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Biomedical subjects

E Nowak

Publications and source records attributed to E Nowak.

At least 37 records · Page 2Linked to original sources
PubMed1996

Modulation of gelsolin-induced actin-filament severing by caldesmon and tropomyosin and the effect of these proteins on the actin activation of myosin Mg(2+)-ATPase activity.

We have investigated the cumulative effects of three smooth-muscle actin-binding proteins, gelsolin, caldesmon and tropomyosin, on actin activation of myosin Mg(2+)-ATPase activity under low-ionic-strength conditions. A combination of tropomyosin (at a stoicheiometric ratio to actin) and gelsolin (at a molar ratio to actin of up to 1:100) showed essentially additive stimulatory effects that were counteracted by caldesmon. Suppression of the gelsolin-induced activation of the ATPase by caldesmon was higher in the presence of tropomyosin although it was not complete even at stoicheiometric amounts of both proteins to actin. Since activation of actin-activated ATPase activity of myosin by gelsolin is related to its severing action, it is concluded that caldesmon and tropomyosin cannot fully protect actin filaments against the severing activity of gelsolin. Direct analysis of the actin-severing activity of gelsolin by a fluorimetric assay using pyrene-labelled actin confirmed this conclusion. Tropomyosin and caldesmon in saturating amounts relative to actin inhibited the activity of gelsolin by between 21 and 40% and 25 and 48% respectively, depending on the molar ratio of gelsolin to actin. The inhibitory effect was increased with a combination of both (up to 67%) although it was evident that even under these conditions the actin filaments were not fully protected from being severed by gelsolin. These findings were corroborated by electron-microscopic investigation of actin filaments with or without tropomyosin and caldesmon after the addition of gelsolin.

Actins↗
PubMed1994

Polymerization of actin induced by actin-binding fragments of caldesmon.

Our earlier studies revealed that caldesmon causes assembly of G-actin into polymers morphologically indistinguishable from those formed in the presence of salt (Gałazkiewicz, B., Belagyi, J. and Dabrowska, R. (1989) Eur. J. Biochem. 181, 607-614). In this work we have investigated the effect of actin-binding fragments of caldesmon on actin polymerization process followed by measurements of the changes in fluorescence of pyrenyl conjugated with G-actin and ATP hydrolysis. The results indicate that C-terminal 34 kDa fragment of caldesmon containing two actin-binding sites and 19 kDa containing high-affinity binding site have similar capability to polymerize actin to that of intact molecule. Binding of each of these fragments to G-actin causes bypassing of nucleation phase. The 11.5 kDa fragment comprising low affinity actin-binding site has much lower potency to polymerize actin. Conformation of actin monomers in filaments formed upon 19 kDa fragment and that formed upon 11.5 kDa fragment differs. The former fragment seems to resemble more conformation of monomers in filaments formed upon intact caldesmon than the latter one.

Actins↗
PubMed1993

The effect of Ca2+ on the conformation of tropomyosin and actin in regulated actin filaments with or without bound myosin subfragment 1.

The effects of Ca2+ and myosin subfragment 1 on the conformation of tropomyosin and actin in regulated actin filaments in ghost fibers were investigated by means of the polarized fluorescence technique. Regulated thin filaments were reconstituted in skeletal muscle ghost fibers by incorporation into the fibers of either skeletal muscle troponin-tropomyosin or smooth-muscle caldesmon-calmodulin-tropomyosin complexes. Tropomyosin and actin were specifically labeled with fluorescent probes, 1,5-IAEDANS and phalloidin-rhodamine, respectively. Analysis of the fluorescence parameters indicated that the binding of Ca2+ to regulated actin filaments induces conformational changes in tropomyosin and actin that lead to the strengthening of the interaction between these two proteins and weakening of the binding of actin monomers in the filament. These changes become larger when regulated actin forms rigor links with myosin subfragment 1. No notable alterations in the position of tropomyosin relative to actin in the frontal plane of the fiber were detected either upon binding of Ca2+ or upon the additional binding of myosin subfragment 1 to regulated actin.

Actin Cytoskeleton↗
PubMed1991

Troponin I and caldesmon restrict alterations in actin structure occurring on binding of myosin subfragment 1.

