Simvastatin coating of TiO2 scaffold induces osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells

AbstractBone tissue engineering requires an osteoconductive scaffold, multipotent cells with regenerative capacity and bioactive molecules. In this study we investigated the osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells (hAD-MSCs) on titanium dioxide (TiO2) scaffold coated with alginate hydrogel containing various concentrations of simvastatin (SIM). The mRNA expression of osteoblast-related genes such as collagen type I alpha 1 (COL1A1), alkaline phosphatase (ALPL), osteopontin (SPP1), osteocalcin (BGLAP) and vascular endothelial growth factor A (VEGFA) was enhanced in hAD-MSCs cultured on scaffolds with SIM in comparison to scaffolds without SIM. Furthermore, the secretion of osteoprotegerin (OPG), vascular endothelial growth factor A (VEGFA), osteopontin (OPN) and osteocalcin (OC) to the cell culture medium was higher from hAD-MSCs cultured on scaffolds with SIM compared to scaffolds without SIM. The TiO2 scaffold coated with alginate hydrogel containing SIM promote osteogenic differentiation of hAD-MSCs in vitro, and demonstrate feasibility as scaffold for hAD-MSC based bone tissue engineering

Analysis of the Effects of Five Factors Relevant to In Vitro Chondrogenesis of Human Mesenchymal Stem Cells Using Factorial Design and High Throughput mRNA-Profiling

The in vitro process of chondrogenic differentiation of mesenchymal stem cells for tissue engineering has been shown to require three-dimensional culture along with the addition of differentiation factors to the culture medium. In general, this leads to a phenotype lacking some of the cardinal features of native articular chondrocytes and their extracellular matrix. The factors used vary, but regularly include members of the transforming growth factor β superfamily and dexamethasone, sometimes in conjunction with fibroblast growth factor 2 and insulin-like growth factor 1, however the use of soluble factors to induce chondrogenesis has largely been studied on a single factor basis. In the present study we combined a factorial quality-by-design experiment with high-throughput mRNA profiling of a customized chondrogenesis related gene set as a tool to study in vitro chondrogenesis of human bone marrow derived mesenchymal stem cells in alginate. 48 different conditions of transforming growth factor β 1, 2 and 3, bone morphogenetic protein 2, 4 and 6, dexamethasone, insulin-like growth factor 1, fibroblast growth factor 2 and cell seeding density were included in the experiment. The analysis revealed that the best of the tested differentiation cocktails included transforming growth factor β 1 and dexamethasone. Dexamethasone acted in synergy with transforming growth factor β 1 by increasing many chondrogenic markers while directly downregulating expression of the pro-osteogenic gene osteocalcin. However, all factors beneficial to the expression of desirable hyaline cartilage markers also induced undesirable molecules, indicating that perfect chondrogenic differentiation is not achievable with the current differentiation protocols

En ny start

Natur- og Landbrugskommissionen lægger med denne rapport op til at give naturen i Danmark et markant løft. Vi anbefaler også at indføre en helt ny miljøregulering af landbruget til gavn for både erhverv, vandmiljø, natur og klima, så Danmark kan sætte nye standarder for bæredygtig landbrugsproduktion. Og ikke mindst ønsker vi med vores anbefalinger at give et væsentligt bidrag til at skabe ny vækst og udvikling i landbrugserhvervet. Jeg håber, at Natur- og Landbrugskommissionen med vores anbefalinger og vores måde at arbejde på vil bidrage til, at der skabes gode, holdbare løsninger, og at konstruktiv dialog bliver den dominerende arbejdsform i de kommende år, når der skal findes svar på de udfordringer, som natur, miljø og landbrug står over for. Vi må væk fra grøftegravning og mistænkeliggørelse og frem til en mere konstruktiv samtale, der skal bane vejen for flere fælles accepterede og afbalancerede løsninger. Samfundet og de grønne interesser må acceptere, at landbruget som alle andre erhverv skal have lov til at udvikle sig. Ligesom landbruget må anerkende, at det omgivende samfund har brug for en rig natur og et rent miljø. Der er mange muligheder for at finde løsninger, som både kan give udvikling i landbrugserhvervet, beskytte miljøet og skabe mere og rigere natur. Natur, miljø og vækst er ikke nødvendigvis modsætninger, men kan og skal arbejde sammen. Gode løsninger kræver et godt samarbejde mellem de forskellige interesser. Ingen kan få det hele. Alle vil vinde ved at tænke i helheder, balancer og kompromisser. Det kræver lederskab og politisk ansvarlighed. Natur- og Landbrugskommissionen mener at have løst sin opgave på den fastsatte tid og inden for de rammer og vilkår, som regeringen udstak i sit kommissorium i foråret 2012. Kommissionens samlede anbefalinger sikrer en balance mellem hensyn til erhverv, vækst, miljø, klima og natur. Og derfor opfordrer vi til, at anbefalingerne vurderes og anvendes samlet. Vi håber, at vores anbefalinger vil danne grundlag for en konstruktiv, politisk proces i den kommende tid. Nu er det op til regering og Folketing. Jeg vil gerne takke Natur- og Landbrugskommissionens medlemmer for et engageret og konstruktivt arbejde i det forløbne år. Og tak til vores dygtige og flittige sekretariat. Og endelig en meget stor tak til vores følgegruppe og de mange mennesker og organisationer, som har bidraget til vores arbejde med ideer, vurderinger og konkrete forslag i det forløbne, meget intense år. Jeg håber, at det gode samarbejde vil fortsætte under nye former

