N-acetyl-L-cysteine abrogates fibrogenic properties of fibroblasts isolated from Dupuytren's disease by blunting TGF-beta signalling

Dupuytren's disease, a benign fibroproliferative disorder of the palmar fascia, represents an ideal model to study tissue fibrosis. Transforming growth factor-beta1 (TGF-beta1) and its downstream Smad signalling system is well established as a key player during fibrogenesis. Thus, targeting this basic pathomechanism seems suitable to establish new treatment strategies. One such promising treatment involves the substance N-acetyl-L-cysteine (NAC), shown to have antifibrotic properties in hepatic stellate cells and rat fibroblasts. In order to investigate antifibrotic effects of N-acetyl-L-cysteine (NAC), fibroblasts were isolated from surgically resected fibrotic palmar tissues (Dupuytren fibroblasts, DF) and exposed to different concentrations of NAC and recombinant TGF-beta1. Fibroblasts isolated from tendon pulleys served as controls (control fibroblasts, CF). Smad signalling was investigated by a Smad binding element driven reporter gene analysis. Both cell types express TGF-beta1, indicating autocrine signalling in DF and CF. This was confirmed by comparing reporter gene activity from LacZ and Smad7 adenovirus infected cells. NAC treatment resulted in abrogation of Smad mediated signalling comparable to ectopically overexpressed Smad7, even when the cells were stimulated with recombinant TGF-beta1 or ectopically expressed a constitutively active TGF-beta receptor type I. Additionally, NAC dose-dependently decreased expression of three major indicators of impaired fibrotic matrix turnover, namely alpha-smooth muscle actin (alpha-SMA), alpha 1 type I procollagen (Col1A1), and plasminogen activator inhibitor-type I (PAI-1). Our results suggest that TGF-beta signalling and subsequent expression of fibrogenesis related proteins in Dupuytren's disease is abrogated by NAC thus providing a basis for a therapeutic strategy in Dupuytren's disease and other fibroproliferative disorders.     

Lack of association between prohibitin 3' untranslated region C-->T polymorphism and breast cancer in a Turkish population

The 3' untranslated region (3'UTR) of the prohibitin gene has a positive effect on arresting cell proliferation between G1 and S phases and inhibits DNA synthesis. A C-to-T transition within this region creates a variant that alters mRNA function and has been shown to be associated with an increased breast cancer risk among young North Americans who are under 50 years and have at least one first-degree relative with breast cancer. We carried out a population-based case-control study to assess whether this association exists in Turkish women. We examined 106 breast cancer patients and 154 healthy controls by PCR and restriction fragment length polymorphism analysis. In the prohibitin 3'UTR, we did not detect a difference in CT/TT genotype frequency (p = 0.694; odds ratio [OR], 1.106; 95% confidence intervals [CI], 0.659-1.86) or in C/T allele frequency (p = 0.850; OR, 1.043; 95% CI, 0.667-1.62) between the all breast cancer patients and the controls. The results did not change in subgroups defined by age or family history. Hence our results do not lend support to the hypothesis that this polymorphism contributes to risk of breast cancer. The prohibitin T variant is not associated with the risk of breast cancer in Turkish women.     

Temporoparietal fascia: an anatomic and histologic reinvestigation with new potential clinical applications

Temporoparietal fascia constitutes a very important structural unit from both an aesthetic and a reconstructive surgical point of view. A histologically supported anatomic study was conducted for the reappraisal of the anatomic relationships and clinical application potentials of the data obtained. Anatomy of the temporoparietal fascia was investigated on 20 sides from 10 cadavers. After dissections, necropsies were obtained to demonstrate histologic features of the temporoparietal fascia. The outer part of the temporoparietal fascia is continuous with the superficial musculoaponeurotic system (SMAS) in the inferior border and with orbicularis oculi and frontalis muscles in the anterior border. Therefore, plication of the temporoparietal fascia can increase tightness of the SMAS, orbicularis oculi, and frontalis muscle in rhytidectomy. The frontal branches of facial nerve were noted to course parallel to the frontal branch of the superficial temporal artery, lying deeper to the temporoparietal fascia within the innominate fascia. In the view of these findings, conventional subfascial dissection, which is performed to protect frontal branches of the facial nerve, is not reasonable during the temporal part of rhytidectomy. Careful subcutaneous dissection just under the hair follicles is more appropriate to avoid nerve injury and also provides excellent exposure of the temporoparietal fascia for plication in rhytidectomy with protection of the auriculotemporal nerve and the superficial temporal vessels. Furthermore, two layered structures of the temporoparietal fascia are very suitable to insert a framework into the temporoparietal fascia for ear reconstruction to eliminate some of the shortcomings of Brent's technique. A thin muscle layer was also noted within the outer part of the temporoparietal fascia below the temporal line; the term "temporoparietal myofascial flap" would, therefore, be more accurate than "temporoparietal fascial flap." Finally, the innominate fascia and the deep temporal fascia can be elevated with the two layers of the temporoparietal myofascial flap to obtain a well-vascularized, four-layered myofascial flap based on the superficial temporal vessels. This multilayered flap can be used to reconstruct all defects when fine, pliable, thin, multilayered flaps are required.     

