A novel function for tissue inhibitor of metalloproteinases-3 (TIMP3): inhibition of angiogenesis by blockage of VEGF binding to VEGF receptor-2

Tissue inhibitor of metalloproteinases-3 (TIMP3) is one of four members of a family of proteins that were originally classified according to their ability to inhibit matrix metalloproteinases (MMP). TIMP3, which encodes a potent angiogenesis inhibitor, is mutated in Sorsby fundus dystrophy, a macular degenerative disease with submacular choroidal neovascularization. In this study we demonstrate the ability of TIMP3 to inhibit vascular endothelial factor (VEGF)-mediated angiogenesis and identify the potential mechanism by which this occurs: TIMP3 blocks the binding of VEGF to VEGF receptor-2 and inhibits downstream signaling and angiogenesis. This property seems to be independent of its MMP-inhibitory activity, indicating a new function for this molecule.     

Seasonal courses of nutrients and heavy metals in water,sediment and above- and below-ground Typha domingensis biomass in Lake Burullus (Egypt): Perspectives for phytoremediation

The present study was carried out in natural stands of Typha domingensis in Lake Burullus, Egypt, to investigate (1) nutrient dynamics and heavy metals accumulation in its organs, (2) the phytoextractive potential of its organs and (3) the amount of nutrients and heavy metals released back into the water after decomposition of the dead tissues. Nitrogen concentrations were higher in the shoot than in the root and rhizome, while P, Ca, Cu, Fe, Zn and ash concentrations were higher in the root than in the rhizome and shoot. Significant differences in the concentrations of Mg, Cd, Cu and ash were assessed during the growing season of T. domingensis. The content of most nutrients and heavy metals in the shoot increased rapidly during the early growing season in February, reached maximal values in July and then decreased again. The nutrient and heavy metal contents in the below-ground portion of the plant showed an opposite trend compared to the shoot; they decreased sharply during the spring, when they were translocated, supporting the heterotrophic phase of shoot growth. However, they increased slightly from July to September and then decreased again. The transfer factors of all nutrients and heavy metals from the sediment to the below-ground organs were greater than unity. The higher translocation ratio of N in T. domingensis shoots makes it suitable for N phytoextraction from water and sediment, while the lower translocation ratios for Cd, Cu, Fe, Pb and Zn make it suitable for metal ion phytostabilisation. The dead shoot biomass of the stands at the end of 2010 amounted to 1950 g DM m⁻², when the seasonal decomposition process began. With a decay rate of 0.0049 day⁻¹, 1624 g DM m⁻² is decomposed in the lake in a year. This is equivalent to releasing the following nutrient and heavy metals into the surrounding water (in g m⁻²): 23.4 N, 0.8 P, 19.2 Ca, 1.8 Mg, 5.6 Na, 32.8 K, 0.01 Cd, 0.01 Cu, 0.84 Fe, 0.12 Pb and 0.03 Zn.     

Ten years primary succession on a newly created landfill at a lagoon of the Mediterranean Sea (Lake Burullus RAMSAR site)

This study was carried out on the transported bed soil dredged from the outlet of Lake Burullus to the Mediterranean Sea and deposited nearby, forming by this way new land that underwent a primary plant succession. The multi-methodological approach comprised floristic inventories, vegetation sampling and soil composition analyses of the study site in order to detect the crucial parameters controlling the plant resettlement on recently deposited soil as related to time, local micro-topography and substrate characteristics. Floristic composition was assessed for the first 10 years of primary succession (2001–2010) on 18 stands of the area, distributed on basement, slope stands and plateau of the landfill, respectively. Vegetation surveys were the basis of multivariate analyses of the vegetation and soil data using TWINSPAN, DCA and CCA. Relationships between the edaphic gradients, floristic composition and species diversity were assessed.     

Bioaccumulation and rhizofiltration potential of Pistia stratiotes L. for mitigating water pollution in the Egyptian wetlands

The bioaccumulation and rhizofiltration potential of P. stratiotes for heavy metals were investigated to mitigate water pollution in the Egyptian wetlands. Plant and water samples were collected monthly through nine quadrats equally distributed along three sites at Al-Sero drain in Giza Province. The annual mean of the shoot biomass was 10 times that of the root. The concentrations of shoot heavy metals fell in the order: Fe < Mn < Cr < Pb < Cu < Zn < Ni < Co < Cd, while that of the roots were: Fe < Mn < Cr < Pb < Zn < Ni < Co < Cu < Cd. The bio-concentration factor (BCF) of most investigated heavy metals, except Cr and Pb, was greater than 1000, while the translocation factor (TF) of most investigated metals, except Pb and Cu, did not exceed one. The rhizofiltration potential (RP) of heavy metals was higher than 1000 for Fe, and 100 for Cr, Pb and Cu. Significant positive correlations between Fe and Cu in water with those in plant roots and leaves, respectively were recorded, which, in addition to the high BCF and RP, indicate the potential use of P. stratiotes in mitigating these toxic metals.     

