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1.
Waste Manag ; 68: 482-489, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28751173

ABSTRACT

Nd-Fe-B permanent magnets are a strategic material for a number of emerging technologies. They are a key component in the most energy efficient electric motors and generators, thus, they are vital for energy technologies, industrial applications and automation, and future forms of mobility. Rare earth elements (REEs) such as neodymium, dysprosium and praseodymium are also found in waste electrical and electronic equipment (WEEE) in volumes that grow with the technological evolution, and are marked as critical elements by the European Commission due to their high economic importance combined with significant supply risks. Recycling could be a good approach to compensate for the lack of rare earths (REs) on the market. However, less than 1% of REs are currently being recycled, mainly because of non-existing collection logistics, lack of information about the quantity of RE materials available for recycling and recycling-unfriendly product designs. To improve these lack of information, different waste streams of electrical and electronic equipment from an industrial recycling plant were analyzed in order to localize, identify and collect RE permanent magnets of the Nd-Fe-B type. This particular type of magnets were mainly found in hard disk drives (HDDs) from laptops and desktop computers, as well as in loudspeakers from compact products such as flat screen TVs, PC screens, and laptops. Since HDDs have been investigated thoroughly by many authors, this study focusses on other potential Nd-Fe-B resources in electronic waste. The study includes a systematic survey of the chemical composition of the Nd-Fe-B magnets found in the selected waste streams, which illustrates the evolution of the Nd-Fe-B alloys over the years. The study also provides an overview over the types of magnets integrated in different waste electric and electronic equipment.


Subject(s)
Electronic Waste , Magnets , Metals, Rare Earth , Neodymium , Recycling , Refuse Disposal
2.
Arch Toxicol ; 88(12): 2135-90, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25370008

ABSTRACT

The exposure of the skin to medical drugs, skin care products, cosmetics, and other chemicals renders information on xenobiotic-metabolizing enzymes (XME) in the skin highly interesting. Since the use of freshly excised human skin for experimental investigations meets with ethical and practical limitations, information on XME in models comes in the focus including non-human mammalian species and in vitro skin models. This review attempts to summarize the information available in the open scientific literature on XME in the skin of human, rat, mouse, guinea pig, and pig as well as human primary skin cells, human cell lines, and reconstructed human skin models. The most salient outcome is that much more research on cutaneous XME is needed for solid metabolism-dependent efficacy and safety predictions, and the cutaneous metabolism comparisons have to be viewed with caution. Keeping this fully in mind at least with respect to some cutaneous XME, some models may tentatively be considered to approximate reasonable closeness to human skin. For dermal absorption and for skin irritation among many contributing XME, esterase activity is of special importance, which in pig skin, some human cell lines, and reconstructed skin models appears reasonably close to human skin. With respect to genotoxicity and sensitization, activating XME are not yet judgeable, but reactive metabolite-reducing XME in primary human keratinocytes and several reconstructed human skin models appear reasonably close to human skin. For a more detailed delineation and discussion of the severe limitations see the "Overview and Conclusions" section in the end of this review.


Subject(s)
Cytochrome P-450 Enzyme System/metabolism , Models, Biological , Skin/enzymology , Xenobiotics/pharmacokinetics , Xenobiotics/toxicity , Animals , Guinea Pigs , Humans , Mice , Rats , Skin/drug effects , Skin Absorption , Species Specificity , Toxicity Tests
3.
SAR QSAR Environ Res ; 25(7): 565-88, 2014.
Article in English | MEDLINE | ID: mdl-24905588

ABSTRACT

Dermal absorption is a critical part in the risk assessment of complex mixtures such as agrochemical formulations. To reduce the number of in vivo or in vitro absorption experiments, the present study aimed to develop an in silico prediction model that considers mixture-related effects. Therefore, an experimental 'real-world' dataset derived from regulatory in vitro studies with human and rat skin was processed. Overall, 56 test substances applied in more than 150 mixtures were used. Descriptors for the substances as well as the mixtures were generated and used for multiple linear regression analysis. Considering the heterogeneity of the underlying data set, the final model provides a good fit (r² = 0.75) and is able to estimate the influence of a newly composed formulation on dermal absorption of a well-known substance (predictivity Q²Ext = 0.73). Application of this model would reduce animal and non-animal testings when used for the optimization of formulations in early developmental stages, or would simplify the registration process, if accepted for read-across.


