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Notes: Light and electron microscopical studies allow a descriptive account to be given of the preadult development of the ovary of Drosophila melanogaster. The lineage of the cell groups which contribute to the tissues of the adult ovary has been determined. The earliest morphologically detectable event in the differeentiation of each ovariole is the formation during the larval period of its terminal filament. Oogonia play no role in the induction of terminal filaments. The developmental events which transform a spherical mass of ovarian cells into a collection of multicellular cylinders is described. The importance in morphogenesis of acellular membranes secreted at the interface separting cells of different prospective significances is stressed. Such membranes may serve to regulate the future migration of cell populations or as sites of attachment for monolayers of cells which later fuse to form multinucleated muscle sheaths. The transformation of oogonia to cystoblasts coincides with and presumably depends upon the same hormonal stimulus which causes metamorphosis. The first oocytes to undergo crossing over do so between 24 and 36 hours after puparium formation.

Notes: A morphological study of the presoma of adult Corynosoma hamanni (Linstow, 1892) was undertaken in an effort to clarify some problems encountered during analysis of a large collection of juveniles of this species assembled from fishes of McMurdo Sound. This study is based on approximately 600 adult specimens recovered from four Weddel seals, Leptonychotes weddelli, collected at McMurdo Sound.Morphologically the proboscis armature is more varied than previously reported and consists of 18 to 23 longitudinal rows each with 11 to 15 hooks of two distinct types. At the apex of the proboscis is an undivided, bi-nucleated apical organ unlike that described for species of Neoechinorhynchus. In the basal third of the proboscis is a thin-walled vesicle which extends into the anterior quarter of the proboscis receptacle. The more posterior of two external folds on the presoma is a cuticular invagination permitting recognition of the neck-trunk border of C. hamanni. Lemnisci originate between the two folds and extend into the trunk cavity between the body wall and extensive neck retractor muscles. Contrary to Linstow's original description, each lemniscus is a single structure.Several limitations in material prevent speculation at this time about the limits of intraspecific variation in C. hamanni.

Notes: The development of intraneural vessels was studied in response to an induced hypermorphosis of neural tissue inthe midbrains of 38 chick embryos ranging in age from three days through 14 days of incubation. The pattern of vascularization was compared with that of normal chick embryos at comparable stages of development. In the experimental embryos, the increase in mitotic figures along the ventricular borders of the mesencephalon is accompanied by the establishment of an endoneural plexus approximately one day earlier than is the case during normal vascularization of the midbrain. This plexus also penetrates more deeply and extensively into the ependymal layer. Surface vessels and intraneural vascular elements are dilated, and the cerebrospinal fluid contains varying amounts of blood released from large intraneural vessels which protrude into the ventricle. The most prominent cerebrovascular effects seem to occur between the fourth and eighth days of incubation. Thereafter, the cerebrovascular pattern becomes more normal except for relatively few isolated hemorrhagic areas.

Notes: The eye of Haideotriton wallacei is more reduced histologically than those of othe troglobitic salamanders. The tiny eye is imbedded in a mass of adipose tissue. No extrinsic eye muscles are present. A rudimentary lens is present in about half of the eyes examined. In two instances the lens is surrounded by a small chamber; most eyes lack a chamber. The retina and iris are relatively undifferentiated. The relatively massive retina lacks rods and cones, an outer plexiform layer and subdivided nuclear layers. A tiny optic nerve runs to the brain.

Notes: The aesthetascs, short thin-walled pegs on the antennule flagella of Coenobita clypeatus, a terrestrial hermit crab, are similar to those of other decapod crustacea in containing the dendrites of many bipolar neurons whose cell bodies are grouped in spindle-shaped masses beneath the bases of each hair. The dendrites contain rootlets, basal bodies, and cilia, which divide dichotomously before entering the aesthetasc, so that within the hair, each cilium becomes represented by a group of slender branches.The aesthetascs themselves are short, blunt, and partially recumbent so that each has an exposed and an unexposed side. The cuticle on the exposed side is thinner and more tenuous than that on the protected side, and the dendrite branches are concentrated just underneath. The protected side, on the other hand, is lined with nondendritic supporting cells, and the cuticle is thicker, more lamellar, and probably less permeable.All dendritic elements proximal to the dendrite branches are enclosed within the main body of the antennular flagellum, and the initial segments of the cilia lie within a vacuole. In these respects, the aesthetascs of Coenobita resemble the thin-walled pegs on insect antennae more than they do those of the marine decapods thus far examined. This convergence in the terrestrial forms may be in response to the need to conserve water.

Notes: The Brittle Star digestive system is composed of buccal, pharyngeal, esophageal and stomach cavities. The buccal and pharyngeal cavities are lined by columnar cells covered by a cuticle, and are apparently concerned with mucous production. Coelomocytes and tall columnar cells are described in the esophagus and stomach epithelia. The columnar cells are adapted for nutrient absorption, enzyme synthesis, and lipid storage. Nerves are found beneath the epithelia within a connective tissue layer. Smooth muscle and coelomic layers lie external to the connective tissue layer. The coelomic layer lines a perivisceral space and has diverse modifications of its perivisceral surface; a pedicle-cuticle modification perhaps having general significance in echinoderms.

Notes: Two complete composite photographs of the optic nerve of Limulus, made by electron microscopy, reveal the presence of neurosecretory granules in the large axons of the rudimentary eye neurons. The number of intermediate sized, (3-7 μ), of eccentric cells corresponds with the number of ommatidia as expected, but only their sheath of Schwann cells show an intimate interfolding. Based on the number of fine axons within the nerve each ommatidium has an average of 12-13 retinular cells. The diameter of their fibers is between 0.2 and 3 μ although the majority are between 1 and 1.5 μ. They are aggregated into bundles of six to seven fibers by the sheath cells although some bundles contain only two, others as many as 181 fibers. There is no indication in these studies that retinular cell axons within a bundle are associated with the same, adjacent, or other pattern of ommatidia. The photographs suggest that physiological activity in retinular cell axons might be detected most easily in the smallest bundles because they contain the fewest, but the larger retinular cell axons.

Notes: The lateral and ventral external surfaces of the third and fourth abdominal segments were described and muscle attachments were correlated with surface indentations of the larva. The proleg of this species has a symmetrical planta with a complete circle of crochets. Furthermore, it differs externally from the grasping type of proleg in having a largely membranous coxal region confluent with the body wall, and a relatively large subcoxal lobe.The body wall musculature and innervation of the third and fourth abdominal segments are similar in many respects to those described for other lepidopteran larvae to which they are here compared, but differ from most because of the simpler structure of the prolegs which lack highly developed adductor muscles. Like most muscles innervated by the ventral nerve, the principal plantar retractors of these two segments cease to function in the first day of the pupal stage and have completely degenerated by the forty-fifth hour of pupal life. The ventral nerve retains its four primary branches in the adult, in which many smaller rami can be traced to the cuticle and to the neoblastic body wall muscles.

Notes: Symmetrical gaits of 37 breeds of dogs were analyzed. Usual walking and trotting gaits resemble those of other carnivores of similar size and conformation. Only certain long-legged dogs pace - usually at the fast walk or slow run. At the moderate walk, long-legged dogs tend to use lateral-couplets gaits, whereas short-legged breeds tend to use single-foot gaits. Many dogs must turn the axis of the body slightly from the line of travel at the trot to prevent interference between fore and hind feet. The relative duration with the ground made by fore and hind feet is discussed, usual support-sequences of the varicus gaits are presented, and the amount of variation is shown.

Notes: Electron microscope autoradiography was used to study cartilage from regenerating limbs of adult newts, Triturus, after intraperitoneal injections of proline-3H. The labeling in the endoplasmic reticulum, small vesicles, Golgi vacuoles, ground cytoplasm and extracellular matrix was compared during the secretion of radioactive products. The data appear to indicate that a large part of the radioactive secretion probably leaves the cell after having been in only one cellular compartment. Although this compartment may be the endoplasmic reticulum, a considerable amount of radioactivity fluxes through the ground cytoplasm and the possibility cannot be excluded that some secretory components leave the cell directly from the ground cytoplasm. The data appear incompatible with the hypothesis that all the radioactivity seen in the extracellular matrix arrived there via a single pathway involving first the endoplasmic reticulum and then the Golgi vacuoles. It is not, however, incompatible with a hypothesis that a fraction of the radioactive product uses this pathway.

