ABSTRACT
Olive oil mill wastewater (OMW) has a high polluting power, with total phenolics (TP) around 2.5 g l(-1) and chemical oxygen demand (COD) 85 g l(-1). Biological systems offer advantages in treating this type of agro-industrial wastewater. The performance of phenol-adapted Ralstonia eutropha for aerobic biotreatment of OMW has been studied, and a TP concentration of 250 mg l(-1) found to be fully degraded within 24 h. This simple procedure may be adopted as a pretreatment prior to the normal aerobic or anaerobic techniques used for treating OMW. The biodegradative capability of this non-pathogenic gram-negative bacterium towards the TP and COD content of OMW has been evaluated. The adapted free cells were found able to decrease TP and COD in the undiluted OMW by 56% and 42%, respectively. The Monod equation was found suitable to describe the capacity of the cells for growing on undiluted OMW, giving micromax 0.083 per day and Ks = 1846 mg l(-1). Using a packed-bed reactor the performance of loofa-immobilized R. eutropha was assessed and the reduction in TP and COD shown to be 73% and 64%, respectively.
Subject(s)
Cupriavidus necator/metabolism , Plant Oils/chemistry , Plant Oils/metabolism , Water Pollutants, Chemical/isolation & purification , Water Purification/methods , Agriculture/methods , Industrial Waste/prevention & control , Olive Oil , Water Pollutants, Chemical/chemistryABSTRACT
Members of the transforming growth factor-beta (TGF-beta) superfamily regulate cell proliferation, differentiation, and apoptosis, controlling the development and maintenance of most tissues. TGF-beta signal is transmitted through the phosphorylation of Smad proteins by TGF-beta receptor serine/threonine kinase. During early tooth development, TGF-beta inhibits proliferation of enamel organ epithelial cells but the underlying molecular mechanisms are largely unknown. Here we tested the hypothesis that antagonistic effects between Smad2 and Smad7 regulate TGF-beta signaling during tooth development. Attenuation of Smad2 gene expression resulted in significant advancement of embryonic tooth development with increased proliferation of enamel organ epithelial cells, while attenuation of Smad7 resulted in significant inhibition of embryonic tooth development with increased apoptotic activity within enamel organ epithelium. These findings suggest that different Smads may have differential activities in regulating TGF-beta-mediated cell proliferation and death. Furthermore, functional haploinsufficiency of Smad2, but not Smad3, altered TGF-beta-mediated tooth development. The results indicate that Smads are critical factors in orchestrating TGF-beta-mediated gene regulation during embryonic tooth development. The effectiveness of TGF-beta signaling is highly sensitive to the level of Smad gene expression.
Subject(s)
DNA-Binding Proteins/physiology , Gene Expression , Tooth/growth & development , Trans-Activators/physiology , Animals , Base Sequence , DNA-Binding Proteins/genetics , Female , In Situ Hybridization , Mice , Molecular Sequence Data , Morphogenesis , Oligonucleotides, Antisense/pharmacology , Phosphorylation , Polymerase Chain Reaction , Pregnancy , Smad2 Protein , Smad7 Protein , Trans-Activators/genetics , Transforming Growth Factor beta/physiologyABSTRACT
Members of the transforming growth factor-beta (TGF-beta) superfamily signal through their cognate receptors to determine cell phenotypes during embryogenesis. Our previous studies on the regulation of first branchial arch morphogenesis have identified critical components of a hierarchy of different TGF-beta isoforms and their possible functions in regulating tooth and cartilage formation during mandibular morphogenesis. Here we tested the hypothesis that TGF-beta type II receptor (TGF-beta IIR) is a critical component in the TGF-beta signaling pathway regulating tooth formation. To establish the precise location of TGF-beta ligand and its cognate receptor, we first performed detailed analyses of the localization of both TGF-beta2 and TGF-beta IIR during initiation and subsequent morphogenesis of developing embryonic mouse tooth organs. A possible autocrine functional role for TGF-beta and its cognate receptor (TGF-beta IIR) was inferred due to the temporal and spatial localization patterns during the early inductive stages of tooth morphogenesis. Second, loss of function of TGF-beta IIR in a mandibular explant culture model resulted in the acceleration of tooth formation to the cap stage while the mandibular explants in the control group only showed bud stage tooth formation. In addition, there was a significant increase in odontogenic epithelial cell proliferation following TGF-beta IIR abrogation. These results demonstrate, for the first time, that abrogation of the TGF-beta IIR stimulates embryonic tooth morphogenesis in culture and reverses the negative regulation of endogenous TGF-beta signaling upon enamel organ epithelial cell proliferation.
Subject(s)
Branchial Region/embryology , Receptors, Transforming Growth Factor beta/metabolism , Tooth/embryology , Tooth/metabolism , Adenoviridae/genetics , Animals , Branchial Region/cytology , Branchial Region/metabolism , Cell Division , Enamel Organ/cytology , Enamel Organ/embryology , Enamel Organ/metabolism , Epithelium/embryology , Epithelium/metabolism , Gene Expression Regulation, Developmental , Gene Transfer Techniques , Mandible/embryology , Mice , Morphogenesis , Mutation , Oligonucleotides, Antisense/genetics , Organ Culture Techniques , Protein Serine-Threonine Kinases , RNA, Messenger/analysis , Receptor, Transforming Growth Factor-beta Type II , Receptors, Transforming Growth Factor beta/genetics , Reverse Transcriptase Polymerase Chain Reaction , Signal TransductionABSTRACT
Platelet-derived growth factor A (PDGF-A) binding to the PDGF receptor alpha (PDGFR-alpha) mediates signal transduction processes related to DNA synthesis, cell migrations, cytodifferentiation, and wound healing. Recent studies indicate that PDGFR-alpha functions during cranial neural crest cell migrations and first branchial arch morphogenesis (Stephenson et al. [1991] Proc. Natl. Acad. Sci. USA 88:6-10; Morrison-Graham et al. [1992] Development 115:133-142; Hu et al. [1995] Int. J. Dev. Biol. 39:939-945; Soriano [1997] Development 124:2691-2700). The present studies were designed to test the hypothesis that PDGF-A, interacts with its cognate receptor PDGFR-alpha via an autocrine mechanism that regulates the timing, rates, and size of embryonic mouse tooth morphogenesis. Both PDGF-A and PDGFR-alpha transcripts were coordinately expressed in mandibular prominences prior to and during tooth formation using reverse transcriptase-polymerase chain reaction (RT-PCR). During the dental lamina stage, ligand and receptor were present in both enamel organ epithelium and adjacent mesenchymal cells. During the bud stage, ligand and receptor were localized mainly to the enamel organ epithelium. Exogenous PDGF-A at 20 ng/ml enhanced tooth development to reach the cap stage with increased tooth size (P < 0.05) using embryonic day (E)10 mandibular explants cultured in serumless, chemically defined medium. A significant increase in DNA synthesis was observed within enamel organ epithelium at E10+4 when the mandibular explants were treated with PDGF-A at 20 ng/ml. These data suggest that PDGF-A and its cognate receptor (PDGFR-alpha) regulate the size and stage of tooth development via an autocrine mechanism during odontogenesis in vitro.
