Improved Lesion Detection by Using Axial T2-Weighted MRI with Full Spinal Cord Coverage in Multiple Sclerosis.

BACKGROUND AND PURPOSE: Identification of lesions in specific locations gains importance in multiple sclerosis imaging diagnostic criteria. In clinical routine, axial scans are usually exclusively obtained to depict the cervical spinal cord or used to confirm suspected lesions on sagittal scans. We sought to evaluate the detection rate for MS lesions on axial T2WI scans with full spinal cord coverage in comparison with sagittal scans. MATERIALS AND METHODS: One hundred fifteen patients with definite or suspected MS underwent an MR imaging examination including 3-mm sagittal and 3.5-mm axial T2-weighted images with full spinal cord coverage. T2WI lesions were identified on axial and sagittal scans independently by 2 raters. Axial diameter, craniocaudal extension, lesion intensity, and location were analyzed. RESULTS: Four hundred forty-nine of 509 (88.2%) lesions were detected on axial and 337/509 (66.2%) on sagittal scans. Only 277/449 (61.7%) axial lesions were also detected on sagittal images. The number of lesions visible on sagittal and axial images was dependent on the axial lesion diameter (P < .001). CONCLUSIONS: Axial T2WI scans with full spinal cord coverage showed 22% more lesions in patients with MS in comparison with sagittal scans, especially for lesions with small axial diameters. We suggest including biplanar spinal MR imaging with full spinal cord coverage for lesion detection in MS in clinical routine and for clinical studies.

Prevalence of dementia in a rural nursing home population in Southern Germany

Background and Objectives: We sought to obtain data about the prevalence of dementia in rural nursing homes in Germany. Methods: We conducted our data between 2007 and 2009 in the diocese of Passau, Germany. By using a questionnaire we asked all nursing homes in the area to provide information about patients with dementia. We obtained detailed data from three nursing homes by personal visits, telephone calls and mail. Results: Out of 72 nursing homes, 40 provided the requested data. The 40 facilities included in our study house a total of 3,928 residents, 1,892 of whom are diagnosed with dementia (48%). Three nursing homes provided us with more detailed information. Conclusions: We conclude that the prevalence of dementia in nursing homes in Germany is comparable to those rates reported in other countries. As there is a lack of information globally on rural populations affected by dementia, further international research in this area will give important insights into the risk factors, the variables influencing the course of the disease, and the special care needs of this population (AU)

Intracellular pH of crab and crayfish muscle fibre types during GABA application and inhibitory nerve stimulation.

The inhibitory motoneurons of crustaceans form synapses both with the sarcolemma of muscle fibres and with the very distal branchings of the excitatory motoneurons. The transmitter of these synapses is GABA (gamma-aminobutyric acid) which is known to open Cl- channels. Studies on the dactyl opener muscle of crayfish suggest that application of GABA not only leads to an increase in the Cl- permeability but also to a considerable HCO3- conductance that causes an intracellular acidification. To investigate possible physiological implications, we measured the intracellular pH of various muscle fibre types of crayfish and crab using pH-sensitive microelectrodes. Independent of the presence or absence of inhibitory innervation, bath application of 10(-5) mol l(-1) GABA led to acidification in all fibre types (pH change: 0.14 +/- 0.08, n = 11). In no preparation was a change in intracellular pH observed upon stimulation of specific or common inhibitory motoneurons with 1040 pulses s(-1) for 2-5 min. The results suggest that HCO3- conductance cannot be activated through synaptic GABA receptors. However, all crustacean muscle fibre types seem to possess extrasynaptic GABA-sensitive channels that exhibit a considerable HCO3- conductance. The physiological importance of these channels remains to be elucidated.

Ca2+_, Sr2+_force relationships and kinetic properties of fast-twitch rat leg muscle fibre subtypes.

Force generation of fast-twitch and slow-twitch fibres exhibits large differences in its sensitivity to Ca2+ and Sr2+ (e.g. Fink et al. 1986). Little is known about fast-twitch fibre subtypes. Thus, a variety of mechanical measurements on segments of rehydrated freeze-dried fast-twitch rat leg muscle fibres were executed in this study. Among these, the Ca2+- and Sr2+-force relationship and the unloaded shortening velocity were determined. The fibres were classified into subtypes according to their kinetics of stretch activation (Galler et al. 1994). In all fibres, the maximal force under Sr2+ activation was about 0.9 of that under Ca2+ activation. The Ca2+- and Sr2+-force relationship exhibited a biphasic shape with a steeper part (Hill coefficient, n1) below 50% and a flatter part (Hill coefficient, n2) above 50% of maximal force. The difference between the Ca2+ - and Sr2+ -sensitivity was independent of the fibre subtypes. The Hill coefficients were only partially correlated with kinetic properties. The correlation was more pronounced for the unloaded shortening velocity than for the kinetics of stretch activation. The data are consistent with the idea that the Ca2+ and Sr2+ sensitivities of fast-twitch fibres are mainly determined by a single isoform of troponin C. Among several protein isoforms, the isoforms of the myosin light chains seem to be involved for determining the slope of the Ca2+- and Sr2+-force relationship of fast-twitch muscle fibres.