Notes: The ultrastructure of lymphatic capillaries in the tail fin of Rana catesbiana larvae was investigated. With the use of a colloidal marker particle (Biological Carbon) the extent that these delicate vessels ramify throughout the fin region was demonstrated. This opaque substance also serves as a marker particle for identification of lymphatics with some degree of certainty at both light and electron microscopic levels. The cytoplasm of the lymphatic endothelial cell is abruptly attenuated beyond the perinuclear region, reaching widths as thin as 300 Å. Lymphatic Anchoring filaments are present, but to a lesser degree than noted for other species studied. Other features of interest include an extensive Golgi complex and electron dense bodies that are surrounded by a smooth surfaced unit membrane. These bodies are somewhat heterogeneous in size (500 Å up to 0.5 μ in diameter) and density. Numerous exit channels are provided by the extensive supply of lymphatics throughout the tail fin region of amphibian larva thus allowing them to serve an important function during metamorphosis. It is suggested that these vessels also act as passageways through which lysed cellular and connective tissue components may be rapidly removed during the process of tail fin resorption.

Notes: The cuticle of Watersipora nigra is at first translucent, but it later becomes black and differentiates into two layers. It is composed, at least in part, of a protein-polysaccharide complex. Calcified parts are three-layered: (1) an outer, cuticular layer, (2) a calcium carbonate skeleton deposited on a matrix of acid mucopolysaccharide, and (3) a “skeletal membrane.” The relationships of these layers indicate that the skeleton is intracuticular. A layer of cuticular material, the “intercalary cuticle” is present in lateral walls, but not transverse walls; it may become calcified in some species. The cuticles of calcified and uncalcified parts of cheilostomes are not necessarily homologous.

Notes: Thin semi-serial ground sections of coronal dentin were examined radiographically. The bulk of the coronal dentin was characterized by the majority of the tubules having a distinct peritubular zone. With the exception of the tubules running from the tip of the cusp to the pulp cornu, the bulk of peritubular matrix forming the walls of the tubules was disposed eccentrically. The matrix was thicker on the cervical sides of the tubule than it was on the incisal sides. In a relatively narrow layer of the coronal dentin between the bulk of the dentin and the predentindentin border area the thickness of the peritubular matrix varied considerably. It was extremely narrow or absent in some tubules and reached its greatest thickness in others. The tubules in the predentin border area showed little or no evidence of peritubular matrix. The area of dentin beneath the central developmental groove differed somewhat from the bulk of the dentin. Many of the tubules at all levels of this area showed little radiographic evidence of peritubular matrix. Obliterated tubules were seen in some of the sections taken immediately above the predentin-dentin border area in the region of the pulp cornu and were always seen at the junction of the mantle dentin and the circumpulpal dentin beneath the central developmental groove.

Notes: Microscopic studies of human adrenal glands from 58 autopsy specimens, ranging in age from one month gestation to 69 years, revealed a pertinent developmental pattern in the establishment of definitive zonation. This pattern was established using the following criteria: (1) relationship of age to the developing zones; (2) times of formation of definitive zonation; and (3) the morphological determination of developmental patterns based on staining characteristics.Using these criteria, development was divided into five phases: (1) condensation of coelomic epithelium; (2) secondary proliferation of coelomic epithelium; (3) finding of PAS-positive material within the fetal cortex; (4) decline and disappearance of the fetal cortex; and (5) establishment and stabilization of the definitive zonular patterns.Significant features occurring in this development were: (1) the origin of both permanent and fetal cortex from proliferation of coelomic epithelium; (2) the appearance of PAS-positive granules surrounding a homogenous mass in the fetal cortex and the zona reticularis during maturation and organization; and (3) the gradual establishment of definitive zones by proliferation of the permanent cortex, maturation of the fetal cortex, and growth of the medulla; with the adult structure of the adrenal gland achieved by the eleventh to fifteenth year without any apparent major involution or hemorrhage.

Notes: A total of 1125 normal chick embryos, representing 25 each of the 45 stages of Hamburger and Hamilton, were removed, fixed in Bouin's solution, stored in 70% ethanol and weighed with a semi-micro analytical balance. Entire blastoderms of stages 1-8 were weighed, whereas only embryos-proper were weighed in stages 9-45. As a consequence, results constituted two groups, each of which showed a geometric rate of growth marked only by minor deviations which were related to specific events of normal growth and development.

Notes: Estimates of the number of ganglionic neurons of superior cervical sympathetic ganglia and the number of preganglionic axons in the trunks just caudal to these ganglia were obtained from a sample of primates that included: man, chimpanzee, baboon, stump-tailed macaque, rhesus monkey, and squirrel monkey. The number of ganglionic neurons ranged from 63,625 in a squirrel monkey ganglion to 1,041,652 neurons in a human ganglion. Estimates of the number of preganglionic fibers varied between 2,285 in a cervical sympathetic trunk of a squirrel monkey and 12,008 in a human specimen. The resulting ratios of preganglionic fibers to ganglionic neurons ranged from 1:28 in a squirrel monkey ganglion to 1:196 in a human ganglion.The data reported in this study reveal considerable variation in the ratio of pre- to post-ganglionic neurons, and as was noted in regard to the number of cells in the ganglion, the ratios of ganglionic to preganglionic neurons appear to increase as a function of body size. In contrast, the number of preganglionic fibers does not increase as strikingly with body size, but varies greatly in the same species. The resulting ratio between the two orders of neurons is, therefore, less predictable than the number of ganglionic neurons in any given ganglion.

Notes: Adaptive explanations for the temporal fenestration in reptiles are briefly reviewed. With few possible exceptions, fenestrate appeared first in the reptiles, and have seemingly evolved independently in several different phyletic lines.The several explanations for fenestration offered by previous authors include speculations that open spaces in the skull permitted bulging of the jaw-closing muscles, and that fenestrae formed in areas of reduced stress where the presence of bone would be functionally useless. The first of these does not readily apply to initial evolutionary stages; the second is more satisfactory.Certain features of muscular attachments to bones are dealt with, and their implications applied to the fenestration problem to add another possible explanation (which need not contradict previously published suggestions).Considerations of cranial strength in tetrapod skulls led to speculations on the lack of fenestration in temnospondyls, anthracosaurs, microsaurs and cotylosaurs.Emargination of the skull roof in turtles is also discussed.

Notes: The down feather of the chick embryo has been examined by electron microscopy during three distinct stages of its early development; the presumptive stage, represented by dorsal skin of an area from which the feather organ will arise; the thickening stage, during which areas of the basal epidermis form spurs projecting into the mesenchyme, and the latter condenses under a thickened area of the epidermis; the elevation stage, at which time the basal epidermis flattens, the entire epidermis increases in thickness, and the underlying mesenchyme becomes more compact.As development proceeds the rough endoplasmic reticulum of the epidermal cells dilates, but during the elevation stage begins to flatten, and Golgi is observed with increasing frequency. The mitochondria do not appear to differ except for those in the periderm during the presumptive stage, in which case they reveal a vacant matrix and irregular cristae.Evidence is presented for actual contact between basal epidermal spurs and filopodia of cells within the mesenchyme, some of which contain numerous vesicles. The basal epidermal spurs are also seen in intimate association with collagen and anchor filaments and a network of reticulin.Evidence is also presented for the presence of neuronal elements within the mesenchyme during the thickening stage. Cross sections of cell processes within the condensations of the mesenchyme resemble unmyelinated nerve fibers, and cross sections of filopodia similar to arborizing axons abound at and within the basal lamina of both the thickening and elevation stages. Further support for the presence of nerve fibers within the mesenchyme comes from positive staining results with Bodian's and Ungewitter's methods.This comparative study of three stages of early development of the feather organ serves as a basis for more detailed investigations of each stage.

Notes: The structure and cell types of the closely similar glands of Calamoichthys and Polypterus are described and a general comparison made with the teleost pituitary. The Polypterine gland shows some unsual features in the anatomical disposition of its parts and in the arrangement of its neurosecetory and vascular supply and an explanation of these differences is suggested, based on relative growth changes in later development, in order to include these glands in the evolutionary pattern of the actinopterygian pituitary.

Notes: About 10,000 sense organs are present on one antenna of a female mantis, Tenodera angustipennis, and nearly 40,000 on that of a male. These are of four kinds: (1) thick-walled pegs, (2) short thin-walled pegs, (3) medium length thin-walled pegs and (4) long thin-walled pegs. All have the structural characteristics of chemoreceptors. The dendrites of the sensory neurons of the thick-walled pegs are exposed to the air in an opening at its distal end and those of the thin-walled pegs terminate at many pores in the surface. The significance of the larger number of sense organs possessed by the male is discussed. No important differences were found between the antenna of Tenodera angustipennis and those of T. aridifolia and T. australasiae.

