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Computed tomographic appearances of the pelvis following hindquarter amputation.

Bilateral and midline symmetry of the normal pelvic anatomy is an aid to the interpretation of computed tomographic (CT) examinations. Following hindquarter amputation (HQA) or partial hemipelvectomy (PHP) the normal anatomical relationships are disturbed. The CT examinations of 15 patients who had undergone either an HQA or a PHP for an advanced musculoskeletal malignancy are reviewed. The new "normal" anatomy revealed displacement of the bladder and small bowel to the side of surgery in one third of patients, more commonly in the PHP cases. There were varying degrees of wasting of the ipsilateral musculature, gluteus maximus muscle flap, erector spinae and psoas muscles, etc., because of partial denervation and disruption of their origin or insertion. Recurrent tumour was identified in eight of 10 cases in which it was clinically suspected prior to the CT examination. Invariably the recurrence arose within the muscle flap at the resection margin. Bone involvement by direct tumour spread was present in three cases. Pitfalls in differentiating recurrent tumour from scar tissue are discussed.

Adult↗

The effect of thin filament activation on the attachment of weak binding cross-bridges: A two-dimensional x-ray diffraction study on single muscle fibers.

To study possible structural changes in weak cross-bridge attachment to actin upon activation of the thin filament, two-dimensional (2D) x-ray diffraction patterns of skinned fibers from rabbit psoas muscle were recorded at low and high calcium concentration in the presence of saturating concentrations of MgATPgammaS, a nucleotide analog for weak binding states. We also studied 2D x-ray diffraction patterns recorded under relaxing conditions at an ionic strength above and below 50 mM, because it had been proposed from solution studies that reducing ionic strength below 50 mM also induces activation of the thin filament. For this project a novel preparation had to be established that allows recording of 2D x-ray diffraction patterns from single muscle fibers instead of natural fiber bundles. This was required to minimize substrate depletion or product accumulation within the fibers. When the calcium concentration was raised, the diffraction patterns recorded with MgATPgammaS revealed small changes in meridional reflections and layer line intensities that could be attributed in part to the effects of calcium binding to the thin filament (increase in I380, decrease in first actin layer line intensity, increase in I59) and in part to small structural changes of weakly attached cross-bridges (e.g., increase in I143 and I72). Calcium-induced small-scale structural rearrangements of cross-bridges weakly attached to actin in the presence of MgATPgammaS are consistent with our previous observation of reduced rate constants for attachment and detachment of cross-bridges with MgATPgammaS at high calcium. Yet, no evidence was found that weakly attached cross-bridges change their mode of attachment toward a stereospecific conformation when the actin filament is activated by adding calcium. Similarly, reducing ionic strength to less than 50 mM does not induce a transition from nonstereospecific to stereospecific attachment.

Actins↗

Identification of troponin C antagonists from a phage-displayed random peptide library.

Affinity purification of a phage-displayed library, expressing random peptide 12-mers at the N terminus of protein III, has identified 10 distinct novel sequences which bind troponin C specifically. The troponin C-selected peptides yield a consensus binding sequence of (V/L)(D/E)XLKXXLXXLA. Sequence comparison revealed as much as a 62.5% similarity between phiT5, the peptide sequence of the phage clone with the highest level of binding to troponin C, and the N-terminal region of troponin I isoforms. Biotinylated peptides corresponding to library-derived sequences and similar sequences from various isoforms of troponin I were synthesized shown to bind troponin C specifically. Alkaline phosphatase fusion proteins of two of the phage clone sequences bound troponin C specifically, and were specifically competed by both library-derived and native troponin I peptides. Measurement of equilibrium dissociation constants of the peptides by surface plasmon resonance yielded dissociation constants for troponin C as low as 0.43 microM for pT5; in contrast, dissociation constants for calmodulin were greater than 6 microM for all peptides studied. Nondenaturing polyacrylamide gel electrophoresis demonstrated that pT5 formed a stable complex with troponin C in the presence of calcium. We also found that the pT5 peptide inhibited the maximal calcium-activated tension of rabbit psoas muscle fibers.

Amino Acid Sequence↗

Inhibition of unloaded shortening velocity in permeabilized muscle fibres by caged ATP compounds.

