CELL STRUCTURE AND THE METABOLISM OF INSECT FLIGHT MUSCLE

The biochemical properties of insect flight muscle were investigated to ascertain the mechanisms whereby energy is made available for the contractile processes. It was found: 1. The endogenous respiration of muscle homogenates was diminished by starving the flies. The substrate for this respiration was probably glycogen. 2. To obtain the maximal rate of oxidation of glucose, the homogenate had to be fortified with inorganic phosphate, Mg ions, ATP, and cytochrome c. The nucleotides, AMP and ADP, were not as effective as ATP. The addition of DPN or TPN was not necessary for this system. 3. Flight muscle homogenates oxidized glycogen, some sugars, and amino acids, as well as the intermediates of the glycolytic and tricarboxylic acid cycles. Other evidence demonstrated the substrate specificity of the muscle. 4. By centrifugation, the muscle homogenate was divided into two fractions: one, a soluble fraction representing the sarcoplasm; the other, the particulate fraction which contained the fibrils and the sarcosomes. 5. The particulate fraction, alone, oxidized all the citric acid cycle intermediates, α-glycerophosphate, phosphopyruvate, and the amino acids, glutamic, proline, and cysteine. Regardless of the substrate, no oxygen uptake was found with the sarcoplasm by itself. 6. A recombination of the sarcoplasm and the particulate component was required for the oxidation of glycogen, the hexoses, and all the phosphorylated intermediates of glycolysis, except phosphopyruvate. 7. Isolated mitochondria accounted for all the enzymatic activity of the particulate fraction. These results demonstrate that the enzymes of intermediate metabolism are localized in the sarcoplasm or sarcosomes. The third cytological entity, the myofibrils, plays no role in the energy-providing scheme. From a functional viewpoint, the sarcoplasm and the mitochondria, in combination, furnish the energy for the actomyosin contraction. The results are discussed in relation to analogous findings in other insects and vertebrates.     

Mitochondria in the flight muscles of insects. I. Chemical composition and enzymatic content

1. The "indirect" thoracic muscles of adult dipterous and hymenopterous insects consist of a unique type of muscle characterized by the presence of numerous spherical, intracytoplasmic bodies termed "sarcosomes." 2. When the muscle is teased or ground, the sarcosomes are liberated as a turbid suspension of bodies ranging from 1 to 4 µ in diameter. A method is described for the isolation of sarcosomes by a simple differential centrifugation. 3. The cytochemical, chemical, and enzymatic properties of sarcosomes were examined for the purpose of appraising their relation to the cytoplasmic bodies of other tissues. 4. Fresh sarcosomes are slowly but selectively stained by the mitochondrial reagents, Janus green B and pinacyanol. Fixed sarcosomes give a positive reaction with Regaud's mitochondrial stain. 5. Chemical analyses show that approximately 29 per cent of the dry weight of sarcosomes consists of lipids and 60 per cent of protein. Microbiological assay indicates the presence of about 1 gamma of riboflavin per milligram of nitrogen. These values resemble those reported for isolated mitochondria of vertebrate liver and kidney. 6. When examined spectroscopically the sarcosomes, like the vertebrate mitochondria, show a high titer of cytochromes a, b, and c. 7. The titer of cytochrome oxidase varies systematically with the adult age of the insect. A similar relation is observed for the enzyme catalase. 8. Isolated sarcosomes show significant titers of succinoxidase, α-glycerophosphate dehydrogenase, malic dehydrogenase, and pyruvic dehydrogenase. The following dehydrogenases could not be demonstrated: xanthine, phenylalanine, glycine, lactic, choline, glutamic, and alcohol. These results are compared with those previously reported for vertebrate mitochondria. 9. In view of their manifold points of biochemical similarity, it is concluded that the sarcosomes are the mitochondria of this highly specialized muscular tissue.     

