RNA interference

RNA interference is a naturally occurring process of gene silencing that reduces the expression of the gene from which the RNA sequence is derived, when added in a double-stranded form. RNA interference works as defense against viruses and transposons. The aim of this study was to optimize a protocol for RNA interference in mouse mammary epithelial HC11 cells by using different controls, and finally to silence the drug transporter BCRP to do an uptake-study with the BCRP-substrate mitoxantrone. The optimization of the protocol was performed with four different lipids, as well as sicontrol tox, negative control and a positive control Cyclophilin B. Sicontrol tox and the negative control were assessed with ATP determination. Real time PCR was performed to check the down-regulation of Cyclophilin B after RNAi treatment of the HC11 cells. The results of the optimization protocol showed that transfection for 48 h in the presence of lipid nr 1 diluted 1:4 times works best for transfection in HC11 cells. Cyclophilin B was down-regulated about 90 % with this protocol, and BCRP was down-regulated 80 %. The uptake-study with mitoxantrone showed that the BCRP gene silenced cells accumulated the double amount of mitoxantrone as compared to controls. In conclusion, the obtained results provide a promising model for screening BCRP substrates, which can be used to examine other transporters in mammary epithelial cells, as well

A model of secreting murine mammary epithelial HC11 cells comprising endogenous Bcrp/Abcg2 expression and function

Breast cancer resistance protein (Bcrp/Abcg2) and multidrug transporter 1 (Mdr1/Abcb1) are efflux proteins located in the apical membrane of mammary epithelial cells (MEC). Bcrp is induced in MEC during gestation and lactation, while Mdr1 is down-regulated during lactation. Numerous drugs and toxic compounds are known to be actively secreted into milk by Bcrp, but most chemicals have not been investigated in this respect, emphasizing the need for functional Bcrp studies in an established cell line with secreting mammary epithelial cells. The present study was undertaken to examine expressions of Bcrp and Mdr1 in mammary epithelial HC11 cells, derived from a mid-gestational murine mammary gland. In addition, Bcrp function was assessed by transport experiments with mitoxantrone (MX) in undifferentiated HC11 cells, in HC11 cells subjected to Bcrp RNA interference (RNAi) as well as in HC11 cells stimulated to differentiate by treatment with lactogenic hormones. Differentiated HC11 cells organized into alveolar-resembling structures and gene expression of the major milk protein B-casein was induced, whereas undifferentiated cells formed monolayers with lower B-casein expression. Bcrp and Mdr1 gene and protein were expressed in both undifferentiated and differentiated HC11 cells. Differentiation of HC11 cells resulted in increased Bcrp protein expression, while Mdr1 gene and protein expressions were reduced. The Bcrp inhibitor elacridar (GF120918) reduced secretion and increased accumulation of MX in both undifferentiated and differentiated HC11 cells. Silencing of the Bcrp gene caused an increased accumulation of MX. The results indicate that the HC11 cell model provides a promising tool to investigate transport of potential Bcrp substrates in mammary epithelial cells

Att sitta ikläm - Hur mellanchefen implementerar en strategi och hanterar direktiv från ledning och personal

Centrala begrepp: Organisationskultur, servicekultur, medarbetarskap, mellanchef, servicefokuserad strategi Syfte: Att med en fallstudie undersöka mellanchefens roll i att implementera en strategi genom sitt ledarskap i en större organisation. Studien fokuserar på butikschefens komplexa roll som mellanchef för att titta närmare på hur direktiv från ledningen och personalen hanteras. Metod: Studien har en kvalitativ ansats där semistrukturerade intervjuer använts som metod för insamlingen av empiri och har ett konstruktionistiskt synsätt. Empiri: Valet av fallstudieföretag gjordes utifrån deras servicefokuserade strategi och för att organisationen var tillräckligt stor för att passa studiens syfte. Empirin utgjordes av tio kvalitativa intervjuer varav sju stycken utfördes med butikschefer, en med regionchef och två från koncernledningen. Teoretiska perspektiv: Organisationskultur presenteras för att se hur det genom rekrytering och socialisation av medarbetare fungerar som ett styrinstrument. För att avgränsa ett bredare begrepp som organisationskultur lyfts servicekultur. Medarbetarskap avser hur ledaren arbetar med sin arbetsgrupp och anlägger på så sätt ett smalare perspektiv än servicekultur. Dessa perspektiv formas av varandra. Slutsatser: Fallstudieföretaget har en servicefokuserad strategi som butikschefen påverkar genom rekrytering, socialisation och användandet av styrinstrument. Dessa faktorer skapar i sin tur en servicekultur. Butikscheferna hanterar resultatorienterade faktorer från företaget och relationsorienterade faktorer från sina medarbetare genom tydlig kommunikation av butikens mål och uppmuntran av sina anställda