The effects of both the P3-1-(2-nitrophenyl)ethyl ester of adenosine 5'-triphosphate (NPE-caged ATP) and its separate diastereoisomers, and the P3-3',5'-dimethoxybenzoin ester of ATP (DMB-caged ATP) were studied on the unloaded shortening velocity of glycerinated rabbit psoas muscle fibres. The unloaded shortening velocities of the active fibres were measured as a function of ATP concentration (0.1-5 mM) using the 'slack-test' with and without 2 mM caged ATP. Shortening velocity followed a Michaelis-Menten relationship with ATP concentration, the Km for ATP being 170 microM. The caged ATP compounds inhibited shortening velocity, in a manner consistent with competitive inhibition, with a Ki of 1-2 mM. The R- and S-diastereoisomers of NPE-caged ATP showed the same degree of competitive inhibition of the shortening velocity, as did DMB-caged ATP. These observations suggest that caged ATP compounds bind to the ATPase site of the actomyosin where they compete with the substrate, Mg2+ ATP.

Adenosine Triphosphate↗

Time course of rise of muscle stiffness at onset of contraction induced by photorelease of ATP.

Isometric contraction (9-10 degrees C) of skinned fibres from rat psoas muscle was elicited by photorelease of ATP (1.3-1.4 mM), and force (F), in-phase (I) and quadrature (Q) stiffness were monitored. The sinusoidal length change for the stiffness measurement was approximately 0.09% fibre length and 200-1000 Hz. On photolysis, F and I initially fell, and then rose. The half-time of the rise of I was shorter than that of F. Contrary to the previous assumption, Q rose even earlier than I. Although the time courses of the rise of F and I were greatly affected by the presence of phosphate ion (Pi; 4 mM), its effect on Q was modest. We propose the following hypothesis: the rigor crossbridges, after binding ATP, initially enter a state characterized by high Q with low I and none or little F, and then, releasing Pi during passing the state with the increased I, reach the final force generating state.

Adenosine Triphosphate↗

Connecting filament mechanics in the relaxed sarcomere.

By examining the mechanical properties of single unactivated myofibrils it has been shown that shortening and stretching of sarcomeres occurs in stepwise fashion, and that steps are seen also in the relaxed state (Yang et al. (1998) Biophys J 74: 1473-1483; Blyakhman et al. (2001) Biophys J 81: 1093-1100; Nagornyak et al. (2004) J. Muscle. Res. Cell Motil. 25: 37-43). The latter are inevitably associated with connecting filaments. Here, we carried out measurements on single myofibrils from rabbit psoas muscle to investigate steps in unactivated specimens in more detail. Myofibrils were stretched and released in ramp-like fashion. For the single sarcomere the time course of length change was consistently stepwise. We found that in the unactivated myofibrils, step size depended on initial sarcomere length, diminishing progressively with increase of initial sarcomere length, whereas in the case of activated sarcomeres, step size was consistently 2.7 nm.

Animals↗

[Femoral neuropathy due to a spontaneous hematoma of the iliopsoas muscle during therapy with heparin-calcium].

With increased use of anticoagulant agents, femoral neuropathy subsequent to spontaneous hemorrhage within the ileo-psoas muscle has become a serious and more frequent clinical problem. The authors describe one case of femoral neuropathy from iliac muscle hematoma during heparin administration. The successful surgical treatment by decompression and drainage is reported. The anatomical and CT scanning features with the suspected pathogenesis are emphasized, as well as the role of early diagnosis in the assessment of prognosis and for successful operative decompression.

Aged↗

Vertebrate muscle Z-line structure: an electron microscopic study of negatively-stained myofibrils.

Structural features of the Z-lines of rabbit psoas muscle myofibrils have been studied in the electron microscope with a negative staining technique. The results obtained suggest the presence of about 20 nm periodicity in the structural organization of the Z-line region: a band pattern of five bands of extra density spaced about 20 nm apart was revealed in the Z-region and the Z-filaments connecting actin filaments from neighbouring sarcomeres often appeared to be positioned at intervals of 17-20 nm. An electron microscopic investigation of the interaction in vitro of two major Z-line proteins, alpha-actinin and F-actin, indicated that the positions of alpha-actinin bridges between actin filaments are defined by relative azimuthal positions of actin subunits. A possible arrangement of actin-linking macromolecular bridges in the Z-region is considered. It is supposed that the arrangement of the Z-filaments is related to the helical symmetry of actin-containing filaments. Also, the banded appearance of the Z-region is interpreted as arising from the arrangement of crossbridges connecting thin filaments of the same sarcomeres.

Actinin↗

Regulation of force development studied by photolysis of caged ADP in rabbit skinned psoas fibers.

