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  • Life and Medical Sciences  (754)
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  • 1995-1999  (629)
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  • 1975-1979
  • 1935-1939  (125)
  • 1910-1914
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938), S. 91-111 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The clam Mya differs from other pelecypods which have been investigated in this respect in having most of the reserve nutritive materials formed within the vacuolated follicle cells of the gonads instead of within the very limited amount of mesenchymatous connective tissue of the visceral mass. The profusely branching tubular gonads originate from two groups of primordial germ cells situated in the position of the future genital apertures. The germinal primordia soon become differentiated into two types of nuclei, one of which becomes associated with the large, vacuolated follicle cells, which form the principal volume of the gonadal tissue, while the other type proliferates to form the primary gonia which become widely scattered along the walls of each alveolus.There is much degeneration and cytolysis during gametogenesis in both sexes, with the accumulation of characteristic inclusions within the follicle cells. Atypical spermatogenesis followed by cytolysis occurs throughout the year but the normal method only in preparation for spawning. No evidence of protandry or change of sex was obtained; only three hermaphrodites were found in the examination of more than 1000 individuals.
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  • 2
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938), S. 1-2 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 3
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The regeneration of muscle in larval Amblystoma punctatum is preceded by an extensive dedifferentiation of the old muscles of the limb stump. The process of muscle dedifferentiation consists in a separation of muscle nuclei, surrounded by a small amount of cytoplasm, from the injured ends of the muscle fibers. The dedifferentiation of the cut muscles of the limb stump progresses proximad as far as the origin of the muscles on the humerus and results in a complete transformation of these muscles into undifferentiated cells which appear to contribute to the formation of the regeneration blastema. Shoulder muscles, which were attached to the humerus, also undergo a partial dedifferentiation when their points of insertion on the humerus are destroyed by the degeneration of the perichondrium. These muscles never dedifferentiate, however, for more than one-fourth their original length. The process of dedifferentiation in the shoulder muscles is similar to that found in the cut muscles of the limb stump.The regeneration of the injured muscles occurs in two ways. The shoulder muscles reconstitute themselves by means of terminal and lateral sarcoplasmic buds formed near the distal regions of the muscle fibers. The muscles of the limb proper, distal to the shoulder, differentiate out of local aggregations of blastema cells. No myoblasts were observed.The regeneration blastema arose chiefly from dedifferentiated cells of muscle, nerve connective tissue sheath, perichondrium and cartilage.
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  • 4
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 5
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938), S. 177-218 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The original innominate bone consisted of ischiopubis only. From this developed a dorsally-directed ilium, upon which the dorsal limb muscles, originally arising from fascia, settled, and which thrust dorsalward between roots of the limb plexus, thus dividing the nerves into prozonal (dorsal and ventral) and metazonal (dorsal and ventral) groups. The primitive muscles of the tetrapod hip and thigh comprised a dorsal mass, soon divisible into sheets, innervated by both prozonal and metazonal dorsal nerves, and a similar ventral mass comparably innervated. The original two elements thus became four basic elements, and probably in early mammals or mammal-like reptiles all dually innervated muscles split into singly innervated units. With this four-group basis as the chief criterion, but considering other factors as well, it is possible to homologize the muscles of urodeles (ventral components only), lacertilians, mammals, and birds in entirely satisfactory manner, except for doubt in several instances in which specialization has secondarily obscured the precise relationships. In different mammals there is shown a tendency toward a final fusion of certain unrelated muscle units (biceps plus gluteus longus, human type of biceps, adductor magnus, and tensor fasciae femoris with gluteus maximus).
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 7
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938), S. 415-443 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In the blastula stage the roof of the subgerminal cavity is composed of an irregular layer of cells, the nuclei of which lie in the upper or middle part of the cytoplasm.On the floor of the subgerminal cavity groups of already degenerating cells occur. They represent the vegetative pole of the blastula. Almost every cell contains glycogen, and mitotic cells show no special orientation.In the gastrula stage the cells of the area pellucida become regularly arranged as a single-layered, cylindrical epithelium with basally situated nuclei.The yolk endoderm cells are formed from the proliferating upper layer of the area opaca.The embryonic endoderm is formed at the posterior end of the area pellucida by outgrowth of single cells from a circumscribed area, the primitive plate.This plate eventually bends inward to form a typical archenteric canal, through which endoderm continues to invaginate from the epiblast.The endoderm spreads in a cranial and lateral direction until it has formed a complete layer.The epiblast cells lose their glycogen as they invaginate to form endoderm, which is free of glycogen.In the area opaca the upper layer and the yolk endoderm contain glycogen.The mitotic cells of the epiblast of the area pellucida are always orientated horizontally, but in the primitive plate and archenteric canal they are orientated vertically as well.
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  • 8
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 62 (1938), S. 559-597 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The larval, metamorphosing and definitive aortic arches of Desmognathus fuscus, Plethodon cinereus, Eurycea bislineata, and Gyrinophilus porphyriticus, described in this paper, were investigated because it was thought possible that, if the fourth or pulmonary arch failed to develop in these lungless forms, a new factor associated with the loss of lungs might be revealed.The salient points of structure disclosed, so far as the problem involved is concerned, are: D. fuscus and P. cinereus develop fourth arches which remain functional in the larvae and adults and supply the pharynx, oesophagus, stomach and skin of the shoulder region. In some instances the fourth arch in the larvae of P. cinereus is reduced in length or entirely lacking in which cases correspondingly less of the fourth arch and more of the third arch is present in the adult. E. bislineata and undoubtedly G. porphyriticus fail to develop fourth arches and hence do not possess these in either the larval or adult states.It is concluded, therefore, that the failure of the fourth arch to develop has evidently not been a factor involved in the advent of lunglessness in plethodontid salamanders. Also, the fact that E. bislineata never develops a fourth arch, yet is able to transform, furnishes additional evidence against Figge's view that Necturus fails to metamorphose because the ventral portion of the fourth arch is absent.
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  • 9
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 10
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938), S. 75-86 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This study is based on serial sections of the occipital and otic regions of a therocephalian from the Tapinocephalus zone. The occipital region as preserved consists of the basioccipitals and exoccipitals. The basioccipital is long and slender and is separated from the more anterior basisphenoid by an unossified zone. The exoccipitals are large and contain a part of the jugular foramen and two foramina for cranial nerve XII.The otic bones are fused together to form a periotic. The most striking feature of the inner ear is the medioventral position of the vestibule. Passing back into the periotic from the vestibule is a deep recessus scala tympani. This recess opens anteriorly into a ventral fenestra in the vestibule, the fenestra rotunda. These structures are similar to those of Dimetrodon and the gorgonopsian.The anterior part of the periotic is projected ventrally to form a basicranial process. This probably arose by intramembranous ossification. The unossified zone between the basioccipital and basisphenoid may represent a persistent basicranial fenestra.
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  • 11
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938), S. 63-73 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A study has been made of the growth of the eye anlage, and of the increase in number of the elements of the dioptric system of larvae of Drosophila melanogaster.In newly hatched larvae the eye rudiment grows fast, but slows down later and becomes nearly stationary during the second half of larval life. The increase in number of elements in the eye disc parallels the growth of the whole anlage and reaches a maximum about 70 hours after hatching. The imaginal disc cells increase in number but not in size, in contrast to other larval cells which increase in size but not in number.
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  • 12
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938), S. 87-117 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The form of the vertebral column is definitely related to its function as a supporting rod, a base for attachment of body and limb muscles, and a protection of the spinal cord and nerves. Primitively composed of a series of simple undifferentiated blocks, it progressively becomes complicated through development of articular processes giving added strength and greater mobility. Simultaneously, the centrum and the neural arch become adapted to withstand tension and compression stresses which vary with the movements possible in different regions of the column. These movements are partially determined by the plane of the zygapophyses and the nature of the intercentral articulation, together with the action of the axial muscles and ligaments.In fish and primitive tetrapods the axial musculature serves as the chief locomotor organ and consists of a series of myomeres extending with little interruption from the head to the tail. In tetrapods the locomotor function is taken over by the limbs and the axial muscles become progressively differentiated into long flexors and extensors of the column and gradually lose their external segmentation.
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  • 13
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Adult salamanders of Ambystoma tigrinum show a high degree of sexual dimorphism. The normal growth of the urogenital ducts system in relation to the development of the gonads is traced from the sexually indifferent period preceding metamorphosis to sexual maturity at 1 year. Differentiation and growth of secondary sex characters is correlated with spermatogenesis in the male and with growth of ovocytes in the female.Testicular hormone manifests its initial appearance in male differentiation of the wolffian and urinary collecting ducts following metamorphosis. Growth of ovocytes and oviducts begins before metamorphosis and may be assumed to indicate the initial appearance of the ovarian hormone. Females retain the larval arrangement of wolffian and urinary ducts. The oviducts of the male regress during the period of rapid male differentiation. This suggests the possibility of antagonistic activity of the male hormones in Ambystoma.
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  • 14
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938), S. 143-161 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A batch of fifty newts was subjected to complete food deprivation for a period of 4 months. During the experiment twenty-two individuals died and twenty-eight were killed at intervals. From the end of the second month the erythrocytes began to show in increasingly greater numbers a progressive alteration in the nucleo-cytoplasmic ratio. The nucleus of these atypical erythrocytes gradually enlarged and ultimately occupied the entire cytosome. Meanwhile the nuclear reticulum became extremely fine and closely meshed, eventually appearing practically homogeneous and relatively chromophobic. The end result was complete dissolution. The leukocytes became greatly reduced in number, the eosinophils disappearing completely. The neutrophils and basophils were agranular in the blood smears. The erythrocyte transformations represent late results of a chain of factors: cytoplasmic hypotonicity, nuclear endosmosis, attenuation and fenestration of nuclear membrane. The subcapsular lymphogranulocytopoietic tissue of the liver had almost completely disappeared. The spleen was greatly reduced in size; it was essentially lymphoid in character and erythrocytopoietic activity had ceased practically completely. Complete restoration of normal conditions in blood, liver and spleen was effected by the end of 2 weeks by feeding with earthworms. The regenerating blood showed many naked nuclei of disintegrating atypical erythrocytes, many immature erythrocytes in mitosis, increased number of small lymphocytes (lymphoid hemoblasts), large but variable numbers of microcytes, many giant thrombocytes and normal granulocytes.
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  • 15
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Colloidal carbon injected into the coelom of the larval lamprey, Ammocoetes, is taken up directly by the pronephric tubules. Due to the absence of nephrostomes the mesonephric tubules do not function in a similar way. The tubules of neither show any intracellular deposition of carbon. The reticular elements which support both these kidneys exhibit pronounced phagocytic and hemocytopoietic activity. Carbon in either a free or included form reaches all the other organs both as a result of direct invasion or secondary distribution by the vascular system. The liver is the only organ whose vascular endothelium exhibits cytopoietic properties. To the diffuse spleen as a site of blood cell formation thus should be added the reticular tissue of the pronephros and mesonephros and the vascular endothelium of the liver. Playing a minor role in a similar way are the intestinal mucosa exclusive of that in the typhlosole and the spongy tissue dorsal to the neural tube.
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  • 16
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 63 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 17
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    Journal of Morphology 63 (1938), S. 181-205 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In an earlier paper (Slifer, '37) experiments were reported which showed that in Melanoplus differentialis eggs the hatching enzyme, which destroys the white cuticle, is secreted by the pleuropodia (appendages of the first abdominal segment of the embryo) during the last few days of incubation. In the present report the development and differentiation of the pleuropodia, together with the cytological changes which occur in them before, during and after they become functional, have been followed. The time at which the secretion granules are discharged from the pleuropodial cells was found to be closely correlated with the time at which the tough, white cuticle begins to disintegrate. The cytological evidence, then, supports the author's earlier conclusion, based upon experimental work, that the pleuropodia secrete the hatching enzyme.
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  • 18
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    Journal of Morphology 63 (1938), S. 207-217 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Comparison of intramembral and trunk/limb proportions in the roadrunner (Geococcyx) and two related genera of cuckoos (Coccyzus and Crotophaga), together with a consideration of their habits of locomotion, lead to the following two generalizations: (1) The incipient cursorial leg of more primitive, arboreal birds, with the metatarsus shorter than the femur, is not an efficient mechanism for bipedal terrestrial locomotion. The direct adaptation to the cursorial habit in terrestrial birds lies in the further elongation of the whole leg, the distal segments undergoing a relatively greater elongation than the femur; the greatest degree of elongation is shown by the most distal segment. (2) When the development of terrestrial, cursorial habits in birds leads to diminished use, or discuse, of the wings as organs of locomotion, the wing skeleton becomes reduced in length; each segment of the wing is reduced, but the degree of reduction is greatest in the more distal segments. It is probable that reduction appears first in the most distal segment and later successively in each segment proceeding proximally therefrom.
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  • 19
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    Journal of Morphology 63 (1938), S. 219-227 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The median cord arises as a hypodermal invagination along the midline from stomodeum to end of tenth abdominal ganglion. It separates from the hypoderm, but does not differentiate into any tissue; apparently it degenerates.
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  • 20
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: There are 19 chromosomes in diploid cells of male Paratylotropidia brunneri; of these, four are V-shaped multiples. There are twenty chromosomes in diploid cells of females; of these, four are V-shaped multiples. If the two arms of each multiple are counted as separate chromosomes, we have the usual number of chromosomes for the Acrididae, i.e., twenty-three in male and twenty-four in female.There are nine chromosomes in the first spermatocyte divisions: seven tetrads, one octad and a decad. The latter is made up of the accessory chromosome associated with an octad.One of the V-shaped multiples in the male is limited to that sex. The homologue of one of the dyads of which it is composed is a free dyad, the homologue of the other forms a V-shaped multiple with the accessory chromosome. The V-shaped multiple limited to the male shows differential heteropycosis in the prophases of the first spermatocyte. There is evidence that its homologous parts are isolated from each other as far as crossingover is concerned.A study of the first maturation division of the heterogametic sex is essential for the identification of the sex chromosome.
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  • 21
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    Journal of Morphology 63 (1938), S. 229-261 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Although thoracic diverticula of the aorta ending dorsally in pulsatile organs were discovered by Brocher as early as 1917 no detailed account of their structure was published. The condition of these structures in the adult was not investigated and their development through larval instars was not followed. Brocher's later papers announced the discovery of similar organs in the orders Orthoptera, Coleoptera and Lepidoptera. In 1931 E. Meyer described in some detail pulsatile organs in the Ephemerida. In general these accounts are either superficial or of doubtful interpretation.The present paper deals with the Odonata and stresses the Anisoptera. Anax junius has been studied as the type both anatomically and histologically in all stages of larva and imago except the first four instars. Other types have been compared with Anax, and some of the other orders mentioned have been checked for presence and nature of these organs.Aortic diverticula and pulsatile organs occur in all Odonata both larval and adult. These are derived from the same origins and their histology is that of the membranes involved in aortic and body walls. Through inference from morphology and from physiological examination it appears that pulsatile organs (1) supplement heart action, perhaps substituting for it during emergence. (2) assist in the functioning of the ostia and ostial glands, and (3) may even be important in production and distribution of hormones.
