Quantitative determination and localization of cathepsin D and its inhibitors.

A literature survey was performed of the methods of quantitative assessment of the activity and concentration of cathepsin D and its inhibitors. Usefulness of non-modified and modified proteins and synthetic peptides as measurement substrates was evaluated. The survey includes also chemical and immunochemical methods used to determine the distribution of cathepsin D and its inhibitors in cells and tissues

Cathepsin A activity of a parietal thrombus of an abdominal aortic aneurysm

We evaluated the cathepsin A activity of a parietal thrombus of an abdominal aortic aneurysm. We compared this activity to that of a retracted blood clot homogenate. Cathepsin A of aneurysm parietal thrombus homogenate and blood clot homogenate showed the highest activity on Z–Phe–Ala. It was lower on Z–Phe–Phe, Z–Glu–Tyr, Z–Glu–Phe, Z–Gly–Phe, and the lowest activity was on Z–Gly–Ala. We conclude that cathepsin A’s activity on a parietal thrombus of an aneurysm is much higher than blood clot cathepsin A activity. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 1, pp. 10–12

Regulatory role of cathepsin D in apoptosis.

Cathepsin D (CTSD, EC 3.4.23.5) is well known aspartyl protease. Among different role in cell physiology, a new function of this enzyme is examined. Cathepsin D is an important regulator of apoptotic pathways in cells. It acts at different stage of intrinsic and extrinsic pathway of apoptosis. Cathepsin D can either induce apoptosis in presence of cytotoxic factors, but in certain studies an inhibitory role in apoptosis was also reviewed. Detailed review of involvement of cathepsin D in cell apoptosis is a purpose of this paper

Human cathepsin D.

A literature survey was performed of human cathepsin D gene, cathepsin D biosynthesis, posttranslatory modifications, transport within the cell, substrate specificity and catalytic effect. Methods used to determine the activity and level of this proteinase as well as its role in the biochemistry and pathobiochemistry of cells, tissues and organs were considered

Czynnik wzrostu śródbłonka naczyniowego i jego receptory w ścianie żylaków kończyn