The present study examined the effects of Ca(2+) and strongly bound cross-bridges on tension development induced by changes in the concentration of MgADP. Addition of MgADP to the bath increased isometric tension over a wide range of [Ca(2+)] in skinned fibers from rabbit psoas muscle. Tension-pCa (pCa is -log [Ca(2+)]) relationships and stiffness measurements indicated that MgADP increased mean force per cross-bridge at maximal Ca(2+) and increased recruitment of cross-bridges at submaximal Ca(2+). Photolysis of caged ADP to cause a 0.5 mM MgADP jump initiated an increase in isometric tension under all conditions examined, even at pCa 6.4 where there was no active tension before ADP release. Tension increased monophasically with an observed rate constant, k(ADP), which was similar in rate and Ca(2+) sensitivity to the rate constant of tension re-development, k(tr), measured in the same fibers by a release-re-stretch protocol. The amplitude of the caged ADP tension transient had a bell-shaped dependence on Ca(2+), reaching a maximum at intermediate Ca(2+) (pCa 6). The role of strong binding cross-bridges in the ADP response was tested by treatment of fibers with a strong binding derivative of myosin subfragment 1 (NEM-S1). In the presence of NEM-S1, the rate and amplitude of the caged ADP response were no longer sensitive to variations in the level of activator Ca(2+). The results are consistent with a model in which ADP-bound cross-bridges cooperatively activate the thin filament regulatory system at submaximal Ca(2+). This cooperative interaction influences both the magnitude and kinetics of force generation in skeletal muscle.

Adenosine Diphosphate↗

Evaluation of a deformable musculoskeletal model for estimating muscle-tendon lengths during crouch gait.

The hamstrings and psoas muscles are often lengthened surgically in an attempt to correct crouch gait in persons with cerebral palsy. The purpose of this study was to determine if, and under what conditions, medial hamstrings and psoas lengths estimated with a "deformable" musculoskeletal model accurately characterize the lengths of the muscles during walking in individuals with crouch gait. Computer models of four subjects with crouch gait were developed from magnetic resonance (MR) images. These models were used in conjunction with the subjects' measured gait kinematics to calculate the muscle-tendon lengths at the body positions corresponding to walking. The lengths calculated with the MR-based models were normalized and were compared to the lengths estimated using a deformable generic model. The deformable model was either left undeformed and unscaled, or was deformed or scaled to more closely approximate the femoral geometry or bone dimensions of each subject. In most cases, differences between the normalized lengths of the medial hamstrings computed with the deformable and MR-based models were less than 5 mm. Differences in the psoas lengths computed with the deformable and MR-based models were also small (<3 mm) when the deformable model was adjusted to represent the femoral geometry of each subject. This work demonstrates that a deformable musculoskeletal model, in combination with a few subject-specific parameters and simple normalization techniques, can provide rapid and accurate estimates of medial hamstrings and psoas lengths in persons with neuromuscular disorders.

Biomechanical Phenomena↗

Lumbar plexus block: an anatomical study.

The anatomy of the lumbar plexus and the various approaches used to perform lumbar plexus blockade are reviewed. A single needle technique for a posterior approach to the plexus at the L2-3 interspace is described. This technique was used bilaterally in six intact cadavers, and the extent of spread of an injected dye was documented photographically during a subsequent detailed dissection of the region. In all cases, dye was confined to the posterior part of the psoas muscle, and tracked down the nerves of the lumbar plexus. No dye was seen anterior to the psoas, around the sympathetic chain, on the sacral plexus or in the extradural or subarachnoid spaces. Further studies in patients with needle position and drug disposition being confirmed using computerised tomography and X ray scanning were in agreement with the results observed in the cadavers. This technique represents a simple approach to the lumbar plexus which does not require needle localisation by X ray screening.

Humans↗

[The use of cryomethods for research on the sarcomere ultrastructure of rabbit skeletal muscles].

The ultrastructure of sarcomeres of glycerinated rabbit psoas muscle was studied using freeze-fracture-etching, freeze-drying and optical diffraction techniques in comparison with the investigation of this muscle by plastic sections and negative staining methods. In frozen and dried myofibrils isolated from the above muscle the stripes of minor proteins location in A- and I-disks were clearly seen. The pivot structure in thick filaments was revealed in longitudinal fractures of the muscle. The ordered arrangement of myosin heads (crossbridges) associated with actin filaments was preserved in frozen longitudinal fractures as evidenced by optical diffraction. Freeze etching technique allowed to revealed some details of Z-line structure: alpha-actinin bridges connecting the ends of actin filaments of neighbouring sarcomeres and to preserve the lateral struts between actin filaments in I-disks.