Expression of Sorsby's fundus dystrophy mutations in human retinal pigment epithelial cells reduces matrix metalloproteinase inhibition and may promote angiogenesis

Sorsby's fundus dystrophy (SFD) is an autosomal dominant degenerative disease of the macula caused by mutations in the tissue inhibitor of metalloproteinase-3 (TIMP-3) gene. Choroidal neovascularization is a hallmark of this disease, which closely resembles the exudative form of age-related macular degeneration. However, the mechanism by which TIMP-3 mutations induce the disease phenotype in SFD remains unknown. To address this question we established human retinal pigment epithelial cell lines expressing wild type or S156C (Ser(156) changed to cysteine) mutant TIMP-3. S156C TIMP-3 had reduced matrix metalloproteinase (MMP) inhibitory activity in retinal pigment epithelial cells and resulted in increased secretion and activation of gelatinase A and B. The conditioned medium from these cells induced angiogenesis in "in vivo" chick chorioallantoic membrane assays that could be reversed with recombinant wild type TIMP-3. Our data indicate that the choroidal neovascularization in SFD may be a result of increased MMP activity, which could lead to the stimulation of angiogenesis. These results also suggest the potential therapeutic use of TIMP-3 or synthetic MMP inhibitors in this disease.     

Tissue Inhibitor of Metalloproteinases-3 Peptides Inhibit Angiogenesis and Choroidal Neovascularization in Mice

Tissue inhibitors of metalloproteinases (TIMPs) while originally characterized as inhibitors of matrix metalloproteinases (MMPs) have recently been shown to have a wide range of functions that are independent of their MMP inhibitory properties. Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a potent inhibitor of VEGF-mediated angiogenesis and neovascularization through its ability to block the binding of VEGF to its receptor VEGFR-2. To identify and characterize the anti-angiogenic domain of TIMP-3, structure function analyses and synthetic peptide studies were performed using VEGF-mediated receptor binding, signaling, migration and proliferation. In addition, the ability of TIMP-3 peptides to inhibit CNV in a mouse model was evaluated. We demonstrate that the anti-angiogenic property resides in the COOH-terminal domain of TIMP-3 protein which can block the binding of VEGF specifically to its receptor VEGFR-2, but not to VEGFR-1 similar to the full-length wild-type protein. Synthetic peptides corresponding to putative loop 6 and tail region of TIMP-3 have anti-angiogenic properties as determined by inhibition of VEGF binding to VEGFR-2, VEGF-induced phosphorylation of VEGFR-2 and downstream signaling pathways as well as endothelial cell proliferation and migration in response to VEGF. In addition, we show that intravitreal administration of TIMP-3 peptide could inhibit the size of laser-induced choroidal neovascularization lesions in mice. Thus, we have identified TIMP-3 peptides to be efficient inhibitors of angiogenesis and have a potential to be used therapeutically in diseases with increased neovascularization.     

Bioaccumulation and translocation of nine heavy metals by Eichhornia crassipes in Nile Delta,Egypt: perspectives for phytoremediation

Abstract

The current research was carried out to estimate the potential of water hyacinth (WH) for removal of nine heavy metals (HMs) from three irrigation canals in Nile Delta. Sampling was achieved in monospecific and homogeneous WH stands at three irrigation canals in the study area, and WH biomass was sampled at monthly intervals from April 2014 to November 2014 using five randomly distributed quadrats (each 0.5?×?0.5?m) at each canal. All HM concentrations were significantly higher in the roots compared with the other WH organs. The WH was recognized by a bioaccumulation factor >1.0 for all HMs. The WH was recognized by translocation factor <1.0 for all HMs (except Pb). In many cases, the concentrations of the HMs in the different organs of WH were correlated with the same HMs in the water. Such correlations indicate that WH reflects the cumulative influences of environmental pollution from the water, and thereby suggesting its potential use in the bio-monitoring of most examined HMs. In conclusion, WH is a promising macrophyte for remediation of irrigation canals polluted with Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn.     