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  • 22
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The germinal epithelium was studied in an effort to determine its role in postpubertal ovogenesis. Active and inactive areas of epithelium were observed regardless of the oestrous periods. The active areas, exemplified by numerous mitoses and a frequent stratification of the epithelium, are foci for egg cell production.Egg cells are produced from the germinal epithelium by: activation, migration and transformation of single germinal epithelial cells; migration and differentiation of nests of cells; ingrowths of cords of cells.Young ova in the cortex of the ovary are at first surrounded by a single layer of flattened follicular cells of germinal epithelial origin. In the subsequent development of the follicle stratified layers make their appearance. The zona pellucida appears as a well-defined structure coincident with the stratification of the follicular layers.The follicular cavity develops as a split between the follicular cells at one side of the follicle. In the mature follicle the cavity is broad and contains liquor folliculi. The theca interna and externa are not distinguishable until the follicle is highly cellular.
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  • 23
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The spermatogenetic cycle of the testis of the musk turtle is limited to the summer months of the year, and closely parallels that of the anuran Amphibia. Spermatozoa are present in the testis from September to May, and breeding may occur in either fall or spring. A pronounced increase in the size of the epididymis and a corresponding decrease in the size of the testis occurs at the end of the cycle (September).During the months of March, April and May, all spermatozoa are eliminated from the seminal tubules, and the germinal epithelium is built up in preparation for spermatogenesis. Spermatogonial divisions occur in small numbers in May, and the division tempo increases during June. Primary spermatocytes and maturation divisions appear after the middle of June, and continue through July and August. Spermiogenesis begins in late July, is in full progress in August, and is practically completed by October. Laboratory specimens usually show an active spermatogenesis in winter, but it is not probable that a second spermatogenetic cycle occurs in specimens under normal hibernating conditions.No seasonal changes are observed in the interstitial cells of the testis, and no seasonally variable secondary sex characters are known for turtles. Special studies of the problem are being conducted.
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  • 24
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    Journal of Morphology 63 (1938), S. 345-361 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This species represents a typical example of protandric consecutive sexuality, the male phase becoming functional at a very early age, when the body has reached a length of only 3 to 7 mm. Those males which become functional earliest usually seek the association of older individuals in the female phase; others, maturing later, are more likely to remain solitary. Mated males have accentuated masculine characteristics and retain the male phase longer than solitary individuals or those that are isolated experimentally. Unfavorable environmental conditions postpone or prevent functional sexuality and terminate the male phase promptly without inaugurating the female phase. The influence of the female in accentuating the sexuality of the male is thought to be due to stimuli received by the latter through sense organs in tentacles and penis and mediated by the nervous system through hormonal secretions. Termination of the male phase and transition to the functional female are comparable to metamorphosis from the immature to the mature condition in other animals and, like metamorphosis, the primary (male) sexual phase can be abbreviated or prolonged experimentally, but the sequence cannot be reversed.
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  • 25
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Data concerning the seasonal distribution of twenty-two species of nudibranchs are recorded for a period of 9 months. The occurrence of copulation and of egg laying for these animals in the laboratory aquaria is recorded for the same period. These data indicate that the seasonal distribution of certain species is well marked, and that there is in many cases a definite breeding season.The characteristics of the egg ribbons of these nudibranchs are described, and figured by means of photographs. These egg ribbons approach the mathematical form of a spiral of Archimedes. In every case observed, the ribbon was deposited in a counter-clockwise direction, viewed dorsally.The relationship between these observations and earlier work on the life histories of the nudibranchs is discussed.
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  • 26
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Salivary gland nuclei in Chironomus regularly show large, conspicuous nucleoli. In Sciara no true nucleoli have been found, but sometimes certain particular chromosome regions expand greatly, forming ‘puffs’ or ‘bulbs,’ somewhat nucleolus-like in nature. Detailed study has been made of the chromosome structure in the affected regions in both genera.In the nucleolar regions of Chironomus the banded structure of the chromosome is distinctly modified. In the case of the large nucleolus the chromosome breaks up into a heavy network in which solid discs are replaced by interconnected chromatic spheres and granules, extending out somewhat into the clear nucleolar substance. In the case of the smaller nucleolus, Balbiani's ring, the banded structure is less disturbed. A chromatic network, resembling a system of rootlets, runs out from the chromatic bands into the clear nucleolar substance.In Sciara ocellaris the ‘puff’ regions are at times normally banded, but at other times in the condition described as ‘puffed.’ The same is true of the ‘bulb’ regions. The relation between nucleoli, puffs and bulbs is discussed. Also that between ‘heterochromatin’ and ‘euchromatin.’ Evidence seems to indicate that the latter are merely extremes in a continuous range involving different relative amounts of chromatic and achromatic materials. The ‘puff’ regions appear to be structurally similar to the chromocenter in Drosophila.
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  • 27
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    Journal of Morphology 63 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 28
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    Journal of Morphology 63 (1938), S. 397-419 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The summary of this paper is as follows:1From an examination of 108 ovaries from eighty-nine individuals it was apparent that, in the cat a certain but variable amount of degeneration of primary follicles occurs in all life epochs.2The primary oocytes undergo the degeneration, while the follicle cells remain for an undetermined time.3The most profound instances of such degeneration were encountered in kittens of 6 to 9 weeks, accompanying or follwing the resolution of the egg cords into primary follicles and the establishment of the cortical or marginal zone. A sporadic degeneration of young oocytes occurs at earlier periods.4No evidence was seen of a new formation of egg cells from the residual follicle cells after the degeneration of the oocytes. The degenerations of the 6 to 9 weeks epoch are not believed to be of universal occurrence.5No adequate evidence was encountered of a new formation of egg cells from the surface epithelium, either before or after sexual maturity.6No evidence linking the degeneration of primary follicles with the estrous cycle was seen.7The stock of primary follicles established in the first few weeks after birth is believed to be adequate for the growth of graafian.
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  • 29
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    Journal of Morphology 63 (1938), S. 421-439 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The maturation of the germ cells of the two hermaphroditic species, Curtisia foremanii (Girard) and Bdelloura candida (Girard) has supplied the material for this chromosome study.Both species of flatworms are believed by the author to possess a diploid number of twelve chromosomes and a haploid number of six, although Curtisia foremanii has previously been reported as having a smaller and variable number of chromosomes.A tendency of the chromatids comprising individual chromosomes to separate from one another at certain times was noted in both species. This action results in giving the appearance of a larger number of chromosomes than the germ cells actually possess. A further source of apparent increase in chromosome number in the Curtisia oocytes, after treatment with the usual Allen's B3 and B15 and Heidenhain's iron haematoxylin, is the presence of some deeply staining, non-chromatin material.No significant differences in number, form and behavior of chromosomes of male and female complexes were noted, with the possible exception of the tendency of the chromatids to separate from one another, to be greater in the female than in male germ cells.
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  • 30
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    Journal of Morphology 63 (1938), S. 441-475 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Four cases of gynandry in American spiders present the following morphologic deviations: The specimen belonging to Neoantistea agilis (Keyserling) externally is a bilateral gynandromorph with distortion of the single male palp, with half of the epigynum nearly normal; internally both testes and ovaries are present. The Drassodes neglectus (Keyserling) is not a completely developed bilateral gynandromorph. One side has a normal male palp, a larger chelicera and longer legs than the other, the epigynum is complete and normal; internally only degenerate ovaries are present. The Linyphiid, near Bathyphantes, is three-quarters female, the only male organ being an imperfectly developed male palp. Ovaries are well developed but immature, whereas the epigynum is complete. In the Pardosa sternalis (Thorell) the anterior part of the body is male, as indicated by the swollen palps and the length of the legs; the epigynum is completely developed, but internal reproductive organs are missing except for a very much aborted ovary. The bisexuality in all cases has caused degeneration or abnormalities in the reproductive structures.Included is a tabulation of all recorded instances of gynandry in spiders, arranged according to the type of abnormality. The term ‘leg-index’ is introduced to express ratio between length of the leg and length of cephalothorax.
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  • 31
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    Notes: Centrifuging the eggs of Rana pipiens in the early gastrula stage prevents the formation of the hypophysis in some of the tadpoles. The absence of the melanophorotropic hormone normally secreted by the hypophysis seems to be responsible for the contraction of the pigment cells. In addition, there are actually fewer pigment cells present in both the dermal and epidermal layers of the light tadpoles than there are in the controls. The paleness, therefore, of the tadpoles seems to be due to both a contraction of the pigment cells present and to an actual reduction in their number. The failure of the hypophysis to develop was brought about by centrifuging at an earlier stage in development of the embryo than in previous extirpation studies. The effects produced by centrifuging that are responsible for the failure of the hypophysis to develop are unknown. However, it is suggested that interference in some way with the presumptive hypophysis-forming tissue has resulted in an inactivation of its inductive potencies.
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  • 32
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    Journal of Morphology 63 (1938), S. 491-529 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This paper is based on evidence obtained from sectioned skull of Galesaurus planiceps, Owen. Width-length index of skull is 67.7; height-width index 47.6. Dental formula is: \documentclass{article}\pagestyle{empty}\begin{document}$ \frac{{{\rm I\; 3\; or\; 4}}}{{{\rm I\; ?}}} $\end{document}, \documentclass{article}\pagestyle{empty}\begin{document}$ \frac{{{\rm C\; 1}}}{{{\rm C\; 1}}} $\end{document}, \documentclass{article}\pagestyle{empty}\begin{document}$ \frac{{{\rm PC\; 7}}}{{{\rm PC\; 9}}} $\end{document}. Length of prevomer is 24.0 mm., that of skull 62.0 mm. Maxilla contains cavity which appears to be equivalen of sinus maxillaris of mammals. Ear structure shows large internal auditory meatus. There is no evidence that horizontal and posterior semicircular canals pass through bone. Fenestra ovalis is large. Parasphenoid, basisphenoid, and basioccipital are fused. Parasphenoid consists of short body and a medial anterior process. Sella turcica lacks floor, probably due to erosion. Detached bone may be sphenethmoid. Dentary contains two canals, probably vascular. Comparisons with related forms lead to following conclusions: (1) Specimen is of young animal. (2) Dental succession of postcanines, distichical replacement of postcanines, long prevomer, short parasphenoid, incomplete osseous incasement of semicircular canals, and other characteristics label Galesaurus as primitive cynodont. (3) Reduction of parasphenoid and development of prevomer support Broom's view that Ictidosaurus is not a cynodont. (4) Prevomer as nasal septum; fusion of parasphenoid, basisphenoid, and basioccipital; large lacrimal, splenial, and articular; and other characteristics indicate that Galesaurus is off line leading to mammals. (5) In certain respects cynodonts are more mammal-like than gorgonopsians.
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  • 33
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    Notes: The development of the cranial musculature of Amblystoma punctatum is described in detail, for both larva and adult. In addition, a brief account of the innervation of each muscle is given. The study of normal development is supplemented by extirpation experiments performed on embryos in early stages of development. These extirpations include the mesodermal head segments, mesoderm of mandibular, hyoid, second branchial, third branchial arches, and somites 1, 2 and 3. The eye muscles are found to develop from the mesodermal head segments in precisely the same manner as in other classes of vertebrates; head segmentation of Amphibia, though less accentuated, is shown to be homologous with that of other classes. The remaining extirpation experiments corroborate, in general, the finding from normal development studies. An attempt is made to summarize in tabular form the prospective fate of the prechordal plate and parachordal mesoderm, the two earliest divisions of the head mesoderm, with regard to their complete muscle derivatives.
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  • 34
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    Journal of Morphology 62 (1938) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 35
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    Journal of Morphology 62 (1938), S. 3-15 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Comparisons have been made of the weights of the suprarenals of 525 mature female dogs in dioestrus, oestrus, pregnancy and lactation. Tables and graphs show that there was a slight increase in the size of the suprarenals during oestrus. There was hypertrophy of both the cortex and the medulla in the oestrous dogs. Only slight increase was shown in the weights of the glands of pregnant animals when the net body weight was considered and no increase when the total body weight was used in the study. There was no hypertrophy of the glands of lactating animals.There was only a slight variation in the seasonal occurrence of oestrus. From 34 to 37% of the total numbers of mature females examined in the seasons were found to be in oestrus. From 13 to 16% of the total numbers were found to be pregnant.
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  • 36
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: Nucleoli in the germ cells of certain Orthoptera, Hemiptera and the crayfish have been studied. In Ceuthophilus (camel cricket) these bodies are found to be definitely connected with chromatin threads and are believed to be products of chromatin.The large structure in the ova of Nemobius (cricket) is interpreted as a nucleolus because of its very irregular behavior and apparent complete disappearance.It is found that some of the nucleoli of Gelastocoris (toad bug) arise within the chromatin threads.Observations on Cambarus (crayfish) fail to show nucleolar extrusion, a phenomenon which has formerly been reported for this form.The unique behavior and fate of the nucleolar material in Pselliopus (true bug) is described. In this species the nucleolar material is added to and becomes part of the general oxychromatin network of the nucleus.This study fully supports the opinion that nucleoli have a chromatin origin.
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  • 37
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: A new limb skeleton will form in regenerating limbs of the larval Amblystoma in which the humerus had been completely removed. The cartilage of the new limb skeleton develops out of a blastema in which there are no cells of cartilage origin. This regeneration blastema is a composite structure made up of cells derived from a dedifferentiation of the injured tissues of the limb. The tissues which have been observed as contributing to the regeneration blastema are: muscles, especially the muscles of the shoulder; connective tissue of the sheath of the brachial nerve plexus; muscle connective tissue; and, to a certain extent, subcutaneous connective tissue. The new cartilage of the limb skeleton develops out of this composite blastema by means of a differentiation of cells in the central axis of the blastema.The amount of cartilage regenerated appears to depend on the mass of the blastema. When the blastema does not extend fully into the glenoid cavity of the scapula, the head of the developing humerus is deficient in size and structure. However, a complete limb skeleton is regenerated when the blastema does extend fully into the glenoid cavity.Since the new cartilage of the regenerated limb has no genetic continuity with the old limb skeleton, it would appear that the limb field exerts some kind of histogenetic determining action.