Notes: Forty-two unselected ovaries from adult mares were examined histologically, and with histochemical methods for mucins. A considerable part of the surface of the ovulation fossa was directly covered by columnar epithelium, with many ciliated cells. This epithelium, which was distributed mainly on the anterior side of the ovulation fossa, closely resembled the contiguous epithelium of the infundibulum of the oviduct, was frequently folded, and gave rise to short clefts projecting into the ovarian substance. The remainder of the ovulation fossa was covered by non-ciliated, low cuboidal or squamous epithelium, lacking folds or clefts.“Fossa cysts,” up to 6.5 mm in diameter, were observed in the ovarian tissue around the ovulation fossa in 27 (64%) of these ovaries. Both simple and branched, tubular and vesicular forms were present, and all were blind-ending. Their epithelial lining cells, which varied from simple squamous to columnar in type, were frequently ciliated. Many fossa cysts contained secretions histochemically similar to those of the columnar epithelium of the ovulation fossa and infundibulum. Both sialic acidcontaining and neutral mucins were present. It is suggested that these cysts were probably derived by ingrowth from the columnar epithelium of the ovulation fossa. This epithelium may be of müllerian duct origin.

Notes: Light microscopic sections of the adult opossum (Didelphis virginiana) spleen were observed to lack venous sinuses; this primitive mammalian spleen may be classified as non-sinusal in nature. In the spleen of the opossum, the capillary segments of the penicillar arteries lacked ellipsoid sheaths characteristic of certain mammalian spleens.Separating the lymphoid nodules from the surrounding red pulp was a distinct band of vascular tissue, the marginal zone. Arising from the central artery within the lymphoid nodule, vessels of capillary dimension were observed to terminate within the marginal zone and the area between lymphoid nodule and marginal zone. In addition to the vascular channels established by the terminal arterial vessels within the red pulp, the system of vessels within the marginal zone has been implicated as an important intermediate vascular channel within the spleen.

Notes: A total of 54 embryos of Chrysemys (Chelonia) and 29 embryos of Aristelliger (Lacertilia) were used in examining septation of the embryonic bulbus cordis. Division of this region of the heart includes a period of cushion or septal primordia formation and a period of physical partitioning. In both reptilian genera, the physical configuration of the early bulbus, the temporal sequence of appearance of the endocardial cushions, the number of major endocardial cushions, the primordia composing the two primary bulbar septa, and the mode of descent of the bulbar septa are strikingly similar. The two genera differ primarily in the pattern of the endocardial cushions and consequently the rotation of the two bulbar septa. In both the turtle and the lizard the aortico-pulmonary septum passes through an angle of about 120° in its descent toward the ventricle. In Aristelliger the aortic septum rotates through an angle of approximately 120°. By contrast, the same partition in Chrysemys spirals through an angle of about 90°. The lesser spiral of this septum in the turtle is interpreted as the result of a decrease in the rotation of distal endocardial ridge 4. The pattern of the two bulbar septa in the turtle appears to represent an advanced phylogenetic feature in terms of the evolution of the reptilian bulbus cordis.

Notes: Oogenesis and the relationships between oocytes and other ovarian tissues have been studied in Sypharochiton septentriones. The ovarian tissues were examined by electron microscopy and by histochemical methods.The sac-like ovary is dorsal, below the aorta, and opens to the exterior by two posterior oviducts. Ventrally, the ovarian epithelium is folded inwards to form a series of plates of tissue, which support the developing ova. Each ovum is attached to a tissue plate by a stalk, the plasma membrane of which is bathed by the blood in the tissue plate sinus. Dorsally, ciliated vessels from the aorta enter the ovary and open into blood sinuses in the top of the plates.After each germinal epithelial cell rounds up to become a primary oogonium, it undergoes four mitotic divisions to give rise to a cluster of 16 secondary oogonia. Of these, the outer ones become follicle cells and the inner ones become oocytes. As in other molluses, the increases in nuclear and nucleolar volume are relatively greatest towards the end of previtellogenesis, when chromosomal and nucleolar activity are most intense. This phase of activity is accompanied by a great increase in cytoplasmic basophilia. Subsequently this basophilia is decreased during vitellogenesis, when chromosomal and nucleolar activity diminish. Fluid filled interstices appear in the cytoplasm during early vitellogenesis. Protein yolk deposition is associated with these interstices, but the lipid yolk appears to arise de novo. The follicle cells do not appear to be directly involved in oocyte nutrition.At times during oogenesis, certain manifestations of polarity can be found in the oocyte. This polarity is based on an apical-basal axis and can be related to the nutritive source of the oocyte, namely the blood which bathes the plasma membrane of the oocyte in the stalk.Numerous granulated cells are present in the ovarian tissue plates and ventral epithelium as storage cells containing lysosomes, and they are capable of phagocytosis and micropinocytosis of extracellular material. A scheme is outlined whereby reserves in these cells may be incorporated into the oocyte cytoplasm. Lysosomal activity is responsible for autolysis of the cells as well as resorption of unspawned ova.

Notes: Among eight species of mammals in this study (cattle, sheep, pig-tail and rhesus monkeys, rabbit, pig, rat, and dog) four basic patterns of anatomical structure at the uterotubal junction are described. The classification of types is based upon the presence or absence of an intramural portion of the oviduct and of isthmal folds or plicae projecting into the lumen of the uterine cornu.Histological variations are reported for three tissues: epithelial and connective of the mucosa and smooth muscle of the tunica muscularis. In the epithelium during the estrous cycle the differences recorded include: (a) absence of ciliated cells in the distal end of the oviduct in rat and dog; (b) variations in ciliated and nonciliated cells in (1) cell height, (2) location, shape and stainability of the nucleus, and (3) in amount and stainability of apical cytoplasm; (c) presence of lymphoblast-like cells which appear to migrate through the epithelium from the lamina propria. The connective tissue of the mucosa, as a circular layer and as cores for the mucosal folds, shows variations in thickness and in relative density of cells and fibers of the matrix. Emphasis is given to the presence of an inner longitudinal layer of smooth muscle in the tunica muscularis of the distal oviduct in six of the eight species.

Notes: Sinistral ovariectomy in the Japanese quail resulted in some hypertrophy of the rudimentary right gonad in about 80% of the cases. The hypertrophied right gonads were composed of cords of epithelial origin, fat laden cells and a connective tissue stroma containing masses of lymphocytes. Neither cortical tissue nor germ cells were found in any of the gonads. In some cases regeneration of a testis-like tissue was seen on the site of removed left ovary. This, however, did not alter the effects of ovariectomy on rudimentary right gonad, accessory sex organs, plumage or sexual behavior. Neither Wolffian nor Müllerian ducts exhibited hormonal stimulation in poulards showing hypertrophy of right gonad with exception of the latter in two poulards. Early orchiectomy inhibited growth and differentiation of the cloacal gland. This organ revealed no noticeable stimulation in poulards showing hypertrophied right gonads. Castration produced no significant changes in plumage of males. Similarly, sinistral ovariectomy did not effect the first juvenile, but the second juvenile, adult winter and summar plumages changed to the male type. However, the plumage of some of these poulards began to revert to the female character as early as 10 to 12 weeks following ovariectomy. The behavior of capons and poulards revealed no conspicuous difference and neither showed any masculine behavior. The average weight of adult females was 20 to 30 gm above that of adult males whereas that of capons was above normal males and that of poulards below normal females. The average weight of capons was somewhat above that of poulards.

Notes: The volumes of a sample of primate superior cervical sympathetic ganglia were measured and related to body weight and to the number of ganglionic neurons. Estimates of volumes of the ganglia varied between 1.956 mm3 in squirrel monkey and 173.530 mm3 in a human specimen. Average cell densities for the ganglia ranged from 4,455 cells/mm3 in a human ganglion to 32,528 cells/mm3 in a squirrel monkey ganglion. Mean cell territories varied from 0.0000307 mm3 in a squirrel monkey ganglion to 0.0002245 mm3 in a human ganglion.Analysis of the data reveals striking trends of correlation between body size, volume of ganglia, and average cell territories. Since similar correlations have been described for other types of neuronal cell aggregates, it is suggested that for any given nucleus, ganglion or cortical area, the neuronal packing density varies as a function of body size.

Notes: Electron microscope observations on the differentiating Drosophila eye show an extensive proliferation of parallel arrays of microtubules at periods preceding, or coinciding with, alterations in cellular morphology. In the retinular cells they are aligned in the direction of elongation and close to the developing rhabdomeres, forming a cylinder around the central ommatidial axis. At a later stage, in the cone cells, they are aligned in the direction of cellular contraction. Thus as in other developing systems microtubules appear to be directly involved in the morphogenesis of the Drosophila eye. In the retinular cells they gradually disappear during elongation, whereas they persist in the cone cells. The pigment cells contain few of these structures. The distribution of two types of specialised cell attachments, adhering zones and septate desmosomes is discussed in relation to intercellular morphogenesis and communication. The rhabdomeres originate from infoldings of the plasma membrane which later grow out into typical microvilli. Unusul cytoplasmic granules are described in the pigment cells of early pupae.

