Skills and Expertise
PCRCell CultureWestern Blot AnalysisCancer BiologyImmunohistochemistryMolecular BiologyCell BiologyBiochemistryCancerCancer ResearchCancer CellsProstate CancerCancer Cell LineCase-Control StudiesChemical BiologyStainingProstateVitamin DTissueAndrogensProgesteroneGrowth FactorsImmunochemistryVitamin A
May 2006 - Aug 2013
- Rauma, Satakunta, Finland
Infections and infectious diseases are a continuous threat to human health. A potential and promising weapon against bacterial growth and possibly the development of multi-drug resistant bacteria has been found in AntiMicrobial (nano)-Coatings (AMC). AMICI brings together stakeholders from different countries and disciplines.The central aim is to evaluate the impact of (introducing) AMC in healthcare on the spread of infections and on the efficacy in fighting HCAI and bacterial resistance to current antibiotics. http://www.cost.eu/COST_Actions/ca/CA15114
To evaluate the impact of (introducing) AntiMicrobial Coatings in healthcare on the spread of infections and on the efficacy in fighting HealthCare Associated Infections and bacterial resistance to current antibiotics. AMiCI secondary objectives: This Action will coordinate international and cross sectoral research efforts, by a variety of activities such as collaborating with existing projects, organizing new events, jointly developing and implementing working groups, so that forces can be joined in combatting AMR. This Action will stimulate collaboration between industry and research institutes using available knowledge, and disclose state of- the- art nanotechnology and biotechnology knowledge, to industrial partners. This will lead to more effective AMC and successful new market applications. More to be found on project website: www.amici-consortium.eu
AMICI brings together stakeholders from different countries and disciplines, including knowledge institutes, producers and processors of antimicrobial coatings, and organizations involved in the compliance with international standards on hygiene. The central aim is to evaluate the impact of (introducing) AMC in healthcare on the spread of infections and on the efficacy in fighting HCAI and bacterial resistance to current antibiotics. http://www.cost.eu/COST_Actions/ca/CA15114
Research Item (27)
- Mar 2018
Recognized issues with poor hand hygiene compliance among healthcare workers, and reports of re-contamination of previously chemically disinfected surfaces through hand contact, emphasize need for novel hygiene methods in addition to those currently available. One such approach involves antimicrobial (nano)-coatings (AMC), whereby integrated active ingredients are responsible for elimination of microorganisms that come into contact with treated surfaces. While widely studied under laboratory conditions with promising results, studies under real life healthcare conditions are scarce. The views of 75 contributors from 30 European countries were collated regarding specialised cleaning associated with antimicrobial coatings for reduction of hospital acquired infection. There was unanimous agreement that generation of scientific guidelines for cleaning of antimicrobial coatings, using traditional or new processes, are needed. Specific topics included: understanding mechanisms of action of cleaning materials and their physical interactions with conventional and antimicrobial coatings; that assessments mimic the life-cycle of coatings to determine the impact of repetitive cleaning and other aspects of ageing (e.g., exposure to sunlight); determining concentrations of AMC-derived biocides in effluents, and development of effective de-activation and sterilisation treatments for cleaning effluents. Further, the consensus opinion was that prior to widespread implementation of AMCs, the varying responsibilities of involved clinical, healthcare management, cleaning services, and environmental safety stakeholders need clarification.
