69 research outputs found
Evaluation of manure management systems in Europe
The gross value of the agricultural
goods in 2014 amounted up to 370 billion Euros.
Almost 50% of all agricultural production is provided by livestock farming.Postprint (published version
Aberrant DNA Methylation of Matrix Remodeling and Cell Adhesion Related Genes in Pterygium
10.1371/journal.pone.0014687PLoS ONE62
Validation of Endogenous Control Genes for Gene Expression Studies on Human Ocular Surface Epithelium
PURPOSE: To evaluate a panel of ten known endogenous control genes (ECG) with quantitative reverse transcription PCR (qPCR), for identification of stably expressed endogenous control genes in the ocular surface (OS) epithelial regions including cornea, limbus, limbal epithelial crypt and conjunctiva to normalise the quantitative reverse transcription PCR data of genes of interest expressed in above-mentioned regions. METHOD: The lasermicrodissected (LMD) OS epithelial regions of cryosectioned corneoscleral buttons from the cadaver eyes were processed for RNA extraction and cDNA synthesis to detect genes of interest with qPCR. Gene expression of 10 known ECG--glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta actin (ACTB), peptidylprolyl isomerase (PPIA), TATA-box binding protein (TBP1), hypoxanthine guanine phosphoribosyl transferase (HPRT1), beta glucuronidase (GUSB), Eucaryotic 18S ribosomal RNA (18S), phosphoglycerate kinase (PGK1), beta-2-microglobulin (B2M), ribosomal protein, large, P0 (RPLP0)--was measured in the OS epithelial regions by qPCR method and the data collected was further analysed using geNorm software. RESULTS: The expression stability of ecgs in the os epithelial regions in increasing order as determined with genorm software is as follows: ACTB<18S<TBP<B2M<PGK1<HPRT1<GUSB<GAPDH<PPIA-RPLP0. In this study, geNorm analysis has shown the following ECGs pairs to be most stably expressed in individual OS epithelial regions: HPRT1-TBP in cornea, GUSB-PPIA in limbus, B2M-PPIA and RPLP0-TBP in LEC and conjunctiva respectively. However, across the entire ocular surface including all the regions mentioned above, PPIA-RPLP0 pair was shown to be most stable. CONCLUSION: This study has identified stably expressed ECGs on the OS epithelial regions for effective qPCR results in genes of interest. The results from this study are broadly applicable to quantitative reverse transcription PCR studies on human OS epithelium and provide evidence for the use of PPIA-RPLP0 ECGs pair in quantitative reverse transcription PCR across the OS epithelium
Advances in exosome therapies in ophthalmology–From bench to clinical trial
During the last decade, the fields of advanced and personalized therapeutics have been constantly evolving, utilizing novel techniques such as gene editing and RNA therapeutic approaches. However, the method of delivery and tissue specificity remain the main hurdles of these approaches. Exosomes are natural carriers of functional small RNAs and proteins, representing an area of increasing interest in the field of drug delivery. It has been demonstrated that the exosome cargo, especially miRNAs, is at least partially responsible for the therapeutic effects of exosomes. Exosomes deliver their luminal content to the recipient cells and can be used as vesicles for the therapeutic delivery of RNAs and proteins. Synthetic therapeutic drugs can also be encapsulated into exosomes as they have a hydrophilic core, which makes them suitable to carry water-soluble drugs. In addition, engineered exosomes can display a variety of surface molecules, such as peptides, to target specific cells in tissues. The exosome properties present an added advantage to the targeted delivery of therapeutics, leading to increased efficacy and minimizing the adverse side effects. Furthermore, exosomes are natural nanoparticles found in all cell types and as a result, they do not elicit an immune response when administered. Exosomes have also demonstrated decreased long-term accumulation in tissues and organs and thus carry a low risk of systemic toxicity. This review aims to discuss all the advances in exosome therapies in ophthalmology and to give insight into the challenges that would need to be overcome before exosome therapies can be translated into clinical practice
Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing
Pterygium is a lesion of the eye surface which involves cell proliferation, migration, angiogenesis, fibrosis, and extracellular matrix remodelling. Surgery is the only approved method to treat this disorder, but high recurrence rates are common. Recently, it has been shown in a mouse model that treatment with doxycycline resulted in reduction of the pterygium lesions. Here we study the mechanism(s) of action by which doxycycline achieves these results, using massive sequencing techniques. Surgically removed pterygia from 10 consecutive patients were set in short term culture and exposed to 0 (control), 50, 200, and 500 µg/ml doxycycline for 24 h, their mRNA was purified, reverse transcribed and sequenced through Illumina’s massive sequencing protocols. Acquired data were subjected to quantile normalization and analyzed using cytoscape plugin software to explore the pathways involved. False discovery rate (FDR) methods were used to identify 332 genes which modified their expression in a dose-dependent manner upon exposure to doxycycline. The more represented cellular pathways included all mitochondrial genes, the endoplasmic reticulum stress response, integrins and extracellular matrix components, and growth factors. A high correlation was obtained when comparing ultrasequencing data with qRT-PCR and ELISA results
Transplantation of human adipose stem cell-derived hepatocyte-like cells with restricted localization to liver using acellular amniotic membrane
Upgrading the temperature-phased anaerobic digestion of waste activated sludge by ultrasonic pretreatment
10 páginas; 5 figuras; 5 tablasBiochemical methane potential assays were performed to assess the influence of ultrasonic pretreatment on the temperature-phased anaerobic digestion (TPAD) of waste-activated sludge (WAS). Ultrasound (specific energy = 3380 kJ kg−1 TS) was applied to the WAS before the thermophilic stage or to the effluent after the thermophilic stage. In addition, a control system without pretreatment was also carried out. No significant differences were found in the overall performance of the TPAD process, but different behaviours were observed between the thermophilic and mesophilic stages. Total methane production was enhanced by more than 50% and the volatile solid removal increased by 13% in comparison to the TPAD control process. Finally, a previously defined kinetic model was applied successfully to the experimental data and showed an excellent fit.The authors wish to express their gratitude to the Spanish Ministry for the Environment, Rural Affairs and Marine Policy (Project 148/PC08/3-04.3) for providing financial support and to the 2008–2011 Research Plan of the University of Cadiz for supporting the first author through the ‘Contrato Puente para Doctores’.Peer reviewe
Manual suction versus femtosecond laser trephination for penetrating keratoplasty: Intraocular pressure, endothelial cell damage, incision geometry, and wound healing responses
10.1167/iovs.11-8403Investigative Ophthalmology and Visual Science5362571-2579IOVS
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