisältävästä lakasta (Rubus chamaemorus) mielenkiintoisen tutkimuskohteen. Tämän työn tavoitteena oli tutkia lakan ja sen komponenttien vaikutuksia paksusuolisyövän kehittymiseen sen kaikissa vaiheissa. Lakan vaikutusta uusien kasvainten muodostumiseen ja jo olemassa olevien kasvainten kasvuun tutkittiin käyttäen ApcMin-hiirikantaa. Näille hiirille muodostuu spontaanisti kasvaimia, koska niillä on geenivirhe kasvainten kasvua estävässä geenissä. Paksusuolisyövän kehityskaaren viimeiselle eli progressiovaiheelle ominaista syöpäsolujen liikkumista ja soluissa tapahtuvaa signalointia tutkittiin ihmisen paksusuolisyövän soluviljelmissä. Ruokavalio, joka sisälsi 10 % painosta pakastekuivattua lakkaa, vähensi merkitsevästi sekä muodostuvien kasvainten lukumäärää että niiden kokoa ApcMin-hiirillä. Sen sijaan hiirillä, joiden ruoassa oli kokonaisena jauhetun lakan sijaan pelkästään lakan siemeniä, hedelmälihaa tai ellagihappoa, kasvainten kehittymisessä ei havaittu muutoksia vertailuryhmään nähden. Vertailun vuoksi tutkittiin myös runsaasti antosyaaneja sisältävän mustikan (Vaccinium myrtillus) vaikutusta kasvainten kehittymiseen. Mustikka pienensi muodostuvien kasvainten lukumäärää, mutta ei estänyt kasvainten koon kasvua vertailuryhmään nähden. Lakan ja mustikan erilaiset vaikutukset kasvainten kokoon selittyivät niiden erilaisilla vaikutuksilla suoliston mikrobiflooraan ja soluvälitteiseen immuunivasteeseen sekä solun energia-aineenvaihduntaan liittyvien geenien ilmentymiseen suolen limakalvossa. Näiden havaintojen perusteella lakka voi mahdollisesti estää paksusuolikasvainten muodostumista ja hidastaa niiden kasvua. Lakkauute hidasti merkitsevästi ihmisen paksusuolen syöpäsolujen (HT29 ja HCA7) liikkumista. Solujen liikkumisen hidastuminen oli yhteydessä siihen, että lakkauute vähensi Met-reseptorin välittämää solusignalointia. Koska syöpäsolujen liikkuminen on yksi syövän leviämisen edellytyksistä, lakka ja sen sisältämät yhdisteet voivat mahdollisesti estää syövän etenemistä. Tähän väitöskirjaan sisältyvien tutkimusten perusteella lakka ja sen sisältämät yhdisteet voivat estää kokeellisen paksusuolisyövän kehittymistä sen kaikissa vaiheissa. Näillä havainnoilla voi olla merkitystä pyrittäessä estämään paksusuolisyövän syntyä ja hidastamaan sen etenemistä sekä mahdollisesti jo hoidetun syövän uusiutumista. Havainnot tulisi kuitenkin pyrkiä varmistamaan pitkäkestoisissa tutkimuksissa ihmisillä

Partial replacement of red and processed meat with legumes: a modelling study of the impact on nutrient intakes and nutrient adequacy on the population level

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Western diet enhances intestinal tumorigenesis in Min/ plus mice, associating with mucosal metabolic and inflammatory stress and loss of Apc heterozygosity

Western-type diet (WD) is a risk factor for colorectal cancer, but the underlying mechanisms are poorly understood. We investigated the interaction of WD and heterozygous mutation in the Apc gene on adenoma formation and metabolic and immunological changes in the histologically normal intestinal mucosa of Apc(Min/+) (Min/ +) mice. The diet used was high in saturated fat and low in calcium, vitamin D, fiber and folate. The number of adenomas was twofold higher in the WD mice compared to controls, but adenoma size, proliferation or apoptosis did not differ. The ratio of the MM to wild-type allele was higher in the WD mice, indicating accelerated loss of Apc heterozygosity (LOH). Densities of intraepithelial CD3 epsilon(+) T lymphocytes and of mucosal FoxP3(+) regulatory T cells were higher in the WD mice, implying inflammatory changes. Western blot analyses from the mucosa of the WD mice showed suppressed activation of the ERK and AKT pathways and a tendency for reduced activation of the mTOR pathway as measured in phosphoS6/S6 levels. The expression of pyruvate dehydrogenase kinase 4 was up-regulated in both mRNA and protein levels. Gene expression analyses showed changes in oxidation/reduction, fatty acid and monosaccharide metabolic pathways, tissue organization, cell fate and regulation of apoptosis. Together, our results suggest that the high-risk Western diet primes the intestine to tumorigenesis through synergistic effects in energy metabolism, inflammation and oxidative stress, which culminate in the acceleration of LOH of the Apc gene. (C) 2016 Elsevier Inc. All rights reserved.Peer reviewe

Cancer-Predicting Gene Expression Changes in Colonic Mucosa of Western Diet Fed Mlh1(+/-) Mice

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COPD-potilaiden kuntouttava hoitotyö sairaalassa : Posteri potilashuoneisiin