- Dec 2017
Aims: To study stability of biofilms and water quality in pilot scale drinking water copper and PEX pipes in changing conditions (extra disinfection, magnetic water treatment MWT). Methods and results: Next-generation sequencing (NGS) of 16S ribosomal RNA genes (rDNA) to describe total bacterial community and ribosomal RNA (rRNA) to describe active bacterial members in addition to traditional microbiological methods were applied. Biofilms from control copper and PEX pipes shared same most abundant bacteria (Methylobacterium spp., Sphingomonas spp., Zymomonas spp.) and average species diversities (Shannon 3.8-4.2) in rDNA and rRNA libraries whereas few of the taxa differed by their abundance such as lower total Mycobacterium spp. occurrence in copper (<0.02%) to PEX (<0.2%) pipes. Extra disinfection (total chlorine increase from ca. 0.5 to 1 mg l-1 ) affected total and active population in biofilms seen as decrease of many bacterial species and diversity (Shannon 2.7, P < 0.01, rRNA) and increase of Sphingomonas spp. as compared to control samples. Further, extra disinfected copper and PEX samples formed separate clusters in unweighted non-metric multidimensional scaling plot (rRNA) similarly to MWT-treated biofilms of copper (but not PEX) pipes that instead showed higher species diversity (Shannon 4.8, P < 0.05 interaction). Conclusions: Minor chlorine dose addition increased selection pressure and many species were sensitive to chlorination. Pipe material seemed to affect mycobacteria occurrence, and bacterial communities with MWT in copper but not in PEX pipes. Significance and impact of the study: This study using rRNA showed that chlorination affects especially active fraction of bacterial communities. Copper and PEX differed by the occurrence of some bacterial members despite similar community profiles. This article is protected by copyright. All rights reserved.
Worldwide, millions of patients are affected annually by healthcare-associated infection (HCAI), impacting up to 80,000 patients in European Hospitals on any given day. This represents not only public health risk, but also an economic burden. Complementing routine hand hygiene practices, cleaning and disinfection, antimicrobial coatings hold promise based, in essence, on the application of materials and chemicals with persistent bactericidal or –static properties onto surfaces or in textiles used in healthcare environments. The focus of considerable commercial investment and academic research energies, such antimicrobial coating-based approaches are widely believed to have potential in reduction of microbial numbers on surfaces in clinical settings. This belief exists despite definitive evidence as to their efficacy and is based somewhat on positive studies involving, for example, copper, silver or gold ions, titanium or organosilane, albeit under laboratory conditions. The literature describes successful delay and/or prevention of recontamination following conventional cleaning and disinfection by problematic microbes such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE), amongst others. However, there is a scarcity of studies assessing antimicrobial surfaces other than copper in the clinical environment, and a complete lack of published data regarding the successful implementation of these materials on clinically significant outcomes (including HCAI). Through its Cooperation in Science and Technology programme (COST), the European Commission has funded a four-year initiative to establish a network of stakeholders involved in development, regulation and use of novel anti-microbial coatings for prevention of HCAI. The network (AMiCI) comprises participants of more than sixty universities, research institutes and companies across twenty-nine European countries and, to-date, represents the most comprehensive consortium targeting use of these emergent technologies in healthcare settings. More specifically, the network will prioritise coordinated research on the effects (both positive and negative) of antimicrobial coatings in healthcare sectors; know-how regarding availability and mechanisms of action of (nano)-coatings; possible adverse effects of such materials (e.g., potential emergence of microbial resistance or emission of toxic agents into the environment); standardised performance assessments for antimicrobial coatings; identification and dissemination of best practices by hospitals, other clinical facilities, regulators and manufacturers.
Infections and infectious diseases are considered a major challenge to human health in healthcare units worldwide. This opinion paper was initiated by EU COST Action network AMiCI (AntiMicrobial Coating Innovations) and focuses on scientific information essential for weighing the risks and benefits of antimicrobial surfaces in healthcare settings. Particular attention is drawn on nanomaterial-based antimicrobial surfaces in frequently-touched areas in healthcare settings and the potential of these nano-enabled coatings to induce (eco)toxicological hazard and antimicrobial resistance. Possibilities to minimize those risks e.g., at the level of safe-by-design are demonstrated.
- Oct 2016
Significance and impact of the study: The efficiency of copper as an antimicrobial material has been noted in laboratory studies and in the hospital environment. The present study further shows that copper exerted an antibacterial effect in different facilities, i.e. in a hospital, a kindergarten, an office building and in a retirement home for the elderly. The study suggests that copper has potential use as an antibacterial material and therefore might serve as a means to lower the incidence of transmission of infectious agents from inanimate surfaces in different facilities, with everyday functions.
