Roland Labas

The Challenge of Mastication: Preparing a Bolus Suitable for Deglutition

The main function of mastication is to transform a solid food into a bolus that can be swallowed safely. The bolus characteristics such as particles size or cohesiveness, are continuously sensed during mastication and they are important in initiating deglutition. This study examined the following question: What is the condition of the bolus just before swallowing? Ten subjects with normal dentition aged 37.5 ± 3.7 years were asked to chew without swallowing six different foods (three nuts and three vegetables) while the number of cycles and the duration of the sequence were recorded. The particle size distribution shown by the expectorated food bolus just before swallowing was examined by image analysis. The results showed that, for a given food, the sizes of the bolus particles just before swallowing were comparable in all subjects. However, the number of cycles and duration of the sequence varied between subjects. Taken together these data strongly suggest that the granularity of the bolus before swallowing has to reach a predetermined state which is obtained by using an individual chewing strategy. This suggests that the bolus structure reflects a key factor for homeostasis and explains the large interindividual variability of the mastication physiologic parameters.

Characterisation of PSE zones in semimembranosus pig muscle.

Pig semimembranosus muscles, sampled from normal hams or from PSE-zones of defective hams, were analysed by histochemistry and electrophoretic techniques. PSE zones were characterised by a disorganisation of fibre alignment and a significant increase of inter fibre spacing (26.2% vs. 16.9%, p<0.05). Protein solubility was significantly lower in defective muscle (55.4 vs. 91.5mg/g, p<0.001). SDS-PAGE evidenced in such samples a lower abundance of the 97, 40 and 26kDa bands in the sarcoplasmic fraction and a higher abundance of the 97, 58, 34, 31, 15 and 11kDa bands in the myofibrillar fraction. Intensity of the MHC band (200kDa) was lower in PSE zone samples. By 2-D electrophoresis, it was shown that troponin T, MLC 1 and alpha-crystallin were less proteolysed in defective muscles, while creatine kinase fragments were more represented. One form of HSP 27 was absent from PSE zone samples. Overall, meat from PSE-zones and fast pH fall-PSE meat show numerous histological and biochemical similarities, particularly in their protein characteristics.

Magnetic resonance imaging of connective tissue: a non‐destructive method for characterising muscle structure

A magnetic resonance imaging technique based on susceptibility-induced contrast was used to visualise the spatial distribution of connective tissue in meat. Magnetic resonance imaging of bovine meat samples was carried out with a high-field 4.7 T imager. Magnetic resonance images obtained with spin-echo and gradient-echo sequences were compared to elucidate the role of connective tissue in the additional signal losses observed in the gradient-echo images. maps were reconstructed from the multiple gradient-echo images, which provide quantitative information. Comparison with histological pictures indicates that these maps exhibit the overall organisation of the primary perimysium at the scale of the whole muscle. The distinct perimysial organisation shown between the Gluteo biceps and Pectoralis profundis muscles illustrates the potential of magnetic resonance imaging for characterising the muscle connective tissue structure. © 2000 Society of Chemical Industry

Curcumin treatment prevents increased proteasome and apoptosome activities in rat skeletal muscle during reloading and improves subsequent recovery.

Immobilization is characterized by activation of the ubiquitin (Ub)-proteasome-dependent proteolytic system (UPS) and of the mitochondrial apoptotic pathway. Increased oxidative stress and inflammatory response occur in immobilized skeletal muscles. Curcumin exhibits antioxidant and anti-inflammatory properties, blocked proteasome activation in intact animals, and may favor skeletal muscle regeneration. We therefore measured the effects of curcumin on immobilization-induced muscle atrophy and subsequent recovery. Rats were subjected to hindlimb immobilization for 8 days (I8) and allowed to recover for 10 days (R10). Fifty percent of the rats were injected daily with either curcumin or vehicle. Proteolytic and apoptotic pathways were studied in gastrocnemius muscles. Curcumin treatment prevented the enhanced proteasome chymotrypsin-like activity and the trend toward increased caspase-9-associated apoptosome activity at I8 in immobilized muscles. By contrast, the increase of these two activities was blunted by curcumin at R10. Curcumin did not reduce muscle atrophy at I8 but improved muscle recovery at R10 and the cross-sectional area of muscle fibers of immobilized muscles. Curcumin reduced the increased protein levels of Smac/DIABLO induced by immobilization and enhanced the elevation of X-linked inhibitory apoptotic protein levels at R10. Ub-conjugate levels and caspase-3 activity increased at I8 and were normalized at R10 without being affected by curcumin treatment. Altogether, the data show that curcumin treatment improved recovery during reloading. The effect of curcumin during the atrophic phase on proteasome activities may facilitate the initiation of muscle recovery after reloading. These data also suggest that this compound may favor the initial steps of muscle regeneration.

Development of image analysis tool for the classification of muscle fibre type using immunohistochemical staining

An accurate characterisation of muscle fibres is essential for studying muscle plasticity. During some transient events such as ageing, myogenesis, physical activity or conversion of muscle to meat, the morphological parameters and/or the fibre type distribution may change. Nowadays, this information is generally obtained using immunohistology techniques, but these analyses are acknowledged to be laborious and time-consuming. In fact, each myofibre, from thousands, must be measured individually and its expression profile in response to different anti-myosin antibodies must be established step by step. In this paper, we describe a new histological approach using double-labelling (laminin, myosin) serial sections, fluorescence microscopy visualisation and, finally, semi-automatic image analysis. The goal of the study was to propose a tool allowing faster fibre type characterisation, including the identification of hybrid fibres from pure ones. The steps in the image processing prone to subjectivity have been fully automated. On the other hand, the expert retained control of all image analysis procedures requiring visual diagnosis. The tool that we developed with the Visilog software allowed a rapid and objective fibre typing and morphometric characterisation of two different bovine muscles. The results were in agreement with our previous histological and densitometric assays. The method and the tool proved to be potentially more efficient than other techniques used in our institute or described in the literature. A more global evaluation will be considered in other laboratories as well as on other animal species.