Subject(s)
Agrochemicals/pharmacokinetics , Computer Simulation , Quantitative Structure-Activity Relationship , Skin Absorption , Skin/metabolism , Animals , Complex Mixtures/pharmacokinetics , Humans , Linear Models , Models, Biological , Rats
4.
Transpl Infect Dis ; 1(4): 284-7, 1999 Dec.
Article in English | MEDLINE | ID: mdl-11428999

ABSTRACT

Listeria monocytogenes has long been known as a pathogen of immunocompromised hosts, including solid organ and bone marrow transplant recipients. Its principal manifestations include bacteremia and meningitis. Endocarditis due to Listeria is far less common and in general affects the left side of the heart. We here report an unusual case of Listeria tricuspid valve endocarditis and septic pulmonary emboli in a sulfa-intolerant liver transplant recipient with a history of relapsing cytomegalovirus (CMV) hepatitis and an indwelling Hickman catheter. The literature on Listeria endocarditis and infections in transplant recipients is reviewed. The possible relationship between susceptibility to Listeria infection and the discontinuation of trimethoprim-sulfamethoxazole prophylaxis is of interest.


Subject(s)
Ampicillin/therapeutic use , Drug Therapy, Combination/therapeutic use , Endocarditis, Bacterial/diagnosis , Gentamicins/therapeutic use , Listeriosis/diagnosis , Liver Transplantation , Postoperative Complications , Pulmonary Embolism/diagnosis , Sepsis/diagnosis , Tricuspid Valve , Adult , Antiviral Agents/therapeutic use , Cytomegalovirus Infections/prevention & control , Echocardiography, Transesophageal , Endocarditis, Bacterial/complications , Endocarditis, Bacterial/drug therapy , Female , Ganciclovir/therapeutic use , Humans , Immunoglobulins, Intravenous/therapeutic use , Immunosuppressive Agents/therapeutic use , Listeria monocytogenes , Listeriosis/complications , Listeriosis/drug therapy , Liver Transplantation/immunology , Penicillins/therapeutic use , Pulmonary Embolism/complications , Sepsis/drug therapy
5.
J Muscle Res Cell Motil ; 20(7): 645-53, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10672512

ABSTRACT

It is well known that cardiac troponin C (cTnC) regulates the association of force-generating myosin cross-bridges. We report here evidence for an additional role for cTnC. This hypothesis states that Ca2+ binds more strongly to cTnC when force-generating myosin cross-bridges are attached to actin and that removal of this bound Ca2+ accelerates the dissociation of force-generating myosin cross-bridges. Intact Fura-2-loaded rat papillary muscles and skinned (permeabilized) ventricular preparations were used. The preparations were mounted in the Guth Muscle Research System which is capable of measuring simultaneously fluorescence and force in response to length perturbations. All mechanical perturbations of muscle length (isotonic shortening, quick stretches and releases, and length vibrations) which cause dissociation of force-generating myosin cross-bridges during a twitch resulted in Ca2+ being released from troponin as judged from changes in the Ca2+ transients (Fura-2 (340/380) fluorescence ratio). Thus dissociation of force-generating myosin cross-bridges cause Ca2+ to be released from cTnC. Conversely, it would be expected that removal of strongly bound Ca2+ from cTnC would result in an increase in the rate of dissociation of force-generating myosin cross-bridges. To test this hypothesis actomyosin ATPase (NADH fluorescence change) and isometric force were measured in skinned cardiac preparations. The ratio of the ATPase/Force is proportional to the rate constant (gapp) for the dissociation of force-generating myosin cross-bridges. The data showed that decreasing the amount of Ca2+ bound to cTnC in skinned cardiac fibers caused an increase in the ratio of ATPase/Force, the rate of dissociation (gapp) of force-generating myosin cross-bridges.