Notes: The structure of the cellular cyst which encapsulates the parasitic copepod, Scolecodes huntsmani, in the subendostylar blood vessel of the ascidian, Styela gibbsii, is described from light and electron microscopic studies.The cells comprising the cyst are contributed by the ascidian. The cells are columnar, contain large central reservoirs of glycogen and lipid, and have a conspicuous Golgi apparatus, many small cisternae of smooth endoplasmic reticulum and peripheral mitochondria. The cells are held together by complex basal interdigitations and a short apical zonula occludens. Long cilia emerge in circular clusters from the cell apices and beat in the lumen of the cyst. As atypical of a columnar epithelial layer, the nuclei are staggered in position in the cells and there is no basal lamina. One end of the cyst is blind, but the other end, which may be either anterior or posterior with respect to the longitudinal axis of the host, narrows to a profusely ciliated duct which opens through the wall of the blood vessel to the atrium of the ascidian by a ciliated funnel. The effective beat of the cilia of the duct and the funnel is outward toward the atrium.The first nauplii of the copepod emerge from the incubatory pouch of the adult and pass to the exterior sea water through the cyst funnel and the atrium and atrial siphon of the ascidian. As in other notodelphyid copepods, the life cycle of this incarcerated form also involves free-living naupliar stages followed by two free-living copepodid stages. The provision of an egress for the first nauplii is, therefore, important to the survival of the species.The adult females of Scolecodes, which range in length from 2 to 14.6 mm, are sluggish when removed from the cyst and fail to survive in sea water for more than 24 hours. The males, which have only been obtained when parasitic fifth copepodids molt in culture, are much smaller, averaging 0.8 mm, and are very active. Since one dead male has been found inside the cyst of an adult female and females are often found with attached spermatophores, it is suggested that the funnel of the cyst may also serve as an entrance for the males.Evidence is presented for the formation of the cyst as an accumulation of totipotent lymphocytes around the copepod. Cysts of parasitic developmental stages (third through fifth copepodids) are also described. All of these cysts and those of immature adult females lack funnels to the atrium. The funnel of the cyst of mature females is formed, in part, by modified cells of the wall of the blood vessel, but is induced after the major portion of the cellular cyst has been formed.Cells in the general circulation of the ascidian and those inside the lumen of the cyst are compared. The cells in the lumen of the mature cyst do not arise by diapedesis of blood cells from the subendostylar blood vessel, but by conversion and migration of cells composing the cyst proper. These cells have been found in the guts of the copepods and they may serve as a nutritive source.The ascidian appears not to be harmed by the association, but the copepod gains in many ways.

Notes: The cervicothoracic musculature of the adult cockroach, Nauphoeta cinerea (Olivier) is described for the first time. The adult thoracic ventral intersegmental muscles are compared with those of the nymph and of the adult cockroach, Periplaneta americana (Linnaeus).

Notes: The thoracic skeleton and musculature of the adult bittacid mecopteron Bittacus strigosus Hagen is described. In its musculature, Bittacus shows only moderate differences from two panorpids (Neopanorpa, Panorpa) that have been studied by Maki ('38) and by Hasken ('39), respectively. Not only are these three genera much alike in their musculature generally, but in all of them, and in Boreus (Boreidae) too, the mesothorax is extremely similar to the metathorax. Functional emphasis (for flight) on either of the two pterothoracic segments has not appeared among neuropteroid insects at the metopteran evolutionary level.Although the “snowfleas” of the genus Boreus possess striking alterations of pterothoracic structure in comparison with other mecopterons (Füller, '54, '55), these are related to their unusual activities and have not, to any great extent, affected the two pterothoracic segments differentially.In terms of thoracic specialization, the overall mecopteran pattern represents a stage somewhat advanced beyond the primitive conditions exemplified by the Megaloptera and certain coleopterous larvae, but one that is in general less highly developed than is charatceristic of such neuropteroid orders as the Siphonaptera, Diptera, Trichoptera, and Lepidoptera.

Notes: The comparative morphology of the gonads and fat bodies of members of 17 genera and 46 species of caecilians (Amphibia: Gymnophiona) is described and analyzed. Comparison is made with the morphology of salamanders and frogs in order to elucidate evolutionary trends and relationships within the order Gymnophiona and within the class Amphibia. The structure of the testis lobes and transverse and longitudinal ducts is described based on gross dissection and histological investigation. The pattern of spermatogenesis and interstitial tissue changes are described and compared with those of other amphibians. A trend toward fusion of testis lobes is analyzed. The characteristics of the seasonal reproductive cycle of male Gymnopis m. proxima are described, and evidence for cyclic reproductive activity in other forms is presented. The morphology of the ovaries and ova is described. Size of ovary and size and number of ova is dependent on the state of maturation of the ova. Some evidence for seasonal ovum production and breeding is presented. Fat body morphology is found to be correlated with size, nutrition, and gonad condition, as in other amphibians.

Notes: The distribution of carbohydrates, proteins, and lipids was studied in the cochleae of kangaroo rat, gerbil, and guinea pig using both fixed paraffin sections and fresh-frozen cryostat sections. Enzyme distribution in the cochleae of the three speices was studied with both EDTA-decalcified and undecalcified fresh-frozen cryostat sections.Although the cochleae of the three species are morphologically different, their distributions of proteins, carbohydrates, and lipids are similar. The zona pectinata of the basilar membrane - which is hypertrophied in the kangaroo rat and gerbil but normal in the guinea pig - stains the same in all three species. The unique, flaskshaped Hensen cells of the kangaroo rat contain more protein than do the normal Hensen cells of the gerbil and guinea pig. At least some of the protein in the kangaroo rat Hensen cells is in the form of carboxylic esterases which are not affected by 10-4 M eserine, but are inhibited by 10-2 M eserine and 10-6 M E600. More than one population of carboxylic esterases is indicated by this reaction to inhibitors and by the results of enzyme distribution tests which used different substrates. A high concentration of malate dehydrogenase in the kangaroo rat Hensen cells may be related to the synthesis of carboxylic esterases. The possible role of these esterases in cochlear functioning is discussed.

Notes: This report presents light microscopic descriptions of normal histology, including innervation, of the lymph glands and jugular bodies, of larval and adult Rana catesbeiana. A brief description of two other adult organs, the propericardial and procoracoid bodies, is also included. The parenchyma was studied by employing the May-Grunwald-Giemsa staining technique for better cytoplasmic differentiation; the Periodic Acid-Schiff technique and hematoxylin and eosin yielded clearer nuclear and cytoplasmic delineations. The intercellular portion of the stroma was studied from sections stained with Masson's trichrome, Weigert's elastic stain, Periodic Acid-Schiff and Wilder's reticulum stain. Demonstration of phagocytes was facilitated by intraperitoneal India ink injections followed by the above staining procedures. Innerrvation was observed in serial sections of silver impregnated whole organs as well as in the other serial sections. These organs are lymphocytopoietic and to a certain extent granulopoietic; they also serve, like the spleen, as graveyards for dead cells and most probably play a role in immunity especially in the synthesis of antibodies as indicated by the presence of plasma cells, macrophages and lymphocytes.

Notes: The cells surrounding a wound in the integument of Rhodnius adults show an increase in RNA content, cytochrome oxidase and esterase activity. An excision in the integument is filled by blood which coagulates and is tanned into an insoluble membrane. The basement membrane of the adjoining epidermis acts as a self-sealing membrane and contracts to cover the excision. The epidermis is attached to the cuticle by the subcuticular layer which it resorbs and by pore canal filaments which are left behind as it migrates. The epidermis migrates as a sheet in contact with the cuticle then with the coagulated blood and basement membrane which cover the excision. Blood cells migrate individually into an excision and do not adhere to a surface in the process. Microtubules cannot be identified with movement. Both epidermal and blood cells remove the cells killed by wounding as evidenced by the appearance of coated vesicles and phagocytic bodies in both cell types. The reconstituted integument consists of a surface membrane in which the layers of the epicuticle are not distinguishable, a nonlamellate cuticle secreted by an epidermis which also appears to secrete the new basement membrane.