THE ORGANIZATION OF THE FLIGHT MUSCLE IN A DRAGONFLY, AESHNA SP. (ODONATA)

The structure of the flight muscle of a dragonfly (Aeshna sp.) has been studied with the light and electron microscopes, and the organization of this specialized tubular muscle is described. This tissue is characterized by the great development of the sarcosomes, which are slab-like and are arranged within the fiber opposite each sarcomere of the radially oriented lamellar myofibrils. A well developed and highly ordered sarcoplasmic reticulum is present, consisting of perforated curtain-like cisternae extending across the face of each fibril, together with tubular invaginations of the fiber plasma membrane situated within indentations in the sarcosomes and traversing the fibril surface midway between the Z and M levels. The structure of these fibers, and notably the organization of the reticulum, is compared with that of other types of muscle, and the possible role of the two components of the sarcoplasmic reticulum in the contraction physiology of the dragonfly muscle fiber is discussed.     

Mitochondria in the flight muscles of insects. III. Mitochondrial cytochrome c in relation to the aging and wing beat frequency of flies

1. In the present study a correlation has been sought between aging, flight muscle mitochrondria (sarcosomes), cytochrome c, and flight ability in the blowfly, Phormia regina. 2. During the 1st week of adult life, individual sarcosomes increase in mass from 2.7 x 10^–7 µg. dry weight at the time of emergence, to 8.5 x 10^–7 µg. by the 7th day. During this period of growth, the number of sarcosomes per fly (6.7 x 10⁸) remains constant. When mature, the sarcosomes account for 32.6 per cent of the total muscle dry weight, or close to 40 per cent on a wet weight basis. 3. It appears probable that the high content of flight muscle cytochromes is entirely localized in the sarcosomes. The cytochromes continue to be synthesized and increase in titer within the sarcosomes for 7 days after adult emergence. 4. As determined spectroscopically, the various cytochrome components at all times maintain a constant ratio both to one another and to the sarcosomal dry weight. This suggests the possibility that the cytochrome system may be synthesized as a single entity. 5. The wing-beat frequency of Drosophila funebris and Phormia varies with the age of these flies, being lowest at the time of emergence and maximum after the 6th day. 6. The relations between wing-beat frequency, respiration during flight, and sarcosomal cytochrome c content are discussed. On the basis of some likely assumptions it is calculated that the cytrochrome c turnover number is over 5,000, and that the cytochrome c turns over once for every two wing-beat cycles.     

Mitochondria in the flight muscles of insects. II. Effects of the medium on the size form,and organization of isolated sarcosomes

1. The sarcosomes of Drosophila and the blowfly, Phormia, are dense, spherical, homogeneous bodies when isolated from flight muscle and promptly examined under the phase contrast, oil immersion objective. 2. Their average diameter in newly emerged flies is about 1 micro. This value increases rapidly during the 1st week of adult life and then becomes constant at approximately 2.5 micro. At each age the variation in sarcosome diameter conforms approximately to a normal distribution. 3. The degree to which isolated sarcosomes retain their initial size and organization is remarkably conditioned by the composition and the hydrogen ion concentration of the medium in which they are teased and suspended. In suboptimal media three major categories of change were encountered: (1) swelling, with or without compaction of the contents (as in distilled water and salt solutions); (2) shrinkage to rod-like, pleomorphic forms (as in blood serum); and (3) fuzzy degeneration (as in sugar solutions). 4. The membrane that surrounds each sarcosome becomes plainly visible in swollen sarcosomes. A continuation of swelling is accompanied by the escape of the sarcosomal contents, the vacated membrane persisting as a spherical, optically empty ghost. 5. Sarcosomes appear to behave like osmometers when suspended in various aqueous solutions. Solutes which penetrate the membrane show only transient effects in preventing the osmotic entry of water. 6. Under this analysis we find the membrane to be more or less freely permeable to the ions of sodium, potassium, calcium, magnesium, chloride, and phosphate, to non-electrolytes smaller than hexoses, to phosphorylated hexoses, and to several intermediates of the citric acid cycle. 7. The sarcosomal membrane appears to be less permeable to non-electrolytes larger than pentoses, provided that such molecules are not phosphorylated. 8. The membrane shows a higher permeability to ATP than to ADP. The significance of this observation is considered with respect to the ADP-ATP shuttle between sarcosomes and muscle fibrils. 9. Simple solutions of electrolytes or non-electrolytes cause more or less conspicuous changes in the microscopic appearance of sarcosomes. Prolonged preservation was achieved only in more complicated media containing protein. It is concluded that the Donnan equilibrium is the source of the principal osmotic forces regulating the movement of water through the sarcosomal membrane. 10. The optimal medium for the preservation of isolated sarcosomes was an intracellular Ringer solution containing 2.5 per cent crystalline bovine albumin in 0.16 M potassium phosphate buffer at pH 7.0.     