Influence of altering magnetic field on chemical properties of water, scale formation and morphology was studied using polyethylene and copper pipes located both in a laboratory pilot-system and in an apartment complex formed by five different buildings each having four or five different homes. In the pilot network, magnetic field decreased calcium scaling by 15% in the both studied piping materials. Also, surface film on the copper pipe appeared to be less compact by the magnetic exposure. In the studies conducted in the apartment complex, magnetic field effectively mobilized earlier accumulated iron and copper from the copper pipe surfaces especially dealing with a hot water circuit.
Vitamin D and its analogues are potent regulators of cell growth and differentiation both in vivo and in vitro. We studied the effects of 25-hydroxyvitamin D(3) [25(OH)D(3)], 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and vitamin D analogue, EB 1089, on the growth of a human ovarian cancer cell line, OVCAR-3. We also studied the expression of vitamin D metabolising enzymes 24-hydroxylase (24OHase) and 1alpha-hydroxylase (1alphaOHase). Our results showed that high concentrations (10 and 100 nM) of 1,25(OH)(2)D(3) inhibited a cell proliferation, whereas low concentration (0.1 nM) stimulated growth of the OVCAR-3 cells. In the concentration range of 10-500 nM a prohormone, 25(OH)D(3), stimulated growth. An amount of 1 nM EB 1089 and 100 nM 1,25(OH)(2)D(3) inhibited growth with an equal magnitude. The expression of 24OHase was strongly induced by 1,25(OH)(2)D(3) and EB 1089 in OVCAR-3 cells, and analysis of vitamin D metabolites showed the functionality of 24OHase. An inhibition of 24OHase activity with a novel 24OHase inhibitor enhanced growth-inhibiting effects of 1,25(OH)(2)D(3) and suppressed the growth stimulation of 100 nM 25(OH)D(3). We also report the expression of a vitamin D activating enzyme, 1alphaOHase, in 7 ovarian cancer cell lines. The production of 1,25(OH)(2)D(3) in OVCAR-3 cells was low, possibly due to an extensive activity of 24OHase or a low 1alphaOHase activity. These results suggest that in ovarian cancer cells vitamin D metabolizing enzymes might play a key role in modulating the growth response to vitamin D. The possible mitogenic effects of vitamin D should be considered when evaluating treatment of ovarian cancer with vitamin D.
Vitamin D inhibits the development and growth of prostate cancer cells. Epidemiologic results on serum vitamin D levels and prostate cancer risk have, however, been inconsistent. We conducted a longitudinal nested case-control study on Nordic men (Norway, Finland and Sweden) using serum banks of 200,000 samples. We studied serum 25(OH)-vitamin D levels of 622 prostate cancer cases and 1,451 matched controls and found that both low (</=19 nmol/l) and high (>/=80 nmol/l) 25(OH)-vitamin D serum concentrations are associated with higher prostate cancer risk. The normal average serum concentration of 25(OH)-vitamin D (40-60 nmol/l) comprises the lowest risk of prostate cancer. The U-shaped risk of prostate cancer might be due to similar 1,25-dihydroxyvitamin D(3) availability within the prostate: low vitamin D serum concentration apparently leads to a low tissue concentration and to weakened mitotic control of target cells, whereas a high vitamin D level might lead to vitamin D resistance through increased inactivation by enhanced expression of 24-hydroxylase. It is recommended that vitamin D deficiency be supplemented, but too high vitamin D serum level might also enhance cancer development.
