Morphological changes in rat rectus abdominis muscle induced by diabetes and pregnancy.

The urethral muscle of diabetic pregnant rats is affected by long-term mild diabetes and short-term severe diabetes, which plays a crucial role in the pathogenesis of pelvic floor disorders. We hypothesized that muscles outside the pelvis are subject to similar changes. The current study aimed at analyzing the effects of long-term mild and short-term severe diabetes on the structure and ultrastructure of fiber muscles and collagen in rats' rectus abdominis (RA) muscle. Therefore, the RA muscle of virgin, pregnant, long-term mild diabetic, short-term severe diabetic, long-term mild diabetic pregnant and short-term severe diabetic pregnant 3-month-old Wistar rats were collected. The structure was analyzed by picrosirius red staining, immunohistochemistry for fast and slow muscle fibers and transmission electron microscopy. We investigated two levels of STZ- induced diabetes: long-term mild diabetes (blood glucose level: 120-200 mg/dL) and short-term severe diabetes (blood glucose level >300 mg/dL). Long-term mild diabetic pregnant and short-term severe diabetic pregnant rats had decreased fast fibers and increased slow fibers, disrupted areas of sarcomere, intermyofibrillar mitochondria and myelin figures in the RA muscle. Both groups enabled us to analyze the specific influence of pregnancy, separately from diabetes. The current study demonstrated that diabetes and pregnancy induced intramuscular transformation and reorganization of RA muscle with a switch of fiber type adjusting their architecture according to intensity and duration of hyperglycemic insult within pregnancy.

Morphological changes in rat rectus abdominis muscle induced by diabetes and pregnancy

The urethral muscle of diabetic pregnant rats is affected by long-term mild diabetes and short-term severe diabetes, which plays a crucial role in the pathogenesis of pelvic floor disorders. We hypothesized that muscles outside the pelvis are subject to similar changes. The current study aimed at analyzing the effects of long-term mild and short-term severe diabetes on the structure and ultrastructure of fiber muscles and collagen in rats' rectus abdominis (RA) muscle. Therefore, the RA muscle of virgin, pregnant, long-term mild diabetic, short-term severe diabetic, long-term mild diabetic pregnant and short-term severe diabetic pregnant 3-month-old Wistar rats were collected. The structure was analyzed by picrosirius red staining, immunohistochemistry for fast and slow muscle fibers and transmission electron microscopy. We investigated two levels of STZ- induced diabetes: long-term mild diabetes (blood glucose level: 120-200 mg/dL) and short-term severe diabetes (blood glucose level >300 mg/dL). Long-term mild diabetic pregnant and short-term severe diabetic pregnant rats had decreased fast fibers and increased slow fibers, disrupted areas of sarcomere, intermyofibrillar mitochondria and myelin figures in the RA muscle. Both groups enabled us to analyze the specific influence of pregnancy, separately from diabetes. The current study demonstrated that diabetes and pregnancy induced intramuscular transformation and reorganization of RA muscle with a switch of fiber type adjusting their architecture according to intensity and duration of hyperglycemic insult within pregnancy.

Ultrasound Assessment of Abdominal Muscle Thickness in Postpartum vs Nulliparous Women.

OBJECTIVE: The purpose of this study was to determine the effect size in measurable change of abdominal musculature morphology using ultrasonography in postpartum women within 1 month of a healthy, vaginal delivery. METHODS: One hundred fifty-six participants were recruited for this study. B-mode ultrasound imaging was used to measure abdominal muscle thickness on 80 nulliparous women and 76 mothers who had delivered within the past 4 weeks. Measures were taken for the upper and lower rectus abdominus, external and internal obliques, and transversus abdominus at rest. RESULTS: Statistically significant differences were found in the thickness of the rectus abdominus muscle at both sites; upper (P < .0001) and lower (P < .0001) as well as the internal oblique (P < .0001). All 3 muscles were thinner in postpartum participants (8.29 ± 1.83 mm, 8.89 ± 2.29 mm, and 7.06 ± 1.82 mm, respectively) within the first month of delivery than in controls (10.82 ± 1.93 mm, 11.13 ± 2.38 mm, and 8.36 ± 1.87 mm, respectively). Large effect sizes were found for the influence of pregnancy on the rectus muscle segments (1.35 for the upper rectus abdominus and 1.00 for the lower rectus abdominus) and a medium effect size for the internal oblique (0.71). No significant differences were observed in the remaining 2 muscles. CONCLUSION: This study showed that there are differences in morphology of the abdominal muscles in pregnant women vs nonpregnant controls. The large effect sizes reported may provide the basis for future studies examining relationships between morphology, functional change, and back pain during pregnancy.