THE FINE STRUCTURE OF THE VENTRAL INTERSEGMENTAL ABDOMINAL MUSCLES OF THE INSECT RHODNIUS PROLIXUS DURING THE MOLTING CYCLE : II. Muscle Changes in Preparation for Molting

The development of the ventral intersegmental abdominal muscles of Rhodnius prolixus is triggered by feeding. The early muscle (1 day after feeding) contains essentially nonstriated fibrils. However, in cross-sections, areas indicating early I bands, Z lines, and A bands can be recognized. Interdigitating thick and thin myofilaments do not assemble into a precise lattice until sometime between 4 and 5 days after feeding. As development continues, the number of fibrils increases, the region corresponding to the Z line increases in density, and the fibrils contain more recognizable striations. The newly formed fibrils broaden as myofilaments are added peripherally. At all stages throughout development, the ratio of thin to thick myofilaments is always 6:1. The formation of fibrils in the abdominal muscles of Rhodnius is different from that in chick embryo skeletal muscle. The major differences are that at all stages in Rhodnius there are (1) a constant ratio of thin to thick myofilaments, and (2) detectable Z-line material. Other findings in Rhodnius suggest (1) that fusion of mononucleated cells with the multinucleated muscle cell occurs, (2) that microtubules develop in the tendon cell concomitantly with development of myofibrils in the associated muscle cell, and (3) that filaments 55A in diameter aggregate into microtubules.     

Neural- and endocrine control of flight muscle degeneration in the adult cricket,Gryllus bimaculatus

Neural- and endocrine mechanisms controlling degeneration of a dorsal longitudinal flight muscle, M112a, have been studied in adult Gryllus bimaculatus (Orthoptera: Gryllidae). Decapitation completely prevented muscle degeneration. Implantation of a pair of corpora allata or injection of juvenile hormone III into decapitated crickets caused muscle degeneration. Denervation of M112a resulted in reduction of muscle mass compared with that in sham-operated crickets. Denervation of M112a in decapitated crickets, however, did not affect muscle mass. Birefringence and ultrastructure of M112a showed an obvious regional difference in the onset of degeneration. Fibrillar structures of M112a always disappeared from the ventral to dorsal part. Distribution of axon terminals of motor neurons and mechanical responses to the motor nerve stimuli showed that M112a is composed of five motor units with similar twitch properties. When M112a was fully denervated, regional differences in degeneration disappeared. Partial denervation resulted in denervated muscle fibers losing birefringence earlier than innervated fibers. These results suggest that juvenile hormone causes breakdown of flight muscles, and neural factors control degeneration of flight muscles to some extent under the presence of the juvenile hormone.     

Ultrastructural changes in sarcosomes after exposure of adult Calliphora erythrocephala to lethal and sub-lethal high temperatures

The effects of lethal and sub-lethal high temperatures on the morphology of intact flight muscle sarcosomes of adult Calliphora erythrocephala are described. Treatment of adult flies to lethal temperatures results in ultrastructural changes in the organisation of the cristae and the appearance of electron opaque inclusions. These changes have not been observed after sub-lethal heat treatment, when 50% of the animals recover. It is suggested that changes in the ultrastructure of the intact sarcosomes may be correlated with changes in their ability to couple oxidative phosphorylation with α-glycerophosphate. Age-dependent changes in the sensitivity of sarcosomes are related to changes in the heat death point of the animal and suggest that the impairment of sarcosomal function may be one of the primary lesions in heat death of adult C. erythrocephala.     