The effect of troponin I and caldesmon on phalloidin-rhodamine- and 1,5-IAEDANS-labelled actin in skeletal muscle ghost fibers was investigated by polarized fluorescence. Both these proteins inhibited the structural alterations in the actin monomer and the increase of flexibility of actin filaments occurring on binding of myosin heads, and their effects were potentiated by tropomyosin. This immobilization of the actin filament through troponin I and caldesmon seems to originate from restriction of the relative motions of the two domains within the monomer.

Actins↗
PubMed1991

Orientation in birds. Satellite tracking: a new method in orientation research.

The study of bird migration by satellite tracking began in the 1980s, after transmitters had become sufficiently small to allow the monitoring of larger migrating species. The initial studies in this direction were all successful with respect to establishing this method (Section 2) of elucidating new migration patterns and various aspects of orientation. These pilot studies are summarized in Section 3, Table 1, Figures 1-4. We believe that studies on avian orientation and navigation will, in general, greatly benefit from satellite tracking if some prerequisites are fulfilled and adequate experiments planned (Section 4). In future studies, smaller transmitters with increased life expectancies are needed, and possible effects of these transmitters on bird behaviour must be examined. Considerable improvement of satellite tracking in avian orientation research is to be expected, along with the development of a new generation of satellites allowing communication between satellites and transmitters (Sections 4, 5).

Animals↗
PubMed1990

Retroperitoneal accessory spleen.

We report a lower abdominal mass found to be a retroperitoneal accessory spleen. The vascular supply originated from retroperitoneal vessels independent of the splenic, testicular, and spermatic vessels.

Adult↗
PubMed1989

Caldesmon weakens the bonding between myosin heads and actin in ghost fibers.

Earlier studies using polarized microphotometry have shown that caldesmon inhibits the alterations in structure and flexibility of actin in ghost fibers that take place upon the binding of myosin heads (Gałazkiewicz et al. (1987) Biochim. Biophys. Acta 916, 368-375). The present investigations, performed with an IAEDANS label attached to myosin subfragment 1 (S-1), revealed that this inhibition results from the weakening of the binding between myosin heads and actin as indicated by the caldesmon-induced increase in the random movement of S-1. Parallel experiments with actin labeled at Cys-374 demonstrated that this effect of caldesmon is transmitted to the C-terminus of the actin molecule resulting in a conformational adjustment in this region of the molecule.

Actins↗
PubMed1989

The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin.

The stoichiometric actin--DNase-I complex was used to study the actin--nucleotide and actin--divalent-cation interactions and its ATPase activity in the presence of MgCl2 and cytochalasin D. Treatment of actin--DNase-I complex with 1 mM EDTA results in almost complete restoration of its otherwise inhibited DNase I activity, although the complex does not dissociate, as verified by size-exclusion chromatography. This effect is due to a loss of actin-bound nucleotide but is prevented by the presence of 0.1-0.5 mM ATP, ADP and certain ATP analogues. In this case no increase in DNase I activity occurs, even in the presence of EDTA. At high salt concentrations and in the presence of Mg2+ ('physiological conditions') the association rate constants for ATP, ADP and epsilon ATP (1,N6-ethenoadenosine 5'-triphosphate) and the dissociation rate constant for epsilon ATP were determined. Both the on and off rates were found to be reduced by a factor of about 10 when compared to uncomplexed actin. Thus the binding constant of epsilon ATP to actin is almost unaltered after complexing to DNase I (2.16 x 10(8) M-1). Titrating the increase in DNase I activity of the actin--DNase I complex against nucleotide concentration in the presence of EDTA, the association constant of ATP to the cation-free form of actin--DNase I complex was found to be 5 x 10(3) M-1, which is many orders of magnitude lower than in the presence of divalent metal ions. The binding constant of Ca2+ to the high-affinity metal-binding site of actin was found not to be altered when complexed to DNase I, although the rate of Ca2+ release decreases by a factor of 8 after actin binding to DNase I. The rate of denaturation of nucleotide-free and metal-ion-free actin--DNase I complex was found to be reduced by a factor of about 15. The ATPase activity of the complex is stimulated by addition of Mg2+ and even more effectively by cytochalasin D, proving that this drug is able to interact with monomeric actin.

