The molecular basis of genistein-induced mitotic arrest and exit of self-renewal in embryonal carcinoma and primary cancer cell lines

<p>Abstract</p> <p>Background</p> <p>Genistein is an isoflavonoid present in soybeans that exhibits anti-carcinogenic properties. The issue of genistein as a potential anti-cancer drug has been addressed in some papers, but comprehensive genomic analysis to elucidate the molecular mechanisms underlying the effect elicited by genistein on cancer cells have not been performed on primary cancer cells, but rather on transformed cell lines. In the present study, we treated primary glioblastoma, rhabdomyosarcoma, hepatocellular carcinoma and human embryonic carcinoma cells (NCCIT) with μ-molar concentrations of genistein and assessed mitotic index, cell morphology, global gene expression, and specific cell-cycle regulating genes. We compared the expression profiles of NCCIT cells with that of the cancer cell lines in order to identify common genistein-dependent transcriptional changes and accompanying signaling cascades.</p> <p>Methods</p> <p>We treated primary cancer cells and NCCIT cells with 50 μM genistein for 48 h. Thereafter, we compared the mitotic index of treated versus untreated cells and investigated the protein expression of key regulatory self renewal factors as OCT4, SOX2 and NANOG. We then used gene expression arrays (Illumina) for genome-wide expression analysis and validated the results for genes of interest by means of Real-Time PCR. Functional annotations were then performed using the DAVID and KEGG online tools.</p> <p>Results</p> <p>We found that cancer cells treated with genistein undergo cell-cycle arrest at different checkpoints. This arrest was associated with a decrease in the mRNA levels of core regulatory genes, <it>PBK</it>, <it>BUB1</it>, and <it>CDC20 </it>as determined by microarray-analysis and verified by Real-Time PCR. In contrast, human NCCIT cells showed over-expression of <it>GADD45 A </it>and <it>G </it>(growth arrest- and DNA-damage-inducible proteins 45A and G), as well as down-regulation of OCT4, and NANOG protein. Furthermore, genistein induced the expression of apoptotic and anti-migratory proteins p53 and p38 in all cell lines. Genistein also up-regulated steady-state levels of both <it>CYCLIN A </it>and <it>B</it>.</p> <p>Conclusion</p> <p>The results of the present study, together with the results of earlier studies show that genistein targets genes involved in the progression of the M-phase of the cell cycle. In this respect it is of particular interest that this conclusion cannot be drawn from comparison of the individual genes found differentially regulated in the datasets, but by the rather global view of the pathways influenced by genistein treatment.</p

Nestedness of Ectoparasite-Vertebrate Host Networks

Determining the structure of ectoparasite-host networks will enable disease ecologists to better understand and predict the spread of vector-borne diseases. If these networks have consistent properties, then studying the structure of well-understood networks could lead to extrapolation of these properties to others, including those that support emerging pathogens. Borrowing a quantitative measure of network structure from studies of mutualistic relationships between plants and their pollinators, we analyzed 29 ectoparasite-vertebrate host networks—including three derived from molecular bloodmeal analysis of mosquito feeding patterns—using measures of nestedness to identify non-random interactions among species. We found significant nestedness in ectoparasite-vertebrate host lists for habitats ranging from tropical rainforests to polar environments. These networks showed non-random patterns of nesting, and did not differ significantly from published estimates of nestedness from mutualistic networks. Mutualistic and antagonistic networks appear to be organized similarly, with generalized ectoparasites interacting with hosts that attract many ectoparasites and more specialized ectoparasites usually interacting with these same “generalized” hosts. This finding has implications for understanding the network dynamics of vector-born pathogens. We suggest that nestedness (rather than random ectoparasite-host associations) can allow rapid transfer of pathogens throughout a network, and expand upon such concepts as the dilution effect, bridge vectors, and host switching in the context of nested ectoparasite-vertebrate host networks

A feasibility pilot study on the use of complementary therapies delivered via mobile technologies on Icelandic surgical patients’ reports of anxiety, pain, and self-efficacy in healing

BACKGROUND: Complementary therapies (CT), such as relaxation technique, massage, guided imagery, and accupuncture have shown to benefit patients undergoing surgery. The aim of this study was to determine the feasibility of using audio relaxation technique (ART), music intervention (MI), nature video application with music (NVAM), and nature video application without music (NVA) delivered via mobile technologies in a clinical setting. Secondary, the effects of ART, MI, NVAM and NVA on patients’ state anxiety, pain perception, and perceived self-efficacy in healing were determined. METHODS: A randomized clinical trial (RCT) involving 105 same day surgery (SDS) patients, who were assigned to an ART (n = 25), MI (n = 25), NVAM (n = 15), NVA (n = 16), or a control group (n = 24) were assessed for state anxiety, self-reported pain, and self-efficacy four days prior to surgery, immediately prior and following a surgical intervention, and day five post-operative. RESULTS: ANOVA found no statistically significant differences in anxiety scores; pain, or perceived self-efficacy between the five groups. Matched pairs t-Test revealed all participants had an increase in anxiety from pre-op to day 10 follow-up; a significant change in pain levels from pre-op to day 10 follow-up; and all participants had a significant increase in general self-efficacy from pre-op to day 10 follow-up. Mean pain level scores from day 1 to pre-op showed a significant decrease in pain for the ART group and NVAM group. Matched pairs t-Test for self-efficacy scores indicated the MI group and the NVA group had significant increases in self-efficacy. A significant decrease in anxiety from pre-op to day 10 for participants reporting a prior history of anxiety and for those reporting prior history of taking anti-anxiety medications. CONCLUSIONS: Despite the non-significant findings between the five groups, at any measurement point, there were valuable trends toward significance and confirmed feasibility in a clinical setting. Among the groups there were statistically significant findings for all interventions on anxiety, pain, and self-efficacy. The feasability of the implementation of novel interventions of NVAM and NVAM adds to clinical practice and the CT literature. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02236455 (September 4, 2014

The role of professionals complementary to dentistry in orthodontic practice as seen by United Kingdom orthodontists

Objective To determine the opinions of UK orthodontists regarding potential clinical tasks to be carried out by orthodontic professionals complementary to dentistry (PCDs). Design and setting Postal questionnaire survey of orthodontists within the UK. Subjects and methods A survey was developed including questions about respondents' current and previous posts, followed by seventeen potential tasks to be carried out by PCDs to prescription. Respondents stated whether a suitably trained PCD should perform each task to direct prescription. Respondents could mention additional clinical tasks that they considered appropriate. The questionnaire was posted to members of the British Orthodontic Society who were eligible to be included on the finalised specialist orthodontist register held by the General Dental Council. Results Five hundred and eighty one (70.6%) of 821 questionnaires were returned. Most procedures were supported but there was less support for four procedures, a majority against the fitting of bonded lingual retainers and respondents were undecided about molar band cementation, direct bonding of brackets and fitting of headgear. Amount or type of clinical practice or the date of qualification did not alter the pattern of responses given. Conclusions There is support for the introduction of a class of orthodontic PCD and agreement regarding the nature of clinical tasks