COMPARATIVE CYTOPHYSIOLOGY OF STRIATED MUSCLE WITH SPECIAL REFERENCE TO THE ROLE OF THE ENDOPLASMIC RETICULUM

1. The structure and distribution of the components of striated muscle cells vary with the species and with the specialization of muscle fiber function. 2. There appear to be two, easily distinguishable, general categories of striated muscle structure. A. High frequency muscle (represented by flight muscle of higher insects and hummingbird, and cicada tympanal muscle) is characterized by widely spaced, non-branching fibrils of large diameter and short period, little endoplasmic reticulum, and large quantities of large mitochondria (low fibril-sarcoplasm ratio). This structure is correlated with heavy tracheolization or vascularization, high oxidative activity, and dark color as compared with other muscles of the same species. B. Low frequency muscle is characterized, in general, by high fibril-sarcoplasm ratio, relatively long period, few mitochondria increasing with activity and decreasing with absolute power of the fiber. Oxidative capacity and color are proportional to the quantity of mitochondria. These fibers are further differentiated into (a) fibrillar arrangement of contractile material which permits a regular pattern of interfibrillar and segmental reticulum, and (b) afibrillar arrangement of contractile material leading to an unsystematic distribution of reticulum. 3. The endoplasmic reticulum appears as a complex coordination system in the muscle fiber. Peripherally, it links the Z and M lines of the fibrils to the sarcolemma and between the fibrils it links the cross-bands, forming the Grundmembran of earlier authors. By longitudinal linkage, it connects with the sarcolemma at the muscle extremity to form a digital arrangement into which the tendon fibrils are spliced. The extent of its development and its position have a definite relationship to the degree and site of fiber shortening. At present the reticulum is the only structure that one can consider to be an internal conducting system. It may distribute the excitation transversely from fibril to fibril, and lengthwise saltatorially to the symmetry centers of the sarcomeres. 4. The nucleus is the mediating element between the cytoplasmic phases within and without the tubular system of the endoplasmic reticulum. A possible mechanism which correlates nucleus, adenylic acid system, ion exchange, and reticulum with the initiation of contraction is postulated.     

Cell structure and the metabolism of insect flight muscle

The biochemical properties of insect flight muscle were investigated to ascertain the mechanisms whereby energy is made available for the contractile processes. It was found: 1. The endogenous respiration of muscle homogenates was diminished by starving the flies. The substrate for this respiration was probably glycogen. 2. To obtain the maximal rate of oxidation of glucose, the homogenate had to be fortified with inorganic phosphate, Mg ions, ATP, and cytochrome c. The nucleotides, AMP and ADP, were not as effective as ATP. The addition of DPN or TPN was not necessary for this system. 3. Flight muscle homogenates oxidized glycogen, some sugars, and amino acids, as well as the intermediates of the glycolytic and tricarboxylic acid cycles. Other evidence demonstrated the substrate specificity of the muscle. 4. By centrifugation, the muscle homogenate was divided into two fractions: one, a soluble fraction representing the sarcoplasm; the other, the particulate fraction which contained the fibrils and the sarcosomes. 5. The particulate fraction, alone, oxidized all the citric acid cycle intermediates, alpha-glycerophosphate, phosphopyruvate, and the amino acids, glutamic, proline, and cysteine. Regardless of the substrate, no oxygen uptake was found with the sarcoplasm by itself. 6. A recombination of the sarcoplasm and the particulate component was required for the oxidation of glycogen, the hexoses, and all the phosphorylated intermediates of glycolysis, except phosphopyruvate. 7. Isolated mitochondria accounted for all the enzymatic activity of the particulate fraction. These results demonstrate that the enzymes of intermediate metabolism are localized in the sarcoplasm or sarcosomes. The third cytological entity, the myofibrils, plays no role in the energy-providing scheme. From a functional viewpoint, the sarcoplasm and the mitochondria, in combination, furnish the energy for the actomyosin contraction. The results are discussed in relation to analogous findings in other insects and vertebrates.     