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  • 38
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    Journal of Morphology 62 (1938), S. 263-297 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Artemia salina, a very widely distributed species in North America, Europe and Asia, breeds freely and rapidly under laboratory conditions, living in all salinities from fresh water to almost saturated brine. Its soft exoskeleton permits of easy sectioning.The morphology and embryology of the segmentally arranged excretory glands - the antennal, mandibular and maxillary glands, are described. The antennal gland, consisting of extracellular end sac and intracellular excretory duct, is located in the second antenna and opens to the exterior on the lateroposterior face of the protopodite. It attains its maximum development by the sixth instar, degenerating by the tenth. The vestigial mandibular gland consists only of an end sac surrounding a schizocoele, and a rudimentary duct. The large maxillary gland in the adult consists of an end sac, an excretory duct of three circular coils, and a terminal duct opening to the exterior. The maxillary gland develops from a mass of mesenchyme cells in the maxillary segment of the nauplius, becoming fully differentiated by the sixth instar.The vestigial second maxilla has two bristles. It serves only as a terminus for the maxillary gland. A pair of small ganglia are accepted as internal evidence for the presence of a maxillular segment.The presence of three pair of schizocoeles in three consecutive segments is interpreted as evidence of homology with the Annelid coelome, with the possibility of further homologizing the excretory ducts with the segmental nephridia.
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  • 39
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    Journal of Morphology 62 (1938), S. 503-521 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In early embryonic stages of Passalus there are, in addition to the brain and suboesophageal ganglion, three thoracic and ten abdominal ganglia; one ganglion to each body segment. Before hatching the tenth, ninth and eighth abdominal ganglia coalesce. During the three larval instars the terminal ganglion remains in the sixth abdominal segment and only minor changes occur. By the end of the third day of pupal life the adult form of the nervous system is practically assumed. All abdominal ganglia are fused into a single, solid, elongated ganglionic mass. Connectives have disappeared between meso- and meta-thoracic and between metathoracic and abdominal ganglia; and with exception of the brain, sub-oesophageal and prothoracic ganglia, the entire ventral chain has come to lie in the mesothorax. The peripheral nerves still arise from the ganglia and ganglionic mass in their same relative positions and still supply the same segments in which they were originally located.
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  • 40
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    Journal of Morphology 62 (1938), S. 599-607 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Analysis of measurements of the trunk region of the vertebral column and of the various segments of the wing and leg skeleton in sixty-four specimens (thirty-four males, thirty females) of Fulica americana shows that: The males average about 7% larger (longer) than females in all parts of the skeleton; the difference is uniform throughout the body. The coefficient of variability ranges from about 3.00 (humerus, et al.) to about 5.00 (hind toe). The correlation between lengths of parts is very high between different segments of the wing skeleton, or of the leg; it is somewhat less between homologous segments of the wing and leg; and it is still less between limbs and trunk. This decreasing order of correlation indicates an increasing order of independence of variability. The wing proportions in Fulica are those characteristic of the type of flight called flatterflug. The proportions of the leg are those of a bird moderately adapted for cursorial locomotion; the very long toes characterize a swimming bird.
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  • 41
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: Cyto-centrosomes containing centrioles but lacking asters originate de novo in great number throughout the ooplasm of Habrobracon eggs during the formation of the first maturation metaphase. These cyto-centrosomes apparently arise from accessory nuclei which are formed from granules extruded by the germinal vesicle. The accessory nuclei react negatively to the Feulgen nucleal test.Asters similar to those present in artificially parthenogenetic eggs appear around many of the cyto-centrosomes, thus forming cytasters. These are manifest only after the female pronucleus establishes an association with a cyto-centrosome, which thus becomes the nuclear centrosome.In the parthenogenetic egg the centriole of the nuclear centrosome is continuous from one cell generation to the next, whereas the centrioles of the cyto-centrosomes which are unassociated with nuclear material are not perpetuated and disappear during early cleavage.Concerning the origin of the first cleavage centrioles in fertilized eggs, evidence shows that in some ova it is entirely maternal, that is, from two cyto-centrosomes, one associated with the female pronucleus and occupying one pole of the first mitotic spindle, and the other associated with the male pronucleus and located at the opposite pole; while in other ova it is maternal-paternal; from two centrosomes, one the sperm centrosome which occupies one pole of the first cleavage spindle, and the other a cyto-centrosome associated with the female pronucleus and situated at the opposite pole.
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  • 42
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    Journal of Morphology 62 (1938), S. 243-261 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Collections of Campeloma rufum were made at all seasons of the year from the Salt Fork River at Homer Park, Illinois, for gross anatomical and histological studies. No males nor male reproductive cells were found, all individuals showing definite female structures. The gonad, found on the columellar surface of the digestive gland, shows ovarian cells. A U-shaped tube from the albumen gland to the uterus receives the oviduct near its proximal end. This duct leads into a glandular pocket, the shell gland, which in turn empties into the sac-like uterus. The uterus terminates near the edge of the mantle in a muscular vaginal tube. The previously undescribed kidney, a glandular, pyramidal organ, lies posterior and to the left of the upper end of the mantle cavity. The kidney possesses two orifices opening into the pericardium and ureter respectively. The ureter opens between the vagina and the anus. Intra-uterine young were observed during all seasons of the year varying progressively from small veliger to fully formed young. A later paper on oogenesis, will attempt to substantiate parthenogentic reproduction for C. rufum.
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  • 43
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A detailed study of Wright's polydactylous monster (produced by a semi-dominant lethal gene) indicates that it belongs to a general type found also in rare human cases. The diagnostic characteristics are: clubbed feet and approximately double the usual number of digits, embryonic posture, microphthalmia and enlargement of the diencephalon, and missing tibia and telescoped sternum; all organ systems in the body except the genital and circulatory are grossly abnormal. The defects appear to be produced by an arrest of morphogenesis and an alteration of relative growth rates. It is indicated that a controlling center of digit formation exists on the lateral (postaxial) side of the foot, that skeletal and dermal structures are controlled by it, but that muscles are differentiated according to the area of the limb in which they lie. The gene itself is not atavistic, although its effects in the heterozygote have that appearance.
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  • 44
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    Journal of Morphology 62 (1938), S. 393-413 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The microscopical examination of the first haploid salamander to live through metamorphosis shows that the transformation to a terrestrial vertebrate was normal and practically completed at the time of its death. All the organs and structures that develop shortly before and during metamorphosis and are typical for the adult salamander, e.g., the integument, the pharyngeal derivatives, and various glands of special function, are present.The nuclei in all organs and tissues of the haploid animal are smaller than normal. This is indirect evidence of the uniformly haploid condition of the animal. The majority of the organs are smaller than in the control but contain a larger number of cells. In some glands, the reduced cell size has been completely compensated by the increase in cell number.
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  • 45
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    Journal of Morphology 62 (1938), S. 375-391 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A study is made of the pancreatic islands in Rana clamitans, R. catesbiana, R. sylvatica and Hyla versicolor during various stages of development. The islands, which were first seen in the 8-mm. R. clamitans larvae, originate from the endodermal cells of the primitive pancreatic anlagen. Following early larval development in all of the species, new islands differentiate from the cells of the pancreatic ductules. During metamorphosis, much of the acinous tissue degenerates, but the islands persist. They become aggregated and the definitive islands are formed from the primary islands before the mature adult stage is reached.With the azure-eosin technique, two types of island cells were distinguishable before, or at the time of metamorphosis, in all of the anurans studied. They are designated as the red and blue types. The blue cells are considered the primitive or embryonic type, and may transform into the red type; or the red type may originate independently.
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  • 46
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    Journal of Morphology 62 (1938), S. 523-557 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ovaries of bats, Myotis lucifugus lucifugus and M. grisescens, have been studied in all phases, except gestation, of the annual reproductive cycle. Oocytes arise in the germinal epithelium and grow with an orderly differentiation of the storage products of metabolism - chondriosomes, fat drops, and yolk vesicles. The evidence indicates that the zona pellucida is derived from the oocyte. Squamous cells of the primary follicle become cuboidal and then columnar as follicular growth begins (unilaminar secondary stage). The secondary follicle becomes multilaminar and in a very few cases gives rise to a tertiary follicle by antrum formation. Most growing follicles undergo retrogression by one of two methods. Degeneration of type I is common in multilaminar follicles and begins in the granulosa which is almost completely obliterated before the oocyte is affected. Spindles occur in some of these oocytes and fragmentation is common; phagocytes eliminate the contents of the zona. In type II, which is rare and has been found only in unilaminar secondary follicles, the oocyte degenerates, leaving an uninjured granulosa. Medullary cords are common embryonic vestiges in these ovaries; epithelial nodules are less frequent. Interstitial cells are very numerous in newborn bats, arise throughout life from the hypertrophied thecae internae of retrogressing follicles of type I, are phagocytized and, also, undergo hyaline degeneration, are hypertrophied in late pregnancy, and continue so during lactation.
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  • 47
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    Notes: Separation of the mantle from the visceral mass on the right side in deeply cupped species of oyster provides a shorter channel for egress of water from the right demibranchs. Correlated with this change is reduction in size of the right epibranchial chamber beneath the adductor together with backward displacement of the adductor itself. This water passage, designated the promyal chamber, develops during the first week after attachment of the oyster larva. The greatest displacement of the adductor and the largest promyal chamber occur in O. frons. The mantle border shows three reduplications of which the innermost, the pallial curtain, controls the amount and the place of entrance and egress of water. The importance of pallial curtains and promyal chamber to survival in turbid waters is discussed; oysters with the promyal chamber have invaded the river mouths, the flat oysters without this chamber have remained in the clear waters of high salinity near the sea. The bearing of these factors on the disappearance of fossil oysters is considered. It is proposed to relate to the genus Ostrea all flat, larviparous oysters which lack the promyal chamber and to raise the subgenus Gryphaea to generic rank to include all deeply cupped oviparous oysters with the promyal chamber. Anatomical and histological features of the pallium and branchial chambers are considered in detail.
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  • 48
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    Journal of Cellular Biochemistry 60 (1996), S. 4-11 
    ISSN: 0730-2312
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Genetic analysis of programmed cell death in Caenorhabditis elegans has led to the identification of 13 genes that constitute a developmental pathway of programmed cell death. Two of the three key genes in this pathway, ced-9, a cell death suppressor, and ced-3, a cell death inducer, were found to encode proteins that share structural and functional similarities with the mammalian proto-oncogene product Bcl-2 and interleukin-1β converting enzyme, respectively. These results suggest that the genetic pathway of programmed cell death may be evolutionarily conserved from worms to mammals. © 1996 Wiley-Liss, Inc.
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  • 49
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    Journal of Cellular Biochemistry 60 (1996), S. 12-17 
    ISSN: 0730-2312
    Keywords: bcl-2 gene ; localization ; apoptosis ; antioxidants ; oxidative stress ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The bcl-2 gene has a unique function among mammalian oncogenes as a negative regulator of apoptosis. Its expression pattern in embryonic and adult tissues is consistent with a role in maintaining in vivo survival of specific cell types.The biochemical function of bcl-2 is unknown, but its localization to mitochondrial and microsomal membranes suggests several possibilities, bcl-2 is protective against oxidative stress in mammalian cells and can be replaced by antioxidants in a factor-deprivation model of apoptosis. These results are consistent with a model of apoptotic death involving oxidative stress in a central pathway.The recent discovery of several bcl-2-related genes, some of which also inhibit apoptosis and others that unexpectedly promote apoptosis, has shed new light on several aspects of bcl-2 action. © 1996 Wiley-Liss, Inc.
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  • 50
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    Journal of Cellular Biochemistry 60 (1996), S. 33-38 
    ISSN: 0730-2312
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: No abstract.
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  • 51
    ISSN: 0730-2312
    Keywords: BCL-2 gene ; Bcl-2 protein ; homologs ; homo- and heterotypic dimers ; cancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The BCL-2 gene was first discovered because of its involvement in the t(14;18) chromosomal translocations commonly found in lymphomas, which result in deregulation of BCL-2 gene expression and cause inappropriately high levels of Bcl-2 protein production. Expression of the BCL-2 gene can also become altered in human cancers through other mechanisms, including loss of the p53 tumor suppressor which normally functions as a repressor of BCL-2 gene expression in some tissues. Bcl-2 is a blocker of programmed cell death and apoptosis that contributes to neoplastic cell expansion by preventing cell turnover caused by physiological cell death mechanisms, as opposed to accelerating rates of cell division. Overproduction of the Bcl-2 protein also prevents cell death induced by nearly all cytotoxic anticancer drugs and radiation, thus contributing to treatment failures in patients with some types of cancer. Several homologs of Bcl-2 have recently been discovered, some of which function as inhibitors of cell death and others as promoters of apoptosis that oppose the actions of the Bcl-2 protein. Many of these Bcl-2 family proteins can interact through formation of homo- and heterotypic dimers. In addition, several nonhomologous proteins have been identified that bind to Bcl-2 and that can modulate apoptosis. These protein-protein interactions may eventual serve as targets for pharmacologically manipulating the physiological cell death pathway for treatment of cancer and several other diseases. © 1996 Wiley-Liss, Inc.
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  • 52
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    Journal of Cellular Biochemistry 60 (1996), S. 61-82 
    ISSN: 0730-2312
    Keywords: protein kinases ; cyclins ; nuclear import ; NLS ; acidic domains ; cell cycle ; phosphatases ; p34cdc2 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Karyophilic and acidic clusters were found in most nonmembrane serine/threonine protein kinases whose primary structure was examined. These karyophilic clusters might mediate the anchoring of the kinase molecules to transporter proteins for their regulated nuclear import and might constitute the nuclear localization signals (NLS) of the kinase molecules. In contrast to protein transcription factors that are exclusively nuclear possessing strong karyophilic peptides composed of at least four arginines (R) and lysines (K) within an hexapeptide flanked by proline and glycine helix-breakers, protein kinases often contain one histidine and three K + R residues; this is proposed to specify a weak NLS structure resulting in the nuclear import of a fraction of the total cytoplasmic kinase molecules as well as in their weak retention in the different ionic strength nuclear environment. Putative NLS peptides in protein kinases may also contain hydrophobic or bulky aromatic amino acids proposed to further diminish their capacity to act as strong NLS. Most kinases lacking karyophilic clusters (c-Mos, v-Mos, sea star MAP, and yeast KIN28, SRA1, SRA3, TPK1, TPK2) also lack acidic clusters, which is in contrast to most kinases containing both acidic and karyophilic peptides; this and the presence of R/K clusters in the transporter proteins supports a role of acidic clusters on kinases in nuclear import. Cyclins B lack karyophilic signals and are proposed to be imported into nuclei via their association with Cdc2. © 1996 Wiley-Liss, Inc.