Subject(s)
Heart/physiology , Myocardial Contraction/physiology , Myosins/physiology , Troponin C/physiology , Animals , Calcium/physiology , Rats
6.
J Mol Cell Cardiol ; 30(8): 1459-70, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9737933

ABSTRACT

Diastolic dysfunction at high heart rates may be associated with increased myocardial energy consumption. Frequency-dependent changes of isometric force and oxygen consumption (MVO2) were investigated in strip preparations from endstage failing human hearts exhibiting various degrees of diastolic dysfunction. MVO2 was determined by a new method which was validated. When stimulation rate was increased from 40 to 200 min-1 (n=7), developed force decreased from 16.5+/-4.3 to 7.9+/-2.9 mN/mm2 (P<0.01), diastolic force increased from 15.9+/-3.2 to 22.0+/-3.0 mN/mm2 (P<0.01), and total MVO2 increased from 2.6+/-0.6 to 4.7+/-0.9 ml/min/100 g (P<0.025). Resting MVO2 and resting force were 1.8+/-0.4 ml/min/100 g and 15.9+/-3.0 mN/mm2, respectively. After addition of 30 mm 2,3-butanedione monoxime (BDM) to inhibit crossbridges, resting MVO2 and resting force decreased by 46% (P<0.05) and 15% (P<0.01), respectively, indicating the presence of active force generation in unstimulated failing human myocardium. In each muscle preparation, there was a significant correlation between force-time integral (FTI) and total MVO2 (r=0.96+/-0.01). The strength of these correlations did not vary with the contribution of diastolic FTI to total FTI. The ratio of activity related MVO2 to developed FTI, an inverse index of the economy of contraction, increased depending on the rise of diastolic FTI at higher stimulation rates. In conclusion, in failing human myocardium, diastolic force development is occurring at the same energy expenditure as systolic force generation. Therefore, in muscle preparations with disturbed diastolic function economy of contraction decreases with higher stimulation rates, depending on the rise of diastolic force.


Subject(s)
Cardiomyopathies/metabolism , Electrophysiology/methods , Myocardium/metabolism , Oxygen Consumption , Biomechanical Phenomena , Diastole , Female , Humans , Male , Middle Aged , Reproducibility of Results
7.
Pediatr Nurs ; 24(6): 578-86, 1998.
Article in English | MEDLINE | ID: mdl-10086002

ABSTRACT

Immigrant visas issued to orphans entering the United States (a proxy measure for international adoptions) totaled 13,620 in 1997, more than double the amount in 1992. As international adoption numbers rise, pediatric nurses encounter more adopted children and adoptive families. Among the many adoption-related issues confronting pediatric health care providers, the one most frequently voiced is: "Should we do or look for anything differently in adopted children? And, if so, what?" To address this issue, a three-fold approach is suggested: (a) recognize the physical conditions and medical problems prevalent in the international countries with high adoption rates; (b) identify the unique adoption-related tasks encountered by children according to their current developmental stage; and (c) determine recommendations for providers, parents, and children to successfully address or cope with these adoption-related tasks and conditions.