A Structural Equation Model Demonstrating the Relationship between Food Safety Background, Knowledge, Attitudes and Behaviour among Swedish Students

Traditionally, food safety knowledge has been seen as a factor in improving food safety behaviour. However, the relationship between knowledge and behavior is complex. The aim of the present study was to investigate self-reported data from 408 university students regarding food safety background, knowledge, attitudes, and behaviour using Structural Equation Model (SEM) to examine the influence of different factors on food safety behaviour. The SEM was applied to four factors derived from the data: Background, Knowledge, Attitude and Behaviour. The novelty of this current investigation is the inclusion of the Background factor (genus; experience of cooking and handling different food items; experience of a food safety education course; the foremost sources of food safety knowledge). The factors were constructed from variables with sufficient factor loadings and set up in a predetermined structure confirmed to be valid in previous studies. The results, demonstrated as regression coefficients between factors, confirm that the Background factor strongly influenced Knowledge (0.842). The Knowledge factor, in turn, strongly affected Attitude (0.605), while it did not directly affect Behaviour (0.301) in the same way as Attitude. Attitude had a stronger influence on Behaviour (0.438) than Knowledge. Thus, the Attitude factor seemed to play a mediating role between Knowledge and Behaviour. This indicates that students ' attitudes towards the importance of food safety may have an impact on their food safety behavior, which should have implications for the development of food safety education. This warrants further investigation and practical development

Workplace Alignment: An evaluation of office worker flexibility and workplace provision

Purpose – The paper aims to explore the relationship between office occupier work activity and workplace provision. It tests the proposition that location-fixed office workers are not as well-supported in the working environment as location-flexible office workers. The research also explores the perceptions of the workplace provision based upon the types of tasks completed at the desk-location, whether this was collaborative or focused. Design/methodology/approach – The research adopts a cross-sectional approach using an online questionnaire to collect data from several offices in the Middles East. The dataset consists of 405 responses. One-way ANOVA was conducted to understand the relationship between location flexibility and perception of productivity. In addition, a series of T-Test were used to evaluate the relationship between work activities and office environment. Findings – The results show that those workers who were location-fixed perceived the workplace provision to have a more negative impact on their productivity than those who had a greater level of location-flexibility, particularly with regards to noise levels and interruptions. In terms of types of activities, those that undertook more collaborative tasks valued the facilitation of creativity and interaction from the workplace provision. Research limitations/implications – The research has limitations as data collection was at one-point in time and therefore lacks the opportunity to undertake longitudinal analysis. However, the research gives greater insights into the alignment of office environments based on flexibility and work activity. Practical implications – The paper identifies implications for the design and development of office environments by identifying the need for office occupier activity profiles. These profiles can underpin data led design which should promote a tailored choice appropriate work setting that can maximise productivity. Originality/value – This paper contributes to the research area of workplace alignment. It establishes that optimal workplace alignment requires a better understanding of office occupier needs based on location-flexibility and work activity

Workplace productivity and office type: an evaluation of office occupier differences based on age and gender

Purpose Open plan office environments are considered to offer workplace productivity benefits because of the opportunities that they create for interaction and knowledge exchange, but more recent research has highlighted noise, distraction and loss of privacy as significant productivity penalties with this office layout. This study aims to investigate if the purported productivity benefits of open plan outweigh the potential productivity penalties. Design/methodology/approach Previous research suggests that office environments are experienced differently according to the gender and age of the occupier across both open-plan and enclosed configurations. Empirical research undertaken with office occupiers in the Middle East (N=220) led to evaluations to establish the impact different offices had on perceived productivity. Factor analysis was used to establish five underlying components of office productivity. The five factors are subsequently used as the basis for comparison between office occupiers based on age, gender and office type. Findings This research shows that benefits and penalties to workplace productivity are experienced equally across open-plan and enclosed office environments. The greatest impact on perceived workplace productivity however was availability of a variety of physical layouts, control over interaction and the 'downtime' offered by social interaction points. Male occupiers and those from younger generations were also found to consider the office environment to have more of a negative impact on their perceived workplace productivity compared to female and older occupiers. Originality/value The originality of this paper is that it develops the concept of profiling office occupiers with the aim of better matching office provision. This paper aims to establish different occupier profiles based on age, gender and office type. Data analysis techniques such as factor analysis and t-test analysis identify the need for different spaces so that occupiers can choose the most appropriate space to best undertake a particular work task. In addition, it emphasises the value that occupiers place on ‘downtime’ leading to the need for appropriate social space

The WID-BC-index identifies women with primary poor prognostic breast cancer based on DNA methylation in cervical samples