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  • 53
    ISSN: 0730-2312
    Keywords: protein kinase FA/GSK-3α ; PKC inhibition ; calphostin C ; down-regulation ; carcinoma dedifferentiation/progression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The signal transduction mechanism of protein kinase FA/GSK-3α by tyrosine phosphorylation in A431 cells was investigated using calphostin C as an inhibitor for protein kinase C (PKC). Kinase Fa/GSK-3α could be tyrosine-dephosphorylated and inactivated to ∼ 10% of control in a concentration-dependent manner by 0.1-10 μM calphostin C (IC50, ∼ 1 μM), as demonstrated by immunoprecipitation of kinase Fa/GSK-3α from cell extracts, followed by phosphoamino acid analysis and by immunodetection in an antikinase Fa/GSK-3α immunoprecipitate kinase assay. In sharp contrast, down-regulation of PKC by 0.05 μM calphostin C (IC50, ∼ 0.05 μM for inhibiting PKC in cells) or by tumor promoter phorbol ester TPA was found to have stimulatory effect on the cellular activity of kinase Fa/GSK-3α, when processed under identical conditions. Furthermore, TPA-mediated down-regulation of PKC was found to have no effect on calphostin C-mediated tyrosine dephosphorylation/inactivation of kinase Fa/GSK-3α. Taken together, the results provide initial evidence that the PKC inhibitor calphostin C may induce tyrosine dephosphorylation/inactivation of kinase Fa/GSK-3α in a pathway independent of TPA-mediated down-regulation of PKC, representing a new mode of signal transduction for the regulation of this multisubstrate/multifunctional protein kinase by calphostin C in cells. Since kinase Fa/GSK-3α is a possible carcinoma dedifferentiation/progression-promoting factor, the results further suggest calphostin C as a potential anticancer drug involved in blocking carcinoma dedifferentiation/progression, possibly via inactivation of protein kinase FA/GSK-3α in tumor cells. © 1996 Wiley-Liss, Inc.
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  • 54
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    Journal of Cellular Biochemistry 60 (1996), S. 363-378 
    ISSN: 0730-2312
    Keywords: cyclin D1 function ; CDK activity ; pRB phosphorylation ; G1 phase ; cell cycle control ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The sequential transcriptional activation of cyclins, the regulatory subunits of cell cycle specific kinases, regulates progress through the cell cycle. In mitogen-stimulated cells cyclin D1 induction in early G1 is followed by induction of cyclin E, activation of the cyclin-dependent kinase Cdk2, and hyperphosphorylation of the retinoblastoma gene product (pRB) in mid-to-late G1 phase. T-47D breast cancer cells expressing cyclin D1 under the control of a metal-responsive metallothionein promoter were used to determine whether Cdk2 activation and pRB hyperphosphorylation are consequences of cyclin D1 induction. A 4-5-fold increase in cyclin D1 protein abundance was followed by approximately 2-fold increases in cyclin E protein abundance and Cdk2 activity and by hyperphosphorylation of pRB. These responses were apparent ∼ 3 h after the increase in cyclin D1 protein, and ∼ 3 h prior to the entry of cyclin D1-stimulated cells into S phase 12 h after zinc treatment. Cyclin D1 immunoprecipitates contained Cdk4 but no detectable Cdk2 and displayed pRb but not histone H1 kinase activity. Cdk2 activation was therefore likely to be due to increased abundance of cyclin E/Cdk2 complexes rather than formation of active cyclin D1/Cdk2 complexes. The sequence of events following zinc induction of cyclin D1 thus mimicked that following mitogen induction of cyclin D1. These data show that cyclin D1 induction is sufficient for Cdk2 activation and pRB hyperphosphorylation in T-47D human breast cancer cells, providing evidence that cyclin D1 induction is a critical event in G1 phase progression. © 1996 Wiley-Liss, Inc.
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  • 55
    ISSN: 0730-2312
    Keywords: heregulin ; transformation ; erb B-2 ; c-Ha-ras ; mammary cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Heregulin β1 was found to stimulate the anchorage-dependent, serum-free growth of nontransformed human MCF-10A mammary epithelial cells. Unlike epidermal growth factor, transforming growth factor α, or amphiregulin, heregulin β1 was also able to induce the anchorage-independent growth of MCF-10A cells. In contrast, the anchorage-dependent, serum-free growth of c-Ha-ras or c-erb B-2 transformed MCF-10A cells was unaffected by heregulin β1, whereas heregulin β1 was able to stimulate the anchorage-independent growth of these cells. c-Ha-ras or c-erb B-2 (c-neu) transformed MCF-10A or mouse NOG-8 mammary epithelial cells express elevated levels of 2.5, 5.0, 6.5, 6.8, and 8.5 kb heregulin mRNA transcripts and/or synthesize cell-associated 25, 29, 50, and 115 kDa isoforms of heregulin. Since the MCF-10A cells and transformants also express c-erb B-3, these data suggest that endogenous heregulin might function as an autocrine growth factor for Ha-ras or erb B-2 transformed mammary epithelial cells. © 1996 Wiley-Liss, Inc. This article is a US Government work and, as such, is in the public domain in the United States of America.
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  • 56
    ISSN: 0730-2312
    Keywords: ecto-enzyme ; ALP inhibitor ; Ca incorporation ; glycosylphosphatidylinositol-anchored proteins ; PI-PLC ; bone differentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Alkaline phosphatase (ALP) activity expressed on the external surface of cultured fetal rat calvaria cells and its relationship with mineral deposition were investigated under pH physiological conditions. After replacement of culture medium by assay buffer and addition of p-nitrophenyl phosphate (pNPP), the rate of substrate hydrolysis catalyzed by whole cells remained constant for up to seven successive incubations of 10 min and was optimal over the pH range 7.6-8.2. It was decreased by levamisole by a 90% inhibition at 1 mM which was reversible within 10 min, dexamisole having no effect. Values of apparent Km for pNPP were close to 0.1 mM, and inhibition of pNPP hydrolysis by levamisole was uncompetitive (Ki = 45 μM). Phosphatidylinositol-specific phospholipase C (PI-PLC) produced the release into the medium of a p-nitrophenyl phosphatase (pNPPase) sensitive to levamisole at pH 7.8. The released activity whose rate was constant up to 75 min represented after 15 min 60% of the value of ecto-pNPPase activity. After 75 min of PI-PLC treatment the ecto-pNPPase activity remained unchanged despite the 30% decrease in Nonidet P-40-extractable ALP activity. High levels of 45Ca incorporation into cell layers used as index of mineral deposition were decreased by levamisole in a stereospecific manner after 4 h, an effect which was reversed within 4 h after inhibitor removal, in accordance with ecto-pNPPase activity variations. These results evidenced the levamisole-sensitive activity of a glycosylphosphatidylinositol-anchored pNPPase consistent with ALP acting as an ecto-enzyme whose functioning under physiological conditions was correlated to 45Ca incorporation and permit the prediction of the physiological importance of the enzyme dynamic equilibrium at the cell surface in cultured fetal calvaria cells. © 1996 Wiley-Liss, Inc.
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  • 57
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    Journal of Cellular Biochemistry 60 (1996), S. 521-528 
    ISSN: 0730-2312
    Keywords: myosin heavy chains ; smooth muscle ; alternative splicing ; contractility ; myosin light chains ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The aim of our study was to determine the relation between alternatively spliced myosin heavy chain (MHC) isoforms and the contractility of smooth muscle. The relative amount of MHC with an alternatively spliced insert in the 5′ (amino terminal) domain was determined on the protein level using a peptide-directed antibody (a25K/50K) raised against the inserted sequence (QGPSFAY). Smooth muscle MHC isoforms of both bladder and myometrium but not nonmuscle MHC reacted with a25/50K. Using a quantitative Western-blot approach the amount of 5′-inserted MHC in rat bladder was detected to be about eightfold higher than in normal rat myometrium. The amount of heavy chain with insert was found to be decreased by about 50% in the myometrium of pregnant rats. Although bladder contained significantly more 5′-inserted MHC than myometrium, apparent maximal shortening velocities (Vmax) were comparable, being 0.138 ± 0.012 and 0.114 ± 0.023 muscle length per second of skinned bladder and normal myometrium fibers, respectively. Phosphorylation of myosin light chain 20 induced by maximal Ca2+/calmodulin activation was the same in bladder and myometrial fibers. These results suggest that the amount of 5′-inserted MHC is not necessarily associated with contractile properties of smooth muscle. © 1996 Wiley-Liss, Inc.
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  • 58
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    Journal of Cellular Biochemistry 61 (1996), S. 9-17 
    ISSN: 0730-2312
    Keywords: antiestrogen ; human breast cancer ; programmed cell death ; tamoxifen ; TGF-β1 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We report here that the antiestrogen tamoxifen (TAM) induces cell death in human breast cancer cell line MCF-7. We assessed the type of cell death induced by TAM in this breast cancer cell line on the basis of morphological and biochemical characteristics. Dying cells showed morphological characteristics of apoptosis, such as chromatin condensation and nuclear disintegration. DNA isolated from these cells revealed a pattern of distinctive DNA bands on agarose gel. The DNA fragmentation in MCF-7 cells induced by TAM could also be detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling. Northern blot hybridization revealed a substantial increase in the amounts of TRPM-2 and TGF-β1 mRNAs in MCF-7 cells after treatment with TAM. In contrast, the mRNA level of the estrogen-induced pS2 gene was strongly suppressed. The biological activity of TGF-β was increased at least fourfold in the media from MCF-7 cells treated with TAM. The results presented in this study suggest that TAM induces apoptosis of MCF-7 cells and it may be mediated by the secretion of active TGF-β. © 1996 Wiley-Liss, Inc.
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  • 59
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    Journal of Cellular Biochemistry 61 (1996), S. 39-47 
    ISSN: 0730-2312
    Keywords: α2M ; PAF ; RBF ; PKC ; lyso-PAF acetyltransferase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The binding of receptor-recognized forms of α2-macroglobulin (α2M) to macrophage α2M signaling receptors increases inositol-1,4,5-triphosphate synthesis and induces Ca2+ mobilization. In this report, we demonstrate that ligation of the macrophage α2M signaling receptor is also associated with synthesis of platelet activating factor (PAF) by both the de novo and remodeling pathways. Both α2M-methylamine and a cloned and expressed 20-kDa receptor binding fragment (RBF) from rat α2M+, stimulated macrophage synthesis of PAF from [3H]acetate, [3H]methylcholine, and 1-O-[3H]alkyl lyso-PAF by two- to threefold. PAF levels reached a peak in 20 min after the cells were exposed to α2M-methylamine or RBF; they remained elevated for about 1 h after ligand addition to the cells. When [3H]methylcholine was the substrate, pertussis toxin did not block PAF synthesis, but the protein kinase C inhibitor staurosporin reduced synthesis by 65-70%. Cycloheximide completely abolished the increase in synthesis of PAF by macrophages exposed to α2M-methylamine. By contrast, when [3H]acetate was employed as a precursor, staurosporin or cycloheximide did not abolish the increase in PAF synthesis. These studies suggest that protein kinase C is necessary for the induction of the de novo pathway by α2M-methylamine. Both α2M-methylamine and RBF stimulated the activity of lyso-PAF acetyltransferase by about fourfold. Both ligands also stimulated the activity of PAF acetylhydrolase by about six- to sevenfold, indicating that ligation of the α2M signaling receptor also regulates the degradation of PAF. The ability of receptor-recognized forms of α2M to regulate levels of PAF suggests that α2M-proteinase complexes not only regulate macrophage function by activating intracellular signaling but also may indirectly regulate the function of other cells that cannot bind α2M-proteinase complexes. © 1996 Wiley-Liss, Inc.
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  • 60
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    Journal of Cellular Biochemistry 61 (1996), S. 109-117 
    ISSN: 0730-2312
    Keywords: aggregin ; chemical modification ; ADP-induced platelet responses ; NBD-Cl ; cAMP ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 61
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    Journal of Cellular Biochemistry 61 (1996), S. 72-80 
    ISSN: 0730-2312
    Keywords: hypoxia ; S-adenosylmethionine ; DNA methylation ; hypomethylation ; t-RNA methyltransferase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Young rats were maintained in a 10% oxygen atmosphere for 2, 6, and 10 days and administered normal rat chow and water ad libitum. Thereafter, their hepatic S-adenosyl-L-methionine (AdoMet) and activity and mRNA levels of AdoMet synthetase were assayed. AdoMet levels decreased by 45% after 10 days; hepatic AdoMet synthetase also declined by ∼40%. In rats with low hepatic AdoMet, the mRNA level of AdoMet synthetase also declined by up to 80%. No significant change in AdoMet or AdoMet synthetase was noted in pair-fed normoxic rats. DNA hypomethylation was determined in terms of incorporation of [3H]methyl of AdoMet incorporated at unmethylated sites in DNA in reactions mediated by methylases Hpall and Sssl. As compared to the normal hepatic DNA, [3H]methyl group incorporation in the 10-day hypoxic DNA was almost double in the Hpall-mediated reaction and ∼10-fold in the Sssl-mediated reaction. Hepatic tRNA methyltransferase activity doubled after 10 days of hypoxia. However, hypoxic rats showed no detectable mRNA transcripts for c-myc and c-fos oncogenes on Northern blot analysis. These observations show that because of subnormal activity of AdoMet synthetase, hypoxic liver is depleted of AdoMet, even when the animals are administered a complete diet. However, unlike rats on chronic lipotrope-deficient diets, hypoxic rats on a complete diet show no aberrant expression of oncogenes. © 1996 Wiley-Liss, Inc.
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  • 62
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    Journal of Cellular Biochemistry 61 (1996), S. 31-38 
    ISSN: 0730-2312
    Keywords: cell density ; DNA synthesis ; Mr receptor substrates ; IRS-1 protein ; tyrosine phosphorylation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In order to examine alterations in the phosphorylation state of proteins involved in insulin action that might accompany the reduced growth state of density-arrested cells, we measured the insulin-stimulated phosphorylation of the receptor and high Mr cellular substrates of the receptor kinase in rat hepatoma cells at different cell densities. As cell density increased from 2 × 105 to 3.2 × 106 per 35-mm well, the rate of DNA synthesis fell to 22% of control, while insulin-stimulated tyrosine phosphorylation of high Mr receptor substrates (“pp185”) was enhanced to 198% of control, without a change in the abundance of insulin receptor substrate (IRS)-1 protein. In anti-IRS-1 immunoprecipitates, tyrosine phosphorylation was increased by only 30%, suggesting that increased tyrosine phosphorylation of additional high Mr proteins (e.g., IRS-2) accounted for much of the observed increase in tyrosine phosphorylation of the receptor substrates. In spite of increased tyrosine phosphorylation of IRS-1 and total pp185-related proteins, however, cells studied at high growth density exhibited a 25% decrease in IRS-1-associated phosphatidylinositol 3′-kinase activity and only a 39% increase in phosphatidylinositol 3′-kinase activity in antiphosphotyrosine immunoprecipitates. To explore the potential role of hepatic protein-tyrosine phosphatases (PTPases) in the hyperphosphorylation of pp185 proteins, we found by immunoblotting that at high cell density the intracellular PTPase PTP18 and the transmembrane PTPase LAR were reduced in abundance by 49% and 55%, respectively, while the abundance of the SH2-domain containing PTPase SH-PTP2 was increased by 48%. These data demonstrate that the attenuation of post-receptor signaling by insulin in hepatoma cells at increasing growth density involves changes in endogenous substrate phosphorylation which may result from alterations in specific PTPases implicated in the regulation of the insulin action pathway. © 1996 Wiley-Liss, Inc.
