Necrotic Muscle Fibers Research Articles

Isolation of influenza virus from muscle in myoglobinuric polymyositis

We report the first isolation of influenza virus from muscle in a man with myoglobinuria and acute polymyositis. Influenza virus was isolated from cultures of Madin Darby bovine kidney and primary rhesus monkey kidney cells inoculated with muscle homogenates in the presence of trypsin; the virus was identified by neutralization and hemagglutination inhibition studies using influenza B/Lee antiserum. Viral plaque assay was performed with Madin Darby canine cells. Viral antigen was also detected by specific immunofluorescence in muscle, and myxovirus-like particles were seen in subsarcolemmal vacuoles by electronmicroscopy. The pathologic findings were similar to those of childhood dermatomyositis, except for a large proportion of necrotic muscle fibers. The evidence suggests that the pathogenesis of influenzal polymyositis in this patient involved direct viral infection of muscle.

Effect of ischemia on contractile and histochemical properties of the rat soleus muscle

Ligature and section of the abdominal aorta results in only minor and temporary functional and metabolic changes in the slow soleus muscle of the rat. A very small decrease in maximal tetanic tension corresponds to a few scattered areas of damaged and necrotic muscle fibres, in which decreased succinic dehydrogenase and loss of phosphorylase activity was observed. A new experimental approach, i.e. ligature and section of the abdominal aorta combined with terminal devascularisation, preserving intact tendons and innervation of the muscle causes maximal muscle ischemia, followed by an almost complete loss of tetanic tension output, marked shortening of contraction time and profound morphological and histochemical changes. The decrease in succinic dehydrogenase and ATPase activities and loss of phosphorylase activity occur in the majority of degenerating muscle fibres except for a thin rim of peripheral fibres during the first 4 days. Subsequently, the contractile properties recover gradually and enzyme activities reappear in the regenerating muscle fibres simultaneously with new revascularisation. Thirty days after the operation all the parameters observed returned to control values.

Further observations on the pathological responses of rat skeletal muscle to toxins isolated from the venom of the Australian tiger snake, Notechis scutatus scutatus.

1. Some aspects of the response of mammalian skeletal muscle following the injection of purified toxins from the venom of the Australian tiger snake, Notechis scutatus scutatus, are described. 2. The toxins used were notexin, notechis II-5, notechis II-1 and a modified form of notexin (PBP-notexin). They were injected into the dorso-lateral aspect of one himd limb so that the soleus muscle would be exposed to the toxins. 3. Within 1 h after the injection of notexin, the soleus muscles were oedematous and by 3--6 h, polymorphonuclear leucocytes had entered the interstitial spaces. The invasion of necrotic muscle fibers was extensive by this time. Muscle spindles appeared relatively unaffected by the toxin. 4. The muscle regenerated via myoblasts at 2--3 days to myotubes at 3--5 days, immature muscle fibers at 7--14 days and fully differentiated muscle fibers by 21--28 days. Even after 6 months, however, the nuclei of many muscle fibres remained in a central position. 5. A second component of Australian tiger snake venom was also found to be myotoxic. It was slightly less potent than notexin, but caused qualitatively similar damage to that caused by notexin. It was identified as notechis II-5. A third fraction, notechis II-1, was found to be inactive. 6. Notexin could be neutralized by incubation with tiger snake antivenene; the simultaneous injection of antivenene with notexin did not afford complete protection against muscle damage.

ヒトの脳動脈および冠状動脈の動脈硬化症における中膜平滑筋細胞の電子顕微鏡的研究

Main branches of the middle cerebral arteries and coronary arteries of normotensive and hypertensive human autopsy cases were investigated electron microscopically.The cerebral arteries showed intimal thickening consisting of increased smooth muscle cells, basement membrane-like substance, elastin and collagen fibrils with aging. The intimal smooth muscle cells underwent necrotic change, and the internal elastic lamina showed destructive change with aging gradually. Medial muscle cells showed necrotic aging change, such change was severe immediately beneath the internal elastic lamina.Intimal thickening of the cerebral arteries was marked in hypertensive cases. In hypertensive cases intimal muscle cell injury was slight but medial muscle cell injury was so severe that all of medial cells showed necrotic change and muscle cells near the internal elastic lamina could not be found because of necrosis.Intimal thickening of the coronary arteries became marked with aging, but intimal and medial muscle cell injuries were extremely rare in both normotensive and hypertensive cases.