Genetic and non-genetic factors contribute to breast cancer development. An epigenome-based signature capturing these components in easily accessible samples could identify women at risk. Here, we analyse the DNA methylome in 2,818 cervical, 357 and 227 matched buccal and blood samples respectively, and 42 breast tissue samples from women with and without breast cancer. Utilising cervical liquid-based cytology samples, we develop the DNA methylation-based Women’s risk IDentification for Breast Cancer index (WID-BC-index) that identifies women with breast cancer with an AUROC (Area Under the Receiver Operator Characteristic) of 0.84 (95% CI: 0.80–0.88) and 0.81 (95% CI: 0.76–0.86) in internal and external validation sets, respectively. CpGs at progesterone receptor binding sites hypomethylated in normal breast tissue of women with breast cancer or in BRCA mutation carriers are also hypomethylated in cervical samples of women with poor prognostic breast cancer. Our data indicate that a systemic epigenetic programming defect is highly prevalent in women who develop breast cancer. Further studies validating the WID-BC-index may enable clinical implementation for monitoring breast cancer risk.publishedVersio

Antiprogestins reduce epigenetic field cancerization in breast tissue of young healthy women

Background: Breast cancer is a leading cause of death in premenopausal women. Progesterone drives expansion of luminal progenitor cells, leading to the development of poor-prognostic breast cancers. However, it is not known if antagonising progesterone can prevent breast cancers in humans. We suggest that targeting progesterone signalling could be a means of reducing features which are known to promote breast cancer formation. Methods: In healthy premenopausal women with and without a BRCA mutation we studied (i) estrogen and progesterone levels in saliva over an entire menstrual cycle (n = 20); (ii) cancer-free normal breast-tissue from a control population who had no family or personal history of breast cancer and equivalently from BRCA1/2 mutation carriers (n = 28); triple negative breast cancer (TNBC) biopsies and healthy breast tissue taken from sites surrounding the TNBC in the same individuals (n = 14); and biopsies of ER+ve/PR+ve stage T1–T2 cancers and healthy breast tissue taken from sites surrounding the cancer in the same individuals (n = 31); and (iii) DNA methylation and DNA mutations in normal breast tissue (before and after treatment) from clinical trials that assessed the potential preventative effects of vitamins and antiprogestins (mifepristone and ulipristal acetate; n = 44). Results: Daily levels of progesterone were higher throughout the menstrual cycle of BRCA1/2 mutation carriers, raising the prospect of targeting progesterone signalling as a means of cancer risk reduction in this population. Furthermore, breast field cancerization DNA methylation signatures reflective of (i) the mitotic age of normal breast epithelium and (ii) the proportion of luminal progenitor cells were increased in breast cancers, indicating that luminal progenitor cells with elevated replicative age are more prone to malignant transformation. The progesterone receptor antagonist mifepristone reduced both the mitotic age and the proportion of luminal progenitor cells in normal breast tissue of all control women and in 64% of BRCA1/2 mutation carriers. These findings were validated by an alternate progesterone receptor antagonist, ulipristal acetate, which yielded similar results. Importantly, mifepristone reduced both the TP53 mutation frequency as well as the number of TP53 mutations in mitotic-age-responders. Conclusions: These data support the potential usage of antiprogestins for primary prevention of poor-prognostic breast cancers

Views on primary prevention of cardiovascular disease - an interview study with Swedish GPs

Background: General practitioners (GPs) have gradually become more involved in the prevention of cardiovascular disease (CVD), both through more frequent prescribing of pharmaceuticals and by giving advice regarding lifestyle factors. Most general practitioners are now faced with decisions about pharmaceutical or non-pharmaceutical treatment for primary prevention every day. The aim of this study was to explore, structure and describe the views on primary prevention of cardiovascular disease in clinical practice among Swedish GPs. Methods: Individual interviews were conducted with 21 GPs in southern Sweden. The interview transcripts were analysed using a qualitative approach, inspired by phenomenography. Results: Two main categories of description emerged during the analysis. One was the degree of reliance on research data regarding the predictability of real risk and the opportunities for primary prevention of CVD. The other was the allocation of responsibility between the patient and the doctor. The GPs showed different views, from being convinced of an actual and predictable risk for the individual to strongly doubting it; from relying firmly on protection from disease by pharmaceutical treatment to strongly questioning its effectiveness in individual cases; and from reliance on prevention of disease by non-pharmaceutical interventions to a total lack of reliance on such measures. Conclusions: The GPs' different views, regarding the rationale for and practical management of primary prevention of CVD, can be interpreted as a reflection of the complexity of patient counselling in primary prevention in clinical practice. The findings have implications for development and implementation of standard treatment guidelines, regarding long-time primary preventive treatment