Background. Disturbances in the regulation of vessel wall homeostasis are a potential factor initiating varicose vein development. Vascular endothelial growth factor A (VEGF-A) may play a hypothetic role in this process. The aim of the study was to evaluate mRNA expression and protein content of VEGF-A and its receptors (VEGF R1, VEGF R2) in varicose veins and varicose veins complicated by thrombophlebitis. Material and methods. Walls of varicose veins and varicose veins complicated by thrombophlebitis were the studied material. Walls of normal saphenous veins, which were harvested from patients with chronic limb ischaemia undergoing infrainguinal by-pass grafting, were the control material. The RT-PCR method was employed to assess mRNA expression of VEGF-A and its receptors, whereas the ELISA method was used to evaluate contents of VEGF-A and its receptors. Results. VEGF-A and VEGF R2 mRNA expression is increased, whereas VEGF R1 mRNA expression is unchanged in the wall of varicose veins in comparison with the wall of normal ones. VEGF-A and VEGF R1 mRNA expression is increased in the wall of varicose veins complicated by thrombophlebitis in comparison with walls of normal and varicose veins. VEGF R2 mRNA expression is also increased in the wall of varicose veins complicated by thrombophlebitis in comparison with control veins, but it is comparable to that in the wall of varicose veins. Changes in mRNA expression are correlated with appropriate changes in protein contents of VEGF-A, VEGF R1, and VEGF R2. Conclusions. The demonstrated changes in mRNA expression, as well as in the contents of VEGF-A, VEGF R1, and VEGF R2, in the wall of varicose veins may be accepted as one of the reasons for the clinical symptoms of the disease and can predispose to its progression. Acta Angiol 2011; 17, 2: 141–149Wstęp. Zaburzenia regulacji homeostazy ściany naczyniowej są potencjalnym czynnikiem inicjującym rozwój żylaków kończyn. Hipotetyczną rolę w tym procesie może odgrywać czynnik wzrostu śródbłonka naczyniowego A (VEGF-A). Celem pracy była ocena ekspresji mRNA i zawartości VEGF-A oraz jego receptorów (VEGF R1, VEGF R2) w ścianie żylaków niepowikłanych i żylaków powikłanych zakrzepowym zapaleniem. Materiał i metody. Materiałem badanym były ściany żylaków niepowikłanych i żylaków w stanie zakrzepowego zapalenia. Materiałem kontrolnym były ściany prawidłowych żył odpiszczelowych, pobrane od chorych na przewlekłe niedokrwienie kończyn, u których wykonano zabieg pomostowania udowo-podkolanowego. Na podstawie metody reakcji polimerazy łańcuchowej w połączeniu z odwrotną transkrypcją (RT-PCR) oceniono ekspresję mRNA VEGF-A i jego receptorów, natomiast wykorzystując metodę ELISA, zbadano zawartość VEGF-A i jego receptorów. Wyniki. W ścianie żylaków w porównaniu ze ścianą żył prawidłowych ekspresja VEGF-A mRNA oraz VEGF R2 mRNA jest większa, podczas gdy ekspresja VEGF R1 mRNA nie ulega istotnym zmianom. W ścianie żylaków powikłanych zakrzepowym zapaleniem ekspresja VEGF-A mRNA i VEGF R1 mRNA jest większa w porównaniu ze ścianą żył prawidłowych i ścianą żylaków niepowikłanych. Natomiast ekspresja VEGF R2 mRNA jest również większa niż w ścianie żył prawidłowych, ale porównywalna z ekspresją w ścianie żylaków niepowikłanych. Zmianom w ekspresji mRNA odpowiadają zmiany w zawartości białka VEGF-A, VEGF R1 i VEGF R2 w ścianie żylnej. Wnioski. Wykazane zmiany w ekspresji mRNA i zawartości VEGF-A, VEGF R1 i VEGF R2 w ścianie żylaków kończyn mogą być jedną z przyczyn objawów klinicznych choroby oraz predysponować do jej postępu. Acta Angiol 2011; 17, 2: 141–14

Cathepsin D inhibitors.

Inhibitors of cathepsin D belong to chemical compounds that estrify carboxyl groups of the Asp33 and Asp231 residues of its catalytic site, penta-peptides containing statin, i.e. the amino acid similar in structure to the tetraedric indirect product, and polypeptides found in the spare organs of many plants and forming permanent noncovalent complexes with cathepsin. Cathepsin D activity is also inhibited by alpha2-macroglobulin and antibodies directed against this enzyme. Methods used to determine the activity and concentration of these inhibitors and their analytical, preparative and therapeutic applications are discussed