Subject(s)
Adoption , Family Health , Foster Home Care , International Cooperation , Pediatric Nursing , Adolescent , Adoption/legislation & jurisprudence , Adoption/psychology , Child , Child, Preschool , Emigration and Immigration/legislation & jurisprudence , Emigration and Immigration/statistics & numerical data , Emigration and Immigration/trends , Foster Home Care/legislation & jurisprudence , Foster Home Care/psychology , Foster Home Care/statistics & numerical data , Humans , Infant , Infant, Newborn , International Agencies , Nursing Assessment , Patient Education as Topic , Pediatric Nursing/methods , Primary Health Care/methods , United States
8.
J Muscle Res Cell Motil ; 12(2): 208-15, 1991 Apr.
Article in English | MEDLINE | ID: mdl-2061413

ABSTRACT

The specific feature of stretch activation of the indirect flight muscle of the tropical waterbug Lethocerus was used to correlate mechanical and structural aspects of muscle contraction. The time courses of the changes in intensities of the strongest equatorial reflections, the (10) and (20) and of the first meridional reflection at 14.5 nm-1 were monitored using synchrotron radiation as a high intensity X-ray source. The ratio of the intensities of the equatorial reflections, (I20/I10), which reflects the mass distribution within the filament lattice array, increases by about 10% relative to the Ca(2+)-activated level when a rapid stretch is imposed, compared with a 200% change seen when fibres change from the relaxed to the rigor state, while the spacing of the lattice planes decreases by about 1%. The intensity of the first meridional reflection at 14.5 nm-1 decreases by about 35% during stretch activation with a slightly faster time course than the delayed tension increase. The results suggest that the average structure of cycling crossbridges is different from that present in the rigor state.


Subject(s)
Coleoptera/physiology , Muscle Contraction , Actin Cytoskeleton/physiology , Actin Cytoskeleton/ultrastructure , Animals , Coleoptera/anatomy & histology , Muscles/physiology , Muscles/ultrastructure , Stress, Mechanical , X-Ray Diffraction/instrumentation
9.
J Muscle Res Cell Motil ; 11(5): 392-402, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2266166

ABSTRACT

In order to identify which steps in the crossbridge are affected by changes in ionic strength, we studied the effect of ionic strength on the rate constants and magnitudes of three exponential processes, the ATP hydrolysis rate and isometric tension during maximal activation (pCa 4.52, 5 mM MgATP). Equatorial X-ray diffraction measurements were also carried out in both relaxing and rigor conditions to examine whether the distance between thick and thin filaments changes with ionic strength (range: 100-300 mM). All experiments were carried out at 20 degrees C and at pH 7.0 on chemically skinned rabbit psoas muscle fibres. Isometric tension and muscle stiffness declined significantly as the ionic strength was increased from 150 mM to 300 mM. The concomitant decrease in the ATP hydrolysis rate was much less than tension, resulting in a large increase in the tension cost. Three rate constants of exponential processes, deduced from sinusoidal analysis, did not change appreciably. The magnitude parameters of all three processes diminished as the ionic strength was increased. During relaxation the filament spacing increased by 5% when the ionic strength was increased from 150 mM to 300 mM. After rigor induction, the spacing did not change with ionic strength. We conclude that a change in ionic strength modifies the rapid equilibrium between the detached state and the 'weakly attached' state, and that this causes considerable effect on isometric tension. We also conclude that other steps in the crossbridge cycle are less sensitive to ionic strength, and that the lattice spacing change is unable to account for the considerable effect of ionic strength on isometric tension.


Subject(s)
Adenosine Triphosphate/metabolism , Muscle Contraction/physiology , Animals , Biomechanical Phenomena , Hydrolysis , In Vitro Techniques , Kinetics , Osmolar Concentration , Rabbits , X-Ray Diffraction
10.
Pflugers Arch ; 416(5): 512-8, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2146588

ABSTRACT

The temporal relationship between myosin phosphorylation, contractile force and ATPase activity was studied in skinned preparations from the guinea-pig Taenia coli. When free Calcium concentration [( Ca2+]) was increased from pCa (-log[Ca2+]) 9 to pCa 4.5 at low calmodulin concentration (0.05 microM), ATPase activity and myosin light-chain phosphorylation rose quickly, while the increase in force and stiffness was delayed. The time-course of tension increase was faster at higher calmodulin concentrations (5 microM), although the maximal level of phosphorylation was unchanged. Lowering the calcium concentration from pCa 4.5 to pCa 9 at the plateau of contraction caused a rapid decrease in ATPase activity and in myosin phosphorylation, while force and stiffness decayed more slowly. The force decay could be accelerated by inorganic phosphate. These results suggest that, during contraction, force may be produced actively by phosphorylated and ATP-splitting crossbridges, but may be maintained by dephosphorylated crossbridges which cycle slowly. However, force could also be modulated by calmodulin and inorganic phosphate in a manner not involving an alteration in the extent of myosin phosphorylation.