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  • 63
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    Journal of Cellular Biochemistry 61 (1996), S. 48-60 
    ISSN: 0730-2312
    Keywords: nuclear pore structure ; digitonin permeabilization ; immunofluorescence ; coiled-coil proteins ; Tpr ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have identified a component of the human nuclear pore complex and have shown that it is the product of a gene involved in oncogenic activation. A monoclonal antibody raised against purified nuclear matrix proteins recognizes a single protein with an electrophoretic mobility of approximately 300 kDa and stains the nuclear envelope in a punctate pattern typical of nuclear pores. The antibody was used to screen λgt11 human cDNA libraries, and the resulting clones were sequenced and compared to sequences in the Genbank database. An exact match was found with the human tpr (for translocated promoter region) gene, a gene shown previously to be involved in the oncogenic activation of several protein kinases. Double-label immunofluorescent microscopy with the anti-Tpr antibody and an antibody to the previously characterized nuclear pore complex protein nup153 confirms that Tpr is localized to the nuclear pore complex. Tpr is located on the cytoplasmic face of the nucleus, as demonstrated by immunofluorescent staining of cells permeabilized with digitonin. Tpr is a 2,349-amino acid protein with extensive coiled-coil domains and an acidic globular C-terminus. The protein contains 10 leucine zipper motifs and numerous sites for phosphorylation by a variety of protein kinases. Immunoprecipitation of Tpr from 32P-orthophosphate-labeled cells shows that it is a phosphoprotein. Potential functions for Tpr and possible mechanisms for the transforming activity of Tpr fusion proteins are discussed. © 1996 Wiley-Liss, Inc.
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  • 64
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    Journal of Cellular Biochemistry 61 (1996), S. 127-138 
    ISSN: 0730-2312
    Keywords: β1 integrin ; β7 integrin ; α/β integrin subunit association ; VLA-4/VCAM adhesion ; integrin surface expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We report here an analysis of the expression and function of the α chain of human VLA-4 in stable mouse L cell transfectants and the requirement for the β chain in these processes. L cells were transfected with human α4 cDNA or α4 and human β1 cDNA. Unexpectedly, human α4 cDNA, when transfected alone, could induce de novo surface expression of host β7 and increased expression of host β1. Induction of mouse β7 and β1 surface expression was not due to de novo gene activation, but instead represented α4/β intracellular subunit association and transport to the cell surface. Transfection with human β1 prevented surface expression of mouse β integrins. Whereas human α4 and human β1 subunits associated very tightly in anti-α4 immunoprecipitates, human α4 and mouse β subunits were only partially associated. Furthermore, binding of human/mouse chimeric receptors to recombinant VCAM, a major ligand for α4β7 and α4β1, was very poor, whereas human α4/human β1 receptors bound strongly to VCAM. One α4 transfectant, which exhibited a tight human α4/mouse β1 association, could be induced, but only after PMA activation, to bind strongly to VCAM. These results indicate that α4 subunits have specific affinity for β7 and β1 integrins and require β subunits for surface expression as well as high affinity ligand binding activity. Our results indicate that a tight association between the α4 and β subunit appears to be critical for ligand binding, consistent with a direct as well as regulatory role for the β subunit in ligand binding. Furthermore, these studies demonstrate that expression of foreign recombinant proteins can alter host cell protein expression resulting in de novo surface protein expression. © 1996 Wiley-Liss, Inc.
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  • 65
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    Journal of Cellular Biochemistry 61 (1996), S. 230-237 
    ISSN: 0730-2312
    Keywords: retinoic acid ; retinol ; binding ; transglutaminase ; hepatic ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: When rat liver epithelial cells were exposed to retinoic acid or retinol for 24 hr, the levels of transforming growth factor-β (TGF-β) receptors were reduced in a dose-dependent way. The decrease appeared after 12 hr of incubation with the retinoids and binding levels remained low until 24 hr after the removal of the molecules. Retinoid treatment induced a fourfold enhancement of transglutaminase (TGase) activity in the cell membranes, and cystamine, an inhibitor of TGase, prevented the decrease of the receptors. Neutralization of TGF-β by a monoclonal antibody did not suppress the decrease of the binding levels, indicating that decreased TGF-β binding capacity was not due merely to the internalization of ligand-bound receptors promoted by a stimulation of TGF-β synthesis. Thus, retinoid treatment resulted in an intense disappearance of the functional receptors from the membranes that seemed to be mediated by increased TGase activity. This phenomenon can represent a strong signal attenuation for TGF-β following retinoid exposure. © 1996 Wiley-Liss, Inc.
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  • 66
    ISSN: 0730-2312
    Keywords: human hepatoma ; dedifferentiation/progression ; PDPK ; overexpression ; kinase FA/GSK-3α ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Computer analysis of protein phosphorylation sites sequence revealed that transcriptional factors and viral oncoproteins are prime targets for regulation of proline-directed protein phosphorylation, suggesting an association of the proline-directed protein kinase (PDPK) family with neoplastic transformation and tumorigenesis. In this report, an immunoprecipitate activity assay of protein kinase FA/glycogen synthase kinase-3α (kinase FA/GSK-3α) (a member of PDPK family) has been optimized for human hepatoma and used to demonstrate for the first time significantly increased (P 〈 0.01) activity in poorly differentiated SK-Hep-1 hepatoma (24.2 ± 2.8 units/mg) and moderately differentiated Mahlavu hepatoma (14.5 ± 2.2 units/mg) when compared to well differentiated Hep 3B hepatoma (8.0 ± 2.4 units/mg). Immunoblotting analysis revealed that increased activity of kinase FA/GSK-3α is due to overexpression of the protein. Elevated kinase FA/GSK-3α expression in human hepatoma biopsies relative to normal liver tissue was found to be even more profound. This kinase appeared to be ∼fivefold overexpressed in well differentiated hepatoma and ∼13-fold overexpressed in poorly differentiated hepatoma when compared to normal liver tissue. Taken together, the results provide initial evidence that overexpression of kinase FA/GSK-3α is involved in human hepatoma dedifferentiation/progression. Since kinase FA/GSK-3α is a PDPK, the results further support a potential role of this kinase in human liver tumorigenesis, especially in its dedifferentiation/progression. © 1996 Wiley-Liss, Inc.
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  • 67
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    Journal of Cellular Biochemistry 61 (1996), S. 246-254 
    ISSN: 0730-2312
    Keywords: marrow stromal cells ; cell morphogenesis ; attachment ; ECM ; mRNA expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Our aim was to study the role of various extracellular matrices (ECM) on growth and differentiation of marrow stromal cells in vitro. Morphology changes, gene expression, and enzymatic activities were monitored in stromal osteoblastic MBA-15 and adipocytic 14F1.1 cells. These stromal cells were plated on dishes precoated with different substrata, such as matrigel (basement membrane), collagen type I, and endothelial ECM, and compared with cells plated on protein-free dishes. Striking morphological differences were observed when the cells grew on these different substrata. Changes in cell shape and growth also led to differential mRNA expression and enzymatic activities. When MBA-15 cells were plated on collagen, there was a decrease in mRNA for alkaline phosphatase (ALK-P), osteopontin (OP), and osteonectin (ON), and an increase in mRNA for procollagen (I). A differential effect was noted on 14F1.1 cells, the mRNA for ALK-P increased, the expressions of OP and ON lowered, and no expression for procollagen (I) was monitored. MBA-15 cells cultured on matrigel had decreased mRNA for ALK-P and OP, while they had increased ON mRNA expression and remained unchanged for procollagen 1. No change in mRNA expression by 14F1.1 cells was monitored when cultured on matrigel. Functional enzymatic activities of ALK-P markedly decreased in MBA-15 cells cultured on various substrata, and increased or were unchanged in 14F1.1 cells. An additional enzyme, neutral endopeptidase (CD10/NEP), altered differentially in both cell types; this enzymatic activity increased or was unchanged when cells were cultured on these matrices. The results indicate a specific role for different ECM on various stromal cell types and their function. © 1996 Wiley-Liss, Inc.
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  • 68
    ISSN: 0730-2312
    Keywords: adhesion ; migration ; protease ; lymphocyte ; immunity ; connective tissue ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Human T lymphoblastoma cells of the CD4+ 8+ Tsup-1 line, that express alpha4 and alpha5 but not alpha6 integrins of the beta1 family, and CD4+ human blood T cells bind vasoactive intestinal peptide (VIP) with high affinity, leading to increased adherence, secretion of matrix metalloproteinases (MMPs), and chemotaxis. VIP-enhanced adherence of T cells to fibronectin was inhibited significantly by neutralizing monoclonal antibodies to beta1 〉 alpha4 〉〉 alpha5, but not to alpha6. Antibodies to beta1 and alpha4 suppressed to a similarly significant extent VIP stimulation of both MMP-dependent T cell chemotaxis through fibronectin-enriched Matrigel and T cell degradation of 3H-type IV collagen in the Matrigel, without affecting VIP-evoked secretion of MMP by suspensions of T cells. The lesser inhibition of VIP-enhanced adherence of T cells to fibronectin by anti-alpha5 antibody, than antibodies to beta1 or alpha4 chains, was associated with lesser or no suppression of MMP-dependent T cell chemotaxis through Matrigel and T cell degradation of type IV collagen in the Matrigel in response to VIP. Specific beta1 integrins thus mediate interactions of stimulated T cells with basement membranes, including adherence, localized digestion by MMPs, and chemotactic passage, that promote entry of T cells into extravascular tissues. © 1996 Wiley-Liss, Inc.
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  • 69
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    Journal of Cellular Biochemistry 61 (1996), S. 459-466 
    ISSN: 0730-2312
    Keywords: adenylyl cyclase ; BAT3 ; cytoskeleton ; RAS ; signaling ; yeast ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We previously identified human CAP, a homolog of the yeast adenylyl cyclase - associated protein. Previous studies suggest that the N-terminal and C-terminal domains of CAP have distinct functions. We have explored the interactions of human CAP with various proteins. First, by performing yeast two-hybrid screens, we have identified peptides from several proteins that interact with the C-terminal and/or the N-terminal domains of human CAP. These peptides include regions derived from CAP and BAT3, a protein with unknown function. We have further shown that MBP fusions with these peptides can associate in vitro with the N-terminal or C-terminal domains of CAP fused to GST. Our observations indicate that CAP contains regions in both the N-terminal and C-terminal domains that are capable of interacting with each other or with themselves. Furthermore, we found that myc-epitope-tagged CAP coimmunoprecipitates with HA-epitope-tagged CAP from either yeast or mammalian cell extracts. Similar results demonstrate that human CAP can also interact with human CAP2. We also show that human CAP interacts with actin, both by the yeast two-hybrid test and by coimmunoprecipitation of epitope-tagged CAP from yeast or mammalian cell extracts. This interaction requires the C-terminal domain of CAP, but not the N-terminal domain. Thus CAP appears to be capable of interacting in vivo with other CAP molecules, CAP2, and actin. We also show that actin co-immunoprecipitates with HA-CAP2 from mammalian cell extracts. © 1996 Wiley-Liss, Inc.
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  • 70
    ISSN: 0730-2312
    Keywords: basic helix-loop-helix ; interleukin-1 ; interleukin-3 ; granulocyte-macrophage colony-stimulating factor ; progenitor ; transcription factor ; c-kit ligand ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The basic helix-loop-helix (bHLH) transcription factors form heterodimers and control steps in cellular differentiation. We have studied four bHLH transcription factors, SCL, lyl-1, E12/E47, and Id-1, in individual lineage-defined progenitors and hematopoietic growth factor - dependent cell lines, evaluating mRNA expression and the effects of growth factors and cell cycle phase on this expression. Single lineage-defined progenitors selected from early murine colony starts and grown under permissive conditions were analyzed by RT-PCR. SCL and E12/E47 were expressed in the vast majority of tri-, bi-, and unilineage progenitors of erythroid, macrophage, megakaryocyte, and neutrophil lineages. Expression for E12/E47 was not seen in unilineage megakaryocyte and erythroid or bilineage neutrophil/mast cell progenitors. Lyl-1 showed a more restricted pattern of expression, although expression was seen in some bi- and unilineage progenitors. No expression was detected in erythroid, erythroid-megakaryocyte-macrophage, macrophage-neutrophil, macrophage, or megakaryocytic progenitors. Id-1, an inhibitory bHLH transcription factor, was also widely expressed in all bi- and unilineage progenitors; only the trilineage erythroid-megakaryocyte-macrophage progenitors failed to show expression. Expression of these factors within a progenitor class was generally heterogeneous, with some progenitors showing expression and some not. This was seen even when two sister cells from the same colony start were analyzed. Id-1, but not E12/E47, mRNA was increased in FDC-P1 and MO7E hematopoietic cell lines after exposure to IL-3 or GM-CSF, Id-1, E12, and lyl-1 showed marked variation at different points in cell cycle in isoleucine-synchronized FDC-P1 cells. These results suggest that SCL, lyl-1, E12/E47, and Id-1 are important in hematopoietic progenitor cell regulation, and that their expression in hematopoietic cells varies in response to cytokines and/or during transit through cell cycle. © 1996 Wiley-Liss, Inc.
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  • 71
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    Journal of Cellular Biochemistry 61 (1996), S. 493-501 
    ISSN: 0730-2312
    Keywords: basement membrane ; cell binding ; epidermolysis bullosa ; extracellular matrix ; gene knock-out ; integrin ; laminin ; muscular dystrophy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Extracellular matrix molecules are often very large and made up of several independent domains, frequently with autonomous activities. Laminin is no exception. A number of globular and rod-like domains can be identified in laminin and its isoforms by sequence analysis as well as by electron microscopy. Here we present the structure-function relations in laminins by examination of their individual domains. This approach to viewing laminin is based on recent results from several laboratories. First, some mutations in laminin genes that cause disease have affected single laminin domains, and some laminin isoforms lack particular domains. These mutants and isoforms are informative with regard to the activities of the mutated and missing domains. Second, laminin-like domains have now been found in a number of other proteins, and data on these proteins may be informative in terms of structure-function relationships in laminin. Finally, a large body of data has accumulated on the structure and activities of proteolytic fragments, recombinant fragments, and synthetic peptides from laminin. The proposed activities of these domains can now be confirmed and extended by in vivo experiments. © 1996 Wiley-Liss, Inc.