- Mar 2001
Our recent epidemiological study (Ahonen et al., Cancer Causes Control 11(2000) (847-852)) suggests that vitamin D deficiency may increase the risk of initiation and progression of prostate cancer. The nested case-control study was based on a 13-year follow-up of about 19000 middle-aged men free of clinically verified prostate cancer. More than one-half of the serum samples had 25OH-vitamin D (25-VD) levels below 50 nmol/l, suggesting VD deficiency. Prostate cancer risk was highest among the group of younger men (40-51 years) with low serum 25-VD, whereas low serum 25-VD appeared not to increase the risk of prostate cancer in older men (>51 years). This suggests that VD has a protective role against prostate cancer only before the andropause, when serum androgen concentrations are higher. The lowest 25-VD concentrations in the younger men were associated with more aggressive prostate cancer. Furthermore, the high 25-VD levels delayed the appearance of clinically verified prostate cancer by 1.8 years. Since these results suggest that vitamin D has a protective role against prostate cancer, we tried to determine whether full spectrum lighting (FSL) during working hours could increase serum 25-VD concentrations. After 1-month exposure, there was no significant increase in the serum 25-VD level, although there was a bias towards slightly increasing values in the test group as opposed to decreasing values in controls. There was no significant change in the skin urocanic acid production. The possibility to use FSL in cancer prevention is discussed. In order to clarify the mechanism of VD action on cell proliferation and differentiation, we performed studies with the rat and human prostates as well prostate cancer cell lines. It is possible that 25-VD may have a direct role in the host anticancer defence activity, but the metabolism of vitamin D in the prostate may also play an important role in its action. We raised antibodies against human 1alpha-hydroxylase and 24-hydroxylase. Our preliminary results suggest that vitamin D is actively metabolised in the prostate. Vitamin D appears to upregulate androgen receptor expression, whereas androgens seem to upregulate vitamin D receptor (VDR). This may at least partially explain the androgen dependence of VD action. VD alone or administered with androgen causes a suppression of epithelial cell proliferation. VD can activate mitogen-activated kinases, erk-1 and erk-2, within minutes and p38 within hours. Also, auto/paracrine regulation might be involved, since keratinocyte growth factor (mRNA and protein) was clearly induced by VD. Based on these studies, a putative model for VD action on cell proliferation and differentiation is presented.
The aim was to evaluate the association between serum vitamin D (25-hydroxyvitamin D) level and risk of prostate cancer. The nested case-control study was based on a 13-year follow-up of about 19,000 middle-aged men who attended the first screening visit within the Helsinki Heart Study and were free of clinically verified prostate cancer at baseline. Through record linkage with the files of the Finnish Cancer Registry, 149 prostate cancer cases were identified in the cohort. They were matched (1:4) to probability density sampled controls for age, time of sample retrieval, and residence. Serum levels of 25-hydroxyvitamin D (25-VD) at entry were measured for cases and controls. The relative risks of prostate cancer were derived using conditional logistic regression analysis. Prostate cancer risk, analyzed by quartiles of the 25-VD levels, was inversely related to 25-VD. Men with 25-VD concentration below the median had an adjusted relative risk (OR) of 1.7 compared to men with 25-VD level above the median. The prostate cancer risk was highest among younger men (< 52 years) at entry and low serum 25-VD (OR 3.1 nonadjusted and 3.5 adjusted). Among those younger men (< 52 years), low 25-VD entailed a higher risk of non-localized cancers (OR 6.3). The mean age at diagnosis of the patients with 25-VD concentration above the median was 1.8 years higher than that of patients with vitamin D below the median (63.1 vs 61.3 years). We conclude that low levels of 25-VD associated with an increased risk for subsequent earlier exposure and more aggressive development of prostate cancer, especially before the andropause.
- Aug 2000
Keratinocyte growth factor (FGF-7/KGF) is a secreted member of the fibroblast growth factor family, which functions primarily as an important paracrine mediator of cell growth and differentiation. Inhibitory pathways of vitamin D may also involve participation of some growth factors. To determine whether vitamin D may play a role in the expression of FGF-7, we investigated FGF-7 expression in human breast cancer cells treated with 1,25-dihydroxyvitamin D3, which inhibited the growth of the cells. By means of cDNA microarray, RT-PCR, and Western blot analysis, we have shown an increase in expression of FGF-7 on both mRNA and protein levels after vitamin D exposure. This is the first demonstration of vitamin D regulation of FGF-7 expression and its possible involvement in mediating growth and differentiation by vitamin D.
