Peter P. Purslow

The effect of ageing on the water-holding capacity of pork : role of cytoskeletal proteins

The water-holding capacity (WHC) of pork decreases post-mortem but has been shown to increase during subsequent ageing. In order to test a hypothesis that water-holding capacity increases during ageing due to degradation of the cytoskeleton, WHC was followed 10 days post-mortem and related to the extent of proteolysis of cytoskeletal proteins. A fast method for measuring WHC in small meat samples was developed by the use of centrifugation. The WHC of fresh pork decreases in the first part of post-mortem storage after which it increases to the level of 1 day PM. No changes in total water content of the meat were observed which could explain changes in WHC during ageing. Vinculin and desmin degrade gradually during ageing while talin degrades rapidly. These observations are consistent with the hypothesis that degradation of the cytoskeleton slowly removes the linkage between lateral shrinkage of myofibrils and shrinkage of entire muscle fibres, so removing the force that causes flow into the extracellular space. Inflow of previously expelled water is then possible, so increasing WHC as observed in later periods of storage.

Strain: induced reorientation of an intramuscular connective tissue network: implications for passive muscle elasticity

The most abundant intramuscular connective tissue component, the perimysium, of bovine M. sternomandibularis muscle was shown to be a crossed-ply arrangement of crimped collagen fibres which reorientate and decrimp on changing muscle fibre sarcomere length. Reorientation of perimysial strands was observed by light microscopy and identification of these strands as collagen fibres was confirmed by high-angle X-ray diffraction. Mean collagen fibre direction with respect to the muscle fibres ranged from approximately 80 degrees at sarcomere length = 1.1 micron to approximately 20 degrees at 3.9 microns. This behaviour was well described by a model of a crimped planar network surrounding a muscle fibre bundle of constant volume but varying length. Modelling of the mechanical properties of the perimysium at different sarcomere lengths produced a load-sarcomere length curve which was in good agreement with the passive elastic properties of the muscle, especially at long sarcomere lengths. It is concluded that the role of the perimysial collagen network is to prevent over-stretching of the muscle fibre bundles.

Physiological and structural events post mortem of importance for drip loss in pork.

Early post mortem metabolism and structural changes from 3 to 24 h, together with pH, temperature and impedance Py development were investigated in 37 Duroc×Landrace×Large White (DLY) pigs covering a range of drip loss from 2.2 to 12.6%. Multivariate statistical analysis was used to assess the impacts of different metabolites, pH and temperature, impedance, cytoskeletal protein degradation and extracellular cross-sectional area on drip loss. Taken as single factors, the concentration of lactate could explain 80% of the variation in drip, inosine monophosphate (IMP) and adenosine triphosphate (ATP) concentration explained 71 and 68%, respectively, whereas inosin and glycogen levels explained only 59 and 60%. The extracellular area was found to explain 39% of the variation in drip. The area between fibres provided more significant information than did the area between fibre bundles. The degradation of the cytoskeletal proteins was not related to drip loss. Impedance Py development over 24 h could explain 66% of the variation in drip, whereas pH and temperature explained 85 and 87%, respectively. A model including all measured variables could explain 83% of the variation in drip. However, only pH, temperature, impedance, [ATP](1 h) and [lactate](1 h and 2 h) were significant in relation to drip. By reducing the variables in the multivariate analysis, 89% of the variation in drip could be explained by a model containing only pH(2 h) and temperature(1 min). To explain variation in drip loss, pH and temperature measurements at significant time points were sufficient. Variation in post-mortem metabolites did, however, explain why variation in pH and temperature occurred. Development of drip channels was ruled by pH and temperature while impedance development was highly correlated to pH. This multi-faceted study shows those parameters, which can best be used to indicate or predict WHC, as well as those indicating the basic mechanism underlying variations in drip.