Natur- og Landbrugskommissionens statusrapport

Regeringen har i foråret 2012 nedsat en uafhængig Natur- og landbrugskommission. Kommissionens opgave er, at udarbejde anbefalinger til løsning af landbrugets økonomiske, strukturelle og miljømæssige udfordringer, herunder hvordan erhvervet kan bidrage til klima-, natur- og miljøindsatsen. Det fremgår af regeringsgrundlaget ’Et Danmark, der står sammen’ fra oktober 2011, at der på Danmarks areal skal være plads til at producere sunde og velsmagende fødevarer af høj kvalitet samt en varieret, mangfoldig og sammenhængende natur. Regeringen ønsker derfor et økonomisk og miljømæssigt bæredygtigt og fremtidsorienteret landbrugserhverv, der fortsat kan bidrage væsentligt til beskæftigelse og eksport i fødevaresektoren. Regeringen ønsker samtidig at stoppe tilbagegangen i og styrke biodiversiteten og forbedre miljøtilstanden i vandmiljøet og bidrage til at reducere udledningen af drivhusgasser. Regeringen peger derfor på, at der er brug for nye afbalancerede løsninger, så landbrugets rammevilkår fremadrettet understøtter en grøn omstilling med vækst og nye udviklingsmuligheder for erhvervet og samtidig styrker natur-, vandmiljø og klimaindsatsen. Natur- og Landbrugskommissionen påbegyndte sit arbejde i marts 2012 og har et år til at komme med forslag til at løse de økonomiske og miljømæssige udfordringer, som landbruget og naturen står overfor. Kommissionens anbefalinger skal præsenteres for regeringen i første halvdel af 2013. Kommissionen skal komme med konkrete forslag til, hvordan dansk landbrug og natur kan udvikles bæredygtigt både på kort og lang sigt. Kommissionens første opgave består i at udarbejde en statusredegørelse over landbrugets økonomiske situation, indtjeningsmuligheder og rammevilkår samt natur-, vandmiljø- og klimatilstanden, herunder status for implementeringen af EU- og andre internationale forpligtelser. Denne rapport udgør statusredegørelsen

Enhanced detection of circulating tumor DNA by fragment size analysis.

Existing methods to improve detection of circulating tumor DNA (ctDNA) have focused on genomic alterations but have rarely considered the biological properties of plasma cell-free DNA (cfDNA). We hypothesized that differences in fragment lengths of circulating DNA could be exploited to enhance sensitivity for detecting the presence of ctDNA and for noninvasive genomic analysis of cancer. We surveyed ctDNA fragment sizes in 344 plasma samples from 200 patients with cancer using low-pass whole-genome sequencing (0.4×). To establish the size distribution of mutant ctDNA, tumor-guided personalized deep sequencing was performed in 19 patients. We detected enrichment of ctDNA in fragment sizes between 90 and 150 bp and developed methods for in vitro and in silico size selection of these fragments. Selecting fragments between 90 and 150 bp improved detection of tumor DNA, with more than twofold median enrichment in >95% of cases and more than fourfold enrichment in >10% of cases. Analysis of size-selected cfDNA identified clinically actionable mutations and copy number alterations that were otherwise not detected. Identification of plasma samples from patients with advanced cancer was improved by predictive models integrating fragment length and copy number analysis of cfDNA, with area under the curve (AUC) >0.99 compared to AUC 0.91 compared to AUC < 0.5 without fragmentation features. Fragment size analysis and selective sequencing of specific fragment sizes can boost ctDNA detection and could complement or provide an alternative to deeper sequencing of cfDNA.We would like to acknowledge the support of The University of Cambridge, Cancer Research UK and the EPSRC (CRUK grant numbers A11906 (NR), A20240 (NR), A22905 (JDB), A15601 (JDB), A25177 (CRUK Cancer Centre Cambridge), A17242 (KMB), A16465 (CRUK-EPSRC Imaging Centre in Cambridge and Manchester)). The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013) / ERC Grant Agreement n. 337905. The research was supported by the National Institute for Health Research Cambridge, National Cancer Research Network, Cambridge Experimental Cancer Medicine Centre and Hutchison Whampoa Limited. This research is also supported by Target Ovarian Cancer and the Medical Research Council through their Joint Clinical Research Training Fellowship for Dr Moore. The CALIBRATE study was supported by funding from AstraZeneca