Subject(s)
Adenosine Triphosphatases/metabolism , Muscle Contraction/physiology , Muscle Relaxation/physiology , Muscle, Smooth/physiology , Myosins/metabolism , Animals , Calcium/pharmacology , Calmodulin/pharmacology , Colon/physiology , Egtazic Acid/pharmacology , Guinea Pigs , Kinetics , Phosphates/pharmacology , Phosphorylation
11.
Pflugers Arch ; 411(2): 200-3, 1988 Feb.
Article in English | MEDLINE | ID: mdl-3357758

ABSTRACT

Novel techniques of flash photolysis experiments require high intensity light sources in the near UV. We describe here a simple and inexpensive flash device which may complete with bulky and expensive laser systems if the experiments do not necessitate very short light pulses. Using a particular optical arrangement and stored electrical energy, a variation of the parameters voltage and capacitance led to a difference in light output by a factor of more than two. The system is used to relax both skeletal and smooth muscle fibres in the rigor state by releasing up to 2 mM ATP from 12.5 mM caged-ATP.


Subject(s)
Adenosine Triphosphate/metabolism , Muscle Contraction , Muscle Relaxation , Photolysis , Animals , Muscle, Smooth/physiology , Muscles/physiology , Technology
12.
Adv Exp Med Biol ; 226: 203-17, 1988.
Article in English | MEDLINE | ID: mdl-2970207

ABSTRACT

The role of phosphate ions (Pi) in crossbridge kinetics is investigated by parallel measurements of the ATP hydrolysis rate and the tension transients in maximally activated, chemically skinned rabbit psoas fibers. The tension transients are induced by sinusoidal length oscillations, and resulting time courses are analyzed in terms of three exponential processes (A), (B), and (C). The ATP hydrolysis rate is measured by the NADH fluorscence method coupled with enzymatic reactions which rephosphorylate ADP and oxidize NADH. The hydrolysis rate of the standard activation at 20 degrees C is measured at 0.61 mM/sec. The isometric tension, stiffness, and the ATPase rate progressively decrease with increasing concentrations of Pi (0-16 mM). The decrease is most notably observed with tension, followed by stiffness and the ATPase rate. Both the apparent rate constant and the magnitude parameters of exponential process (B) increase with Pi concentration resulting in a large increase in the oscillatory power output. The active species of Pi which causes this effect is found to be the monovalent anion H2PO4-. The effects of Pi on processes (A) and (C) are only marginal. When fibers are oscillated at 1 Hz, no increase in the ATP hydrolysis rate is observed; a small increase is noticed at 10 Hz (1%), and at 100 Hz (6%). We interpret these results in terms of a crossbridge scheme which adds a branch pathway to the conventional hydrolysis cycle: the number of crossbridges entering the branch pathway increases at higher Pi concentrations and in the presence of oscillations.


Subject(s)
Adenosine Triphosphatases/metabolism , Muscles/physiology , Phosphates/pharmacology , Adenosine Triphosphate/metabolism , Animals , Anions , Elasticity , Hydrolysis , In Vitro Techniques , Kinetics , Models, Biological , Muscle Contraction , Muscles/drug effects , Rabbits
13.
J Biol Chem ; 262(28): 13627-35, 1987 Oct 05.
Article in English | MEDLINE | ID: mdl-3654633