Animals↗

Isometric force redevelopment of skinned muscle fibers from rabbit activated with and without Ca2+.

Fiber isometric tension redevelopment rate (kTR) was measured during submaximal and maximal activations in glycerinated fibers from rabbit psoas muscle. In fibers either containing endogenous skeletal troponin C (sTnC) or reconstituted with either purified cardiac troponin C (cTnC) or sTnC, graded activation was achieved by varying [Ca2+]. Some fibers were first partially, then fully, reconstituted with a modified form of cTnC (aTnC) that enables active force generation and shortening in the absence of Ca2+. kTR was derived from the half-time of tension redevelopment. In control fibers with endogenous sTnC, kTR increased nonlinearly with [Ca2+], and maximal kTR was 15.3 +/- 3.6 s-1 (mean +/- SD; n = 26 determinations on 25 fibers) at pCa 4.0. During submaximal activations by Ca2+, kTR in cTnC reconstituted fibers was approximately threefold faster than control, despite the lower (60%) maximum Ca(2+)-activated force after reconstitution. To obtain submaximal force with aTnC, eight fibers were treated to fully extract endogenous sTnC, then reconstituted with a mixture of a TnC and cTnC (aTnC:cTnC molar ratio 1:8.5). A second extraction selectively removed cTnC. In such fibers containing aTnC only, neither force nor kTR was affected by changes in [Ca2+]. Force was 22 +/- 7% of maximum control (mean +/- SD; n = 15) at pCa 9.2 vs. 24 +/- 8% (mean +/- SD; n = 8) at pCa 4.0, whereas kTR was 98 +/- 14% of maximum control (mean +/- SD; n = 15) at pCa 9.2 vs. 96 +/- 15% (mean +/- SD; n = 8) at pCa 4.0. Maximal reconstitution of fibers with aTnC alone increased force at pCa 9.2 to 69 +/- 5% of maximum control (mean + SD; n = 22 determinations on 13 fibers) and caused a small but significant reduction of kTR to 78 +/- 8% of maximum control (mean +/- SD; n = 22 determinations on 13 fibers); neither force nor krR was significantly affected by Ca>2(pCa 4.0). Taken together, we interpret our results to indicate that kTR reflects the dynamics of activation of individual thin filament regulatory units and that modulation of kTR by Ca> is effected primarily by Ca>+ binding to TnC.

Animals↗

Ca2+ and cross-bridge-induced changes in troponin C in skinned skeletal muscle fibers: effects of force inhibition.

Changes in skeletal troponin C (sTnC) structure during thin filament activation by Ca2+ and strongly bound cross-bridge states were monitored by measuring the linear dichroism of the 5' isomer of iodoacetamidotetramethylrhodamine (5'IATR), attached to Cys98 (sTnC-5'ATR), in sTnC-5'ATR reconstituted single skinned fibers from rabbit psoas muscle. To isolate the effects of Ca2+ and cross-bridge binding on sTnC structure, maximum Ca2+-activated force was inhibited with 0.5 mM AlF4- or with 30 mM 2,3 butanedione-monoxime (BDM) during measurements of the Ca2+ dependence of force and dichroism. Dichroism was 0.08 +/- 0.01 (+/- SEM, n = 9) in relaxing solution (pCa 9.2) and decreased to 0.004 +/- 0.002 (+/- SEM, n = 9) at pCa 4.0. Force and dichroism had similar Ca2+ sensitivities. Force inhibition with BDM caused no change in the amplitude and Ca2+ sensitivity of dichroism. Similarly, inhibition of force at pCa 4.0 with 0.5 mM AlF4- decreased force to 0.04 +/- 0.01 of maximum (+/- SEM, n = 3), and dichroism was 0.04 +/- 0.03 (+/- SEM, n = 3) of the value at pCa 9.2 and unchanged relative to the corresponding normalized value at pCa 4.0 (0.11 +/- 0.05, +/- SEM; n = 3). Inhibition of force with AlF4- also had no effect when sTnC structure was monitored by labeling with either 5-dimethylamino-1-napthalenylsulfonylaziridine (DANZ) or 4-(N-(iodoacetoxy)ethyl-N-methyl)amino-7-nitrobenz-2-oxa-1,3-diazole (NBD). Increasing sarcomere length from 2.5 to 3.6 microm caused force (pCa 4.0) to decrease, but had no effect on dichroism. In contrast, rigor cross-bridge attachment caused dichroism at pCa 9.2 to decrease to 0.56 +/- 0.03 (+/- SEM, n = 5) of the value at pCa 9. 2, and force was 0.51 +/- 0.04 (+/- SEM, n = 6) of pCa 4.0 control. At pCa 4.0 in rigor, dichroism decreased further to 0.19 +/- 0.03 (+/- SEM, n = 6), slightly above the pCa 4.0 control level; force was 0.66 +/- 0.04 of pCa 4.0 control. These results indicate that cross-bridge binding in the rigor state alters sTnC structure, whereas cycling cross-bridges have little influence at either submaximum or maximum activating [Ca2+].