Force responses of skinned molluscan catch muscle following photoliberation of ATP.

Isometric force responses following flash photolysis of caged-ATP were measured from skinned preparations of the catch muscle anterior byssus retractor of Mytilus (ABRM). When fibres were transferred from Ca(2+)-free to Ca(2+)-containing rigor solution (pCa < 4) the force remained low, but flash photolysis produced an extended force increase (half-time, 0.30 +/- 0.07 s, n = 6). When Ca(2+)-activated fibres were transferred to a Ca(2+)-free rigor solution, their force remained at a high level. Flash photolysis produced a rapid force decay (half-time, 0.28 +/- 0.06 s, n = 9) to about 19% of the initial Ca(2+)-activated force. In the presence of 0.5 mM MgADP, the force increase was slowed down by a factor of 3 and the force decay by a factor of 5. These effects of MgADP on crossbridge kinetics are comparable to those observed in vertebrate smooth muscle and are thought to cause "latch", a catch-like state (Fuglsang et al. J Muscle Res Cell Motil 14:666-677, 1993). They are consistent with a model implicating competition between MgADP and MgATP for the nucleotide-binding site on crossbridges. Considering the relatively fast force responses induced by caged-ATP photolysis, even in the presence of MgADP, it appears unlikely that the detachment of crossbridges from the rigor state can account for catch-related processes. In view of the low myosin ATPase under maximal activating conditions (0.6 s-1, Butler et al. Biophys J 75:1904-1914, 1998), neither crossbridge attachment nor detachment of rigor crossbridges seems to be the rate-limiting processes of the crossbridge cycle.

Shortening properties of two biochemically defined muscle fibre types of the Norway lobster Nephrops norvegicus L.

Mechanical properties of myofibrillar bundles from single chemically skinned fibres from the superficial abdominal flexor muscle of the Norway lobster Nephrops norvegicus were measured, and the protein content of these fibres was analysed by SDS-PAGE. Two slow fibre phenotypes (S1, S2) were distinguished on the basis of their myofibrillar protein assemblages. Data from 9 S1 and 8 S2 fibres obtained at similar sarcomere length demonstrate significant differences between the fibre types in maximal tension (N cm-2, S1: 10.5 +/- 3.9; S2: 3.1 +/- 0.8), in the delay of the peak of stretch activation (ms, S1: 122 +/- 18; S2: 412 +/- 202), in fibre stiffness (N cm-2 per nm half sarcomere, S1: 0.36 +/- 0.19; S2: 0.09 +/- 0.03) and in maximal shortening velocity (fibre length s-1, S1: 0.53 +/- 0.10; S2: 0.27 +/- 0.06). Furthermore, the maximal power output of the type S1 fibres was about five times larger than that of S2 fibres. The power output was maximal at lower loads in S1 fibres (relative load = 0.37 +/- 0.04) than in S2 fibres (relative load = 0.44 +/- 0.05). This study represents a comprehensive investigation of two slow muscle fibre types which are thought to be specialized for slow movements (S1 fibres) and for the postural control of the abdomen (S2 fibres).

Kinetic properties of myosin heavy chain isoforms in single fibers from human skeletal muscle.

The head portion of the myosin heavy chain is essential in force generation. As previously shown, Ca2+-activated muscle fibers from rat and rabbit display a strong correlation between their myosin heavy chain isoform composition and the kinetics of stretch activation, corresponding to an order of velocity: myosin heavy chain Ib > myosin heavy chain IId(x) > myosin heavy chain IIa >> myosin heavy chain I. Here, we show a similar correlation for human muscle fibers (myosin heavy chain IIb > myosin heavy chain IIa >> myosin heavy chain I), suggesting isoform-specific differences between the kinetics of force-generating power strokes. The kinetics of myosin heavy chain I are similar in human and rodents. This holds also true for myosin heavy chain IIa, but human myosin heavy chain IIb is slower than rodent myosin heavy chain IIb. It is similar to rodent myosin heavy chain IId(x).