Ultrastructural Alterations in Gizzard Smooth Muscle of Selenium-Vitamin-E-Deficient Ducklings

Lesions of gizzard smooth muscle were studied by light and electron microscopy in newly hatched ducklings fed a selenium-vitamin-E-deficient diet for 13-21 days. Histopathologic alterations included initial hyaline change in damaged smooth-muscle cells, subsequent mineralization of sarcoplasmic debris in necrotic smooth-muscle cells, macrophagic invasion and phagocytosis of sarcoplasmic debris, and eventual fibroblastic proliferation and scarring of damaged areas in the gizzard wall. Ultrastructurally, mild damage in smooth-muscle cells was manifested by altered mitochondria with matrical densities and disrupted membranes and dilated elements of sarcoplasmic reticulum. In smooth-muscle cells with severe injury, the alterations of mitochondria and sarcoplasmic reticulum were accompanied by myofibrillar lysis and disruption of plasma membranes and external laminae. Expanding zones of mineralization of sarcoplasmic debris in necrotic smooth muscle cells were present at dense masses of lysed myofilaments that surrounded mineralized and disrupted mitochondria. Numerous macrophages infiltrated the areas of necrosis of smooth muscle and phagocytosed sarcoplasmic debris.

Sites and mechanisms of localization of technetium-99m phosphorus radiopharmaceuticals in acute myocardial infarcts and other tissues.

This study was performed to elucidate the localization at the cellular level of technetium-99m phosphorus ((99m)Tc-P) radiopharmaceuticals in acute myocardial infarcts and the mechanisms responsible for (99m)Tc-P uptake in acute myocardial infarcts and other tissues. In 20 dogs with proximal left anterior descending coronary arterial ligation for 1-3 days, elevated calcium levels were measured at all sites of increased (99m)Tc-P uptake (acute myocardial infarcts, necrotic thoracotomy muscle, lactating breast, and normal bone); however, a consistent linear relationship between (99m)Tc-P and calcium levels was not observed. A strong correlation (r = 0.95 and 0.99, n = 2 dogs) was demonstrated between levels of (3)H-diphosphonate and (99m)Tc-P in infarcted myocardium. Autoradiographic studies with (3)H-diphosphonate revealed extensive labeling in the infarct periphery which contained necrotic muscle cells with features of severe calcium overloading, including widespread hypercontraction as well as more selective formation of mitochondrial calcific deposits. Autoradiography also demonstrated labeling of a small population of damaged border zone muscle cells which exhibited prominent accumulation of lipid droplets and focal, early mitochondrial calcification. Cell fractionation studies revealed major localization of both (99m)Tc-P and calcium in the soluble supernate and membrane-debris fractions of infarcted myocardium and less than 2% of total (99m)Tc-P and calcium in the mitochondrial fractions; however, electron microscopic examination showed that mitochondria with calcific deposits were not preserved in the mitochondrial fractions. In vitro studies evaluating the role of serum protein binding on tissue uptake of (99m)Tc-P agents demonstrated that, in spite of significant complexing with serum proteins, serum (99m)Tc-P activity retained the ability to adsorp to calcium hydroxyapatite and amorphous calcium phosphate. In vivo studies showed that concentration of human serum albumin (labeled with iodine-131) in infarcted myocardium reached a maximum of only 3.8 times normal after a circulation time of 96 h, whereas (99m)Tc-P uptake was at least 10 times normal after a circulation time as short as 1 h. It is concluded that: (a) (99m)Tc-P uptake in acutely infarcted myocardium, and possibly other types of soft tissue damage, is limited to necrotic and severely injured cells; (b) concentration of (99m)Tc-P results from selective adsorption of (99m)Tc-P with various forms of tissue calcium stores, including amorphous calcium phosphate, crystalline hydroxyapatite, and calcium complexed with myofibrils and other macromolecules, possibly supplemented by calcium-independent complexing with organic macromolecules; and (c) lack of a linear relationship between (99m)Tc-P and tissue calcium levels mainly results from local differences in composition and physicochemical properties of tissue calcium stores and from local variations in levels of blood flow for delivery of (99m)Tc-P agents.