ABSTRACT

Intrinsic troponin C (TnC) was extracted from small bundles of rabbit psoas fibers and replaced with TnC labeled with dansylaziridine (5-dimethylaminonaphthalene-1-sulfonyl). The flourescence of incorporated dansylaziridine-labeled TnC was enhanced by the binding of Ca2+ to the Ca2+-specific (regulatory) sites of TnC and was measured simultaneously with force (Zot, H.G., Güth, K., and Potter, J.D. (1986) J. Biol. Chem. 261, 15883-15890). Various myosin cross-bridge states also altered the fluorescence of dansylaziridine-labeled TnC in the filament, with cycling cross-bridges having a greater effect than rigor cross-bridges; and in both cases, there was an additional effect of Ca2+. The paired fluorescence and tension data were used to calculate the apparent Ca2+ affinity of the regulatory sites in the thin filament and were shown to increase at least 10-fold during muscle activation presumably due to the interaction of cycling cross-bridges with the thin filament. The cross-bridge state responsible for this enhanced Ca2+ affinity was shown to be the myosin-ADP state present only when cross-bridges are cycling. The steepness of the pCa force curves (where pCa represents the -log of the free Ca2+ concentration) obtained in the presence of ATP at short and long sarcomere lengths was the same, suggesting that cooperative interactions between adjacent troponin-tropomyosin units may spread along much of the actin filament when cross-bridges are attached to it. In contrast to the cycling cross-bridges, rigor bridges only increased the Ca2+ affinity of the regulatory sites 2-fold. Taken together, the results presented here indicate a strong coupling between the Ca2+ regulatory sites and cross-bridge interactions with the thin filament.


Subject(s)
Aziridines/pharmacology , Azirines/pharmacology , Calcium/metabolism , Dansyl Compounds/pharmacology , Muscle Contraction , Muscles/metabolism , Troponin/metabolism , Animals , Binding Sites , Fluorescent Dyes/pharmacology , In Vitro Techniques , Kinetics , Muscle Contraction/drug effects , Muscles/drug effects , Protein Binding , Rabbits , Troponin C
14.
Biophys J ; 51(3): 457-63, 1987 Mar.
Article in English | MEDLINE | ID: mdl-2952177

ABSTRACT

Inorganic phosphate (Pi) decreases maximal tension in contracted skeletal and heart muscle fibers. We investigated the effects of 10 mM Pi on the force-calcium relationship in Triton X-100-skinned Taenia coli smooth muscle fibers. Isometric force measurements show that the calcium sensitivity of the force depends on the phosphate concentration. Furthermore 10 mM Pi relaxes the fibers more at intermediate than at high calcium ion concentrations: At pCa 4.5 tension decreases in the presence of 10 mM Pi by approximately 12% but it decreases 70% at pCa 6.17. Removal of phosphate partially reverses the relaxation. Simultaneous determination of actomyosin ATPase activity and force (Güth, K., and J. Junge, 1982, Nature (Lond.), 300:775-776) shows that the ATPase activity does not correlate with the changes in force. In the presence of Pi, tension decreases more than the ATPase activity. The level of phosphorylation of the 20,000-D regulatory myosin light chain is not changed in the presence or absence of 10 mM Pi. The results are discussed in terms of slowly or noncycling myosin crossbridges formed at lower calcium concentrations, which contribute to the force development but not to the ATPase activity. These crossbridges are considered to be dissociated in the presence of phosphate.


Subject(s)
Calcium-Transporting ATPases/metabolism , Calcium/pharmacology , Muscle, Smooth/physiology , Phosphates/pharmacology , Animals , Colon/drug effects , Colon/physiology , Guinea Pigs , In Vitro Techniques , Kinetics , Muscle, Smooth/drug effects , Phosphorylation
15.
Pflugers Arch ; 408(1): 1-9, 1987 Jan.
Article in English | MEDLINE | ID: mdl-3822768