Activation of skinned muscle fibres from the Norway lobster Nephrops norvegicus L. by manganese ions.

Effects of Mn2+ and Ca2+ on the mechanical properties of glycerinated myofibrillar bundles originating from slow S1 type muscle fibres of superficial flexor muscles of the lobster Nephrops norvegicus were investigated. Mn2+ (5-20 microM) activated the preparations in a dose-dependent manner. The sensitivity of myofibrillar force generation for Mn2+ was around 30 times lower than that for Ca2+. The maximal tension produced under Mn2+ activation was about 75% of that under Ca2+ activation. At higher free Mn2+ concentrations (>2 mM), the steady-state force decreased; it was completely abolished at 30 mM free Mn2+. These high Mn2+ solutions were accompanied by changed in MgATP and MnATP concentrations, and in the ionic strength. Control experiments have shown that none of these parameters seemed fo account fully for the observed force depression in high Mn2+ solutions. It is likely that direct effects of Mn2+ such as a change of the myofilament surface charges are responsible. The maximal unloaded shortening velocity of the myofibrillar preparations was shown to be similar under maximal Mn2+ and Ca2+ activation. Conversely, the kinetics of stretch-induced delayed force increase were about two to three times faster under Mn2+ activation. These results suggest that certain steps of the cross-bridge cycle depend on the ion species bound to the regulatory proteins.

Improvement of the measurements on skinned muscle fibres by fixation of the fibre ends with glutaraldehyde.

Experiments with activated skinned muscle fibre segments are limited by the structural and mechanical instability of the preparations. The present study shows that fixation of the muscle fibre ends with glutaraldehyde significantly improves the reliability of such experiments. We tested the effects of a specific glutaraldehyde fixation technique on the structural stability and the mechanical properties of skinned rat and rabbit skeletal muscle fibres in an approach where the fibre segments are attached to the apparatus by gluing. Preparations with fixed and unfixed ends were compared. During the first few minutes of maximal activation, fibres with fixed and unfixed ends exhibited similar mechanical properties to one another, suggesting that our fixation procedure selectively impregnates the fibre ends without contaminating the remaining active fibre part. During prolonged maximal activations (3-60 min), preparations with fixed ends exhibited a better stability, both in the sarcomere length signal (detected by laser diffraction) and in the unloaded shortening velocity. Thus, our technique of muscle fibre end fixation caused a substantial improvement in the mechanical measurements on skinned muscle preparations.

Tension/stiffness ratio of skinned rat skeletal muscle fibre types at various temperatures.

It is well known that shortening velocity and maximal tension of muscle preparations are strongly dependent on experimental temperature. Conversely, studies about temperature effects on muscle fibre stiffness are scarce. In the present study, we measured tension and stiffness of maximally Ca2+-activated skinned rat skeletal muscle fibres of different types over a wide temperature range. All fibre types exhibited a similar tension/stiffness ratio at each experimental temperature. This ratio increased almost linearly from 6 to 18 nm when the temperature was raised from 6 to 34 degrees C. Our results are discussed in the light of the drastic discrepancies reported for the amount of compliance inside and outside the attached myosin cross-bridges of activated muscle fibres (Ford et al. 1981, Huxley et al. 1994, Kojima et al. 1994, Wakabayashi et al. 1994, Higuchi et al. 1995). The relation between these compliances had been deduced from various experimental approaches executed at different temperatures. The large temperature sensitivity of the tension/stiffness ratio found in this study provides evidence for the assumption that the compliance outside the cross-bridges increases with rising temperature. This view would reconcile the contrasting results reported for the relation of compliances inside and outside the attached cross-bridges.

Effects of myofibrillar bundle diameter on the unloaded shortening velocity of skinned skeletal muscle fibres.

Using both slack tests and force clamp experiments, the velocity of unloaded shortening (Vu; Vu(st), slack test; Vu(fc), force clamp) was determined for maximally Ca(2+)-activated myofibrillar bundles. These were obtained by mechanically splitting single muscle fibres of rat, rabbit, crab and lobster skeletal muscles. A comparison was made between the Vu of thick (mammalian: 45-70 microns mean diameter; crustacean: 90-175 microns) and thin (mammalian: 25-40 microns; crustacean: 35-85 microns) preparations of the same muscle fibre. The bundle diameter had opposite effects on Vu in mammalian and crustacean muscle fibres. The Vu of thin mammalian bundles was about 0.6 times that of the thick ones, whereas in crustacean preparations this ratio was about 1.5. The kinetics of stretch-induced delayed force increase of maximally Ca(2+)-activated fibres (stretch activation) appeared not to differ between the thick and thin bundles from any animal preparation. Control experiments showed that the observed diameter effects on Vu are not due to differences in the chemical environment of the myofilaments. One possible explanation is that the intrinsic physical factors of the myofibrils modify Vu differently during progressive shortening in mammalian and crustacean preparations.