Actins↗
PubMed1989

Two-dimensional manikins of children - models for design.

The paper discusses anthropometric research for design for children and youth. Two-dimensional dimensional manikins are one of the forms of presenting anthropometric data. Manikins of children aged 1-6 are the object of the paper. The set consists of eight plexiglass models scaled 1:5. The models were prepared with regard to 5th and 95th percentile values and two views - i e, side view when the figure is in the sagittal plane, and top view when the figure is in the transverse plane. the models are used in design and ergonomics assessment of products including equipment and furnishings earmarked for children.

Journal Article↗
PubMed1989

Workspace for disabled people.

The paper aims at determining the workspace for disabled young people. The investigation embraced a group of young people with motor dysfunction of lower extremities. Seventeen measurements were taken of 32 boys and 45 girls aged 15-18. There are significant differences in functional dimensions of the arm between the healthy and the disabled populations. The differences clearly indicate that there is a need for different shaping of space in which people of limited motor efficiency must function. A method based on Das and Grady's concept was used in determining workspace. This method was modified by the author. The method allows determination of workspace by means of several simple anthropometric measurements. Workspace for young people investigated was determined for the sitting position in transverse and sagittal planes. The results of the investigation constitute data for design and estimation of workplaces for disabled young people with motor dysfunction of lower extremities. The results can also be used to determine functional space in classrooms, school workshops, educational and rehabilitation centres.

Adolescent↗
PubMed1988

Kinetics of adenosine 5'-triphosphate and adenosine 5'-diphosphate interaction with G-actin.

Double mixing experiments using a three-syringe stopped-flow apparatus have given values of the second-order rate constants for association of epsilon ATP, ATP, and ADP to G-actin of 6.8 X 10(6) M-1 s-1, 6.1 X 10(6) M-1 s-1, and 6.3 X 10(6) M-1 s-1, respectively, at pH 7.6, 20 degrees C, and 0.65 mM free Ca2+. The previously established ca. 100-fold weaker binding of ADP than ATP to G-actin is due to a much faster dissociation rate of ADP than ATP, rather than to a slower association rate as was previously reported. This difference between ADP and ATP largely disappears under more nearly physiological conditions (0.8 mM Mg2+ and 100 mM KCl). Association rate constants for the three nucleotides under these conditions are 2.13 X 10(6) M-1 s-1, 1.1 X 10(6) M-1 s-1, and 1.2 X 10(6) M-1 s-1, respectively, for epsilon ATP, ATP, and ADP. The rate constant for association of epsilon ATP is only slightly affected by reducing the Mg2+ concentration from 0.8 to 0.2 mM, whereas that for ADP association is reduced by a factor of ca. 3. This, together with the observed increase in the apparent association rate constant of epsilon ATP on increasing the Ca2+ concentration in the 10-100 microM range, suggests that G-actin recognizes and binds the metal-nucleotide complexes.

Actins↗
PubMed1988

Comparison of Ca2+-dependent effects of caldesmon-tropomyosin-calmodulin and troponin-tropomyosin complexes on the structure of F-actin in ghost fibers and its interaction with myosin heads.

Comparison of two types of Ca2+-regulated thin filament, reconstructed in ghost fibers by incorporating either caldesmon-gizzard tropomyosin-calmodulin or skeletal muscle troponin-tropomyosin complex, was performed by polarized microphotometry. The changes in actin structure under the influence of these regulatory complexes, as well as those upon the binding of the myosin heads, were followed by measurements of F-actin intrinsic tryptophan fluorescence and the fluorescence of phalloidin-rhodamine complex attached to F-actin. The results show that in the presence of smooth muscle tropomyosin and calmodulin, caldesmon causes Ca2+-dependent alterations of actin conformation and flexibility similar to those induced by skeletal muscle troponin-tropomyosin complex. In both cases, transferring of the fiber from '-Ca2+' to '+Ca2+' solution increases the number of turned-on actin monomers. However, whereas troponin in the absence of Ca2+ potentiates the effect of skeletal muscle tropomyosin, caldesmon-calmodulin complex inhibits the effect of smooth muscle tropomyosin. This difference seems to be due to the qualitatively different alterations in the structure and flexibility of F-actin in ghost fibers evoked by smooth and skeletal muscle tropomyosins. Troponin can bind to F-actin-smooth muscle tropomyosin-caldesmon complex and, in the presence of Ca2+, release the restraint by caldesmon for S-1-induced alterations of conformation, and reduce that for flexibility of actin in ghost fibers. This effect seems to be related to the abolishment by troponin of the potentiating effect of tropomyosin on caldesmon-induced inhibition of actomyosin ATPase activity.

