PlGF-MMP-9-expressing cells restore microcirculation and efficacy of cell therapy in aged dystrophic muscle

Sclerosis and reduced microvessel density characterize advanced stages of muscular dystrophy and hamper cell or gene delivery, precluding treatment of most individuals with Duchenne muscular dystrophy. Modified tendon fibroblasts expressing an angiogenic factor (placenta growth factor, PlGF) and a metalloproteinase (matrix metalloproteinase-9, MMP-9) are able to restore a vascular network and reduce collagen deposition, allowing efficient cell therapy in aged dystrophic mice. These data open the possibility of extending new therapies to currently untreatable individuals.     

Removal of municipal solid waste COD and NH4–N by phyto-reduction: A laboratory–scale comparison of terrestrial and aquatic species at different organic loads

Laboratory scale tests on phytodepuration of raw and pre-treated leachate from municipal sanitary waste were carried out with four vegetable aquatic and terrestrial species at different organic loads. We used the terrestrial species Stenotaphrum secundatum and the free-floating aquatic species Lemna minor, Eichhornia crassipes and Myriophyllum verticellatum to purify leachate from municipal solid waste. The organic load characterized by COD varied from 2–30 g m⁻² day⁻¹. Blanks using tap water served as controls. Duration of the experiments varied from 9–90 days. Maximum concentrations in the experiments were 1600 mg l⁻¹ COD and 300 mg l⁻¹ NH₄–N for S. secundatum. Best results in terms of COD, BOD, and ammonia removal were obtained for raw leachate with COD=2 g m⁻² day⁻¹ in free water surface (FWS) wetlands, and with 2 and 5 g m⁻² day⁻¹ in subsurface flow (SSF) wetlands. Results show that for pretreated leachate (labeled c) low in BOD and NH₄–N, the aquatic species showed low removal and stress even at the lowest load of COD=2 g m⁻² day⁻¹. We cannot say if this is due to the pretreatment itself or the chemical or microbial composition of this leachate. The Stenotaphrum system operated well with this load of leachate c. For untreated leachate (type a and b) the removal and plant growing conditions seemed good at COD=2 g m⁻² day⁻¹. For S. secundatum a load of COD=5 g m⁻² day⁻¹ operated well. All loads above COD=5 g m⁻² day⁻¹ caused low removal and stress, and the green parts of the plants disappeared. Oxygen was, however, consumed throughout the experimental period. For pretreated leachate (type c), the removal of COD were low (−24 to 17%) but good for NH₄–N (52–91%). This leachate also experienced high ammonia removal from the beginning of the experiments, probably due to existing consortia of nitrifying bacteria in it. Statistical analysis shows that the S. secundatum and L. minor systems maintained higher oxygen levels than the M. verticellatum and E. crassipes systems, when operated with tap water. For Lemna minor, this may be due to a better capacity for transporting oxygen into the water. With leachate all S. secundatum systems have higher oxygen levels than the aquatic systems, basically because the water content of the soil has been kept well below saturation. S. secundatum shows a significantly lower removal of COD than did the aquatic systems at a loading of COD=2 g m⁻² day⁻¹ of raw leachate. There is no significant difference between the systems in the removal of NH₄–N at a loading of COD=2 g m⁻² day⁻¹ of both types of leachate. E. crassipes has a lower removal of NH₄–N than M. verticellatum and S. secundatum at a loading of 5 g m⁻² day⁻¹ of COD of both types of leachate. In our experiments, it appears that the amount of free ammonia explains the toxicity of the leachate to the plants. This, however, does not exclude other possible toxic factors.     

REN: a novel,developmentally regulated gene that promotes neural cell differentiation

Expansion and fate choice of pluripotent stem cells along the neuroectodermal lineage is regulated by a number of signals, including EGF, retinoic acid, and NGF, which also control the proliferation and differentiation of central nervous system (CNS) and peripheral nervous system (PNS) neural progenitor cells. We report here the identification of a novel gene, REN, upregulated by neurogenic signals (retinoic acid, EGF, and NGF) in pluripotent embryonal stem (ES) cells and neural progenitor cell lines in association with neurotypic differentiation. Consistent with a role in neural promotion, REN overexpression induced neuronal differentiation as well as growth arrest and p27Kip1 expression in CNS and PNS neural progenitor cell lines, and its inhibition impaired retinoic acid induction of neurogenin-1 and NeuroD expression. REN expression is developmentally regulated, initially detected in the neural fold epithelium of the mouse embryo during gastrulation, and subsequently throughout the ventral neural tube, the outer layer of the ventricular encephalic neuroepithelium and in neural crest derivatives including dorsal root ganglia. We propose that REN represents a novel component of the neurogenic signaling cascade induced by retinoic acid, EGF, and NGF, and is both a marker and a regulator of neuronal differentiation.     

The meso-angioblast: a multipotent,self-renewing cell that originates from the dorsal aorta and differentiates into most mesodermal tissues

We have previously reported the origin of a class of skeletal myogenic cells from explants of dorsal aorta. This finding disagrees with the known origin of all skeletal muscle from somites and has therefore led us to investigate the in vivo origin of these cells and, moreover, whether their fate is restricted to skeletal muscle, as observed in vitro under the experimental conditions used. To address these issues, we grafted quail or mouse embryonic aorta into host chick embryos. Donor cells, initially incorporated into the host vessels, were later integrated into mesodermal tissues, including blood, cartilage, bone, smooth, skeletal and cardiac muscle. When expanded on a feeder layer of embryonic fibroblasts, the clonal progeny of a single cell from the mouse dorsal aorta acquired unlimited lifespan, expressed hemo-angioblastic markers (CD34, Flk1 and Kit) at both early and late passages, and maintained multipotency in culture or when transplanted into a chick embryo. We conclude that these newly identified vessel-associated stem cells, the meso-angioblasts, participate in postembryonic development of the mesoderm, and we speculate that postnatal mesodermal stem cells may be derived from a vascular developmental origin.     

Subcellular localization of epidermal growth factor in human submandibular gland

Epidermal growth factor in human submandibular gland was localized at the subcellular level by means of an immunogold staining method. Labelling was observed in serous acini and ducts. In the acini, gold particles were found within secretory granules, indicating that the growth factor is released into the saliva through granule exocytosis. In the ductal system, the most intense reactivity was revealed in the principal cells of striated ducts. In these cells, an abundant population of small cytoplasmic vesicles was specifically stained. Immunoreactive vesicles were found both apically and basally, suggesting that ductal cells can release their products not only into the saliva but also into the interstitium.     

Cellular heterogeneity during vertebrate skeletal muscle development

Although skeletal muscles appear superficially alike at different anatomical locations, in reality there is considerably more diversity than previously anticipated. Heterogeneity is not only restricted to completely developed fibers, but is clearly apparent during development at the molecular, cellular and anatomical level. Multiple waves of muscle precursors with different features appear before birth and contribute to muscular diversification. Recent cell lineage and gene expression studies have expanded our knowledge on how skeletal muscle is formed and how its heterogeneity is generated. This review will present a comprehensive view of relevant findings in this field.