- May 2000
The data suggest that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and androgens are essential for regulation of growth and differentiation in, e.g., human reproductive tissues. We investigated the possible cross-talk between 1,25(OH)2D3 and androgens in the human ovarian cancer cell line OVCAR-3. Our data demonstrate that 1,25(OH)2D3 and androgen (dihydrotestosterone, DHT) regulate the growth of OVCAR-3 cells. Nine days' treatment of OVCAR-3 cells with 100 nM DHT resulted in 48% stimulation of growth, whereas growth inhibition (73%) was observed after treatment with 100 nM 1,25(OH)2D3. The combination of 1,25(OH)2D3 and DHT showed that 1,25(OH)2D3 clearly reduces the growth-stimulatory effect of DHT on OVCAR-3 cells. Moreover, Western blot analysis revealed that these cells contain receptors for 1,25(OH)2D3 (VDR) and androgen (AR). Expression of VDR and AR was up-regulated by their cognate ligands. Up-regulation of AR by 1,25(OH)2D3 and of VDR by DHT provides evidence of cross-talk between 2 signaling pathways in OVCAR-3 cells. We also studied the immuno-histochemical distribution of VDRs and ARs in rat ovaries and human ovarian cancer cases. In rat ovaries, VDRs were observed mainly in granulosa and theca cells and ARs in granulosa cells and surface epithelium. In the human ovarian cancer cases studied, 43% were VDR-positive and 64% AR-positive. Combining the results suggests that the growth of ovarian tissue might be regulated by 1,25(OH)2D3 and androgen.
- Oct 1999
Two novel antibodies against the mammalian progesterone receptor (PR) were raised and characterized to study the distribution of PR and the effect of estrogen on PR expression in various female murine tissues by immunohistochemistry. There were estrogen-independent constitutive PR expressions in the smooth muscle cells of uterus, uterine blood vessels, urinary bladder, duodenum, and jejunum of ovariectomized mice. Uterine stromal cells, capsular cells of kidney and adrenal gland, and the epithelial cells of submandibular gland expressed PR constitutively. PR expression was detected in some thymic cells and the number of PR-positive thymic cells increased markedly after estrogen treatment. Estrogen induced PR expression in the epithelial cells of uterus, vagina, urethra, and skin and the stromal cells of vagina, urethra, and pancreatic ducts, as well as the smooth muscle cells of some blood vessels. These results suggest cell-specific progesterone actions in the urinary tract, skin, and gastrointestinal organs, on the immune functions, and on the regulation of local blood flow.
- Jul 1999
Genetic polymorphisms and expression of steroid receptors may explain why some individuals are more at risk of developing prostate cancer. Some risk factors often discussed are androgen stimulation, and vitamin A and D deficiency. Long CAG-repeats in exon 1 of the androgen receptor (AR) gene on the X chromosome seem to have a protective role against androgen overstimulation. Likewise, long vitamin D receptor alleles in the poly-A tract may prevent vitamin D stimulation. Blood samples from 59 Swedish patients with sporadic prostate cancers, 59 with hereditary prostate cancer, and 34 Japanese prostate cancer patients were compared with benign controls. Tissue specimens from 37 Swedish and 23 Japanese prostate cancer patients with matching blood samples were investigated by immunohistochemical techniques. The number of CAG-repeats was identical in sporadic and hereditary prostate cancer patients, but the repeats were significantly shorter than in benign controls. Benign Japanese controls were similar to Swedish controls, but Japanese prostate cancers had longer repeats than did controls. Both the vitamin D and A receptor staining was stronger in Japanese than in Swedish prostate cancer specimens. Prostate cancer occurs approximately 5 years later in Japanese compared with Swedish men. Varying lengths of CAG-repeats of the androgen receptor cannot fully explain racial differences in clinical prostate cancer incidence. A larger content of vitamin A and D receptors may be linked to a delayed onset of clinical prostate cancer in Japanese men.
