miR-145 Contributes to Hypertrophic Scarring of the Skin by Inducing Myofibroblast Activity

Hyperthrophic scarring of the skin is caused by excessive activity of skin myofibroblasts after wound healing and often leads to functional and/or aesthetic disturbance with significant impairment of patient quality of life. MicroRNA (miRNA) gene therapies have recently been proposed for complex processes such as fibrosis and scarring. In this study, we focused on the role of miR-145 in skin scarring and its influence in myofibroblast function. Our data showed not only a threefold increase of miR-145 levels in skin hypertrophic scar tissue but also in transforming growth factor β1 (TGF-β1)-induced skin myofibroblasts compared with healthy skin or nontreated fibroblasts (p < 0.001). Consistent with the upregulation of miR-145 induced by TGF-β1 stimulation of fibroblasts, the expression of Kruppel-like factor 4 (KLF4) was decreased by 50% and α-smooth muscle actin (α-SMA) protein expression showed a threefold increase. Both could be reversed by miR-145 inhibition (p < 0.05). Restoration of KLF4 levels equally abrogated TGF-β1–induced α-SMA expression. These data demonstrate that TGF-β1 induces miR-145 expression in fibroblasts, which in turn inhibits KLF4, a known inhibitor of α-SMA, hence upregulating α-SMA expression. Furthermore, treatment of myofibroblasts with a miR-145 inhibitor strongly decreased their α-1 type I collagen expression, TGF-β1 secretion, contractile force generation and migration. These data demonstrate that upregulation of miR-145 plays an important role in the differentiation and function of skin myofibroblasts. Additionally, inhibition of miR-145 significantly reduces skin myofibroblast activity. Taken together, these results suggest that miR-145 is a promising therapeutic target to prevent or reduce hypertrophic scarring of the skin.     

Removal of heavy metals from contaminated sewage sludge using Aspergillus niger fermented raw liquid from pineapple wastes

The environmental benefits derived from using citric acid in the removal of heavy metals from contaminated sewage sludge have made it promising as an extracting agent in the chemical extraction process. At present, citric acid is produced commercially by fermentation of sucrose using mutant strains of Aspergillus niger (A. niger), and chemical synthesis. In recent years, various carbohydrates and wastes (such as pineapple wastes) have been considered experimentally, to produce citric acid by A. niger. This study investigated the potential of using A. niger fermented raw liquid from pineapple wastes as a source of citric acid, in extracting chromium (Cr), copper (Cu), lead (Pb), nickel (Ni) and zinc (Zn) from anaerobically digested sewage sludge. Results of the study revealed that metal removal efficiencies varied with pH, forms of metals in sludge and contact time. At pH approaching 4, and contact time of 11 days, A. niger fermented liquid seemed to remove all Cr and Zn while removing 94% of Ni. Moreover, chemical speciation studies revealed that metals which are predominantly in the exchangeable and oxidizable phases seemed to exhibit ease of leachability (e.g., Zn). The by-products of the process such as pineapple pulp and mycelium which are rich in protein, can still be used as animal feed. It can be said therefore that this novel process provides a sustainable way of managing contaminated sewage sludge.     

Effect of Drying Process on Heavy Metals Fractionation of Contaminated Sludge for Land Application

Metal fractionation studies on metals in sludge are usually done on dried sludge. Although there are advantages in this form of sludge, it is possible that fractionation of metals in sludge may be influenced by the drying process, which consequently affects mobility of metals at disposal. In this study, sequential chemical extraction was done to assess the effect of drying on fractionation of cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb), nickel (Ni), and zinc (Zn) on wet and dewatered (air-dried and oven-dried) anaerobically digested sludge samples in Bangkok, Thailand. Results revealed an insignificant difference in the fractionation profile of metals for both wet and dewatered sludge. The higher percentage of residual fraction for Cu and oxidizable fraction for Zn in the wet sludge, however, makes this form more suitable for direct land application. In the case of oven-dried and air-dried sludge, the forms of some metals (e.g., Pb and Zn) seem to vary significantly as the sludge is oven-dried, with Pb decreasing its residual phase by 15%, and Zn increasing in oxidizable phase by 41%. The results seem to indicate that drying affects bioavailability of metals in sludge and air-drying seems to favor sludge for land application.     

Helen House--a hospice for children: analysis of the first year.

Helen House, a hospice for children, opened in Oxford in November 1982. From then until December 1983 care was provided for 52 children with terminal illness, progressive and incurable illness, and very severe handicap. The children came from a wide area of the United Kingdom, and eight died during the year. The style of care provided in Helen House is based on that of the families in their own homes, with a minimum of rules, regulations, and routines. Terminal care follows the principles developed by hospices experienced in the care of adults. Despite the great sadness and distress associated with every child admitted, Helen House is neither a sombre nor a depressing place.     

Purification and kinetic properties of 6-phosphofructo-1-kinase from gilthead sea bream muscle

The kinetic properties of 6-phosphofructo-1-kinase (PFK) from skeletal muscle (PFKM) of gilthead sea bream (Sparus aurata) were studied, after 10,900-fold purification to homogeneity. The native enzyme had an apparent molecular mass of 662 kDa and is composed of 81 kDa subunits, suggesting a homooctameric structure. At physiological pH, S. aurata PFKM exhibited sigmoidal kinetics for the substrates, fructose-6-phosphate (fru-6-P) and ATP. Fructose-2,6-bisphosphate (fru-2,6-P(2)) converted the saturation curves for fru-6-P to hyperbolic, activated PFKM synergistically with other positive effectors of the enzyme such as AMP and ADP, and counteracted ATP and citrate inhibition. The fish enzyme showed differences regarding other animal PFKs: it is active as a homooctamer, and fru-2,6-P(2) and pH affected affinity for ATP. By monitoring incorporation of (32)P from ATP, we show that fish PFKM is a substrate for the cAMP-dependent protein kinase. The mechanism involved in PFKM activation by phosphorylation contrasts with previous observations in other species: it increased V(max) and did not affect affinity for fru-6-P. Unlike the mammalian muscle enzyme, our findings support that phosphorylation of PFKM may exert a major role during starvation in fish muscle.