Ultrastructural differences in rectus sheath of hernia patients and healthy controls.

BACKGROUND: The etiology of inguinal hernia remains unclear. Research data indicate the presence of pathologic alterations within the connective tissue; their exact character remains the subject of dispute. The search for new methods to diagnose connective tissue abnormalities, and thoroughly explain the character of the ultrastructural alterations, continues. MATERIALS AND METHODS: The study group included 10 male patients aged 18-60 y (five with primary inguinal hernia and five with acute appendicitis with no history of hernia). A specimen of the rectus muscle sheath was harvested from all of them upon surgery. The tissue samples were fixed and examined by spectrofluorometry and fluorescence microscopy, yielding fluorescence spectra and microscopic fluorescence images. RESULTS: Both techniques have demonstrated significant differences between the biopsy samples harvested from hernia patients and healthy controls. The groups of fluorescence spectra were shifted relative to each other and showed maximum emission at different wavelengths after excitation with 350 nm light (arbitrarily chosen for one of the cross-link proteins). The spectra obtained for healthy controls were more homogenous, while the spectra of the hernia samples differed even between each other. In microscopic images, the difference was a more chaotic distribution of fluorophores in the samples obtained from hernia patients. CONCLUSIONS: The evidence of significant differences between the samples harvested from the same location from hernia patients and healthy controls, found by fluorescence techniques, indicates the presence of abnormalities in the connective tissue forming the rectus muscle sheath. This area is not a part of the hernial defect, therefore, we can assume that the changes can be attributed to a generalized process.

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.

The gross morphology and histochemistry of respiratory muscles in bottlenose dolphins, Tursiops truncatus.

Most mammals possess stamina because their locomotor and respiratory (i.e., ventilatory) systems are mechanically coupled. These systems are decoupled, however, in bottlenose dolphins (Tursiops truncatus) as they swim on a breath hold. Locomotion and ventilation are coupled only during their brief surfacing event, when they respire explosively (up to 90% of total lung volume in approximately 0.3 s) (Ridgway et al. 1969 Science 166:1651-1654). The predominantly slow-twitch fiber profile of their diaphragm (Dearolf 2003 J Morphol 256:79-88) suggests that this muscle does not likely power their rapid ventilatory event. Based on Bramble's (1989 Amer Zool 29:171-186) biomechanical model of locomotor-respiratory coupling in galloping mammals, it was hypothesized that locomotor muscles function to power ventilation in bottlenose dolphins. It was further hypothesized that these muscles would be composed predominantly of fast-twitch fibers to facilitate the bottlenose dolphin's rapid ventilation. The gross morphology of craniocervical (scalenus, sternocephalicus, sternohyoid), thoracic (intercostals, transverse thoracis), and lumbopelvic (hypaxialis, rectus abdominis, abdominal obliques) muscles (n = 7) and the fiber-type profiles (n = 6) of selected muscles (scalenus, sternocephalicus, sternohyoid, rectus abdominis) of bottlenose dolphins were investigated. Physical manipulations of excised thoracic units were carried out to investigate potential actions of these muscles. Results suggest that the craniocervical muscles act to draw the sternum and associated ribs craniodorsally, which flares the ribs laterally, and increases the thoracic cavity volume required for inspiration. The lumbopelvic muscles act to draw the sternum and caudal ribs caudally, which decreases the volumes of the thoracic and abdominal cavities required for expiration. All muscles investigated were composed predominantly of fast-twitch fibers (range 61-88% by area) and appear histochemically poised for rapid contraction. These combined results suggest that dolphins utilize muscles, similar to those used by galloping mammals, to power their explosive ventilation.

Etiology of inguinal hernia: ultrastructure of rectus sheath revisited.

In the last decade, in the search for abdominal-wall hernia etiology, attention has been brought to alterations in the connective tissue ultrastructure as the probable etiological factor. These may cause weakening of connective tissue, which in turn may form ground for hernia formation. To investigate this hypothesis in depth, we compared the ultrastructure of the connective tissue in hernia patients and the control group. The study group consisted of five patients with primary inguinal hernia (Nyhus II = 4, Nyhus IIIa = 1). Another five patients posted for emergency appendectomy created the control group. Tissue specimens, harvested intraoperatively from the rectus muscle sheath (RAMS) and fixed in 4% glutaraldehyde, underwent staining by the Masson, H-E and methylene blue techniques and were assessed by microscopy (light and scanning electron). The examinations showed significant differences in the rectus sheath ultrastructure. They included altered architecture, placement and quantity of collagen and elastic fibers, differences in the caliber of individual fibers and disrupted ground matter-to-fiber ratio. In patients with hernias, chaotic arrangement of collagen fibers was seen, as well as their thinning and a decrease in the general amount of elastic fibers, replaced by ground matter. Our research has shown significant differences in the structure of the RAMS between patients with hernias and healthy individuals. This supports the theory linking connective tissue alterations with the etiology of hernia, and stating that these alterations include connective tissue at locations distant from the hernia site as well, as the rectus sheath itself does not form a hernial defect.