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  • 72
    ISSN: 0730-2312
    Keywords: bone resorption ; tyrphostins ; genistein ; herbimycin ; osteoporosis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We compared the effects of the tyrosine kinase inhibitor genistein, a naturally occurring isoflavone, to those of tyrphostin A25, tyrphostin A47, and herbimycin on avian osteoclasts in vitro. Inactive analogs daidzein and tyrphostin A1 were used to control for nonspecific effects. None of the tyrosine kinase inhibitors inhibited bone attachment. However, bone resorption was inhibited by genistein and herbimycin with ID50s of 3 μM and 0.1 μM, respectively; tyrphostins and daidzein were inactive at concentrations below 30 μM, where nonspecific effects were noted. Genistein and herbimycin thus inhibit osteoclastic activity via a mechanism independent of cellular attachment, and at doses approximating those inhibiting tyrosine kinase autophosphorylation in vitro; the tyrphostins were inactive at meaningful doses. Because tyrosine kinase inhibitors vary widely in activity spectrum, effects of genistein on cellular metabolic processes were compared to herbimycin. Unlike previously reported osteoclast metabolic inhibitors which achieve a measure of selectivity by concentrating on bone, neither genistein nor herbimycin bound significantly to bone. Osteoclastic protein synthesis, measured as incorporation of 3H-leucine, was significantly inhibited at 10 μM genistein, a concentration greater than that inhibiting bone degradation, while herbimycin reduced protein synthesis at 10 nM. These data suggested that genistein may reduce osteoclastic activity at pharmacologically attainable levels, and that toxic potential was lower than that of herbimycin. To test this hypothesis in a mammalian system, bone mass was measured in 200 g ovariectomized rats treated with 44 μmol/day genistein, relative to untreated controls. During 30 d of treatment, weights of treated and control group animals were indistinguishable, indicating no toxicity, but femoral weight in the treated group was 12% greater than controls (P 〈 0.05). Our data indicate that the isoflavone inhibitor genistein suppresses osteoclastic activity in vitro and in vivo at concentrations consistent with its ID50s on tyrosine kinases, with a low potential for toxicity. © 1996 Wiley-Liss, Inc.
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  • 73
    ISSN: 0730-2312
    Keywords: CoA-independent transacylase ; phospholipase D ; subcellular localization ; neutrophils ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Subcellular localizations of CoA-independent transacylase and phospholipase D enzymes have been investigated in human neutrophils performing a two-step gradient system to separate plasma membranes from internal membranes and from the bulk of granules. The internal membranes were constituted by endoplasmic reticulum and by a subpopulation of specific and tertiary granules. The enzymes activities were assayed in vitro on gradient fractions using exogenous substrates. Following cell prelabelling with [3H]alkyllyso-GPC, we also analyzed the in situ localization of labelled products involving the action of both enzymes. The CoA-independent transacylase activity, together with the CoA-dependent transacylase and acyltransferase activities were only located in the internal membranes. Following 15 min cell labelling, part of the [3H]alkylacyl-GPC was recovered in plasma membranes indicating a rapid redistribution of the acylated compound. Very high contents in arachidonate containing [3H]alkylacyl-GPC were recovered both in plasma membranes and internal membranes. Phospholipase D activity being assayed in the presence of cytosol, GTPγS and gradient fractions, only the plasma membrane fractions from resting or stimulated cells allowed the enzyme to be active. The [3H]alkylacyl-GP and [3H]alkylacyl-GPethanol, phospholipase D breakdown products from [3H]alkylacyl-GPC, obtained after neutrophil prelabelling and activation by phorbol myristate acetate, were exclusively present in the plasma membranes. In contrast, the secondary generated [3H]alkylacylglycerols were equally distributed between plasma and internal membranes. No labelled product was recovered on azurophil granules. These data demonstrate that internal membranes are the site of action of the CoA-independent transacylase and plasma membranes are the site of action of the phospholipase D. This topographical separation between CoA-independent transacylase which generated substrate and phospholipase D which degraded it, suggested that subcellular localisation and traffic of substrates within the cell can be important to regulate the enzymes. © 1996 Wiley-Liss, Inc.
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  • 74
    ISSN: 0730-2312
    Keywords: FGF ; receptors ; internalization ; photoactivable cross-linker ; heparan sulfate proteoglycans ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The internalization of basic fibroblast growth factor (FGF-2) was studied in Chinese hamster lung fibroblasts (CCL39). Recombinant FGF-2 was derivatized with a photoactivable agent, N-hydroxysuccinimidyl-4-azido-benzoate (HSAB), iodinated, and used to visualize intracellular FGF-2-affinity-labeled molecules after internalization at 37°C. Iodinated HSAB-FGF-2 maintained the properties of natural FGF-2 such as affinity for heparin, binding to Bek and Flg receptors, interaction with high- and low-affinity binding sites, and reinitiating of DNA synthesis in CCL39 cells. Affinity-labeling experiments at 4°C with 125I-HSAB-FGF-2 led to the detection of several FGF-cell surface complexes with apparent molecular mass of 80, 100, 125, 150, 170-180, 220, 260, and about 320 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), whereas two specific bands at 80 and 130-160 kDa were obtained using the homobifunctional cross-linking reagent, disuccinimidyl suberate. When the cells, preincubated with 125I-HSAB-FGF-2 at 4°C and then washed, were shifted to 37°C, irradiation of the internalized labeled FGF-2 led to detection of a similar but fainted profile with one major specific band at 80 kDa. Heparitinase II treatment of the cells reduced binding of 125I-HSAB-FGF-2 to its cell surface sites by 80% and internalization by 55%, indicating the involvement of heparan sulfate proteoglycans in these processes. Among the heparitinase-sensitive bands was the 80-kDa complex. © 1996 Wiley-Liss, Inc.
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  • 75
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    Journal of Cellular Biochemistry 62 (1996), S. 275-289 
    ISSN: 0730-2312
    Keywords: nuclear matrix ; HeLa S3 cells ; 2-D gel electrophoresis ; heterogeneous nuclear ribonucleoproteins ; B23 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The nuclear matrix is the structure that persists after removal of chromatin and loosely bound components from the nucleus. It consists of a peripheral lamina-pore complex and an intricate internal fibrogranular structure. Little is known about the molecular structure of this proteinaceous internal network. Our aim is to identify the major proteins of the internal nuclear matrix of HeLa S3 cells. To this end, a cell fraction containing the internal fibrogranular structure was compared with one from which this structure had been selectively dissociated. Protein compositions were quantitatively analyzed after high-resolution two-dimensional gel electrophoresis. We have identified the 21 most abundant polypeptides that are present exclusively in the internal nuclear matrix. Sixteen of these proteins are heterogeneous nuclear ribonucleoprotein (hnRNP) proteins. B23 (numatrin) is another abundant protein of the internal nuclear matrix. Our results show that most of the quantitatively major polypeptides of the internal nuclear matrix are proteins involved in RNA metabolism, including packaging and transport of RNA. © 1996 Wiley-Liss, Inc.
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  • 76
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    Journal of Cellular Biochemistry 62 (1996), S. 314-324 
    ISSN: 0730-2312
    Keywords: M1 cell ; heme oxygenase ; transcription ; H2O2 ; TPA ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: It has long been known that heme oxygenase (HO) is a key enzyme in heme catabolism and recently it was also found to acts as an oxidative stress protein to produce carbon monoxide (CO), which has similar actions to those of nitrogen monoxide (NO). Therefore, we examined transcriptional control of the HO gene in mouse M1 (myeloleukemia) cells during their differentiation into macrophages. Since the promoter region of this gene is known to have a TPA-responsive element (TRE), its expression might be regulated by a C-kinase signal transduction pathway. Then we investigated the activation of the HO gene after treatment of M1 cells with TPA and inhibitors of C-kinase. When M1 cells were treated with TPA, they differentiated into macrophage-like cells. Upon treatment with TPA, H2O2 was produced first, the nuclear proto-oncogenes fos and jun were activated, and then the HO gene was activated. The extent of transcriptional activation of the fos, jun, and HO genes in M1 cells treated with TPA was reduced by a specific inhibitor of C-kinase and a scavenger of oxygen radicals. When M1 cells were treated with H2O2 essentially the same level of transcription of the HO gene was observed, but the extent of transcriptional activation of the fos and jun genes was about half of the treatment with TPA. Super-shift assays using the TRE of the HO gene revealed that the Fos and Jun proteins from nuclei of M1 cells treated with TPA bound to the TRE, and same assays using DNA with the NF-kB motif also revealed that the active NF-kB protein from M1 cells treated with H2O2 or TPA also bound to the corresponding motif. These results strongly suggest that the HO gene in M1 cells is activated by TPA through a production of H2O2, an oxidative activation pathway of NF-kB, and a signal-transduction pathway that involves C-kinase during the differentiation of macrophages that occurs upon treatment with TPA. © 1996 Wiley-Liss, Inc.
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  • 77
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    Journal of Cellular Biochemistry 62 (1996), S. 172-180 
    ISSN: 0730-2312
    Keywords: chromatin structure ; nuclear matrix ; transcriptional activation ; replication ; recombination ; differentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The tumor suppressor p53 is a multifunctional protein whose main duty is to preserve the integrety of the genome. This function of wild-type p53 as “guardian of the genome” is achieved at different levels, as a cell cycle checkpoint protein, halting the cell cycle upon DNA damage, and via a direct involvement in processes of DNA repair. Alternatively, p53 can induce apoptosis. Mutations in the p53 gene occur in about 50% of all human tumors and eliminate the tumor suppressor functions of p53. However, many mutant p53 proteins have not simply lost tumor suppressor functions but have gained oncogenic properties which contribute to the progression of tumor cells to a more malignant phenotype. The molecular basis for this gain of function of mutant p53 is still unknown. However, mutant (mut) p53 specifically binds to nuclear matrix attachment region (MAR) DNA elements. MAR elements constitute important higher order regulatory elements of chromatin structure and function. By binding to these elements, mut p53 could modulate important cellular processes, like gene expression, replication, and recombination, resulting in phenotypic alterations of the tumor cells. Mut p53 thus could be the first representative of a new class of oncogenes, which exert their functions via long-range alterations or perturbation of chromatin structure and function. © 1996 Wiley-Liss, Inc.
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  • 78
    ISSN: 0730-2312
    Keywords: dexamethasone ; actin ; polymerization ; Ishikawa cells ; cAMP ; actinomycin D ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Glucocorticoids, in addition to their well characterized effects on the genome, may affect cell function in a manner not involving genomic pathways. The mechanisms by which the latter is achieved are not yet clear. A possible means for this action may involve the actin cytoskeleton, since the dynamic equilibrium of actin polymerization changes rapidly following exposure to several stimuli, including hormones. The aim of the present work was to find out if glucocorticoids exert rapid, nongenomic effects on actin polymerization in Ishikawa human endometrial cells, which represent a well characterized in vitro cell model expressing functional glucocorticoid receptors. Short term exposure of the cells to the synthetic glucocorticoid dexamethasone resulted in an overall decrease of the G/total-actin ratio in a time- and dose-dependent manner. Specifically, in untreated Ishikawa cells the G/total-actin ratio was 0.48 ± 0.01 (n = 26). It became 0.35 ± 0.01 (n = 13, P 〈 0.01) following exposure to 10-7 M dexamethasone for 15 min. This was induced by a significant decrease of the cellular G-actin level, without affecting the total actin content, indicating a rapid actin polymerization. This conclusion was fully confirmed by direct fluorimetry measurements, that showed a significant increase of the F-actin content by 44% (n = 6, P 〈 0.001) in cells treated with dexamethasone (10-7 M, 15 min). The rapid dexamethasone-induced alterations of the state of actin polymerization were further supported by fluorescence microscopy. The latter studies showed that the microfilaments of cells pretreated with 10-7 M dexamethasone for 15 min were more resistant to various concentrations of the antimicrofilament drug cytochalasin B, compared to untreated cells, implying microfilament stabilization. The action of dexamethasone on actin polymerization seems to be mediated via specific glucocorticoid binding sites, since the addition of the glucocorticoid antagonist RU486 completely abolished its effect. Moreover, it appears to act via non-transcriptional pathways, since actinomycin D did not block the dexamethasone-induced actin polymerization. In addition, cell treatment with 10-7 M dexamethasone for 15 min fully reversed the forskolin-, but not the 8-bromo-cAMP-induced actin depolymerization. In line with these findings, the cAMP content of Ishikawa cells was decreased by 29.2% after a 15 min treatment with 10-7 M dexamethasone (n = 4, P 〈 0.01). In conclusion, our results showed that dexamethasone induces rapid, time-, and dose-dependent changes in actin polymerization dynamics in Ishikawa cells. This action seems to be mediated via cAMP, involving probably nongenomic pathways. The above findings offer new perspectives for the understanding of the early cellular responses to glucocorticoids. © 1996 Wiley-Liss, Inc.
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  • 79
    ISSN: 0730-2312
    Keywords: monocyte chemoattractant protein-1 ; gene expression ; pig artery ; balloon injury ; monocyte/macrophages ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) are potent chemokines which attract circulating monocytes and neutrophils respectively to inflamed tissues. JE/MCP-1 gene expression has been previously studied in rabbit aortae after endothelial denudation and the rapid appearance of this transcript was thought to precede emigration of phagocytes. We now report MCP-1 gene expression following de-endothelialization of iliac arteries in the pig, a species which can develop spontaneous atherosclerosis. Using Northern blot analysis, we demonstrated that MCP-1 mRNA was rapidly induced in pig arteries at 2 h and continued to increase to reach a maximum at 8 h before returning to low levels at 16-24 h after injury. The increase seen for MCP-1 mRNA at 8 h was also observed for IL-8 mRNA but was not apparent for growth-related gene expressions, urokinase-type plasminogen activator (u-PA), and plasminogen activator inhibitor-1 (PAI-1). Since smooth muscle cells, endothelial cells, and phagocytes are all capable of expressing MCP-1, we examined pig arteries for immunostaining using a monoclonal antibody to human MCP-1 (5D3-F7). At 8 h after injury, the predominant cell type staining positive for MCP-1 was the monocyte/macrophage. Staining was also observed in occasional scattered neutrophils, but MCP-1 protein could not be detected in smooth muscle cells or on extracellular matrix within the sensitivity constraints posed by our methodology. Our results are consistent with invading monocyte/macrophages having a major input into the production of this chemokine in the arterial wall following injury. The fact that MCP-1 expression accompanied monocyte/macrophage presence in damaged artery, rather than preceding it, is suggestive that continued MCP-1 expression is required for functions other than chemoattraction. © 1996 Wiley-Liss, Inc.
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  • 80
    ISSN: 0730-2312
    Keywords: GLRP ; T-lymphocyte ; immune response ; central nervous system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Coordination of the immune response to injury or disease in the brain is postulated to involve bi-directional discourse between the immune system and the central nervous system. This cross communication involves soluble mediators, including various growth factors, cytokines, and neuropeptides. In this report, we demonstrate that the supernatant from activated T-lymphocytes is able to induce the transcription of a potent cytokine, TGF-β2 in glial cells. The activating stimulus invokes signaling mechanisms distinct from known kinase or protease pathways. Activation of TGF-β2 transcription correlates with the loss of binding activity for an 80 kDa glial labile repressor protein, GLRP, to a responsive region within the TGF-β2 promoter. Although GLRP shares some characteristics with the inducible transcription factor AP-1, it appears to be distinct from known AP-1 family members. These data along with previous observations demonstrating the potent immunosuppressive activity of TGF-β2, support a model for a feedback mechanism between the activated T-lymphocytes and astrocytes via TGF-β2 to regulate the immune response. © 1996 Wiley-Liss, Inc.
