ABSTRACT
Age-related macular degeneration (AMD) in its various forms is a leading cause of blindness in industrialized countries. Here, we provide evidence that ligands for neuropilin-1 (NRP1), such as Semaphorin 3A and VEGF-A, are elevated in the vitreous of patients with AMD at times of active choroidal neovascularization (CNV). We further demonstrate that NRP1-expressing myeloid cells promote and maintain CNV. Expression of NRP1 on cells of myeloid lineage is critical for mitigating production of inflammatory factors such as IL6 and IL1ß. Therapeutically trapping ligands of NRP1 with an NRP1-derived trap reduces CNV. Collectively, our findings identify a role for NRP1-expressing myeloid cells in promoting pathological angiogenesis during CNV and introduce a therapeutic approach to counter neovascular AMD.
Subject(s)
Choroidal Neovascularization , Wet Macular Degeneration , Angiogenesis Inhibitors , Humans , Inflammation , Neuropilin-1/genetics , Vascular Endothelial Growth Factor A , Visual AcuityABSTRACT
Two trains (A and B) of four-stage moving bed biofilm reactors (MBBRs) were developed for the degradation of thiocyanate (SCN(-)), cyanate (OCN(-)) and ammonia (NH3). A pre-denitrification configuration was established in the first-stage reactor of the B train using SCN(-) and OCN(-) as the sole carbon source. SCN(-), OCN(-) and NH3 were completely removed in both trains. The highest removal of total nitrogen equivalent (total-N) occurred at a loading rate of 5.6 mg-N L(-1) h(-1). The pre-denitrification configuration resulted in increased total-N removal in the B train (62.6%) compared to the A train (38.5%). Thiobacillus spp. were the predominant bacteria in all MBBRs. Bacteria related to bioprocesses involving anaerobic ammonium oxidation were present in the B train, suggesting that part of nitrogen removal occurs via this pathway. Our results showed that the pre-denitrification configuration increases the efficiency of removal of total-N compounds in the SCN(-)/OCN(-)-degrading MBBR process.
Subject(s)
Biofilms , Bioreactors/microbiology , Cyanates/isolation & purification , Denitrification , Environmental Restoration and Remediation/instrumentation , Environmental Restoration and Remediation/methods , Thiocyanates/isolation & purification , Ammonia/analysis , Base Sequence , Biodegradation, Environmental , Biodiversity , Equipment Design , Nitrates/analysis , Nitrogen/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , Thiobacillus/metabolism , WastewaterABSTRACT
With the aim of developing a biotechnological tool for the production of foreign proteins in plants, we first engineered an infectious turnip mosaic virus (TuMV) cDNA that contained the jellyfish green fluorescent protein (GFP) gene or the bacterial beta-glucuronidase (GUS) gene (uidA). Two insertion sites were assessed, either between P1 and HCPro cistrons or Pol and CP cistrons. In each construct, the junctions flanking the inserted gene coded for P1 and/or VPg-Pro cleavage recognition site sequences, to produce free GUS or GFP. After transfection by particle bombardment on Brassica perviridis, characteristic symptoms for TuMV infection appeared and Western blot analyses showed that GFP and GUS had been excised from the viral polyprotein. No significant differences in expression level were noticed between the two insertion sites. By RT-PCR, gfp was found to be stable over 30 days post-transfection (dpt) while uidA was gradually lost at 15 dpt. We also created two constructs containing either gene at each insertion sites on the same molecule. Attenuated systemic symptoms were observed after particle bombardment on B. perviridis and Western blot analyses showed that both foreign proteins were produced. Also, the same stability/instability as for the single-gene constructs were observed. These results indicate that it is possible to produce at least two foreign proteins simultaneously in a TuMV-based vector.