Microarray profiling for differential gene expression in PMSG-hCG stimulated preovulatory ovarian follicles of Chinese Taihu and Large White sows

<p>Abstract</p> <p>Background</p> <p>The Chinese Taihu is one of the most prolific pig breeds in the world, which farrows at least five more piglets per litter than Western pig breeds partly due to a greater ovulation rate. Variation of ovulation rate maybe associated with the differences in the transcriptome of Chinese Taihu and Large White ovaries. In order to understand the molecular basis of the greater ovulation rate of Chinese Taihu sows, expression profiling experiments were conducted to identify differentially expressed genes in ovarian follicles at the preovulatory stage of a PMSG-hCG stimulated estrous cycle from 3 Chinese Taihu and 3 Large White cycling sows by using the Affymetrix Porcine Genechip™.</p> <p>Results</p> <p>One hundred and thirty-three differentially expressed genes were identified between Chinese Taihu and Large White sows by using Affymetrix porcine GeneChip (<it>p </it>≤ 0.05, Fold change ≥ 2 or ≤ 0.5). Gene Ontology (GO) analysis revealed that these genes belonged to the class of genes that participated in regulation of cellular process, regulation of biological process, biological regulation, developmental process, cell communication and signal transduction and so on. Significant differential expression of 6 genes including <it>WNT10B </it>and <it>DKK2 </it>in the WNT signaling pathway was detected. Real-time RT-PCR confirmed the expression pattern in seven of eight selected genes. A search of chromosomal location revealed that 92 differentially expressed transcripts located to the intervals of quantitative trait loci (QTLs) for reproduction traits. Furthermore, SNPs of two differentially expressed genes- <it>BAX </it>and <it>BMPR1B </it>were showed to be associated with litter size traits in Large White pigs and Chinese DIV line pigs (<it>p </it>≤ 0.1 or <it>p </it>≤ 0.05).</p> <p>Conclusions</p> <p>Our study detected many genes that showed differential expression between ovary follicles of two divergent breeds of pigs. Genes involved with regulation of cellular process, regulation of biological process, in addition to several genes not previously associated with ovarian physiology or with unknown function, were differentially expressed between two breeds. The suggestive or significant associations of <it>BAX </it>and <it>BMPR1B </it>gene with litter size indicated these genetic markers had the potentials to be used in pig industry after further validation of their genetic effects. Taken together, this study reveals many potential avenues of investigation for seeking new insights into ovarian physiology and the genetic control of reproduction.</p

Ten golden rules for optimal antibiotic use in hospital settings: the WARNING call to action

Antibiotics are recognized widely for their benefits when used appropriately. However, they are often used inappropriately despite the importance of responsible use within good clinical practice. Effective antibiotic treatment is an essential component of universal healthcare, and it is a global responsibility to ensure appropriate use. Currently, pharmaceutical companies have little incentive to develop new antibiotics due to scientific, regulatory, and financial barriers, further emphasizing the importance of appropriate antibiotic use. To address this issue, the Global Alliance for Infections in Surgery established an international multidisciplinary task force of 295 experts from 115 countries with different backgrounds. The task force developed a position statement called WARNING (Worldwide Antimicrobial Resistance National/International Network Group) aimed at raising awareness of antimicrobial resistance and improving antibiotic prescribing practices worldwide. The statement outlined is 10 axioms, or “golden rules,” for the appropriate use of antibiotics that all healthcare workers should consistently adhere in clinical practice