Cardiac pathologic findings in patients treated with bone marrow transplantation

Cardiac pathologic findings were analyzed in 22 necropsy cases from a series of 29 patients with leukemia, aplastic anemia, or metastatic cancer who had been treated with ablative therapy followed by bone marrow transplantation. Some cardiac alterations were similar to those that occur in patients with hematologic and neoplastic diseases not treated with bone marrow transplantation, and consisted of cardiomegaly, cardiac atrophy, hemorrhage, foci of necrosis due to shock associated with sepsis or hepatic failure, myocardial abscesses secondary to systemic candidiasis or staphylococcal infection, fibrinous pericarditis, and hemosiderosis. Other cardiac alterations were more specifically related to factors associated with the transplantation procedure. Six patients exhibited a distinctive interstitial reactive change characterized by the presence of (1) moderate to large numbers of Anitschkow cells, occurring alone or in small cellular aggregates with histiocytes, histiocytic cells with nuclei of the Anitschkow type, lymphoid cells, and plasma cells, and (2) nuclei of the Anitschkow type in cardiac vascular and endocardial smooth muscle, endothelial and Schwann cells, and occasional cardiac muscle cells. This alteration may have been induced by abnormal immune mechanisms, as suggested by the observation that five of the six patients with interstitial change had clinical evidence of graft-versus-host disease. Two patients developed fatal congestive cardiac failure in the early post-transplant period and exhibited myocardial damage with histologic and ultrastructural features indicative of severe acute injury. Findings in these two patients consisted of necrotic muscle cells, which exhibited multiple contraction bands, diastase-resistant PAS staining, and intracellular fibrin deposits; microthrombi, which were composed of fibrin and occasionally of fibrin and platelets; and extravasated erythrocytes and fibrin strands in the interstitium. One of the two patients also exhibited unusual nuclear alterations, which were characterized by replacement of normal chromatin by palely stained fibrous and filamentous material. Clinicopathologic analysis strongly suggested that the fatal cardiotoxicity in both patients resulted primarily from effects of high doses of cyclophosphamide, which were administered as part of a four drug regimen that provided tumor ablation and immunosuppression for bone marrow transplantation. Our findings emphasize the need for less toxic antineoplastic and immunosuppressive therapy for use in bone marrow transplantation procedures.

AGING-AND HYPERTENSION-ASSOCIATED CHANGES OF MEDIAL SMOOTH MUSCLE CELLS OF THE LATERAL STRIATE ARTERIES

Histometric and histological studies were made on aging-and hypertension-associated changes in the lateral striate arteries from 88 autopsy cases, including 34 normotensive cases, aged 9 fetal months to 72 years, 20 hypertensive cases, aged 23 to 80 years, and 34 others, with the following results : 1. In the lateral striate arteries of normotensive cases, smooth muscle cell counts per unit areas of the media decreased with aging while the interstitial tissue increased. The decrease in smooth muscle cells resulted from their necrosis.2. Medial smooth muscle cell counts in the arteries were smaller in hypertensives than in normotensives of the corresponding age, and the difference between them became greater with age. The necrosis and disappearance of the medial smooth muscle cells were most remarkable in aged hypertensive cases.3. In old normotensive or hypertensive cases, the necrosis and disapperance of the medial muscle cells resulted in interstitial widening, where fine granular substance derived from necrotic muscle cells, PAS-positive basement membrane-like substance and collagen fibers were increased. These changes were more intensive in hypertensive cases than in old ones.4. Medial muscle cell counts in the putaminal segment of the lateral striate arteries of hypertensive cases with intracerebral microaneurysms were smaller by 20-40% than those in hypertensive cases without the aneurysms.5. The cross-section areas and external diameters at various sites of the lateral striate arteries were greater in hypertensives than in normotensives, and the differences between them were greater in old cases.6. In the lateral striate arteries of hypertensive cases, cellulofibrous intimal thickening, swelling and duplication of the internal elastic lamina, and blood plasma and foam cell infiltration of the intima were observed from the third decade, and their frequencies and intensities increased with age.