Notes: Adult sloths (Bradypus tridactylus) were studied by electrocardiography and by light and electron microscopy under normal conditions and under experimental conditions as provided for by injection of 2,4-dinitrophenol (DNP) and ether anesthesia. ECG's of the animals indicated heart rates of 45-71/min, which can be considered as the normal heart rate of the sloth under laboratory conditions. Under normal conditions, the contracted ventricular myocardium of the sloth exhibited (a) a wrinkled sarcolemma, (b) the usual pattern of myofibrils and of sarcoplasmic reticulum, (c) small mitochondria with spiked and branched, often anastomosed cristae, including a few small intramitochondrial dark bodies, (d) an amount of sacrcosomes smaller than the amount of myofibrils, (e) many glycogen granules, isolated, in the form of a chain, or as clusters, in subsarcolemmal, intermyofilamentous and perimitochondrial positions, (f) few multivesicular bodies and (g) large flat sections of the transverse tubular system.Injection of DNP (1 mg/kg) caused tachycardia. With ether anesthesia, the ECG showed monophasic action potential of myocardial injury and prolongation of inter or intraventricular condition. Electrically, the sloth's heart responded to hypoxia as do other mammalian hearts.The administration of DNP produced (a) derangement and reduction in number and length of the mitochondrial cristae, (b) disappearance of spikes, connections between the cristae and, consequently, the honeycombed arrangement, (c) increased matricial space in the center of mitochondria which was often filled with a grayish substance, (d) disappearance of small dark intramitochondrial granules, (e) depletion of glycogen particles and (f) few dilations in the sarcoplasmic reticulum.

Notes: Latex endocranial casts, which may be made without damaging the skull, reveal much of the information provided by a hemisected skull. Examination of drawings of endocasts superimposed on skulls may provide insight into the biological significance of skull and brain morphology. The high degree of cranial flexion and the globose brain shape of Daubentonia appear to be related to the functional demands of its gnawing mechanism. The broad frontal lobes of indriids are correlated with orbital orientation; differences in frontal lobe sulcal pattern suggest greater elaboration of the motor filed for the hand in indriids than in lemurids. Several features of lorisid cranial anatomy are discussed. It is suggested that, as a first approximation, increased splanchnocranial declination in small prosimians results from the necessity of accommodating relatively large eyes in a skull with a relatively small splanchnocranium.

Notes: The formation of cortical alveoli and yolk has been investigated in the pipe-fish, Syngnathus fuscus, and the killifish, Fundulus heteroclitus by techniques of light and electron microscopy. In addition to cortical alveoli and yolk components the ooplasm contains many mitochondria, numerous Golgi complexes, copious quantities of the endoplasmic reticulum of the rough variety, ribosomes and particulate glycogen. While the formation of cortical alveoli and yolk may proceed simultaneously, the cortical alveoli are the first to develop. Staining procedures indicate that cortical alveoli, like some of the yolk bodies contain a polysaccharide component and protein. It is suggested that the protein portion is made by the endoplasmic reticulum and is subsequently transferred to the Golgi complex via vesicles. Within the saccules of the Golgi complex the polysaccharide component is fabricated after which time the Golgi produce vesicles containing the products of either the cortical alveoli or yolk bodies.The precursors used in the production of the yolk are produced by the oocyte (endogenous) and by an organ other than the ovary (exogenous). The precursors made exogenously become associated with the morphologically and physiologically specialized oolemma and are subsequently internalized by the process of micropinocytosis.

Notes: In a variety of amphibians examined the stratum corneum was one cell in depth, although in Xenopus it was up to three cells deep. The flattened horny cells were closely fused together along their lateral membranes to form a continuous sheet. Disulphide bonds of keratin were most concentrated in the peripheral cytoplasm, but the interiors of the cornified cells were sufficiently well keratinized to prevent more than slight enzymatic cytolysis of the normal cell components. Characteristically large, weakly stainable, non-shrunken nuclear remnants were found in the salamander and frog horny layers, but the clawed toad had small pyknotic (parakeratotic) nuclei. The mature amphibian keratinocytes contained free fats, bound phospholipids, calcium and sulphydryl groups, together with acid phosphatase and non-specific esterase. Cornification appears to begin by a process of separate individual cell keratinization and lateral membranes of neighbouring cells only later become fused together. This differs from the process in higher vertebrates in which the cells undergoing keratinization form a uniform transitional layer in the epidermis. In the amphibian epidermis neighbouring cells occur in different stages of keratinization.

Notes: Eggs of the common snapping turtle, Chelydra serpentina serpentina were incubated at 30°C and at 20°C. The incubation period at the higher temperature was about 63 days. At the lower temperature, the period was estimated to be 140 days. Lengths of the embryos at various times of development were recorded. A series of 26 stages is described. The staging is based on timed intervals at a constant temperature, 20°C.

Notes: The postnatal development of the pelage and ventral gland of male Mongolian gerbils ranging from newborn to 86 days of age was studied. The development of the gerbil pelage follows a pattern similar to that observed for other rodents. The length of the dorsal and ventral skin juvenile hair cycle was found to be 26 to 28 days with a 15 to 18 day anagen and a ten to 11 day catagen and telogen. Hair follicles in the ventral gland began growth ten days later than those of the general pelage and secondary follicles budded from the sides of primary follicles. The ventral gland area differed from the general pelage in that it lacked a panniculus carnosus. The ventral gland is a complex of pilosebaceous glands which, in the adult, fill the entire hypodermis. The length and width of the pilosebaceous canals of the gland units are greater than those of the dorsum. The period of telogen of the hair follicles in the ventral gland is very short. The mid-ventral gland of the male gerbil appears to be a secondary sexual characteristic.

Notes: The structure of cells in the colleterial glands of the Cecropia silkmoth was examined. Morphologically and functionally the gland is divided into two regions, a tubular one in which columnar protein-synthesizing cells are located, and an expanded region in which flattened cells with very different structure are most prominent. The fine structure of the latter cells which are presumed to secrete a phenolic glucoside, closely resembles that of cells described in the colleterial glands of orthopterans. The protein-secreting cells have many features normally associated with pancreatic acinar, and other cells of similar function. Among these are extensive rough endoplasmic reticulum, an elaborate Golgi complex, and a modest number of mitochondria. Other features which are less usual in cells of this type are an elaborate secretory apparatus consisting of a cuticular tubule inserted into a microvilli-lined cavity at the apical end of the cell, and large numbers of cytolysomes, myelin figures, and lipid droplets. A chitogenous cell with a very distinct and specific type of ultrastructure is found associated with the secretory cell. This cell type is attached to the cuticular elements of the gland, and the main features of its cytoplasm are extensive bundles of microtubules which presumably serve as supportive elements for the secretory cells.

Notes: Embryonic chick myocardium (stages 8+ to 12-) was studied by light and electron microscopy. The myocardium, which is initially comprised of radially oriented cells with large intercellular spaces gradually becomes more tightly packed. Intercellular spaces decrease and the cells assume a circumferential orientation. Myocardial cells remain epithelial throughout formation of the functional tubular heart and specialized epithelial junctions (apical junctional complex or terminal bars) undergo modification to form intercalated discs. Embryonic myocardial cells contain large amounts of free ribosomes and particulate glycogen, the latter often associated with portions of granular reticulum. Unlike developing skeletal muscle. The amount of granular reticulum contained in the myocardial cell cytoplasm is large and, along with a hypertrophied Golgi apparatus, suggests that these cells may have a secretory function. These organelles persist during the initial period of fibril formation. Myofibrils apparently form from non filamentous precursor material and not by alignment of sequentially synthesized components.

Notes: The characteristic anoline climbing organ consists of a number of lamellar scales, on whose outer scale surface are numerous keratinized setae which contact the substrate. These setae are derived from the Oberhautchen of the epidermal generation, and as such are renewed and shed periodically along with the rest of the epidermal material. The histological development of the setae is described, and modifications of the surrounding elements are noted. The relative lengths of the setae and their congregation to form a pad unit poses certain mechanical problems during morphogenesis, simply in terms of accommodation between the functional outer epidermal generation and dermal core of each lamella. Regression of the dermal core and a distal migration of some cells permits accommodation within the lamella for the distal aspect of the Oberhautchen layer, or free margin. Additionally, changes in the gross shape of the lamella occur throughout the sloughing cycle, and a swelling of the cells of the lacunar tissue results in a gap between the stratum corneum of inner and outer epidermal generations. There is a considerable amount of variation in mitotic activity between the germinal layers of opposite sides of the lamella.