Ultrastructure of host tissues exposed to the calyx fluid of the parasitoid,Campoletis sonorensis (Cameron) (Hymenoptera: Ichneumonidae)

Campoletis sonorensis (Cameron) (Hymenoptera: Ichneumonidae) is a solitary endoparasitoid of Heliothis virescens. The lateral oviducts of the female parasitoid contain a particulate suspension called calyx fluid. The particles in calyx fluid are a polydnavirus (CsV) which, when injected into last-instar H. virescens, stimulates degeneration of the host's prothoracic glands. In order to determine if CsV-induced degeneration is specific to prothoracic glands, last-instar H. virescens larvae were injected with C. sonorensis calyx fluid. After 4 days, a variety of host tissues were dissected from both calyx fluid-injected and uninjected control larvae and fixed for transmission electron microscopy. Prothoracic glands from injected larvae were ultrastructurally degenerated by 4 days post-injection, whereas control glands remained intact. Other tissues from calyx fluid-injected larvae (tracheal epithelia, corpora allata, Malpighian tubules, fat body, skeletal muscle, and the brain) showed no signs of ultrastructural degeneration or gross abnormalities as compared with control tissues. These observations suggested that CsV-induced degeneration is specific to the host's prothoracic glands.     

STUDIES ON THE CROSS-STRIATION OF THE INDIRECT FLIGHT MYOFIBRILS OF THE BLOWFLY CALLIPHORA

1. The cross-striation in the indirect flight myofibrils of Calliphora has been studied by phase contrast and polarised light microscopy. The band pattern at rest-length has been determined in flies killed in osmium tetroxide vapour while their wings remained in the resting position. All other observations have been made on unfixed fibrils. Although length changes in situ are probably very slight (about 2 per cent), isolated fibrils, by treatment with crude muscle extract or with ATP, can be induced to elongate to 104 per cent rest-length, or to shorten by 8 per cent but no more. Over the range 98 to 104 per cent rest-length, experimentally induced length changes are reversible. The fibrils can also be stretched beyond 104 per cent rest-length, but the process is irreversible. During the course of glycerol extraction the fibrils elongate to 104 per cent rest-length. 2. The changes in band pattern observed over the range 104 to 92 per cent rest-length are qualitatively the same as the changes observed over a wider range (about 130 to 40 per cent rest-length) in the skeletal myofibrils of rabbits. The earlier stages of shortening appear to be effected by retraction of the I bands into the A bands where they fill up the H zones. No evidence has been found that any changes in band pattern are due to a migration of the A substance. 3. Two components of the sarcomere can be extracted from it and a third component remains behind. These three components, which have also been demonstrated in skeletal myofibrils of the rabbit, where they behave in the same way, are: (a) the A substance which does not change its position as the fibril changes its length, and which can be extracted by the same procedures as remove myosin (shown elsewhere to be the A substance) from rabbit fibrils; (b) a material which extends from the Z lines to the borders of the H zone and which moves inwards during contraction and outwards during elongation; it can capture rabbit myosin from solution and form with it a contractile system, and it is thought to be actin; (c) a "backbone" or stroma bearing Z and M lines. 4. Since all these features of the cross-striation are the same in the insect fibrils as in rabbit fibrils, it is considered very probable that the sarcomere is similarly organised in both types of muscle and contracts by essentially the same mechanism.     