ABSTRACT

The role of orthophosphate ions (Pi) in crossbridge kinetics was investigated by parallel measurements of the ATP hydrolysis rate and tension transients in maximally activated, chemically skinned rabbit psoas fibers. The hydrolysis rate of the standard activation at 20 degrees C was measured at 1.25 nmole X s-1 X m-1 X fiber-1, which corresponds to the hydrolysis of 3 moles ATP per mole of myosin head per second. The isometric tension, stiffness extrapolated to the infinite frequency, and the ATPase rate progressively decreased when increasing concentrations of Pi (0-16 mM) were added to the activating saline. The decrease was greatest with tension, followed by stiffness and the ATPase rate. Both the apparent rate constant and the magnitude parameters of exponential process (B) increased with Pi concentration resulting in a significant increase in the oscillatory power output. The effects of Pi on processes (A) and (C) were only marginal. When fibers were oscillated at 1 Hz [close to the characteristic frequency of process (A)], no significant increase in the ATP hydrolysis rate was observed. However, a small increase was noticed at 10 Hz [1%, process (B)], and at 100 Hz [6%, process (C)]. We interpret these results in terms of a crossbridge scheme which adds a branch pathway to the conventional hydrolysis cycle. In the proposed scheme, the number of crossbridges entering the branch pathway increases at higher Pi concentrations and in the presence of imposed oscillations at the proper frequency.


Subject(s)
Adenosine Triphosphate/metabolism , Muscles/drug effects , Phosphates/pharmacology , Animals , Binding Sites , Hydrolysis , In Vitro Techniques , Kinetics , Muscle Contraction/drug effects , Muscles/physiology , Rabbits
16.
Biophys J ; 52(6): 1039-45, 1987 Dec.
Article in English | MEDLINE | ID: mdl-19431712

ABSTRACT

The ATPase activity of single fibers of small fiber bundles (one to three fibers) of insect flight muscle was measured when fibers were repetitively released and restretched by 1.5% of their initial length. The ATPase activity increased with increasing duration of release-restretch pulses applied at a constant repetition frequency, reaching a maximum at a duration of approximately 20 ms. For a given duration, the average ATPase activity also increased with increasing frequency of applied length changes and reached a maximum (200% of the isometric ATPase) at a frequency of approximately 50 Hz. The data could be fitted to a two-state model in which the apparent rate of crossbridge detachment is enhanced when the crossbridges are mechanically released. Estimates of the apparent rates of attachment and detachment in the isometrically contracting state and of the enhanced detachment rate of unloaded crossbridges were derived from fits to the two-state model. After short pulses of releasing and restretching the fiber the force was low and increased after the restretch in a roughly exponential manner to the initial level. The rate at which force increased after a release-restretch pulse was similar to the sum of the apparent attachment and detachment rates for the isometrically contracting muscle derived from the ATPase activity measurements.

17.
J Biol Chem ; 261(34): 15883-90, 1986 Dec 05.
Article in English | MEDLINE | ID: mdl-2946678

ABSTRACT

Glycerinated rabbit fast skeletal muscle fibers were chemically skinned with 1% Brij 35 and partially depleted of endogenous troponin C subunit (TnC) by exposure of the fibers to EDTA (Zot, H. G., and Potter, J. D. (1982) J. Biol. Chem. 257, 7678-7683). The TnC-depleted fibers exhibited a decrease in maximal tension that was mostly restored by readdition of TnC or by the addition of the fluorescent 5-dimethylaminonaphthalene-1-sulfonyl aziridine analogue, TnCDanz. TnCDanz is known to undergo an increase in fluorescence intensity when Ca2+ binds to the two low affinity Ca2+-specific regulatory sites of TnC. Steady-state fractional fluorescence and tension changes were measured simultaneously as a function of Ca2+. The Ca2+ sensitivity of the fluorescence curve was about 0.6 log unit greater than the tension curve. This difference in sensitivity could be explained if separate conformational states of TnC, brought about by Ca2+ binding to the Ca2+-specific sites, produce the fluorescence and tension changes. TnC-depleted fibers were also reconstituted with the fluorescent 2-[(4'-iodoacetamido)analino]naphthalene-6-sulfonic acid analogue, cardiac TnCIaans, which undergoes an increase in fluorescence intensity when Ca2+ binds to the single Ca2+- specific regulatory site. The steady-state fractional fluorescence and tension curves for fibers reconstituted with cardiac TnCIaans had nearly the same Ca2+ sensitivity. The steady-state fractional fluorescence of myofibrils reconstituted with TnCDanz was found to have a greater sensitivity to Ca2+ than the simultaneously measured ATPase. In all cases paired fractional fluorescence and activity curves tended to have parallel dependence on Ca2+. These procedures make it possible to study the Ca2+ binding properties of the Ca2+- specific sites in intact myofibrils and skinned fibers; the results presented suggest that the Ca2+ affinity of the Ca2+-specific sites of troponin are reduced in the thin filament compared to that of troponin in solution.