Notes: Function and ultrastructure of the excretory organs (antennal glands) of the shore crab Uca mordax were investigated. The crabs were maintained at three different salinities: 50%, 100% and 200% seawater. In spite of previous reports to the contrary, the investigation showed that the powerful osmoregulatory ability found in Uca mordax is not due to participation of the antennal glands. Freezing point depression of urine under all conditions was found to be slightly less than that of the hemolymph, indicating a slightly hypoosmotic urine. It was further found that the antennal gland is extremely effective in resorbing sodium from the filtrate. The higher the salinity to which the crabs were acclimated the lower the sodium concentration in the urine. No water was resorbed from the filtrate as shown by the fact that the inulin U/P ratio remained unity regardless of the salinity to which the crabs were adapted. Electronmicroscopy of the antennal glands revealed that the coelomosac cells are similar to the podocytes described in the crayfish by Kümmel ('64), and the coelomosac appears to be a typical filtration organ. The cells of the labyrinth showed brush border and very elaborate basal infoldings with numerous mitochondria. The deep cytoplasmic infoldings which represent interdigitations with neighboring cells may be correlated with the effective sodium reabsorption in the labyrinth, but apparently not with water movement.

Notes: In a hydrozoan jellyfish, the female gonad is differentiated from a specialized region of the epidermis near the manubrium. Changes in the oocytes during growth and vitellogenesis are described as observed with electron microscopic and cytochemical techniques. Three major types of yolk are formed; these include lipid, glycogen, and membrane-bound granules consisting of both protein and carbohydrate. The latter first appear evident within vesicular and cisternal elements of the numerous Golgi complexes. The orientation and structural variations noted between the endoplasmic reticulum and forming face of the Golgi complexes suggest that the protein component of the yolk granules may be transferred from the cisternae of the endoplasmic reticulum to the Golgi complex where it is joined to carbohydrate perhaps synthesized by the Golgi complexes. Stages in the release of the precursor yolk material sequestered in cisternal elements of the Golgi complexes are illustrated. The presence of coated and uncoated vesicles in the Golgi regions and their possible role in intracellular transport are described and discussed. The presence and possible method of morphogenesis of vesiculate yolk bodies are also described. What appear to represent invaginations of the oolemma extend into the ooplasm and display a special orientation with respect to lamellae of the rough-surfaced endoplasmic reticulum. Intraooplasmic synthesis appears to constitute the major pathway for protein-carbohydrate yolk deposition.

Notes: The dipnoan heart is only in part structurally developed to support a separated circulation in pulmonary and systemic circuits. In the present investigation biplane angiocardiography has been used to describe the extent of such a double circulation and the factors which may modify it in the African lungfish, Protopterus aethiopicus.Contrast injections in the pulmonary vein revealed a clear tendency for aerated blood returing from the lungs to be selectively dispatched to the anterior branchial arteries giving rise to the major systemic circulation. Contrast injections in the vena cava delineated the sinus venosus as a large receiving chamber for systemic venous blood. Contraction of the sinus venosus discharged blood into the right, posterior part of the partially divided atrial space. Contrast injection in the pulmonary vein showed that vessel to pass obliquely from right to left such that blood was emptied distinctly into the left side of the atrium. During contraction the atrial space tended to retain a residual volume in its anterior undivided part which minized mixing.Ventricular filling occurred through separate right and left atrio-ventricular connections. Right-left separation in most of the ventricle was maintained by the partial ventricular septum, the trabeculated, spongelike myocardium and the mode of inflow from the atria. Mixing in the anterior undivided portion of the ventricle during the ejection phase was slight due to a streamlined ejection pattern.The outflow through the bulbus cordis occurred in discrete streams which in part were structurally separated by well developed spiral folds. In the anterior bulbus segment the spiral folds are fused and make completely separate dorsal and ventral outflow tracts. The ventral bulbus channel provides blood to the three anterior branchial arteries. The second and third branchial arteries are large and represent direct shunts to the dorsal aorta. The fourth and fifth branchial arteries are gill bearing and receive blood form the dorsal bulbus channel. The most posterior epibranchial vessels give rise to the pulmonary arteries.

Notes: Variants resistant to 2-deoxy-D-glucose have been isolated from a clonal line of pig kidney cells by serial cultivation in the presence of inhibitor. Hexokinase activity may be affected directly in this system, since the oxidation of glucose to 6-phosphogluconate by extracts from sensitive and resistant cells is blocked by the addition of 2-deoxy-glucose to the reaction mixture. This blockage was removed by the addition of glucose-6-phosphate to the system, but not by ATP. Resistant cells were found to accumulate significantly less 2-deoxyglucose-6-phosphate than sensitive cells. The rate of phosphorylation of 2-deoxyglucose, however, was higher in extracts from the resistant line. Alkaline phosphatase does not account for the reduced level of 2-deoxyglucose-6-phosphate since this enzyme is not detectable in sensitive or resistant pig kidney cells. Increased acid phosphatase activity was observed in resistant cells, but extracts with high acid phosphatase activity proved incapable of hydrolyzing either 2-deoxyglucose-6-phosphate or glucose-6-phosphate. In comparative growth studies, cells resistant to 2-deoxyglucose proliferated more extensively than sensitive cells in a low glucose nutrient. They removed glucose more effectively from this medium, and were less stimulated by the addition of intermediates from the tricarboxylic acid cycle. The evidence suggests that resistance to 2-deoxyglucose in the cells under study may be based on the ability of the resistant cells to proliferate at concentrations of glucose too low to support the growth of sensitive cells.

Notes: The rates of synthesis of various species of RNA were examined in synchronously growing HeLa cells as a function of stage in the cell generation cycle. The synthesis of each RNA species examined occurs throughout interphase, but undergoes a two-fold increase in rate during early S which is dependent on the duplication of DNA; blocking the initiation of DNA synthesis also blocks the acceleration of RNA synthesis. To explain the data, a model is discussed in which the acceleration of RNA synthesis during early S is regulated by the number of active cistrons present; thus, as the genome is duplicated, more template is available for transcription, and the rate of RNA synthesis increases. Some implications of the model, and experimental evidence bearing on them, are also discussed.

Notes: A sulfated glycosaminoglycan has been isolated from the acid-soluble fraction of an established line of Chinese hamster fibroblasts grown in suspension culture. This material has a molecular weight between 5000 and 10,000, contains equimolar amounts of hexosamine and uronic acid (orcinol method), and about 0.6 sulfate groups per hexosamine residue. About 80% of the sulfate groups are N-sulfates on the basis of lability of the sulfate and the formation of equivalent numbers of free amino groups upon mild acid hydrolysis. The material is completely resistant to testicular hyaluronidase but is degraded to reducing monosaccharides and small oligosaccharides upon treatment with lyophilized cells of Flavobacterium heparinum that were grown on heparin. It is thought, therefore, to be related to the known N-sulfated glycosaminoglycans heparin and heparitin sulfate.

Notes: Human diploid cell strains develop progressively higher levels of specific catalase activity as they grow. Following subculture activity falls again. A diploid cell strain heterozygous for the gene for acatalasia I (acatalasemia) was found to develop specific catalase activity at proportionately the same rate as normal cell strains. Yet the mutant gene reduced the absolute level of specific catalase activity which the culture attained at any given point in time. In this respect the heterozygous acatalasia I strain resembles the homozygous acatalasia II strain previously reported.

Notes: The interferon mechanism offers the hope for moderate to high level prophylactic immunity of broad antiviral spectrum but of relatively short duration. Economic and biological considerations offer little hope for utilization of exogenous interferon as a prophylactic or therapeutic substance, unless but a small part of the total molecule be found to carry the activity. The real promise for interferon application is in the administration of suitable inducers so as to cause the body to produce and distribute its own interferon. Certain ribonucleic acids (RNA's) offer hope for high level potency as inducers without adverse effect. The condition for interferon induction by ribonucleic acids appears to be double- or multistrandedness and freedom from inhibitors. These can be of biologic or synthetic origin. The mechanism of action of interferon is not fully understood but appears to fit into the Jacob-Monod model involving two phases: first, a derepression by the inducer to cause the cell to form interferon and second, a derepression by interferon to cause recipient cells to form the active substance which acts by preventing translation from viral messenger RNA. Double or multistranded RNA of viral or other origin appears to be unique to the cell and serves as the alert to it to produce interferon in phase 1. Greatest need for interferon is clearly for those diseases in which there is a multiplicity of immunologic types in excess of the numbers which could be put into a vaccine as, e.g., the common cold and enteric viruses. There might be some overall therapeutic benefit also if inducer were given early enough in infection. Special value for interferon induction might derive by administration in early life before the development of immunologic maturity, as a means for preventing infection with oncogenic or other viruses. Additionally, suitable inducers might be capable of interrupting the reinfection cycle in virus-dependent malignancies.The favorable outlook for interferon utilization must always be tempered with the realization that under certain as yet undiscovered situations, adverse rather than beneficial effects might result from indution of interferon. It is not impossible that in certain special circumstances, as in ordinary immunologic responses, it might be more beneficial to negate rather than to promote the effect.