Age related compliance of the lamina cribrosa in human eyes

AIMS To investigate changes in the mechanical compliance of ex vivo human lamina cribrosa with age. METHODS A laser scanning confocal microscope was used to image the surface of the fluorescently labelled lamina cribrosa in cadaver eyes. A method was developed to determine changes in the volume and strain of the lamina cribrosa created by increases in pressure. The ability of the lamina cribrosa to reverse its deformation on removal of pressure was also measured. RESULTS Volume and strain measurements both demonstrated that the lamina cribrosa increased in stiffness with age and the level of pressure applied. The ability of the lamina cribrosa to regain its original shape and size on removal of pressure appeared to decrease with age, demonstrating an age related decrease in resilience of the lamina cribrosa. CONCLUSIONS The mechanical compliance of the human lamina cribrosa decreased with age. Misalignment of compliant cribriform plates in a young eye may exert a lesser stress on nerve axons, than that exerted by the rigid plates of an elderly lamina cribrosa. The resilience of the lamina cribrosa also decreased with age, suggesting an increased susceptibility to plastic flow and permanent deformation. Such changes may be of importance in the explanation of age related optic neuropathy in primary open angle glaucoma.

The morphology and mechanical properties of endomysium in series-fibred muscles: variations with muscle length

SummaryIn the series-fibred muscle architecture commonly found in large muscles of mammals and birds, the intrafasciculary-terminating muscle fibres have no direct tendinous attachments. Contractile force produced in these fibres must be transmitted between adjacent muscle fibres via the endomysial connective tissue which separates them. The endomysium is thus an essential mechanical component in such muscles. Studies of motor end-plate banding patterns and the frequent occurrence of tapering ends of fibres within the fascicles of the bovine sternomandibularis muscle show it to be a series-fibred muscle. Sodium hydroxide digestion of fixed samples of this muscle to remove the myofibrillar apparatus revealed the endomysium to be a disordered planar network of mainly curvilinear collagen fibrils.The orientation distribution of the collagen fibrils in the endomysial network was measured by image analysis of scanning electron micrographs. Analysis of endomysial preparations from muscle fixed at sarcomere lengths between 1–4 μm showed that the orientation distribution of collagen fibrils is quantitatively related to muscle length. At rest sarcomere length the collagen fibril network is not completely random, but has a slight circumferential bias. The orientation distribution shows a progressive shift towards the circumferential direction at short sarcomere lengths and towards the longitudinal direction at long sarcomere lengths. The relationship between the number-weighted mean collagen orientation and sarcomere length was compared to two geometric models of network behaviour, the isoareal and constant shape models. both fitted the data reasonably, although the constant shape model described the rate of change of mean orientation more closely.From fibrous composites theory, the reinforcement efficiency factor, η was calculated from the measured collagen fibril orientation distributions. These calculations predict a non-linearly increasing longitudinal tensile modulus for the endomysium with increasing sarcomere length, in agreement with its known non-linear properties, but confirm that the tensile properties of the endomysium are unsuitable for transmission of tensile force from muscle fibres contracting near rest length. This reinforces a previous interpretation that contractile force is transmitted between neighbouring muscle fibres by trans-laminar shear through the endomysium rather than by in-plane tension.

The ¤effect of cooking temperature on mechanical properties of whole meat, single muscle fibres and perimysial connective tissue

The structural changes in beef semitendinosus caused by cooking were studied by performing tensile tests of the isolated meat components (i.e. single muscle fibres and perimysial connective tissue) and related to the toughness of the whole meat. Whole meat toughness was found to increase in two separate phases upon cooking from 40-50°C, and again from 60 to 80°C with a decrease in meat toughness between 50 and 60°C, in agreement with previous studies. The changes in whole meat toughness at temperatures below 60°C were found to correspond to changes in the mechanical properties of the perimysial connective tissue, whereas changes of whole meat toughness at temperatures above 60°C were found to correspond to increased breaking strength of single muscle fibres. The myofibrillar component explained approximately 47% of the variation in whole meat toughness upon cooking whereas inclusion of the connective tissue component increased the goodness of fit.

The physical basis of meat texture: Observations on the fracture behaviour of cooked bovine M. Semitendinosus.