Reconstruction of ablated rat rectus abdominis by muscle regeneration.

Skeletal muscle regeneration is a powerful, naturally occurring process of tissue reconstruction that follows myofiber damage secondary to myotoxic injury that does not normally affect the tissue circulation and scaffold. The ablated tissue, in traumatology and free muscle grafts, is frequently replaced by scars. The final outcome is poor even after in situ myoblast seeding of the harvested muscle. The goal of this study was to identify protocols to reconstruct muscle tissue, even in such adverse environments. The authors applied a step-by-step approach to identify factors favoring the survival of autologous satellite cells and, thus, muscle regeneration. In a rat model of full-thickness rectus abdominis muscle ablation, autologous myoblasts were isolated from the explanted rectus abdominis and seeded in a homologous acellular matrix immediately after wall reconstruction (group 1, five animals). In group 2 (five animals), the ablated rectus abdominis was autografted in situ. In a third group of five rats, Marcaine was injected into both the autograft and the surrounding abdominal wall muscle. Three weeks after surgery, serial cross-sections of the reconstructed abdominal wall were stained with hematoxylin and eosin or embryonic myosin antibody, a well-characterized molecular marker of early myogenesis in development and regeneration. Percentages of the patch area covered by regenerated myofibers were determined by morphometry. When autologous myoblasts were seeded in a homologous acellular matrix, the only myofibers observed to regenerate were those along the border of the patch. Autografting of the middle third of the rectus abdominis muscle similarly resulted in scar formation. The few muscle cells in the graft core were scanty myoblasts that could be detected only by monoclonal embryonic myosin antibody. Although negative for myofiber regeneration, the results in both cases confirmed the mechanical patency of the patches with regard to abdominal organ support. Myofibers were successfully regenerated in the graft by injecting Marcaine into both the autograft and the surrounding muscles. Three weeks after surgery, the patch was paved with young, centrally nucleated myofibers intermixed with young myofibers and myotubes expressing embryonic myosin. The difference in percentage of patch area covered by regenerated myofibers in group 3 (Marcaine injection around the patch, 81.6 +/- 3.0 percent) (mean +/- SD) versus either group 1 (Myoblast-seeded acellular patch, 18.0 +/- 3.0 percent) or group 2 (Autograft, 25.8 +/- 7.0 percent) was statistically significant on independent t test analysis (p < 0.0001). Even an acellular matrix showed some myofiber regeneration after surrounding muscles had been injected with Marcaine. This is the first successful evidence of muscle reconstruction after full-thickness ablation of the middle third of the rectus abdominis. Muscle regeneration seems to be the result of successive waves of migration of angioblasts and then satellite cell-derived myoblasts from the muscles surrounding the patch. The results strongly suggest that vascularization of the scaffold and successive coordinate proliferation of the seeded cells are required for myoblasts to be able to migrate into the patch and differentiate up to myofiber stage.

Immunohistochemical analysis of bFGF, TGF-beta1 and catalase in rectus abdominis muscle from cattle foetuses at 180 and 260 days post-conception.

The potential for muscle growth depends on myoblast proliferation, which occurs essentially during the first two thirds of the foetal period in cattle. Thereafter, myofibres acquire their contractile and metabolic properties. Proliferation is regulated by molecular growth factors and by the tissue oxidative activity. The aim of this study was the quantification by immunochemistry of basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-beta1) and also of enzyme catalase (CAT) activity in rectus abdominis muscle. Samples were collected from cattle foetuses of different growth potential at 180 and 260 days post-conception (dpc). One major conclusion from this work is that protein contents of the muscle tissue bFGF and, to a lower extent, CAT activity decreased with increasing age during the foetal life. No differences were found between the different genotypes of cattle. However, the CAT to bFGF ratio tended to be lower in fast-growing cattle and increased with foetal age. TGF-beta1 did not change with age and was localised mostly at the vascular bed. CAT was detected in smooth and rough reticulum in striated muscles at 180dpc, and additionally in mitochondria at 260dpc. In conclusion, the balance between intracellular growth factors (bFGF and TGF-beta1) and the activity of antioxidant enzyme CAT may participate in the regulation of the transition from myoblast proliferation to differentiation. Thus, increased ratio of CAT to bFGF might be a good index indicating initiation of muscle maturation in cattle foetus prior to birth.