TIIVISTELMÄ  Toteutimme toiminnallisen opinnäytetyömme yhteistyössä Päijät-Hämeen keskussairaalan keuhkosairauksien osaston henkilökunnan kanssa. Opinnäytetyömme tarkoituksena oli edistää kuntouttavan työotteen lisäämistä toimeksiantajamme yksikössä.  Toiminnallisena tuotoksena toteutimme yhteistyönä posterin jokaiseen potilashuoneeseen. Rajasimme opinnäytetyömme aiheeksi keuhkoahtaumapotilaiden toimintakykyä ylläpitävän hoitotyön, koska saimme selkeämmän tuotoksen keskittymällä yhteen potilas ryhmään ja kyseinen potilasryhmä on iso osa osaston potilaita.  Posterin on tarkoitus tukea osaston jokapäiväistä kuntouttavan työotteen toteuttamista ja se on suunnattu potilaille sekä henkilökunnalle. Posterin tarkoituksena on kertoa esimerkein, miten osastolla toteutetaan toimintakykyä ylläpitävää hoitotyötä keuhkoahtaumapotilaille. Posteri tuotettiin A3-kokoisena, jossa kerrottiin konkreettisia ohjeita, miten potilaiden voimavaroja voidaan tukea keuhkosairauksien osastolla. Posteri tukee osaston kuntouttavan hoitotyön toimintamallia. ABSTRACT This functional thesis was conducted in collaboration with the pulmonary disease department of the Päijät-Häme Central Hospital. The purpose of the thesis was to promote rehabilitative nursing in the client unit. The functional output was a poster for each patient room. The thesis was limited to nursing care for chronic obstructive pulmonary disease (COPD) patients because it was clearer to focus on one patient group and that patient group is a large part of the department's patients. The poster is intended to support the implementation of the daily rehabilitation work of the department and is aimed at patients and staff. The purpose of the poster is to give examples of how the department conducts functional nursing care for COPD patients. The A3-sized poster provides concrete instructions on how to support the patients' resources in the pulmonary disease department. The poster supports the department's rehabilitative nursing model

Changes in intestinal immunity, gut microbiota and expression of energy metabolism-related genes explain adenoma growth in bilberry and cloudberry-fed ApcMin mice

We showed previously that ellagitannin-rich cloudberries and anthocyanin-rich bilberries reduce the number of intestinal adenomas in multiple intestinal neoplasia/+ (ApcMin) mice. We also found that cloudberries decreased the size of adenomas, while bilberries increased it. Here we hypothesized that the difference in adenoma growth could be explained by dissimilar effects of the berries on intestinal immune responses and gut microbiota, potentially driven by the distinct polyphenol compositions of the two berries. Our objective was to investigate lymphocyte subtypes and the predominant cecal bacterial diversity in mice fed with bilberries and cloudberries, and to analyze global gene expression profiles in the intestinal mucosa. Immunostainings of CD3+ T lymphocytes, FoxP3+ regulatory T lymphocytes and CD45R+ B lymphocytes revealed a smaller ratio of intraepithelial to all mucosal CD3+ T lymphocytes in the cloudberry-fed mice compared to controls, suggesting an attenuation of inflammation. Bilberry feeding induced no changes in the density of any of the lymphocyte subtypes. The predominant bacterial diversity in cecal contents, analyzed using PCR-denaturating gradient gel electrophoresis, was higher in the bilberry group than in the control or cloudberry groups. The microbial profiles of cloudberry-fed mice clustered together and were associated with small adenoma size. Pathway analyses of gene expression data showed that cloudberry downregulated and bilberry upregulated the expression of energy metabolism-related genes in the intestinal mucosa. In conclusion, attenuation of intestinal inflammation, changes in microbial profiles and downregulation of mucosal energy metabolism may account for the smaller adenoma size in cloudberry-fed mice in comparison to bilberry-fed mice

Effects of partial replacement of red and processed meat with non-soya legumes on bone and mineral metabolism and amino acid intakes in BeanMan randomised clinical trial

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Ellagitannin-rich cloudberry inhibits hepatocyte growth factorinduced cell migration and phosphatidylinositol 3-kinase/AKT activation in colon carcinoma cells and tumors in Min mice

Berries have been found to inhibit colon carcinogenesis in animal models, and thus represent a potential source of compounds for prevention and treatment of colorectal cancer. The mechanistic basis for their effects is not well understood. We used human colon carcinoma cells and Min mice to investigate the effects of ellagitannin-rich cloudberry (Rubus chamaemorus) extract on cancer cell migration and underlying cell signaling. Intrinsic and hepatocyte growth factor (HGF) -induced cell motility in human HT29 and HCA7 colon carcinoma cells was assessed carrying out cell scattering and scratch wound healing assays using time-lapse microscopy. Activation of Met, AKT, and ERK in cell lines and tumors of cloudberry-fed Min mice were determined using immunoprecipitation, Western blot and immunohistochemical analyses. Cloudberry extract significantly inhibited particularly HGF-induced cancer cell migration in both cell lines. Cloudberry extract inhibited the Met receptor tyrosine phosphorylation by HGF and strongly suppressed HGF-induced AKT and ERK activation in both HT29 and HCA7 cells. Consistently, cloudberry feeding (10% w/w freezedried berries in diet for 10 weeks) reduced the level of active AKT and prevented phosphoMet localization at the edges in tumors of Min mice. These results indicate that cloudberry reduces tumor growth and cancer cell motility by inhibiting Met signaling and consequent activation of phosphatidylinositol 3-kinase/AKT in vitro and in tumors in vivo. As the Met receptor is recognized to be a major target in cancer treatment, our results suggest that dietary phytochemicals may have therapeutic value in reducing cancer progression and metastasis.peerReviewe

Partial substitution of red or processed meat with plant‑based foods and the risk of type 2 diabetes

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