Microstructural changes in m. rectus abdominis bovine muscle after heating.

A histological and ultrastructural study was conducted to characterize changes in beef muscle structure after heating. Pieces of rectus abdominis muscle were heated at 100 degrees C over varying time frames from 15 min to 60 min and at 270 degrees C for 1 min; samples were then prepared for optical and transmission electron microscopy. After 15 min of heating, at 100 degrees C, a lateral shrinkage in fibre of 48% and an increase in gaps between the myofibrillar masses of 27% was noted. No more significant evolution was observed as heating duration escalated. The ultrastructure showed strong myofibril to sarcolemma detachments in which granular aggregates, coming in part from myofibrillar proteins, are stored. Neighbouring muscle fibres showed strong heterogeneity in morphological behaviour after thermal treatment, suggesting that differences in composition and structure of the cytoskeleton proteins in the different fibres can cause denaturation/shrinkage of the proteins at different times along the timescale of microstructural changes during heating. Short heating at high temperatures expanded the gaps between myofibrillar mass, but the overall changes in the ultrastructure were similar to those obtained when heating at 100 degrees C.

Ultrastructural changes during aging in M. longissimus thoracis from moose and reindeer.

Game meat is commonly consumed in Europe but few studies have examined the quality related parameters. In this study we examined the changes in ultrastructure at four times postmortem in M. longissimus from moose (Alces alces) and reindeer (Rangifer tarandus tarandus). The moose were slaughtered during a hunt and reindeer by Swedish standard practices for this semi-domestic animal. Ultrastructural changes occurring in all animals included separation of the sarcolemma from myofibrils, I band breaks, and cytoskeleton breaks; however both I band breaks and cytoskeletal breaks were less common in moose and reindeer than values reported for sheep and beef. Fiber area in the longissimus thoracis muscle was approximately 3270 μm(2) for moose and 1170 μm(2) for reindeer indicating that the tenderness of reindeer meat may be largely determined by fiber size.

In situ thermal denaturation of myofibre sub-type proteins studied by immunohistofluorescence and synchrotron radiation FT-IR microspectroscopy.

The thermal denaturation of proteins in skeletal muscle was studied and characterised for the first time taking into account the in situ metabolic and contractile fibre types. From serial histological sections, collagen, elastin, various type I, IIa and IIx fibres and type I-IIa and IIa-IIx hybrids were identified by immunohistofluorescence. Histological sections were incubated in buffer solutions at increasing temperatures (40, 50, 60, 70 and 80°C). Protein secondary structure was investigated by synchrotron radiation FT-IR microspectroscopy on connective tissue and in muscle fibres rigorously identified for sub-type. Whatever the target protein components, increasing temperature resulted in a decrease in α-helix secondary structure and an increase in β-sheet structure. This phenomenon was more pronounced for intracellular proteins than for connective tissue. Although hybrid fibres were generally somewhat less sensitive to unfolding than the pure types, the amplitude of the thermal denaturation of intracellular proteins was practically independent of fibre type.

Influence of straw feeding and growth-implant on veal meat quality

Trials have been carried out to examine the effects of a food additive supply (Maturex) and/or of anabolic implant (Revalor) on carcass characteristics and on veal meat qualities. Twelve Holstein × Française Frisonne Pie Noire calves acted as controls, twelve calves were implanted with Revalor, approximately 60 days prior to slaughter, twelve others were fed with Maturex, whilst the remaining twelve animals were fed with Maturex and implanted with Revalor. All animals were slaughtered at 4 months. Revalor implantation had a significant effect on carcass weight and conformation. Feeding with Maturex had no significant effect on different carcass characteristics. Neither anabolic treatment nor Maturex supplementation significantly affected pH value, haematocrit, pigment content, FOP, colour characteristics, collagen content and solubility, or dry matter. Although Revalor had a negative effect on weight losses during cooking, Maturex showed no significant effect. While Maturex showed a positive effect on tenderness and no effect on juiciness, Revalor showed a negative effect on tenderness and juiciness. For all sensory characteristics, no effect was noted when both Maturex and Revalor were used together.

Relationship between histochemical, structural characteristics and oxidative stability of rhea limb muscles

Histochemical and structural characteristics were investigated in Gastrocnemius pars interna (GN) and Iliofiburalis (IF) limb muscles of Rhea americana. The average myofibre area cross-section was greater in GN than IF muscle (p<0.001), whereas the fibre density per section was higher in IF than GN muscle. The only type of myofibre found in both the rhea limb muscles analysed in this study was fast-twitch oxidative-glycolytic fibres (FOG). Immunolabelling analysis and ultrastructural observation of myofibres confirmed the contractile and metabolic characteristics of rhea myofibres, revealing the absolute fast isoform of myosin heavy chain and the abundance of glycogen and mitochondria inside the cells, mainly in IF muscle. These findings converged with previous results on the biochemical and physicochemical characteristics of rhea meat to provide further evidence that myofibre composition substantially influences the oxidative reactions of the muscle and therefore the meat quality, but more in-depth examination is needed to establish the links between myofibre characteristics, myofibre glycogen concentration and meat stability during storage.