Amyloid toxicity in skeletal myoblasts: Implications for inclusion-body myositis

Skeletal muscle disorder, inclusion-body myositis (IBM) has been known for accumulation of amyloid characteristic proteins in muscle. To understand the biophysical basis of IBM, the interaction of amyloid fibrils with skeletal myoblast cells (SMC) has been studied in vitro. Synthetic insulin fibrils and Aβ_25-35 fibrils were used for this investigation. From the saturation binding analysis, the calculated dissociation constant (K_d) for insulin fibril and Aβ_25-35 fibrils were 69.37 ± 11.17 nM and 115.60 ± 12.17 nM, respectively. The fibrillar insulin comparatively has higher affinity binding to SMC than Aβ fibrils. The competitive binding studies with native insulin showed that the amount of bound insulin fibril was significantly decreased due to displacement of native insulin. However, the presence of native insulin is not altered the binding of β-amyloid fibril. The cytotoxicity of insulin amyloid intermediates was measured. The pre-fibrillar intermediates of insulin showed significant toxicity (35%) as compared to matured fibrils. Myoblast treated with β-amyloid fibrils showed more oxidative damage than the insulin fibril. Cell differentiating action of amyloidic insulin was assayed by creatine kinase activity. The insulin fibril treated cells differentiated more slowly compared to native insulin. However, β-amyloid fibrils do not show cell differentiation property. These findings reinforce the hypothesis that accumulation of amyloid related proteins is significant for the pathological events that could lead to muscle degeneration and weakness in IBM.     

Changes in the functional efficiency of flight muscle sarcosomes during heat death of adult Calliphora erythrocephala

The temperature sensitivity of flight muscle mitochondria from adult Calliphora erythrocephala has been studied. Intact flies were treated to sublethal and lethal high temperatures, their sarcosomes were isolated and the efficiency of the coupling of oxidation and phosphorylation was measured. The temperature sensitivity of sarcosomes is correlated with the heat death point of the intact animal and the impairment of oxidative phosphorylation with α-glycerophosphate suggests that a breakdown in ATP synthesis may be one of the causes of heat death. Restitution of enzyme activity was observed in mitochondria isolated from flies recovering from sublethal heat treatment. The results suggest that the impairment of membrane-enzyme complexes may be important lesions in heat death.     

THE STRUCTURE OF THE SMOOTH MUSCLE FIBRES IN THE BODY WALL OF THE EARTHWORM

1. The structure of the smooth muscle fibres in the longitudinal muscle coat of the body wall of Lumbricus terrestris has been investigated by phase contrast light microscopy and electron microscopy. 2. The muscle fibre is ribbon-shaped, and attached to each of its two surfaces is a set of myofibrils. These are also ribbon-shaped, and they lie with their surfaces perpendicular to the surfaces of the fibre, and their inner edges nearly meeting in the middle of the fibre. These fibrils are oriented at an angle to the fibre axis, and diminish greatly in width as they approach the edge of the fibre. The orientation of the set of fibrils belonging to one surface of the fibre is the mirror image of that of the set belonging to the other surface; thus, when both sets are in view in a fibre lying flat on one face, the fibre exhibits double oblique striation. A comparison of extended and contracted fibres indicates that as the fibre contracts, the angle made between fibre and fibril axes increases (e.g. from 5 to 30°) and so does the angle made between the two sets of fibrils (e.g. from 10 to 60°). 3. The myofibril, throughout its length, contains irregularly packed filaments, commonly 250 A in diameter, which are parallel to its long axis and remain straight in contracted muscles. Between them is material which probably consists of much finer filaments. Thus A and I bands are absent. 4. Bound to one face of each fibril, but not penetrating inside it, is a regularly spaced series of transverse stripes. They are of two kinds, alternating along the length of the fibril, and it is suggested that they are comparable to the Z and M lines of a cross-striated fibril. The spacing of these stripes is about 0.5 µ ("Z" to "Z") in extended muscles, and 0.25 µ in contracted muscles. A bridge extends from each stripe across to the stripeless surface of the next fibril.     

Morphology and ultrastructure of an isolated cell type corpora allata in adults of the bertha armyworm,Mamestra configurata Wlk. (Lepidoptera : Noctuidae)