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  • 81
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    Journal of Cellular Biochemistry 62 (1996), S. 454-466 
    ISSN: 0730-2312
    Keywords: nuclear matrix ; histone H5 ; transcription ; transcription factors ; erythroid development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The nuclear matrix has roles in organizing nuclear DNA and in controlling transcription. Transcription factors are associated with the nuclear matrix, with the spectra of transcription factors differing from one cell type to another. In this study we identified the transcription factors and enzymes functioning in the regulation of gene expression that were associated with nuclear matrix and nonmatrix nuclear fractions in erythrocytes isolated from chick embryos at different stages of development, anemic and normal adult birds. We found that the primitive erythroid nuclear matrix had the greatest histone deacetylase activity and highest levels of several transcription factors, including GATA-1, CACCC-binding proteins, and NF1. These transcription factors have key roles in erythroid-specific gene expression. The levels of these transcription factors were lower in the nonmatrix and matrix fractions isolated from definitive erythrocytes. For primitive and definitive erythrocytes, the level of CACCC-binding proteins in the nuclear matrix fraction was greater than that of Sp1. The relative levels of these transcription factors were reversed in the nonmatrix fraction. Casein kinase II was not found in erythroid nuclear matrices. The observed erythroid lineage specific alterations in erythroid nuclear matrix transcription factor composition and abundance may be involved in erythroid-specific gene expression. © 1996 Wiley-Liss, Inc.
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  • 82
    ISSN: 0730-2312
    Keywords: adhesion ; breast cancer cells ; thrombospondin ; receptors ; proteoglycans ; heparin-binding peptides ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Thrombospondin is an adhesive glycoprotein that promotes breast cancer cell adhesion to human vascular endothelial cells (Incardona et al., 1995). In this study, we have identified the molecular domains of thrombospondin that mediate its binding to specific receptors on the human breast adenocarcinoma cell line, MDA-MB-231. Two recombinant fragments from the amino-terminus (TSPN18 and TSPN28), and the fusion proteins of the type 1 and type 2 repeats of human thrombospondin, inhibited binding of radiolabeled thrombospondin to MDA-MB-231 cells in suspension by 40-60% at 50 μg/ml whereas the type 3 repeat, carboxy-terminus and unfused glutathione-S-transferase as well as the synthetic peptide Gly-Arg-Gly-Asp-Ser (500 μg/ml) had little or no effect. Herapin and various glycosaminoglycans as heparan sulfate, chondroitin sulfates A, B or C, and fucoidan inhibited thrombospondin binding to MDA-MB-231 cells by more than 60% whereas dextran sulfate had only little effect. Treatment of cells with heparitinase, chondroitinase ABC, and hyaluronidase, but not with neuraminidase, induced 30-50% inhibition of thrombospondin binding suggesting the participation of both heparan sulfate and chondroitin sulfate cell surface-associated molecules. Inhibition of proteoglycan sulfation by chlorate or inhibition of glycosaminoglycan chain formation by two β-D-xylosides also led to a substantial inhibition of thrombospondin binding. Our results indicate that several domains within the thrombospondin molecule, namely the amino-terminus, type 1 and type 2 repeats, participate in its binding to specific receptors bearing sulfated glycosaminoglycans on MDA-MB-231 cells. Biological assays have indicated that, in addition to these domains, the peptide Gly-Arg-Gly-Asp-Ser inhibited MDA-MB-231 cell attachment to thrombospondin suggesting that the last type 3 repeat of the molecule may also contribute to its cell adhesive activity. © 1996 Wiley-Liss, Inc.
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  • 83
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    Journal of Cellular Biochemistry 62 (1996), S. 506-515 
    ISSN: 0730-2312
    Keywords: heat shock ; pre-rRNA processing ; S-100 extract ; U3 snoRNA ; 3′ processing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The effect of heat shock on pre-rRNA processing at the primary site within external transcribed spacer region 1 (ETS1) was studied in S-100 extract derived from mouse lymphosarcoma cells. In vivo labeling with [32P]orthophosphate showed that the synthesis of the rRNA precursor and its processing to 28S and 18S rRNAs were inhibited significantly due to heat shock. The processing activity was reduced by 50% at 1 h and was completely blocked following 2-h exposure of cells at 42°C. Mixing S-100 extracts from the control and heat-treated cells did not affect the processing activity in the control extract, which proves the absence of a nuclease or other inhibitor(s) of processing in the extract from the heat-shocked cells. Heat shock did not affect interaction between pre-rRNA and U3 snoRNA, a prerequisite for the processing at the primary site, but significantly altered RNA-protein interaction. Three polypeptides of 200, 110, 52 kDa that specifically cross-link to pre-rRNA spanning the primary processing site were inactivated after heat shock. Hyperthermia did not alter 3′ end processing of SV40L pre-mRNA. © 1996 Wiley-Liss, Inc.
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  • 84
    ISSN: 0730-2312
    Keywords: osteosarcoma ; chondrosarcoma ; GCT ; oncogene alterations ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We investigated the structure and the expression of various oncogenes in three of the most common human bone tumors - osteosarcoma (36 samples from 34 patients), giant cell tumor (10 patients), and chondrosarcoma (18 patients) - in an attempt to identify the genetic alterations associated with these malignancies. Alterations of RB and p53 were detected only in osteosarcomas. Alterations of c-myc, N-myc, and c-fos were detected in osteosarcomas and giant cell tumors. Ras alterations (H-ras, Ki-ras, N-ras) were rare. Chondrosarcomas did not contain any detectable genetic alterations. Our results suggest that alterations of c-myc, N-myc, and c-fos oncogenes occur in osteosarcomas, in addition to those previously described for the tumor suppressor genes RB and p53. Moreover, statistical analyses indicate that c-fos alterations occur more frequently in osteosarcoma patients with recurrent or metastatic disease. © 1996 Wiley-Liss, Inc.
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  • 85
    ISSN: 0730-2312
    Keywords: Src kinase ; mercuric chloride ; redox ; sulfhydryl group ; receptor polymerization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Little is known about the regulatory mechanism of c-Src kinase in cells except the suggested regulation through phosphorylation and dephosphorylation of its carboxyl terminal tyrosine residue (Y527). We here demonstrated that exposure of NIH3T3 cells to mercuric chloride (HgCl2) induces both aggregation and activation of Src kinase protein through a redox-linked mechanism. The aggregation of Src proteins was suggested to be induced by the sulfhydryl groups-to-Hg2+ reaction-mediated polymerization of cell membrane proteins to which the Src proteins associate noncovalently. The possibility was ruled out that the aggregation occurred secondarily to the promotion of protein tyrosine phosphorylation. Further study revealed that the Src kinase was activated by HgCl2 at least in part independent of the known Csk kinase-linked or Y527-phosphorylation/dephosphorylation-mediated control. Correspondingly, CNBr cleavage mapping of phosphopeptides for autophosphorylated c-Src protein demonstrated selective promotion of phosphorylation at Y416 in HgCl2-treated cells without obvious change in the phosphorylation level at Y527. These results suggest a unique protein sulfhydryl modification-based pathway of signal transduction for activating Src kinase in NIH3T3 cells. © 1996 Wiley-Liss, Inc.
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  • 86
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    Journal of Cellular Biochemistry 63 (1996), S. 162-173 
    ISSN: 0730-2312
    Keywords: Topo IIα ; Topo IIβ ; interphase ; mitosis ; mitogenic stimulation ; nucleoplasm ; nucleolus ; lymphocytes ; HeLa ; immunofluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have carried out immunofluorescence labelling of two human cell types, HeLa cells and peripheral blood lymphocytes, prepared by several different fixation/permeabilization protocols using a variety of antibodies against DNA Topoisomerase II (Topo II). We have found that the distribution of Topo IIα was overall similar during interphase and mitosis to that previously reported, regardless of antibody and of sample preparation. On the other hand, the interphase distribution of Topo IIβ was quite variable, depending both on the antibody and on the method used to prepare the sample. Our interpretation of the data is that, like Topo IIα, Topo IIβ is primarily a nucleoplasmic protein, but that unlike Topo IIα, small amounts are also associated with intranucleolar chromatin. © 1996 Wiley-Liss, Inc.
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  • 87
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    Journal of Cellular Biochemistry 63 (1996), S. 185-198 
    ISSN: 0730-2312
    Keywords: extracellular matrix ; remodeling ; collagenase ; collagen ; dilated cardiomyopathy ; congestive heart disease ; end-stage heart failure ; matrix metalloproteinase ; tissue inhibitor of metalloproteinase ; differential display mRNA analysis ; gene expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Extracellular matrix metalloproteinases (MMPs) are activated in dilated cardiomyopathic (DCM) hearts [Tyagi et al. (1996): Mol Cell Biochem 155:13-21]. To examine whether the MMP activation is occurring at the gene expression level, we performed differential display mRNA analysis on tissue from six dilated cardiomyopathy (DCM) explanted and five normal human hearts. Specifically, we identified three genes to be induced and several other genes to be repressed following DCM. Southern blot analysis of isolated cDNA using a collagenase cDNA probe indicated that one of the genes induced during DCM was interstitial collagenase (MMP-1). Northern blot analysis using MMP-1 cDNA probe indicated that MMP-1 was induced three- to fourfold in the DCM heart as compared to normal tissue. To analyze posttranslational expression of MMP and tissue inhibitor of matrix metalloproteinase (TIMP) we performed immunoblot, immunoassay, and substrate zymographic assays. TIMP-1 and MMP-1 levels were 37 ± 8 ng/mg and 9 ± 2 ng/mg in normal tissue specimens (P 〈 0.01) and 2 ± 1 ng/mg and 45 ± 11 ng/mg in DCM tissue (P 〈 0.01), respectively. Zymographic analysis demonstrated lytic bands at 66 kDa and 54 kDa in DCM tissue as compared to one band at 66 kDa in normal tissue. Incubation of zymographic gel with metal chelator (phenanthroline) abolished both bands suggesting activation of neutral MMP in DCM heart tissue. TIMP-1 was repressed approximately twentyfold in DCM hearts when compared with normal heart tissue. In situ immunolabeling of MMP-1 indicated phenotypic differences in the fibroblast cells isolated from the DCM heart as compared to normal heart. These results suggest disruption in the balance of myopathic-fibroblast cell ECM-proteinase and antiproteinase in ECM remodeling which is followed by dilated cardiomyopathy. © 1996 Wiley-Liss, Inc.
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  • 88
    ISSN: 0730-2312
    Keywords: regulation of transcription ; control of proliferation ; vitamin D3 analogues ; vitamin D3 receptor ; limited protease digestion assay ; lymphocytes ; breast cancer cells ; promoter selectivity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The biological active form of vitamin D3, 1,25-dihydroxyvitamin D3 (VD), regulates cellular growth and differentiation. This provides the hormone with an interesting therapeutic potential. However, hypercalcemia is a side effect, which is caused by VD's classical action, the regulation of calcium homeostasis. This made the need for VD analogues with selectively increased cell regulatory properties. Studies with 20-epi analogues pointed out the importance of the carbon-20 position and led to the development of 20-methyl derivatives of VD. In this report the biological properties of the compounds ZK161422 and ZK157202, which are 20-methyl- and 20-methyl-23-eneanalogues, respectively, have been analyzed in comparison with VD. Both compounds show about 2-fold lower affinity to the VD receptor (VDR) than VD. However, compared to VD, their antiproliferative effect is up to 30-fold higher on human peripheral blood mononuclear cells and even up to 300-fold higher on human breast cancer MCF-7 cells. Whereas the hypercalcemic effect for ZK157202 is also increased 10-fold, ZK161422 has the same calcium-mobilizing potency as VD. Moreover, ZK161422, but not ZK157202, showed preference for gene activation from a promoter carrying a VD response element with a palindromic arrangement of two hexameric receptor binding sites spaced by 9 nucleotides (IP9) rather than for activation from a response element formed by a direct repeat spaced by 3 nucleotides (DR3). This observation supports a model, in which promoter selectivity reflects the selectively increased antiproliferative effect of VD analogues. © 1996 Wiley-Liss, Inc.
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  • 89
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    Journal of Cellular Biochemistry 63 (1996), S. 239-251 
    ISSN: 0730-2312
    Keywords: lymphocyte activation ; Krebs cycle ; energy metabolism ; immunosuppressives ; cell cycle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Rapamycin (RAPA) strongly inhibits lymphocyte activation and proliferation, but does not affect most of the activation-related gene expression at the mRNA level. In order to understand the mechanism of action of RAPA and to gain further insights in lymphocyte signalling which is impaired by RAPA, we screened for RAPA-sensitive genes using differential hybridization. The expression of human aldolase A gene was found to be inducible during T and B cell activation, and the induction was repressed by RAPA at both the mRNA and enzymatic levels. The other two important immunosuppressants, cyclosporin A and FK506, also inhibited the mitogen-induced upregulation. However, none of these three drugs inhibited the constitutive expression. There was no fluctuation of aldolase A expression during the cell cycle, and RAPA failed to block the first cell cycle after synchronization in Jurkat cells. However, the second cycle was hampered by RAPA, and this was correlated with the inhibition of aldolase A expression during this later stage. Since aldolase A is a key enzyme in glycolysis and lymphocytes mainly depend on glycolysis for energy supply, the data from this study suggest that aldolase A might be one of the downstream targets of RAPA. The inhibition of the enzyme upregulation might deprive the cells of additional supply of energy, and prevent the cells from entering an optimal status for proliferation. © 1996 Wiley-Liss, Inc.
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  • 90
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    Journal of Cellular Biochemistry 63 (1996), S. 268-279 
    ISSN: 0730-2312
    Keywords: nuclear matrix ; mitosis ; Drosophila embryo ; monoclonal antibody ; spindle formation ; nucleus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Little is known about what determines the nuclear matrix or how its reorganization is regulated during mitosis. In this study we report on a monoclonal antibody, mAb2A, which identifies a novel nuclear structure in Drosophila embryos which forms a diffuse meshwork at interphase but which undergoes a striking reorganization into a spindle-like structure during pro- and metaphase. Double labelings with α-tubulin and mAb2A antibodies demonstrate that the microtubules of the mitotic apparatus co-localize with this mAb2A labeled structure during metaphase, suggesting it may serve a role in microtubule spindle assembly and/or function during nuclear division. That the mAb2A-labeled nuclear structure is essential for cell division and/or maintenance of nuclear integrity was directly demonstrated by microinjection of mAb2A into early syncytial embryos which resulted in a disintegration of nuclear morphology and perturbation of mitosis. © 1996 Wiley-Liss, Inc.