- Jun 1999
BACKGROUND Genetic polymorphisms and expression of steroid receptors may explain why some individuals are more at risk of developing prostate cancer. Some risk factors often discussed are androgen stimulation, and vitamin A and D deficiency. Long CAG-repeats in exon 1 of the androgen receptor (AR) gene on the X chromosome seem to have a protective role against androgen overstimulation. Likewise, long vitamin D receptor alleles in the poly-A tract may prevent vitamin D stimulation.METHODS Blood samples from 59 Swedish patients with sporadic prostate cancers, 59 with hereditary prostate cancer, and 34 Japanese prostate cancer patients were compared with benign controls. Tissue specimens from 37 Swedish and 23 Japanese prostate cancer patients with matching blood samples were investigated by immunohistochemical techniques.RESULTSThe number of CAG-repeats was identical in sporadic and hereditary prostate cancer patients, but the repeats were significantly shorter than in benign controls. Benign Japanese controls were similar to Swedish controls, but Japanese prostate cancers had longer repeats than did controls. Both the vitamin D and A receptor staining was stronger in Japanese than in Swedish prostate cancer specimens. Prostate cancer occurs approximately 5 years later in Japanese compared with Swedish men.CONCLUSIONS Varying lengths of CAG-repeats of the androgen receptor cannot fully explain racial differences in clinical prostate cancer incidence. A larger content of vitamin A and D receptors may be linked to a delayed onset of clinical prostate cancer in Japanese men. Prostate 39:262–268, 1999. © 1999 Wiley-Liss, Inc.
- Sep 1998
1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has been found to have a variety of physiological functions, including effects on growth and differentiation in normal and malignant cells. The antiproliferative effects of 1,25(OH)2D3 are reported to be mediated through the genomic signaling pathway by binding to a specific high affinity receptor protein, the 1,25-dihydroxyvitamin D3 receptor (VDR). VDR has been localized in a variety of tissues, but little is known about VDR distribution in human prostate. In this study, we raised an antibody against a synthetic peptide corresponding to amino acids 10-24 of human vitamin D receptor. The sequence selected for immunization is identical in human, rat and mouse VDR. Based on this antibody, we developed an immunohistochemical method suitable for studying VDR expression in paraffin-embedded tissue. The immunohistochemical staining was verified using classical target organs for vitamin D (kidney, intestine, skin). With this method, we studied VDR localization on paraffin-embedded human prostatic tissue obtained from 8 patients undergoing radical prostatectomy for urinary bladder cancer and demonstrate VDR expression in the secretory epithelial and few stromal cells of human prostate. The nuclear staining in the secretory epithelial cells was concentrated near the nuclear membrane and in discrete foci in the nucleoplasm. This suggests that effects of 1,25-dihydroxyvitamin D3 are mediated through VDR in these cells. Moreover our result indicates that there are strong variations in VDR expression between prostatic samples.
- Feb 1998
Genomic actions of progesterone are mediated via A and B isoforms of the progesterone receptor (PR). One major factor controlling PR level is progesterone causing negative autoregulation (down-regulation) of the receptor protein. In this work we studied the mechanism whereby progesterone exerts its effects on PR level in the chicken oviduct. We found that progesterone does not markedly regulate PR mRNA expression. Furthermore, we demonstrate here for the first time that PR is a target for ubiquitylation and that the proportion of ubiquitylated PR is increased by progesterone treatment. Our data suggest that ligand-induced down-regulation of PR involves enhanced degradation of receptor protein by ubiquitin-proteasome system in vivo.
- Jun 1996
An immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) method for a highly sensitive analysis of raspberry bushy dwarf virus (RBDV) in infected plants is described. In the method, preliminary purification of virus particles or viral RNA from the plant material is not necessary. Viruses are enriched during the assay by antibodies bound in the PCR microplate wells, followed by lysis of the viral particles, and RT-PCR of the viral RNA. The reaction mixtures, including reverse transcriptase and DNA polymerase, have been selected so that both enzymes are active in the lysis and amplification conditions; by this way, it is possible to conduct the whole procedure in a single step. Using the method, four fragments from RNA-3 of RBDV have been amplified with various combinations of four primers. The procedure is sensitive enough to allow a simple detection of RBDV in in vitro cultured plants in which the detection of viruses by conventional immunological methods is difficult or even impossible.