Radiation-induced changes in skeletal muscle. An electron microscopic study.

Ultrastructural aspects of skeletal muscle changes in rabbits after local exposure to l,130r and l,300r of X-ray irradiation are presented. A spectrum of multifocal lesions of muscle cells and microvasculature appearing simultaneously as early as 24 hrs. and followed up to day 40 post-irradiation is described. Muscle cell changes were characterized by myofibrillolysis, myonecrosis followed by appearance of muscle buds, focal increase in glycogen content of relatively intact fibers, and myofibcr atrophy and degeneration. Suggestive evidence for intracellular regeneration following myofbrillolysis is presented. Microvasculature showed progressive endothelial cell swelling with increased pinocytotic activity and accelerated transport of ferritin followed by a peculiar metaplastic change characterized by well developed Golgi complex and endoplasmic reticulum. Delayed fibrinoid necrosis of capillaries observed on day 28 probably resulted from toxic products released from degenerating and necrotic muscle and mesenchymal cells, and its probable role in delayed myonecrosis is discussed. Radiation induced myopathic changes seem to follow direct radiation injury and complex phenomena of interaction from changes in individual structural components of muscle.

Response of serum enzymes and other biochemical constituents to strenuous exercise in control subjects and patients with motor neurone disease

Serum enzymes were estimated in 10 patients with motor neurone disease (MND) and in 5 control subjects at rest and after strenuous exercise. Creatine phosphokinase (CPK) levels were abnormally increased at rest in 6 of the MND patients, 1 of whom had the only abnormal increase in resting level of malate dehydrogenase (MDH). MDH and CPK increased, sometimes to abnormal levels, in response to exercise in control subjects. The mean rise in MDH after exercise in the MND patients closely paralleled that of the control group. In spite of higher resting CPK levels in MND patients compared to controls, the percentage increase in CPK activity immediately after exercise in this group was significantly lower than for controls. No significant changes in aspartate aminotransferase (GOT), sodium, total protein or urea were observed in either group before or after exercise, but significant differences were noted in the response patterns of potassium and bicarbonate. We suggest that abnormal resting CPK activities recorded in MND patients are caused by near maximal efflux from actively necrotic muscle cells, and any superimposed rise in CPK levels after exercise may be contributed by enzyme efflux from the remaining normal muscle cell mass.

An ultrastructural study of skeletal muscle in polymyositis

Biopsy material from the quadriceps femoris muscle in a case of subacute polymyositis associated with polyarthropathy and transient cardiac involvement has been examined with the electron microscope. Observations have been made on the ultrastructural changes in necrotic muscle fibres as well as on the origin and differentiation of myoblasts participating in the regenerative processes in damaged fibres. An unexpected finding was the presence of granular cytoplasmic inclusions and nuclear abnormalities in some regenerating muscle fibres. Although the particles in the inclusions are similar to those seen in cells infected with Coxsackie virus, the possibility that the inclusions are unusually large aggregates of glycogen granules cannot be excluded. Thickening of endothelial cells and thickening and reduplication of the basement membrane of capillaries and venules was also found but is not considered to be of primary importance in the pathogenesis of the muscle fibre damage.

Central nervous system and skeletal muscle involvement in systemic candidiasis.

Two necropsy cases of systemicCandida albicans infection with involvement of the kidney, lung and brain are reported. Both patients died from renal failure. In the brain there was a miliary spread of microabscesses and granuloma-like lesions, and Candida was evident within both types of lesion. In case 2, there was additional involvement of skleletal muscle and diaphragma with focal-grouped and single-scattered necrotic muscle fibres and small granulomas containing fungus elements accompanied by mild neurogenic atrophy of skeletal muscle due to “uremic” neuropathy.