Mg2+-independent hairpin ribozyme catalysis in hydrated RNA films

The hairpin ribozyme catalyzes RNA cleavage in partially hydrated RNA films in the absence of added divalent cations. This reaction exhibits the characteristics associated with the RNA cleavage reaction observed under standard conditions in solution. Catalysis is a site-specific intramolecular transesterification reaction, requires the 2'-hydroxyl group of substrate nucleotide A(-1), and generates 2',3'-cyclic phosphate and 5'-hydroxyl termini. Mutations in both ribozyme and substrate abolish catalysis in hydrated films. The reaction is accelerated by cations that may enhance binding, conformational stability, and catalytic activity, and is inhibited by Tb3+. The reaction has an apparent temperature optimum of 4 degrees C. At this temperature, cleavage is slow (k(obs): 2 d(-1)) and progressive, with accumulation of cleavage products to an extent of 40%. The use of synthetic RNAs, chelators, and analysis of all reaction components by inductively coupled plasma-optical spectrophotometry (ICPOES) effectively rules out the possibility of contaminating divalent metals in the reactions. Catalysis is minimal under conditions of extreme dehydration, indicating that the reaction requires hydration of RNA by atmospheric water. Our results provide a further caution for those studying the biochemical activity of ribozymes in vitro and in cells, as unanticipated catalysis could occur during RNA manipulation and lead to misinterpretation of data.     

Application of remote sensing techniques for water quality monitoring

A detailed study and experimental work performed to establish the properties of multispectral remote sensing techniques as applied to the monitoring of water quality, is hihhly desirable since direct synoptic field measurements concerning the quantity and type of pollutants in water is not possible at present.The multispectral characteristics of the signal received from water are a function of hydrological, biological and chemical characteristics of water, and the physical properties of the remote sensor related environment. Spectral data encompassing of the spectral region 0.36 m and 2.36 m showed that the detectibility of water pollutants, using spectral signatures in the 0.4 to 0.9 m range, is promising and feasible.     

Satiation meal and the effects of meal and body sizes on gastric evacuation rate in brook trout Salvelinus fontinalis fed commercial pellets

Gastric evacuation (GE) experiments were performed on brook trout Salvelinus fontinalis fed commercial food pellets. The experiments included small fish (36 g; 15 cm total length, L_T) fed meals of 0·2, 0·4 and 0·8 g and large fish (152 g; 23 cm) fed meals of 0·8, 2·0 and 4·0 g at temperatures ranging from 15·1 to 18·2° C. The stomach contents were thereafter sampled and weighed at 3 h intervals until the first empty stomach was observed. The course of GE was examined by use of a general power function of the data that revealed that the square‐root function described the GE rate (GER) by the current stomach content mass independently of original meal size. Using the square‐root function, the relationship between GER and fish size was described by a power function of fish length, whereas the effect of temperature was described by a simple exponential function. GER of the commercial pellets fed to S. fontinalis could thus be described by (g h^?1), where S_t is stomach mass (g) at time t (h), L is total fish length (cm) and T is temperature (° C). The result of this study should provide a useful tool for planning of feeding regimes in production of S. fontinalis by optimizing growth and minimizing food waste.     

Dual effect of formycin a upon the hydrolysis of phosphoinositides in perifused pancreatic islets

Formycin A (1.0 mM) caused a rapid, sustained and rapidly reversible inhibition of effluent radioactivity in rat pancreatic islets prelabelled with myo-[2-³H]inositol and perifused in the presence of 8.3 mM -glucose. This coincided with a progressive decrease in islet ATP content and transient inhibition of insulin release. Theraafter, however, formycin A increased glucose-induced insulin release. Moreover, in islets that were preincubated with myo-[2-³H]inositol and then exposed during perifusion to a rise in -glucose concentration from 2.8 to 16.7 mM, the release of insulin and ³H fractional outflow rate at both the low and high hexose concentrations were much higher when both the preincubation and perifusion were conducted in the presence, rather than absence, of formycin A. It is concluded that formycin A first inhibits and later enhances both the hydrolysis of phosphoinositides and release of insulin, these effects being possibly related to changes in the islet cell content of adenosine and/or formycin A triphosphates.