A comparative study of aging of the elastic fiber system of the diaphragm and the rectus abdominis muscles in rats.

In the present study the age-related changes of the striated muscle elastic fiber system were investigated in the diaphragm and rectus abdominis muscles of 1-, 4-, 8- and 18-month-old rats. The activation patterns of these muscles differ in that the diaphragm is regularly mobilized tens of times every minute during the entire life of the animal whereas the rectus abdominis, although mobilized in respiration, is much less and more irregularly activated. The elastic fibers were stained by the Verhoeff technique for mature elastic fibers. Weigert stain was used to stain mature and elaunin elastic fibers, and Weigert-oxone to stain mature, elaunin and oxytalan elastic fibers. The density of mature and elaunin elastic fibers showed a progressive increase with age, whereas the amount of oxytalan elastic fibers decreased in both diaphragm and rectus abdominis muscles and their muscular fasciae. These age-related quantitative and structural changes of the elastic fiber system may reduce the viscoelastic properties of the diaphragm and rectus abdominis muscles, which may compromise the transmission of tensile muscle strength to the tendons and may affect maximum total strength.

Skeletal muscle ultrastructure in normal pregnancy and preeclampsia.

The aim of this study was to assess by quantitative methods whether the assumed metabolic disturbance underlying preeclampsia would be reflected in muscle cell composition of lipid, mitochondria, or glycogen. We have reported mitochondrial dysfunction in preeclampsia, and since accumulation of lipid in skeletal muscle is a feature in mitochondrial disorders, our hypothesis was that preeclamptic women would have an increased content of triglyceride droplets. Quantitative investigation of the skeletal muscle ultrastructure was performed in 10 women with severe preeclampsia and in 6 normotensive pregnant women. Biopsy specimens from musculus rectus abdominis were taken during cesarean section and prepared for electron microscopy. Random pictures were taken by transmission electron microscopy, and point-counting stereology was performed. Preeclamptic women did not have a higher lipid volume fraction than normotensive pregnant women, and we had to reject our hypothesis. On the contrary, there was a tendency towards a lower triglyceride volume fraction in pre eclampsia. We did not detect differences in relative volumes of mitochondria or glycogen in skeletal muscle between the two groups.

Fibre type classification and myosin isoforms in the human masseter muscle.

Human masseter muscle is highly unusual since it contains relatively large numbers of fibres with variable myofibrillar ATPase staining as well as fibres that express neonatal and alpha-cardiac myosin heavy chain (MHC). These findings however, have not been organised together into a fibre type classification scheme. Biopsies from the anterior superficial area of masseter were collected from a large sample of healthy young adults. Biopsies were sectioned and stained for myofibrillar ATPase reactivity and the presence of MHC isoforms as detected by a series of antibodies. The MHC composition of the same biopsies was also analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). A series of rectus abdominis muscle biopsies were analysed similarly to serve as a control for type I, IIA and IIB fibres and isoforms. From the histochemical, immunohistochemical and biochemical experiments we found the masseter to contain type I, IM, IIC, IIA and IIB fibres as previously classified, but in addition there were type neonatal, alpha-cardiac, and 'other' (three or more myosins including neonatal and alpha-cardiac). The percentage of each fibre type was highly variable in masseter biopsies, but generally type I fibres were most common, and the proportion of IIB, neonatal, alpha-cardiac and 'other' fibres was low. Even in biopsies that contained relatively large amounts of these last three fibre types, the amount of neonatal and/or alpha-cardiac MHC detected on SDS-PAGE was limited, suggesting that these MHCs are a minor component in the fibres in which they are expressed.

The extracellular space of voluntary muscle tissues.

The volume occupied by the extracellular space has been investigated in six types of voluntary muscles: sartorius (frog), semitendinosus (frog), tibialis anticus longus (frog), iliofibularis (frog), rectus abdominis (frog), and diaphragm (rat). With the aid of four types of probe material, three of which are conventionally employed (inulin, sorbitol, sucrose) and one of which is newly introduced (poly-L-glutamate), and a different experimental method, we have demonstrated that the "true" extracellular space of frog sartorius, semitendinosus, tibialis anticus longus, and iliofibularis muscle and of rat diaphragm muscle is equal to, or probably less than, 8-9% (v/w) of the tissue. The frog rectus muscle shows a somewhat higher ceiling value of 14%.