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  • 91
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    Journal of Cellular Biochemistry 63 (1996), S. 311-319 
    ISSN: 0730-2312
    Keywords: protein phosphatase 2A ; endothelial cells ; cyclic strain ; proliferation ; okadaic acid ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We previously proposed that activation of protein kinase C is a key mechanism for control of cell growth enhanced by cyclic strain [Rosales and Sumpio (1992): Surgery 112:459-466]. Here we examined protein phosphatase 1 and 2A activity in bovine aortic endothelial cells exposed to cyclic strain. Protein phosphatase 2A activity in the cytosol was decreased by 36.1% in response to cyclic strain for 60 min, whereas the activity in the membrane did not change. Treatment with low concentration (0.1 nM) of okadaic acid enhanced proliferation of both static and stretched endothelial cells in 10% fetal bovine serum. These data suggest that protein phosphatase 2A acts as a growth suppressor and cyclic strain may enhance cellular proliferation by inhibiting protein phosphatase 2A as well as stimulating protein kinase C. © 1996 Wiley-Liss, Inc.
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  • 92
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    Journal of Cellular Biochemistry 63 (1996), S. 366-373 
    ISSN: 0730-2312
    Keywords: capsule ; lipid droplet ; Leydig cell ; monoclonal antibody ; immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In this report, we provide direct evidence for the presence of a lipid droplet-associated capsule in hamster steroidogenic Leydig cells by using a monoclonal antibody A2. Leydig cells are characterized by containing many lipid droplets and having 3β-hydroxysteroid dehydrogenase activity. Immunofluorescence staining with this antibody demonstrated a rim or capsule surrounding the lipid droplets in Leydig cells, a pattern not seen with anti-vimentin antibody. Immunogold labelling confirmed ultrastructurally that antibody binding was distributed on the lipid droplet surface. In order to investigate the possible function of the capsule, we examined the morphological changes induced in the capsule following stimulation with LH or dibutyryl cAMP; the fluorescent intensity of the capsule was seen to gradually decrease, accompanied by a decrease in number and size of lipid droplets, and the response to both reagents was time- and concentration-dependent. We thus conclude that hormonal stimulation resulting in the detachment of certain capsular proteins from the surface of lipid droplets is mediated via the cAMP signaling pathway and may allow cholesterol ester hydrolytic enzyme direct access to its substrate in the lipid droplet. © 1996 Wiley-Liss, Inc.
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  • 93
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    Journal of Cellular Biochemistry 63 (1996), S. 453-462 
    ISSN: 0730-2312
    Keywords: FBPase ; gluconeogenesis ; perinuclear association ; metabolic zonation ; immunolocalization ; subcellular fractionation ; confocal microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The localization of fructose 1,6-bisphosphatase (D-Fru-1,6-P2-1-phosphohydrolase, EC 3.1.3.11) in rat kidney and liver was determined immunohistochemically using a polyclonal antibody raised against the enzyme purified from pig kidney. The immunohistochemical analysis revealed that the bisphosphatase was preferentially localized in hepatocytes of the periportal region of the liver and was absent from the perivenous region. Fructose-1,6-bisphosphatase was also preferentially localized in the cortex of the kidney proximal tubules and was absent in the glomeruli, loops of Henle, collecting and distal tubules, and in the renal medulla. As indicated by immunocytochemistry using light microscopy and confirmed with the use of reflection confocal microscopy, the enzyme was preferentially localized in a perinuclear position in the liver and the renal cells. Subcellular fractionation studies followed by enzyme activity assays revealed that a majority of the cellular fructose-1,6-bisphosphatase activity was associated to subcellular particulate structures. Overall, the data support the concept of metabolic zonation in liver as well as in kidney, and establish the concept that the Fructose-1,6-bisphosphatase is a particulate enzyme that can not be considered a soluble enzyme in the classical sense. © 1996 Wiley-Liss, Inc.
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  • 94
    ISSN: 0730-2312
    Keywords: transcription initiation ; CpG island ; transcription factor AP2 ; transcription factor Sp1 ; osteoblasts ; differentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Transforming growth factor (TGF-β) binds several discrete membrane proteins. Of these, a type I receptor appears indispensable for signal transduction. Previous examination of TGF-β receptor expression has been limited to changes in cell surface protein, and more recently, mRNA abundance. In order to learn more about TGF-β function and receptor expression during osteogenesis, we have now cloned a 4 kilobase (kb) DNA fragment 5' proximal to the coding region of the rat TGF-β type I receptor gene. Sequence analysis revealed multiple elements compatible with transcription initiation, including a properly positioned and oriented CCAAT box, six Sp1 binding sites (three defining GC boxes), and two strong AP2 binding sites within a 0.7 kb span directly upstream of the coding region. The 3' terminal 0.3 kb span comprises a GC-enriched (77%) so-called CpG island that, like other similarly organized promoters, lacks a TATA box. Primer extension and RNase protection studies with cRNAs from this area show multiple initiation sites within 220 bp 5' proximal to the initial methionine codon. Transient transfections using nested, deleted, and inverted promoter sequences demonstrated maximal reporter expression by a 1 kb fragment encompassing all of these elements. Truncation of the 1 kb fragment from the 5' and 3' ends indicated the need for several elements for peak promoter activity. These results, and transfections in fetal rat bone and dermal cells, suggest that this promoter contains elements that specify basal and conditional expression of the TGF-β type I receptor in bone. © 1996 Wiley-Liss, Inc.
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  • 95
    ISSN: 0730-2312
    Keywords: protein kinase C ; Drosophila melanogaster ; embryonic neurons ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Embryonic neurons were cultured from transgenic Drosophila melanogaster expressing a highly specific pseudosubstrate inhibitor of protein kinase C (PKC). Flies homozygous for this transgene, which is under the control of the yeast UAS promoter, were crossed to flies homozygous for the yeast heat shock inducible transcription factor GAL 4. Following heat shock, the progeny express the pseudosubstrate inhibitor at high levels. This strategy, which has the advantage of avoiding the non-specific effects of drugs, was used to study the role of PKC in process growth of cultured, differentiating neuroblasts. An external gold particle labeling procedure using a cell surface antigen expressed by mature neurons and processes was used to visualize neuronal processes directly in the scanning electron microscope. We observed that cell cultures expressing a low concentration of the pseudosubstrate inhibitor showed a significant decrease in the number of type I and II processes as compared to control cultures, while the proportions of neuroblasts, ganglion mother cells (GMCs), and mature neurons in the clusters were little affected. © 1996 Wiley-Liss, Inc.
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  • 96
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    Journal of Cellular Biochemistry 61 (1996), S. 18-25 
    ISSN: 0730-2312
    Keywords: osteoblasts ; calvaria ; invasion ; prostate ; PC-3 cells ; differentiation ; metastasis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Bone metastasis is a common event and a major cause of morbidity in prostate cancer patients. After colonization of bone, prostate cells induce an osteoblastic reaction which is not associated with marrow fibrosis (i.e., osteoblast but not fibroblast proliferation). In the present study we test the hypothesis that the tumoral prostatic cell line (PC-3) secretes factors that block the osteoblast differentiation process, resulting in an increase of the relative size of the proliferative cell pool. Our results, using fetal rat calvaria cells in culture, show that conditioned medium from PC-3 cells (PC-3 CM) stimulates osteoblast proliferation and inhibits both alkaline phosphatase (AP) activity (an early differentiation marker) and the mineralization process, measured as calcium accumulation (late differentiation marker). The inhibition of the expression of AP and mineralization depends on the presence of PC-3 CM during the proliferative phase of culture and suggests that both processes occur in a nonsimultaneous fashion. The inhibitory effect of PC-3 CM was not reverted by dexamethasone, which would indicate that prostatic-derived factors and the glucocorticoid do not share a common site of action. Measurement of the proliferative capacity of subcultures from control and treated cells demonstrates that PC-3 CM treatment induces the maintenance of the proliferative potential that characterizes undifferentiated precursor cells. © 1996 Wiley-Liss, Inc.
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  • 97
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    Journal of Cellular Biochemistry 61 (1996), S. 97-108 
    ISSN: 0730-2312
    Keywords: aggregin ; chemical modification ; ADP-induced platelet responses ; NBD-Cl ; cAMP ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: ADP-induced platelet responses play an important role in the maintenance of hemostasis. There has been disagreement concerning the identity of an ADP receptor on the platelet surface. The chemical structure of 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD-Cl) shows considerable resemblance to that of the adenine moiety of adenine-based nucleotides. The reagent has been previously used by other investigators as an affinity label for adenine nucleotide-requiring enzymes, such as mitochondrial ATPase and the catalytic subunit of cAMP-dependent protein kinase. Since ADP-induced platelet responses depend on the binding of ADP to its receptor, we investigated the effect on ADP-induced platelet responses and the nature of ADP-binding protein modified by NBD-Cl. NBD-Cl inhibited ADP-induced shape change and aggregation of platelets in platelet-rich plasma in a concentration- and time-dependent manner. NBD-Cl also inhibited ADP-induced shape change, aggregation, exposure of fibrinogen binding sites, secretion, and calcium mobilization in washed platelets. NBD-Cl did not act as an agonist for platelet shape change and aggregation. Covalent modification of platelets by NBD-Cl blocked the ability of ADP to antagonize the increase in intracellular levels of cAMP mediated by iloprost (a stable analogue of prostaglandin I2). NBD-Cl was quite specific in inhibiting platelet aggregation by those agonists, e.g., ADP, collagen, and U44619 (a thromboxane mimetic), that completely or partially depend on the binding of ADP to its receptor. Autoradiogram of the gel obtained by SDS-PAGE of solubilized platelets modified by [14C]-NBD-Cl showed the presence of a predominant radiolabeled protein band at 100 kDa corresponding to aggregin, a putative ADP receptor. The intensity of this band was considerably decreased when platelets were either preincubated with ADP and ATP or covalently modified by a sulfhydryl group modifying reagent before modification by [14C]-NBD-Cl. These results (1) indicate that covalent modification of aggregin by NBD-Cl contributed to loss of the ADP-induced platelet responses, and (2) suggest that there is a sulfhydryl group in the ADP-binding domain of aggregin. © 1996 Wiley-Liss, Inc.
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  • 98
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    Journal of Cellular Biochemistry 63 (1996), S. 108-111 
    ISSN: 0730-2312
    Keywords: duct carcinoma in situ ; nuclear grade necrosis ; prognostic features ; local recurrence ; invasive transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In the last 6 years a number of non-randomized, predominantly single institutional trials of breast conservation therapy (BCT) with DCIS, have demonstrated that it constitutes a very heterogeneous group of diseases with markedly different risks of local recurrence and invasive transformation. There has been a consensus that DCIS, which exhibits a “comedo” morphology, generally defines a high risk group. Most studies, moreover, have identified the same two features, nuclear grade and necrosis, as contributing most significantly to prognosis [4-6]. Nuclear grade and necrosis have been identified as independent prognostic variables in several studies [5,6]. High nuclear grade DCIS which exhibits comedo necrosis defines the majority of all DCIS which will result in local recurrence and invasive transformation after BCT.Studies utilizing image cytometry, to determine ploidy and S-phase fraction and immunohistochemical studies of proliferation and oncogene distribution have shown a significant association with morphologically identified high nuclear grade and aneuploidy, high S-phase fraction or proliferation rate, presence of HER-2/neu and P53 oncogenes and absence of estrogen receptors. Generally the inverse of this association is seen with low nuclear grade DCIS. However, initial hopes that these adjunctive studies would identify subsets within the high nuclear grade group which might be more likely to recur have not been fulfilled. J. Cell. Biochem. 25S:108-111. © 1997 Wiley-Liss, Inc.
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  • 99
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    Journal of Cellular Biochemistry 63 (1996), S. 123-130 
    ISSN: 0730-2312
    Keywords: carcinogenesis ; predisposing mutation ; malignancy ; DNA testing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Identification of cohorts at genetic risk for cancer offers unique research opportunities to explore the steps in carcinogenesis, from the inheritance of a predisposing mutation to the development of preinvasive lesions or overt malignancy, and to evaluate interventions to modulate the carcinogenic process. However, cancer prevention strategies for most inherited cancer predisposition syndromes are of unproven benefit, and the potential for adverse psychosocial effects and employment or insurance discrimination associated with genetic testing is substantial. Thus testing for genetic cancer risk remains highly controversial, and the National Center for Human Genome Research and the American Society of Human Genetics advise DNA testing for presymptomatic identification of cancer risk only in the setting of a carefully monitored research environment.The commercial availability of predictive genetic testing, particularly for inherited susceptibility to cancer, has focused attention not only on the urgent need for research in cancer prevention for cohorts at genetic cancer risk but also on ethical considerations surrounding clinical prevention research in genetic risk groups. This paper addresses the interrelationship of ethical and scientific issues in conducting chemoprevention research in these cohorts, especially for those studies which require presymptomatic testing for specific gene mutations as a study entry criterion or as a criterion for stratification. Practical approaches to study design and implementation issues for chemoprevention research in genetic risk cohorts are discussed, emphasizing the interactions of ethical and scientific considerations at all levels of the research process. J. Cell. Biochem. 25S:123-130. © 1997 Wiley-Liss, Inc. This article is a U.S. Government work and, as such, is in the public domain in the United States of America.
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  • 100
    ISSN: 0730-2312
    Keywords: acquired risk ; chemoprevention ; colon ; genetic risk ; neoplasia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The progressively abnormal development of epithelial cells prior to tumor development leads to widely differing chemopreventive approaches. The diversity of these approaches has resulted in different assays to measure the activities of the agents. To apply these assays to preclinical studies, we have developed rodent models in which different stages of evolution of colonic neoplasia are expressed. In one model mice carrying a truncated Apc allele with a nonsense mutation in exon 15 have been generated by gene targeting and embryonic stem cell technology (Apc1638 mice). These mice develop multiple gastrointestinal lesions including adenomas and carcinomas, focal areas of high grade dysplasia (FAD) and polypoid hyperplasias with FADS.The incidence of inherited colonic neoplasms has now been modulated by a chemopreventive regimen. Colonic lesions significantly increased in Apc1638 mice on a Western-style diet, compared to Apc1638 mice on AIN-76A diet which has lower fat content and higher calcium and vitamin D. These studies have also been carried out in normal mice, and have demonstrated without any chemical carcinogen that a Western-style diet induced colonic tumorigenesis. Modulation of cell proliferation has also been induced by Western-style diets in other organs including mammary gland, pancreas and prostate. These findings are leading to the development of new preclinical models for evaluating the efficacy of many classes of chemopreventive agents. J. Cell. Biochem. 25S:136-141. © 1997 Wiley-Liss, Inc.
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