Actomyosin↗

The use of patient-controlled boluses of local anaesthetic via a psoas sheath catheter in the management of malignant pain.

Patients who develop malignant infiltration of the psoas muscle and the lumbar plexus often experience a severe complex pain syndrome characterised by deep somatic pain, neuropathic pain and psoas spasm. Conventional analgesic regimes may not relieve these symptoms adequately. We describe the use of patient-controlled boluses of local anaesthetic via a psoas sheath catheter in this scenario. The recent availability of portable infusion pumps with the capability to deliver large volume boluses with long lockout times made this intervention possible and allowed the patient to be discharged home with effective relief of pain.

Analgesia, Patient-Controlled↗

[Paravertebral block of the lumbar plexus. Variations of the original technic].

After studying a hundred or so abdominal CT scans carried out at L5 level and having verified that the interfascial compartment between the quadrate muscle of the lumbi and the psoas muscle is relatively fixed (1.8-2.3 cm), it is proposed to utilise this datum (introduction of the 2 cm) needle after contact with the transverse apophysis of L5) to identify the psoas compartment which contains most of the nerves making up the lumbar plexus. Study of the series and results obtained confirm the value and simplicity of the proposed technique.

Adult↗

Effects of inorganic phosphate on cross-bridge behavior after photorelease of ATP in permeabilized cells of rat skeletal muscle.

Effects of 20 mM inorganic phosphate on the cross-bridge behavior after photorelease of ATP from caged ATP was studied by X-ray diffraction in rat skinned psoas muscle fibers at 16 degrees C. In the first 30 ms after the photorelease, tension was similar in the presence and absence of phosphate. The tension development was then suppressed in the presence of phosphate. At 500 ms after the photolysis, it was lower by about 30% in the presence of phosphate. In the presence of phosphate, the intensity of the third meridional reflection from the thick filament at 1/14.4 nm(-1) increased more slowly and was 60% of the level without phosphate at 500 ms after the photolysis. The intensities of the equatorial (1,1) reflection and the actin layer-line at 1/36 nm(-1) decreased to a lower level in the presence of phosphate. These results suggest that phosphate does not affect dissociation of myosin heads from actin, but decreases the number of myosin heads in force-generating conformation and reduces tension. Phosphate may also shift the equilibrium between the detached and attached states towards the former.

Actins↗

Half-sarcomere dynamics in myofibrils during activation and relaxation studied by tracking fluorescent markers.

To study the dynamics of individual half-sarcomeres in striated muscle contraction, myofibrils prepared from rabbit psoas muscle and left ventricles of guinea pig were immunostained with two conjugated antibody complexes consisting of a primary antibody against either alpha-actinin or myomesin and a secondary fluorescently labeled Fab-fragment. We simultaneously measured force kinetics and determined the positions of the Z-line and M-band signals by fluorescence video microscopy and sophisticated computer vision (tracking) algorithms. Upon calcium activation, sarcomeres and half-sarcomeres shortened nonuniformly. Shortening occurred first rapidly and exponentially during the force rise and then slowly during the force plateau. In psoas myofibrils, time-resolved displacements of the A-band in sarcomeres were observed, i.e., the two halves of individual sarcomeres behaved nonuniformly. Nonuniformity in length changes between the two halves of sarcomeres was comparable to that between two adjacent half-sarcomeres of neighboring sarcomeres. Sequential lengthening of half-sarcomeres was observed in cardiac myofibrils during the rapid phase of force relaxation. The independent dynamics of the halves in a sarcomere reveals the half-sarcomere as the functional unit rather than the structural unit, the sarcomere. The technique will facilitate the study of filament sliding within individual half-sarcomeres and the mechanics of intersegmental chemomechanical coupling in multisegmental striated muscles.

Actinin↗