Stretch activation and isoforms of myosin heavy chain and troponin-T of rat skeletal muscle fibres.

Recent studies on single mammalian skeletal muscle fibres revealed a correlation between the kinetics of stretch-induced delayed force increase (stretch activation) and the isoforms of the myosin heavy chain. This observation suggests a causal relation between stretch activation and myosin heavy chain. However, the assumption is weakened by the fact that isoforms of other myofibrillar proteins tend to be coexpressed with myosin heavy chain isoforms. The relation between the isoforms of the tropomyosin-binding troponin subunit and myosin heavy chain is unknown. For a variety of reasons, tropomyosin-binding troponin subunit is a possible candidate for being involved in stretch activation. Therefore, we measured stretch activation of single, maximally Ca(2+)-activated skinned rat skeletal muscle fibres and characterized them by their myosin heavy chain composition, as well as by the isoform species of tropomyosin-binding troponin subunit. Four myosin heavy chain isoforms (I, IIa, IId or IIx and IIb) and six tropomyosin-binding troponin subunit isoforms (TnT1s, TnT2s, TnT1f, TnT2f, TnT3f, TnT4f) were distinguished. The following preferential coexpression patterns of the myosin heavy chain and tropomyosin-binding troponin subunit isoforms were observed: MHCI-TnT1s, MHCIIa-TnT3f, MHCIId-TnT1f, and MHCIIb-TnT4f. Stretch activation kinetics was found to be correlated with the myosin heavy chain isoform complement also in fibres not displaying one of the preferential MHC-TnTf isoform coexpression patterns. This corroborates the assumption of a causal relation between myosin heavy chain and stretch activation.

Mechanical properties and myosin heavy chain isoform composition of skinned skeletal muscle fibres from a human biopsy sample.

Experiments were conducted to investigate the mechanics of contraction of chemically skinned muscle fibre segments of a biopsied sample of single human quadriceps muscle. Subsequently, the isoforms of the myosin heavy chain (MHC) were analysed by sodium dodecyl sulphate (SDS) gel electrophoresis. Of the 41 fibres, 26 contained MHCI (type I), 11 of the fibres contained MHCIIa (type IIA), and 4 of the fibres contained both MHCI and MHCIIa (of which MHCIIa was always slightly predominant (type IIC)). Distinct differences between fibre types were found in terms of the kinetics of force responses following stepwise length changes (order of velocity: IIA > IIC > I). The differences in maximal shortening velocity and in the kinetics of Ca(2+)-dependent activation were of the same order, but much less pronounced. Type I fibres had significantly greater fibre diameters than type IIA fibres. No significant differences were found among different fibre types in terms of isometric tension, resting sarcomere length or the length change needed to discharge the elasticity of maximally Ca(2+)-activated fibres (V0 value). The distribution of shortening velocity and kinetics of stretch activation values suggest that two muscle fibre subtypes may exist in human type I fibres.

Stretch activation and myosin heavy chain isoforms of rat, rabbit and human skeletal muscle fibres.

The underlying mechanism of stretch-induced delayed force increase (stretch activation) of activated muscles is unknown. To assess the molecular correlate of this phenomenon, we measured stretch activation of single, Ca2+-activated skinned muscle fibres from rat, rabbit and the human and analysed their myosin heavy chain complement by SDS gradient gel electrophoresis. Stretch activation kinetics was found to be closely correlated with the myosin heavy chain isoform complement (I, IIa, IId/x and IIb). In hybrid fibres containing two myosin heavy chain isoforms (especially IId and IIb), the kinetics of stretch activation depended on the percentage distribution of the two isoforms. Muscle fibres of the same type but originating from different mammalian species exhibited similar kinetics of stretch activation. Considering the differing unloaded shortening velocities of these fibres, the time-limiting factors for stretch activation and unloaded shortening velocity appear not to be the same. The stretch activation kinetics of the fibre types IIB, IID and IIA more likely seemed to follow a Normal Gaussian distribution than that of type I fibres. Several type I fibres had extraordinarily slow kinetics. This observation corroborates biochemical data indicating the possible existence of more than one slow myosin heavy chain isoform.

Effects of nitric oxide on force-generating proteins of skeletal muscle.