Decomposition dynamics of Phragmites australis litter in Lake Burullus,Egypt

This study estimated the decomposition rate and nutrient dynamics of Phragmites australis litter in Lake Burullus (Egypt) and investigated the amount of nutrients released back into the water after the decomposition of the dead tissues. Phragmites australis detritus decomposition was studied from April to September 2003 utilizing the leaf, stem, and rhizome litterbags technique with coarse mesh (5 mm) bags on five sampling dates and with nine replicate packs per sample. All samples were dried, weighed and analyzed for N, P, Ca, Mg, Na, and K concentrations. The exponential breakdown rate of leaves (?0.0117/day) was significantly higher than that of rhizomes (?0.0040/day) and stems (?0.0036/day). N, Na and K mineralization were the highest from leaf litter, followed by rhizomes and stems, while P, Ca and Mg mineralization were the highest from rhizomes, followed by leaves and stems. The dead shoot biomass at the end of 2003 amounted to 4550 g DM/m² which enters the decomposition process. By using the decay rate of 0.0117 and 0.0036/day for the leaves and stems, 3487 g DM/m² is decomposed in a year, leaving only 1063 g DM/m² after 1 year. This is mainly equivalent to releasing the following nutrients into surrounding water (in g/m²): 24.4 N, 1.1 P, 15.5 Ca, 3.5 Mg, 11.3 Na and 16.7 K. In conclusion, the present study indicates a significant difference in relation to the type of litter; these breakdown rates were generally greater than most rates reported in previous studies that used the same technique and mesh size.     

Bioaccumulation and translocation of heavy metals by nine native plant species grown at a sewage sludge dump site

In the present study, nine native plant species were collected to determine their potential to clean up nine heavy metals from soil of a sewage sludge dump site. Almost all nine plant species grown at sewage sludge dump site showed multifold higher concentrations of heavy metals as compared to plants grown at the reference site. All the investigated species were characterized by a bioaccumulation factor (BF) > 1.0 for some heavy metals. BF was generally higher for Cd, followed by Pb, Co, Cr, Cu, Ni, Mn, Zn, and Fe. The translocation factor (TF) varied among plant species, and among heavy metals. For most studied heavy metals, TFs were <1.0. The present study proved that the concentrations of all heavy metals (except Cd, Co, and Pb) in most studied species were positively correlated with those in soil. Such correlations indicate that these species reflect the cumulative effects of environmental pollution from soil, and thereby suggesting their potential use in the biomonitoring of most heavy metals examined. In conclusion, all tissues of nine plant species could act as bioindicators, biomonitors, and remediates of most examined heavy metals. Moreover, Bassia indica, Solanum nigrum, and Pluchea dioscoridis are considered hyperaccumulators of Fe; Amaranthus viridis and Bassia indica are considered hyperaccumulators of Pb; and Portulaca oleracea is considered hyperaccumulator of Mn.     

Seasonal variation in the phytomass,chemical composition and nutritional value of Azolla filiculoides Lam. along the water courses in the Nile Delta,Egypt

The present study aims to assess phenological behaviour, phytomass production, chemical composition and nutritional value of Azolla filiculoides in the water courses of the Nile Delta, Egypt. The sampling process was carried out seasonally using twenty‐five plots (each of 1 × 1 m) distributed along 15 irrigation canals and 10 drainage canals in the study area. Sprouting of A. filiculoides had its maximum activity during winter, vegetative growth during spring and summer and withering during autumn. Maximum seasonal phytomass (285.2 g DW m^‐2) was found during spring. The annual mean was significantly higher in drainage canals (278.3 g DW m^‐2) than in irrigation canals (144.4 g DW m^‐2). The concentrations of Ca, Mg and Na in A. filiculoides from drainage canals were significantly higher than in those from irrigation canals. Regarding the type of water courses, there were no significant differences in organic contents and nutritive values between the irrigation and drainage canals. The analysis of growth characteristics indicated that A. filiculoides can grow quite well in drainage canals; its macronutrient (N, Ca, Mg, Na and Fe) contents are high enough to allow it to be used as a bio‐fertilizer. On the other hand, A. filiculoides could be considered as a ‘nutrients and heavy metals remover’ especially in drainage canals for wastewater treatment if the plant is harvested during its maximum phytomass. As feed supplement for animals, this plant could be considered as excellent forage because its high levels of carbohydrate, protein, crude fat, total digestible nutrients and the lower crude fiber contents (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)