Ocena zawartości insulinopodobnego czynnika wzrostu I w ścianie żylaków kończyn

Background. Extensive extracellular matrix remodelling is present in the wall of varicose veins. The process is controlled by numerous factors, including peptide growth factors. The aim of the study was to evaluate the content of insulin-like growth factor I (IGF-I), insulin-like growth factor binding protein 3 (IGFBP-3), and insulin-like growth factor-1 receptor (IGF-1R) in the wall of varicose veins and varicose veins complicated by thrombophlebitis. Material and methods. Walls of varicose veins and varicose veins complicated by thrombophlebitis made up the studied material. Walls of normal saphenous veins, which were harvested from patients with chronic limb ischaemia undergoing infrainguinal by-pass grafting, made up the control material. Contents of IGF-I, IGFBP-3, and IGF-1R in the investigated tissues were evaluated using the ELISA method. Results. IGF-I content in walls of varicose veins and varicose veins complicated by thrombophlebitis were comparable, and were increased compared to the walls of normal veins. IGFBP-3 content was significantly increased only in the walls of varicose veins complicated by thrombophlebitis. IGF-1R level were increased in the walls of varicose veins compared to the walls normal of veins; whereas, in the course of thrombophlebitis, it was increased compared to the walls of normal veins, as well as uncomplicated varicose veins. Conclusions. The study results indicate differences in IGF-I, IGFBP-3, and IGF-1R content between walls of healthy veins, varicose veins, and varicose veins complicated by thrombophlebitis. Acta Angiol 2011; 17, 4: 264–270Wstęp. W ścianie żylaków kończyn wykazano rozległą przebudowę macierzy pozakomórkowej tkanki łącznej. Proces ten jest kontrolowany przez wiele czynników, w tym przez peptydowe czynniki wzrostu. Celem pracy jest ocena zawartości insulinopodobnego czynnika wzrostu I (IGF-I), białka 3 wiążącego insulinopodobny czynnik wzrostu (IGFBP-3) oraz receptora typu 1 dla IGF (IGF-1R) w ścianie żylaków i żylaków powikłanych zakrzepowym zapaleniem. Materiał i metody. Materiałem badanym były ściany żylaków i żylaków w stanie zakrzepowego zapalenia. Materiał kontrolny stanowiły ściany prawidłowych żył odpiszczelowych, pobrane od chorych na przewlekłe niedokrwienie kończyn, u których wykonano pomostowanie udowo-podkolanowe. Metodą ELISA oceniono zawartość IGF-I, IGFBP-3 oraz IGF-1R w badanych tkankach. Wyniki. Zawartość IGF-I w ścianie żylaków oraz żylaków powikłanych zakrzepowym zapaleniem jest porównywalna i jest większa niż w ścianie żył prawidłowych. Zawartość IGFBP-3 jest większa wyłącznie w ścianie żylaków powikłanych zakrzepowym zapaleniem. Zawartość IGF-1R w ścianie żylaków jest większa niż w ścianie żył prawidłowych. Ponadto w przebiegu zakrzepowego zapalenia zawartość tego receptora jest większa także w porównaniu ze ścianą żylaków niepowikłanych. Wnioski. Wyniki badań wskazują na różnice w zawartości IGF-I, IGFBP-3 i IGF-1R pomiędzy ścianami żył zdrowych, żylaków kończyn oraz żylaków kończyn powikłanych zakrzepowym zapaleniem. Acta Angiol 2011; 17, 4: 264–27

Role of cathepsin A and cathepsin C in the regulation of glycosidase activity

Increased tissue activity of cathepsin A and cathepsin C can be observed in many pathological conditions. It is associated with an enhanced degradation of glycosaminoglycans, proteoglycans, and glycoproteins, and results in their decreased tissue content. Cathepsin C releases the glycosidases from complexes formed with cathepsin A, and reinstates their activity. In this review a current state of knowledge is presented concerning the regulation of selected glycosidases activity by cathepsin A (EC 3.4.16.1) and C (EC 3.4.14.1)

Regulatory role of cathepsin D in apoptosis.

Cathepsin D (CTSD, EC 3.4.23.5) is well known aspartyl protease. Among different role in cell physiology, a new function of this enzyme is examined. Cathepsin D is an important regulator of apoptotic pathways in cells. It acts at different stage of intrinsic and extrinsic pathway of apoptosis. Cathepsin D can either induce apoptosis in presence of cytotoxic factors, but in certain studies an inhibitory role in apoptosis was also reviewed. Detailed review of involvement of cathepsin D in cell apoptosis is a purpose of this paper

Thiocyanate concentration in saliva of cystic fibrosis patients.

Thiocyanates (SCN-) are ubiquitous in nature. There are indispensable part of host defense system that act as a substrate for lactoperoxidase (LPO). In our study we present initial data on SCN- concentration in saliva of CF patients in comparison to healthy non-smokers and healthy smokers. 5 ml of saliva was collected from each subject to a sterile tube and thiocyanate concentration was measured in each sample. The results of the measurements are presented on Fig. 1. Mean concentration of SCN- in saliva of CF patients was 0.031 +/- 0.0052 g/l, in healthy non-smokers 0.039 +/- 0.0048 g/l and in healthy smokers 0.048 +/- 0.0161 g/l. The differences between each group were statistically significant. Studies on larger group of patients and probably on different material (BALF or induced sputum) should present interesting data complementing the in vitro studies