Nitric oxide (NO) has recently been identified as a physiologically important intracellular messenger modulating the contractile activity of skeletal muscle [Kobzik L, Reid MB, Bredt DS, Stamler JS (1994) Nature 372: 546-548]. However, the mechanism of action of NO is not yet known. We used skinned (demembranated) muscle fibres to investigate the mechanism of NO function in muscle contraction. Maximally Ca2+-activated single fibres of rat skeletal muscle were exposed to physiologically relevant NO concentrations by adding NO donor molecules into the bath solution. Donor application caused a decline both in the contractile properties and in the myofibrillar adenosine triphosphatase (ATPase) activity. These results reveal a novel molecular mechanism of NO action: a direct inhibition of the force-generating proteins in skeletal muscle.

Two functionally distinct myosin heavy chain isoforms in slow skeletal muscle fibres.

The head part of the myosin heavy chain (MHC) represents the essential component of the molecular force-generating system of muscle [1-3]. To date, three fast but only one slow MHC isoforms have been identified in adult mammalian limb muscles [4,5]. We show here two functionally different slow MHC isoforms, MHCIbeta and MHCIa, coexisting in a considerable fraction of slow fibres of rabbit plantaris muscle. The two isoforms exhibit distinct electrophoretic mobilities and different kinetic properties. Thus, as it is known for the fast muscle, also the slow muscle seems to use different MHC isoforms in order to fulfil different functional demands.

[The effect of compression therapy on the microcirculation of the skin in patients with chronic venous insufficiency (CVI)]. / Einfluss einer Kompressionstherapie auf die Mikrozirkulation der Haut bei Patienten mit chronischer Veneninsuffizienz (CVI).

Compression therapy was employed for 4 weeks 20 patients with chronic venous insufficiency stage CVI I and CVI II according to Widmer's classification. Compression bandaging for 2 weeks was followed by compression stockings for 2 more weeks. The cutaneous microcirculation was evaluated before therapy, after 2 weeks and after finishing compression therapy after 4 weeks. Marked improvement in symptoms such as pain and itching was observed after 4 weeks, along with a significant reduction in lower limb volume. Video capillary microscopy revealed an increase in capillary density associated with a decrease in capillary diameter and pericapillary halo diameter. Compression treatment achieves at least part of its effect by improving the function of the skin microcirculation. The efficacy of bandaging and stockings was similar.

Force responses following stepwise length changes of rat skeletal muscle fibre types.

1. Force responses following stepwise length changes of Ca(2+)-activated skinned leg muscle fibres (6 degrees C) of the rat were correlated with their myosin heavy chain (HC) isoforms (myosin HC I, fibre type I; myosin HC IIA, type IIA; myosin HC IID (HC IIX), type IID (type IIX); myosin HC IIB, type IIB) in order to study the mechanical properties of these molecules. 2. Marked differences in the time behaviour of force transients following quick releases of fibre length existed between various muscle fibres, and a conspicuous correlation with their myosin HC complement was noticed (order of velocity: IIB > IID > IIA > > I). No differences were found in the relationship between the applied length step and the resulting force (T1, T2 curves). 3. Our results suggest that the heads of various myosin heavy chain isoforms exhibit different kinetic properties. The differences concern the kinetics of the myosin head movements and the duration of cyclic interactions between myosin heads and thin filaments. The extent of force-generating movements and the mean elongation of attached heads in the isometric state seem to be independent of the isoform.

Stretch activation, unloaded shortening velocity, and myosin heavy chain isoforms of rat skeletal muscle fibres.

1. Contractile properties were investigated on single skinned-fibre preparations from rat leg muscles. Following the mechanical measurements, the myosin heavy chain (HC) composition of the same fibre was analysed by gradient gel electrophoresis. 2. Fibres were typed according to their myosin HC isoform composition (HCI, type I; HCIIA, type IIA; HCIID, type IID; HCIIB, type IIB). Many fibres showed the co-existence of two myosin HC isoforms (hybrid fibres). 3. A strong correlation was found between fibre type and time characteristics of stretch-induced delayed force increase (stretch activation) of fully Ca(2+)-activated fibres. 4. The maximal unloaded shortening velocity (Vmax), as measured with the slack test, was lowest in type I fibres. Within the type II group, a continuum of Vmax values was found, with large overlaps of the different fibre types. 5. The results suggest that the kinetics of stretch activation is determined by the myosin HCs whereas unloaded fibre shortening seems to be determined by other myofibrillar proteins in addition to the myosin HCs. Assuming that stretch activation represents certain steps of the cross-bridge turnover under isometric conditions and Vmax reflects cross-bridge detachment under unloaded conditions it can be deduced that different myofibrillar proteins are responsible for different steps within the cross-bridge turnover.