The histochemistry of Stellantchasmus falcatus Onji and Nishio, 1915 (Trematoda: Heterophyidae) metacercarial cyst in the mullet Mugil cephalus L. and histopathological alterations in the host†

The metacercarial cyst of the heterophyid trematode Stellantchasmus falcatus in the striated muscles of the mullet Mugil cephalus consists of three layers. Histochemical studies have indicated that these layers are chemically distinct. The acellular inner layer is comprised of complex carbohydrate polyanions rich in acidic groups while the medial cellular layer is comprised of a neutral mucopolysaccharide including a moeity of carbohydrate poiyanions. These two layers are believed to be of parasite origin. The outer layer is cellular and is of host origin. It is the most chemically complex and is believed to include one or more of the following: mucopolysaccharides, glycolipids and phospholipids. In addition, the carbohydrate moeity associated with the mucopolysaccharides and/or glycolipids is in the form of sulphated carbohydrate polyanions with undissociated carboxyls as well as acidic groups.The presence of the parasite results in the necrotic degeneration of the host's myofibers situated in the proximity of the parasite. Large fat cells fill the spaces vacated by necrotic muscle cells and many blood cells occur along the periphery of the parasite.

The distribution of lipids in malignant hypertensive fibrinoid necrosis

The fibrinoid necrosis developing in the course of experimental malignant hypertension was examined for the presence, localization and appearance of lipids. In the initial stage of hypertension, only sudanophilic droplets were demonstrable in the necrotic muscle cells of the media, but later diffuse sudanophilia was apparent over nearly the entire cell body. The subendothelial fibrinoid developing during the second stage was Sudan negative, whereas the necrotic medial cells exhibited a distinct sudanophilia. At the same time a considerable extra- and intracellular sudanophilia also appeared in the perivascular granulation tissue, but it tended to decrease or entirely disappear during the third stage. At this time, foam cells appeared in the subendothelial fibrinoid. Based on our findings, the sudanophilia of the media has been attributed to lipids rendered visible by phanerosis.

Regeneration of skeletal muscle in the Rottnest quokka.

SummaryThe reversal of the nutritional myopathy of the captive Rottnest quokka (Setonix brachyurus) with vitamin E provided a model for the study of regenerative processes in skeletal muscle. Seventeen quokkas recovering from myopathy following treatment with α‐to opherol were studied by means of 2 to 7 serial biopsies from each animal. As the debris within necrotic muscle fibres was removed by macrophages, proliferation of sarcolemmal nuclei occurred. Cells with basophilic cytoplasm, due to a high content of ribonucleic acid (RNA), appeared and showed mitotic activity. Myofibrillar markings identified these cells as myoblasts. Later in the sequence multi‐nucleated masses and young muscle fibres developed, probably by fusion of myoblasts. Cross striations became obvious and the sarcoplasm eosinophilic, due to increased content of protein. Lastly, sarcolemmal nuclei centrally or randomly distributed underwent marginal migration and the muscle fibre became fully reconstituted. With acrolein‐Schiff staining necrotic fibres were rarefied, due to loss of protein, but during maturation staining became deeper as the protein content increased. The early phases occupied several days, while complete restoration required 5 to 8 weeks. There were many features of similarity between the morphology of regeneration in vivo and the growth and differentiation of skeletal muscle in vitro. Regenerative activity is prominent in the histopathological lesion of human polymyositis, and increased attention has been recently given to regenerative phenomena in case of human progressive muscular dystrophy, so that clarification of this property of muscle may improve understanding of these diseases.

Macroscopic identification of early myocardial infarction by dehydrogenase alterations.

A method utilizing a general dehydrogenase reaction has been used to demonstrate early gross myocardial infarctions. The procedure takes advantage of substrate and enzyme loss from the damaged myocardium. In the viable muscle, where endogenous substrates, coenzymes, and dehydrogenases are present, reduction of Nitro-BT yields a dark blue formazan. Necrotic muscle fibres remain unstained by this technique.A survey of 31 human hearts obtained at necropsy disclosed that there is no alteration in the Nitro-BT reaction following acute coronary insufficiency with sudden death or severe congestive heart failure. The earliest myocardial infarct to show loss of dehydrogenase activity was of eight hours' duration. Post-infarction scars and patchy interstitial fibrosis provided very precise information concerning topographic relationships when this method was applied to heart slices.