Notes: The fusion in vitro of embryonic myoblasts to form multinucleated myotubes requires the addition of serum to a basal nutrient medium. The serum requirement for fusion can be satisfied by insulin with somatotropin potentiating its effect. Myotubes formed under these conditions fail to differentiate to cross-striated, spontaneously contractile muscle fibers. This arrest of development is reversible if serum is restored to the medium.Development of the enzyme glycogen synthetase was studied as an additional indicator of muscle differentiation. In cultures developing in the presence of serum, this enzyme was demonstrated by autoradiography to be highly concentrated in myotubes as compared to mononuclear cells. The activity of the enzyme remains low in (1) cultures formed in response to insulin and somatotropin in the absence of serum, as well as (2) in cultures formed in unsupplemented basal medium which are virtually lacking in myotubes. The addition of serum to (1) restores the development of this enzyme. Serum which has been extensively digested with the proteolytic preparation, pronase, and subjected to boiling temperature, when combined with insulin and somatotropin is also capable of promoting the development of glycogen synthetase to a specific activity which exceeds the control. The serum factor is not lost on exhaustive dialysis, nor can enzyme promoting activity be liberated by heat denaturation of serum proteins.

Notes: The specific catalase activity of human diploid cell strains increases with progressive growth of the culture, and falls again following subculture. Although the increase is small, it is readily demonstrable, and is exponential with time.The response of catalase activity to proggressive growth of the culture was studied in three abnormal human cell lines. A diploid cell strain, developed from a patient homozygous for the gene causing acatalasia I, had no detectable catalase activity throughout the life cycle of the culture. Another diploid cell strain, developed from a patient homozygous for the gene causing acatalasia II, had about 5% normal catalase activity, but the proportionate increase in specific activity as the culture grew was the same as for normal cells. Thus the mutation causing acatalasia II does not change the responsiveness of the cell in terms of catalase activity to progressive growth of the culture. The behavior of a heteroploid line was similar to that of the normal diploid strains, but when the growth of the heteroploid cultures reached a plateau, their population densities were four times higher than those of the diploid strains and they had about twice the specific catalase activity.

Notes: Exogenous ATP can induce a marked cell enlargement in TA3 tumor cells which can be reversed or prevented by Ca and Mg. This regulatory effect on cell volume is specific for ATP. The mechanism probably involves changes in cell ionic content.

Notes: Filtration of mouse marrow cell suspensions over columns of glass wool increased the frequency of small and medium-sized lymphocytes (SML) and of erythropoietic progenitor units (EPU) by about the same factor. Identical results were obtained when erythropoiesis was assayed by isotope uptake (59FeCl3 and 125IUdR) or by the spleen-colony techniques.Transfusion of prospective donor mice with erythrocytes virtually eliminated morphologically recognizable erythroid cells from marrow without affecting the frequency of EPU. Injection of prospective donors with cortisol decreased the frequency of SML in marrow but not that of EPU or erythropoietin-sensitive cells. However, glass wool filtration of lymphocyte-poor marrow taken from mice pretreated with cortisol resulted in a similar increase in frequency of residual SML and of EPU. Therefore, it appears that a subpopulation of marrow SML are EPU.Whereas glass wool filtration increased the frequency of erythropoietic progenitor and colony-forming units, the filtration failed to change the frequency of leukopoietic progenitor or colony-forming units (assayed in mice hypertransfused with erythrocytes to suppress erythropoiesis). It follows that separate progenitor cells for erythropoiesis and leukopoiesis are present in bone marrow of adult mice, in addition to pluripotent stem cells.

Notes: The pyrimidine requirements for growth of T. pyriformis and for reversal of the growth inhibition caused by folate deprivation have been studied. The effects of thymidine and 5-fluorodeoxyuridine have been shown to be quantitatively different from the effects of these compounds on growth and the rate of DNA synthesis in mammalian cells. Labelled nucleosides added to the medium have been found to be converted to the corresponding bases with the exception of deoxycytidine, which is first deaminated to deoxyuridine. As a result no deoxynucleosides other than thymidine specifically label DNA.The results allow deductions to be made concerning the enzymes involved in pyrimidine utilization by this organism. It is suggested that pyrimidine utilization is always channeled through uracil in the case of those compounds that can supply the pyrimidine requirement for growth.

Notes: The probability of a colony which originated as a single stem cell to become extinct due to differentiation of all of its stem cells in any generation is closely connected to stem cell self renewal probability p. p can be determined from the coefficient of variation of the colony numbers received by reinjecting single colonies of the same age. Whole spleens containing a known average colony number can also be used with advantage for this purpose. The results of both procedures indicate a stem cell self renewal probability p =0.62 ± 0.04, which does not change significantly between the sixth and the fourteenth day of colony development, and an extinction probability ω = 0.63 ± 0.12.

Notes: The rabbit lens can be maintained in culture under conditions which retain the block to proliferation characteristic of many of the cells in the normal tissue in vivo. Proliferation and cell migration subsequently can be triggered at will by the addition of serum to the medium. Serum is not required throughout the whole period of culture in order to elicit this response but only during an initial period which ends prior to the onset of both DNA synthesis and mitosis. The duration of this period of serum-dependency is specifically related to the location (and hence the state of organization) of the cells within the epithelial layer. Some of the characteristics of the serum factors which promote these reactions and the conditions of culture which affect their activity are reported.

Notes: Cell preparations rich in polymorphonuclear neutrophils (PMN) were obtained from peritoneal exudates of rats without the use of any anticoagulant. The adhesiveness of these PMN to glass bead columns coated with rat serum were studied quantitatively using suspending solutions free of added serum protein. A dependence of the PMN adhesiveness upon divalent cations was demonstrated. Added singly Mg2+, Ni2+, Zn2+, Co2+, Mn2+, Cu2+, or Cd2+ were found to be effective whereas Ca2+, Sr2+ and Ba2+ were ineffective. A possible auxilliary role for Ca2+ when added with Mg2+ is suggested by the data. The ineffectiveness of the ions Ca2+, Sr2+ and Ba2+ was shown by use of an ion electrode not to be due to the unavailability of the ionized species. Procedures are described for obtaining highly reproducible results with the Orion Divalent Cation Electrode. The ineffectiveness of the ions Ca2+, Sr2+ and Ba2+ were also shown not to be due to action as general protoplasmic poisons. The effective ions are distinguished from the ineffective ones by characteristic ranges of ionic radii, coordination number, second ionization potentials, electronegativities and affinity constants. Removal of components of complement from the cells by washing in 0.05 M EDTA, and heating all serum used for 30 minutes at 56°C had no significant effect on the adhesiveness of the PMN. A role for complement, therefore appears largely excluded.

Notes: Because its differentiation can be channeled into different pathways, amphibian gastrula ectoderm is a convenient test system for studying factors that control embryonic differentiation. 1Chemical nature of inducing factors: A substance that induces muscle and notochord in ectoderm has been isolated from chick embryos and other sources. The factor is protein in nature (mol. wt. in 6 m urea 25,000--30,000). Neural tissue is induced by a crude ribonucleoprotein fraction. Purified RNA has only a very weak inducing activity. The inducing factors are preferentially located in cytoplasmic particles.2Mechanism of action: Embryonic induction has to be considered as a derepression. Preliminary experiments have shown that a high-molecular-weight, water-soluble substance takes part in the inhibition of mesodermal differentiation. The inhibition of differentiation is released by the inducing factors. A close relationship between differentiation and RNA synthesis has been revealed by experiments with actinomycin D (0.5--2.5 μg/ml), which inhibits RNA synthesis. If RNA synthesis is completely stopped in the gastrula stage, the mesodermal area, which is already determined to differentiate into muscle and notochord, still forms some notochordal cells and myoblasts. The differentiation of neural tissue, however, is completely inhibited. DNA-RNA hybridization experiments at the saturation level suggest that new messenger RNA species are synthesized if differentiation proceeds. But this does not exclude that the inducing factors exert control primarily at the level of translation.

Notes: Non-dividing mouse embryo fibroblasts which grew to a confluent cell density on one side of an ultra-thin filter did not inhibit the active multiplication of the same type of cells growing at low cell density on the other side of the filter directly opposite the confluent side. The close proximity of the cells across the filter was not sufficient to cause inhibition of cell division. The phenomenon of “contact” or “density dependent” inhibition of cell division is therefore probably not mediated by a cellular product which remains concentrated near the cell surface.The degree of contact inhibition of cell division was correlated with the local cell density on the same side of the filter. This relationship was found to be influenced strongly by the surface on which the cells were growing.