There are 2 broad morphological types of corpora allata in adult Lepidoptera, a capsular type gland and an isolated cell type gland. Compared with the ubiquitous capsular corpus allatum, the isolated cell type has a narrow distribution in Lepidoptera and appears to be restricted to adult noctuids of the sub-family Hadeninae. The isolated cell corpora allata of adults of Mamestra configurata (Lepidoptera : Noctuidae) are composed of 30 – 40 large (ca 0.1 mm), semi-transparent, spherical, isolated cells held in 2 clusters by fine trachae and nerve fibers. These clusters lie directly beneath and in close proximity to the brain. Many similarities exist between the ultrastructure of the isolated cell corpora allata of M. configurata and the more common capsular gland. A high density of mitochondria, the arrangement of smooth endoplasmic reticulum in concentric “finger print” whorls, an interconnecting lacunae system of lipid vacuoles, and the presence of neurosecretory nerve endings are features commonly seen in active corpora allata of both types. The corpora allata of M. configurata are larger in the male, the calculated gland volume of males being more than 4 times that of females. Gland activity may change in senescent males as judged by the degeneration of the mitochondria, the reduction of the lacunae system and the dissolution of the smooth endoplasmic reticulum.     

THE ELONGATION OF MYOFIBRILS FROM THE INDIRECT FLIGHT MUSCLE OF DROSOPHILA

Myofibrils which lengthen by several per cent in the presence of ATP and magnesium ions were prepared by teasing indirect flight muscle of Drosophila in solutions containing ethylenediaminetetraacetate. A study was made of the hydrogen ion, magnesium ion, ATP, and potassium chloride concentrations with which this effect could be observed. The lack of elongation with pyrophosphate and several nucleoside triphosphates suggests that the lengthening is ATP specific. A relaxing factor system comparable to that described for rabbit muscle was not demonstrable, as elongated fibrils did not shorten with calcium ions, carnosine, or digitonin.     

ELECTRON MICROSCOPIC AND HISTOCHEMICAL OBSERVATIONS OF MUSCLE DEGENERATION AFTER TOURNIQUET

As an experimental model for the different forms of muscle degeneration, injury caused by 2 hours' ischemia has been studied from 20 minutes to 16 hours after release of the tourniquet. Discoid degeneration developed in stretched fibers by dissolution of the I bands (Z substances and actin). The discs represented the Q bands (A-H-A). In fibers which passively maintained contraction lengths during degeneration, the Z substances were dissolved, but the continuity of the fibrils was preserved, since the filaments are continuous over all sarcomeres under these conditions. Mitochondria and the tubules of the endoplasmic reticulum swelled, ruptured, and disintegrated. Granular degeneration developed in fibers where mitochondria were abundant. Unstretched degenerating fibers with few mitochondria gave a homogeneous or hyaline appearance. The different forms of degeneration therefore were dependent on the status of stretch and the fiber type. The extent of degeneration was not a function of time after ischemia, there being both nearly normal and severely damaged fibers at 20 minutes and 16 hours after the release of tourniquets. When degeneration occurred, however, the basic alterations were the same in all fibers; there was mitochondrial and reticular swelling, dissolution of the Z substances, and finally disintegration of the contractile material. Some damage developed in the sarcolemmas and capillaries. The mitochondrial disintegration was not linked with inactivation of the succinic dehydrogenase system.     

Histopathological study of corpora amylacea pulmonum

In this paper, we present a rare disorder which is known as corpora amylacea pulmonum. X-ray CT scanning showed an abnormal focus of the lung as a solitary mass with high density and spicular features around the surface. The resected lung tissue was characterized by the appearance of round, concentrically laminated acellular bodies about 40-80 microns in diameter. The bodies were usually found lying free in the alveolar space and surrounded by the exudate alveolar macrophages or multinuclear giant cells. Some of these macrophages were in a state of progressive degeneration. The bodies showed an affinity for Congo red and exhibited partial birefringence. Moreover, all the bodies had a strong positivity for the PAS reaction and anti lysozyme antibodies. The exudate alveolar macrophages and multinuclear giant cells also displayed reactivity for PAS and lysozyme in a similar manner to that of the bodies. Electron microscopically the bodies were fundamentally composed of fibrillar elements, which bore some resemblance to amyloid fibrils and probably accounted for the partial affinity of the bodies for Congo red. These amyloid-like fibrils were also found in the cytoplasm of the macrophages. This suggested that the concentrically laminated bodies in corpora amylacea pulmonum might be formed by sequential aggregation, fusion, coalescence and compaction of degenerated alveolar macrophages.