OXIDATIVE ENZYMES OF INTERMEDIARY METABOLISM IN HEALING BONE FRACTURES; A HISTOCHEMICAL STUDY.

AbstractThe fibulae of young adult rats were cut at the mid‐diaphysis with scissors and examined at various stages of repair. The distribution of enzymes participating in glycolysis, in the citric acid cycle, and in the hexose monophosphate shunt was studied histochemically in healing bone fractures.Three days after the fracture there was marked proliferation and an increased enzyme activity in the osteoprogenitor cells in the inner layer of the periosteum along the entire diaphysis. Remarkable enzyme activity was also observed in the newly formed osteocytes. No qualitative differences were noticed among osteoprogenitor cells, osteoblasts, and osteocytes. Necrotic muscle fibers, periosteum, and bone cells failed to react.Seven days after the fracture, a bulky cartilaginous callus developed. The proliferating osteoprogenitor cells of the periosteum and of the callus had a similar enzyme pattern. As the fibroblast‐like osteoprogenitor cells of the callus developed into chondrocytes, enzyme activity increased. This was especially striking for isocitric dehydrogenase and glucose‐6‐phosphate dehydrogenase. When the chondrocytes became hypertrophic, they gradually lost enzyme activity. Mononuclear and multinuclear osteoclasts, macrophages and foreign body giant cells showed marked activity of most demonstrated enzymes. Glucose‐6‐phosphate dehydrogenase activity was marked in foreign body giant cells, but appeared weak in osteoclasts.At 14 days the enzyme pattern of the callus was essentially the same as at seven days.At 30 days bone union was well established, but there was still considerable osteoblastic and osteoclastic activity.The histochemical findings have been described in detail and discussed in correlation with pertinent morphological and biochemical data. Histochemically, bone regeneration proved to be essentially a recapitulation of normal bone growth.

Observations in Wringer Injuries

An experimental study of the effect of a wringer injury to the hind limbs of immature rats has been presented. The findings in this study indicate that injuries produced by wringers extending over various periods of time (for example, sixty seconds versus 300 seconds) are similar histologically and vary only in degree. The histological preparations indicate that hemorrhage, edema, venous dilatation, leukocyte infiltration, and muscle necrosis may occur and that the injury is more severe near the bone. The changes first appear in from three to five hours and reach their maximum intensity in from sixteen to twenty-four hours. During this period the animals lose their ability to perform in a rat wheel. Necrosis of striated muscle has been observed as well as late replacement of the necrotic muscle fibers by fibrous tissue. Regeneration of muscle fibers may occur after a wringer injury although the evidence in the present material is inadequate for this conclusion to be reached. We have postulated that the factors giving rise to increased capillary pressure produce a condition of reduced oxygenation and ionic exchange at the fiber level and that these changes may produce muscle necrosis.

Evidence for heart damage in association with systemic hypothermia in dogs

Twenty-four dogs were subjected to systemic hypothermia. Nine dogs were autopsied at the onset of fatal cardiac irregularities or at the termination of moderate (26° to 27.5° C.) or deep (21° to 22.5° C.) hypothermia of one to four hours' duration. In all cases the myocardium showed foci of necrotic muscle fibers with an occasional cellular reaction. Fifteen dogs were sacrificed and autopsied three days to three years after survival of moderate or deep hypothermia of the same duration. In thirteen of fifteen cases distinct areas of necrosis showing various stages or organization were detected. The nature and the etiology of the lesions as well as their functional significance are discussed.

The effect of cortisone on experimentally produced myocardial infarcts.

Administration of a moderately large or large dose of cortisone to dogs having acute myocardial infarctions inhibited slightly the rate of removal of necrotic muscle fibers in these animals as compared with untreated animals. Delay was appreciable at four and six days after production of the infarct in animals receiving 2.5 mg. of cortisone per kilogram of body weight and at 4, 6, 12 and 21 days in animals receiving 10 mg. per kilogram. At all other periods up to essentially complete healing of the infarcts as defined at 60 days, no appreciable differences existed between treated and untreated animals.