Subject(s)
Muscles/metabolism , Myofibrils/metabolism , Troponin/metabolism , Adenosine Triphosphatases/analysis , Animals , Binding Sites , Calcium/metabolism , Edetic Acid/pharmacology , Female , Fluorescence , In Vitro Techniques , Muscle Tonus , Protein Conformation , Rabbits , Troponin C
18.
Pflugers Arch ; 407(5): 552-7, 1986 Nov.
Article in English | MEDLINE | ID: mdl-2947040

ABSTRACT

A micro cuvette is described, which allows to measure mechanical, optical and energetic parameters of single skinned muscle fibres or small fibre bundles. Two examples for possible applications are given: The measurement of ATPase activity of single muscle fibres. The detection of fluorescence changes due to Ca binding to fluorescence labeled TnC, which was incorporated into small bundles of rabbit psoas fibres before the experiment.


Subject(s)
Muscles/physiology , Physiology/instrumentation , Adenosine Triphosphatases/analysis , Animals , Biomechanical Phenomena , Fluorescence , Muscles/enzymology , Optics and Photonics , Photometry , Rabbits , Transducers
19.
Life Sci ; 38(2): 191-6, 1986 Jan 13.
Article in English | MEDLINE | ID: mdl-3945163

ABSTRACT

The aim of this study was to clarify whether the increased vascular tone in spontaneous hypertension of rats is due to a change of the calcium-sensitivity of the contractile proteins themselves. In skinned rat tail artery rings from SHRSP and WKY rats the calcium-requirement for half maximal activation (3.6 X 10(-6)M Ca++ for both rat strains) as well as relaxation half times (1.45 +/- 0.43 min, SHRSP and 1.63 +/- 0.48 min, WKY) were found to be identical. The extent of myosin phosphorylation in the contracted and in the relaxed state did not differ between SHRSP and WKY. It is concluded that changes at the level of the contractile proteins are not involved in the increased vascular tone of SHRSP essential hypertension.


Subject(s)
Calcium/pharmacology , Hypertension/physiopathology , Muscle, Smooth, Vascular/physiology , Animals , In Vitro Techniques , Myosins/metabolism , Phosphorylation , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Vasoconstriction/drug effects
20.
Biophys J ; 50(5): 993-7, 1986 Nov.
Article in English | MEDLINE | ID: mdl-19431691

ABSTRACT

The time course of structural changes occurring on ATP-induced relaxation of glycerinated insect flight muscle from the rigor state has been investigated using synchrotron radiation as a source of high intensity x rays and photolysis of caged-ATP to produce a rapid rise in ATP concentration. Temporal resolutions of 1 ms for the strongest equatorial reflections and 5 ms for the 14.5 nm meridional reflection are attainable from single events (i.e., without averaging over several cycles). The equatorial intensity changes completely, the meridional intensity partially, towards their respective relaxed values on a much faster time scale than relaxation of tension. The results suggest that actively cycling bridges present shortly after ATP-release are either too few in number to be detected in the equatorial diffraction pattern or that their structure is different from that of rigor bridges.

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