The fracture behaviour of cooked strips of beef M. semitendinosus was studied by qualitative observation of the manner in which fracture occurred and by quantitative measurements of ultimate tensile strength, work of fracture and notch sensitivity. Qualitative observations showed that fracture started in the perimysial connective tissue in all test configurations used, resulting initially in the separation of intact muscle fibre bundles. The ultimate tensile strength along and across the fibres was ∼-300kNm(-2) and ∼-25kNm(-2), respectively. The qualitative aspects of fracture were explained on the basis of a uniaxial fibrous composite of strong muscle fibre bundles in a weak connective tissue matrix, with poor interfacial strength. Work of fracture through the perimysium was in the range 0·4 to 1·8kJm(-2). The difficulty in propagating fracture across the muscle fibre bundles was explained in terms of the materials complete insensitivity to notches running across the fibres. The results imply that the muscle fibre bundle is an important level of structural organisation as far as fracture is concerned and that the strength of the perimysium, or perimysium/muscle fibre bundle interface, is likely to have a major influence on the toughness ofthe cooked meat.

Immunolocalisation of intermediate filament proteins in porcine meat. Fibre type and muscle-specific variations during conditioning

Two intermediate filament proteins, desmin and vinculin, were immunofluorescently localised in porcine longissimus dorsi(∗∗) and iliocostalis muscles up to 7 days post mortem. Changes in tenderness of the m. longissimus and the water-holding capacity of both muscles were also monitored throughout this storage period. Longissimus muscle shows a rapid decline in the overall intensity of labelling for both desmin and vinculin. In contrast to the vinculin labelling, desmin labelling is preferentially lost from type IIB muscle fibres in the longissimus muscle. In the redder iliocostalis muscle, the loss of desmin and vinculin labelling was less rapid and did not show an obvious relation to muscle fibre type. In one sample with very high water loss, there were indications of greater extracellular space development and delayed loss of intermediate filament labelling. The time course of diminishing intermediate filament labelling is consistent with previous suggestions that degradation of these proteins is involved in post-mortem conditioning. The variations within and between muscles shown here may explain some of the variability in their mechanical properties. Additionally, it is suggested that intermediate filament integrity is necessary for the expulsion of water from the muscle cells during drip formation.

Connective tissue differences in the strength of cooked meat across the muscle fibre direction due to test specimen size

Systematic variations in the tensile strenght of cooked beef M. semitendinosus across the muscle fibre direction due to the cross-sectional size of specimens are demonstrated in specimens from (a) longitudinal and (b) transverse slices. The strength perpendicular to the fibre direction of longitudinal slices of thickness 0·25-5·75 mm varied by a factor of 2, thicker slices being stronger. This factor of 2 is in approximate agreement with the difference in strength of transverse versus longitudinal slices across the fibre direction. These variations of strength due to specimen geometry are explained on the basis of the increasing likelihood of including a ribbon of the perimysial connective tissue network which is continuous along the whole length of the test piece in larger samples. The breaking strength of small cross-sectional area specimens is likely to be dominated by the strength of the endomysial-perimysial junction. Larger cross-sectioned specimens, by including continuous strands of the perimysial network, have higher strengths resulting from the necessity to break these strands. These findings highlight the need to specify specimen dimensions in tensile test results. They also show that by manipulating specimen geometry, the relative magnitude of the two mechanisms of connective tissue fracture (endomysial-perimysial separation and perimysial strand fracture) may be assessed.

Structure and Function of Intramuscular Connective Tissue

In our bodies there are literally hundreds of muscles, working singly or in concerted groups to fulfil many different functions. Leaving aside the smooth, involuntary muscles and the very special cardiac muscle, the voluntary, skeletal or striated muscles perform tasks as diverse as maintaining the posture of the spine, producing rapid and powerful movements of the limbs and executing the precise and finely judged movements of the eye. The structure of individual muscles, therefore, not surprisingly differs from anatomical location to location, from the grossest considerations of size and shape down to the composition and properties of the molecules involved in contraction.