Actins↗
PubMed1988

Kinetics of nucleotide and metal ion interaction with G-actin.

The kinetics of interaction of Ca2+ ions and nucleotides with G-actin have been investigated by making use of the enhancement of 1,N6-ethenoadenosine 5'-triphosphate (epsilon ATP) fluorescence on binding to actin, the enhancement of 2-[[2-[bis(carboxymethyl)amino]-5-methylphenoxy] methyl]-6-methoxy-8-[bis(carboxymethyl)amino]quinoline (Quin-2) fluorescence on binding to Ca2+, and the sensitivity of the fluorescence of an N-(iodoacetyl)-N'-(5-sulfo-1-naphthyl)ethylenediamine (1,5-AEDANS) group on Cys-374 to metal ion binding. It is concluded that metal ion dissociation is the rate-limiting step in nucleotide dissociation (0.016 s-1 for Ca2+ at pH 7.2 and 21 degrees C) and that earlier conclusions that metal ion release is relatively fast and subsequent nucleotide release slow are incorrect. Results presented here and obtained by others on the metal ion concentration dependence of the effective rate of nucleotide exchange can be interpreted in the light of this conclusion in terms of a limiting rate which corresponds to that of metal ion release and an "apparent" dissociation constant for Ca2+ which is without direct physical significance. This apparent dissociation constant is more than 2 orders of magnitude greater than the real dissociation constant of Ca2+ from the Ca-actin-ATP complex, which was estimated to be 2 X 10(-9) M from a titration with Quin-2. Confirmation that the rate of Ca2+ release is rate limiting both in nucleotide dissociation reactions and in replacement of Ca2+ by Mg2+ was obtained with 1,5-AEDANS-actin, since both the replacement of Ca2+ by Mg2+ and the removal of Ca2+ to give the actin-ATP complex occurred at the same (slow) rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗
PubMed1988

Scalpel versus electrocautery in modified radical mastectomy.

Forty nine consecutive mastectomy patients were randomized to one of two groups. Twenty four patients had skin flaps created with the cold scalpel and twenty five with the electrocautery. The two groups were similar with respect to age, stage of disease, size of tumor, and weight. Cautery patients had significantly less operative blood loss when compared with scalpel patients, 352 versus 507 milliliters, respectively, P less than 0.05. No cautery patient required transfusion compared with three transfused scalpel patients, P less than .005. Total postoperative hemovac drainage and hospital stay were not significantly different between the two groups. Although the number of fever days and wound complications were slightly higher in the cautery group, the difference was not statistically significant. The electrocautery may be superior to the scalpel for mastectomy.

Electrocoagulation↗
PubMed1985

Properties of carboxypeptidase A-treated chicken gizzard tropomyosin.

Chicken gizzard tropomyosin was digested with carboxypeptidase A at the weight ratios of enzyme to substrate 1:200 and 1:50. Removal of about 16 C-terminal amino acid residues per tropomyosin molecule, at lower enzyme concentration, caused reversion of the effect on skeletal actomyosin ATPase activity from activating to inhibiting without an influence on polymerizability and actin-binding ability. Removal of about 26 C-terminal amino acid residues per molecule, at higher enzyme concentration, resulted in loss of polymerizability and actin binding ability. Digestion of gizzard tropomyosin with carboxypeptidase A has no dramatic effect on its binding to troponin T. The results show that not only the existence of head-to-tail overlapping regions but also their length is important for the functional properties of chicken gizzard tropomyosin.

Actins↗