Notes: The effects of various ions on the resting membrane potential of the giant axons of Myxicola were determined. The mean resting potential in artificial sea water is 69 mv, inside negative. The membrane potential decreases with incresing external potassium concentrations, while changes in the sodium and chloride concentrations have little or no effect. For potassium concentrations greater than 50 mM/L the relation between membrane potential and concentration approximates that of a perfectly selective potassium electrode. The data for the whole range of concentrations examined can be well fitted by the equation: \documentclass{article}\pagestyle{empty}\begin{document}$$ {\rm E} = \frac{{{\rm RT}}}{{\rm F}}\,{\rm log}_{\rm e} \,\frac{{[{\rm K}^ +]_{\rm o}\, + 0.031\,[{\rm Na}^ +]_{\rm o}}}{{[{\rm K}^ +]_{\rm i}\, + \,0.031\,[{\rm Na}]_{\rm i}}} $$\end{document}It was pointed out that the Myxicola giant axons can be studied under space voltage clamp and can be made available in the laboratory for 12 months out of the year. Myxicola then should become a very useful preparation for the study of membrane phenomena.

Notes: Cultures of several cell lines convert cytidine, if present in their medium, to uridine. The reaction is rapid, being virtually complete within one hour. The enzymatic activity is that of cytidine aminohydrolase (EC 3.5.4.5). The activity is exhibited by the intact cell  -  substrate and products being found in the medium bathing the cells. The activity should be taken into account in studies involving cytidine, and other 6-amino pyrimidine nuclecsides.Of nine transformed cell lines, and nine primary and secondary cell strains screened for the presence of cytidine aminohydrolase activity, six cultures were positive. All the positive cultures were heteroploid transformed lines. No diploid strain cultures were positive, and two near diploid mouse lymphoid neoplasms were negative. PPLO contamination was not detected in the positive cultures.Within the limited series of cell cultures screened in this study, there appears to be a correlation between heteroploidy and the ability of the intact cultures to deaminate cytidine in excess of the cell requirement for the precursor.

Notes: Certain cells, such as 3T3 mouse embryo fibroblasts, are inhibited from dividing when they grow to a characteristic cell density on a surface in tissue culture. We asked whether the inhibition of cell division could be attributed to the inert chemical composition of neighboring cells, that is, whether the residues of lyophilized cells retained the ability to inhibit the division of normal cells. In addition, we wanted to know whether cells in which DNA synthesis was imparied by irradiation would retain the capacity to effectively inhibit normal cells.To answer these questions, confluent and non-confluent layers of 3T3 cells were prepared in tissue culture dishes and the cells were either lyophilized or irrariated in situ. Fresh 3T3 cells were then added to these prepared layers and their growth was followed using radioactive label. There was no growth of added cells on the confluent monolayers of either untreated or irradiated cells. Growth was unimpeded on the monolayers of lyophilized cells. When cells were added to non-confluent cultures of either normal or irradiated cells the added cells grew until they had covered the remaining surface of the culture dish and had come into contact with the pre-existing cells. In the discussion, consideration is given to the role of available surface over which the cells can spread as well as to the possible interactions between neighboring cells.

Notes: The rate of RNA synthesis in synchronously growing HeLa S3 cells was determined as a function of position in the cell generation cycle. Measurements throughout the cycle of both the rate of incorporation of radioactively-labeled uridine and of the total amount of RNA indicate that (1) the rate of RNA synthesis is constant (or increases only slightly) during G1, approximately doubles during the first half of S, and then remains constant during the remainder of S and G2, and (2) cells attain the average G1 rate of RNA synthesis very early in G1, and maintain the average G2 rate until mitosis.If the initiation of DNA synthesis is blocked, the acceleration of RNA synthesis is markedly reduced or eliminated. Further experiments in which DNA synthesis was inhibited at different times in S, or to varying degrees from the beginning of S, suggest that the extent to which RNA synthesis is accelerated depends on the amount of DNA duplicated. These data also indicate that duplication of the first half, and in particular the first few per cent, of the DNA complement results in a disproportionate acceleration of RNA synthesis.The possibility that fluctuations in the sizes of precursor pools may lead to misinterpretation of labeled-uridine incorporation data was examined. Experiments indicate that in this system pool fluctuations do not cause invalid measures of RNA synthesis.It is concluded that RNA synthesis occurs throughout interphase, but undergoes a two-fold increase in rate which is dependent on the duplication of DNA.

Notes: The distribution of in vivo injected 45Ca++ in the subcellular fractions of rat heart has been studied. Most of the radioactivity of the cell was found to be associated with the subcellular organelles; only a small fraction was recovered in the soluble phase. Mitochondria contained the greatest part of the total radioactivity associated with the subcellular organelles. After injection of 45Ca++ the specific activity of the mitochondrial calcium pool was several times higher than that of the calcium of the sarcoplasmic reticulum. Pentachlorophenol has been administered to rats to uncouple oxidative phosphorylation in heart mitochondria in vivo and its effect on the distribution of 45Ca++ in the heart studied. Under these conditions, it has been found that mitochondria contained much less 45Ca++ than the controls; this decrease was paralleled by an increase of the radioactivity associated with the microsomes and with the final supernatant. Experiments in which 45Ca++ was added to heart homogenates at 0° indicated that 45Ca++ also became bound to mitochondria and the other subcellular structures at 0°. However, PCP had no effect on the distribution of radioactivity among the subcellular fractions under these conditions. The results suggest that (1) energy-linked movements of Ca++ take place in mitochondria of the intact rat heart, (2) a part of the uptake of 45Ca++ by mitochondria does not depend on metabolism, and, (3) the movements of Ca++ in heart mitochondria in vivo are probably more active than those in the sarcoplasmic reticulum.

Notes: Aerobic incubation of the spermatozoa of freshwater mussel (Hyriopsis schlegelii) caused a reduction in acyl-ester content and a corresponding diminution in lipid-phosphorus content. Anaerobic incubation also caused a reduction in acyl ester but a smaller reduction in lipid phosphorus, while it brought on an accumulation of free fatty acids. It was mainly palmitic and stearic acids which were accumulated during the anaerobic incubation, and they were preferentially metabolized during the aerobic incubation. Complex lipids from the spermatozoa consisted mainly of diacyl glycerophospholipids, in which phosphatidylethanolamine was predominant, followed by lecithin, while plasmalogen was a minor component. After aerobic incubation of the spermatozoa, there was a marked decrease in ethanolamine-containing phospholipid fraction. However, no diminution was observed in the plasmalogen content. These results lead to the conclusion that mussel spermatozoa utilize as a primary energy source fatty acids which are derived from the breakdown of intracellular diacyl glycerophospholipids.

Notes: An investigation of the physiological effects of phenethyl alcohol (PEA) on exponentially growing yeast cells is reported. RNA, DNA, protein and aminoimidazole ribotide syntheses and glucose uptake and incorporation are inhibited by PEA at concentrations of 0.1% to 0.3%. Two classes of response curves are found and the sensitivities of processes in each class to PEA differ. Glucose incorporation and RNA synthesis are the most sensitive processes in their respective classes. The effects of PEA at 0.3% or less are largely or completely reversible. It is deduced that PEA inhibits intracellular processes as well as the cell membrane.

Notes: Somatic hybrids between pigmented Syrian hamster cells and unpigmented mouse cells were isolated and propagated in vitro. These hybrids are unpigmented and lack dopa oxidase (and tyrosinase) activity, which is correlated with the pigmentation of the Syrian hamster cells. In contrast, the presence of three other enzymes (LDH, MDH, and thymidine kinase) also specified by the hamster genome but unrelated to pigment synthesis is observed in the hybrid cells. This suggests that the repression of dopa oxidase in these cells is a specific effect on the enzyme associated with the differentiated state of pigment cells. It is concluded that the genetic control of differentiation in this case involves a diffusible regulator substance which functions negatively.

Notes: Balb/c mouse embryo cultures that are carried under a rigid transfer schedule that minimizes cell-cell contact evolve into permanent lines that possess properties very similar to 3T3. From the same embryo cultures, a transfer schedule where cell-cell contact is extensive leads to lines (Balb/3T12) that form multiple cell layers and have saturation densities up to 25-fold higher than the Balb/3T3 lines. All the lines are hypotetraploid.Comparison of the Balb/3T3 lines with 3T3 reveals similar morphology, ability to grow at high dilution, low saturation density, and high susceptibility to transformation by SV40 virus. The Balb/3T12 lines, which are continually maintained at high cell density, show little improvement in cloning efficiency. Infection with SV40 produces transformants that can be selected by their ability to form colonies at low cell densities. Using the appropriate selective system for each line, it appears that SV40 T antigen induction and transformation in Balb/3T3 and Balb/3T12 are comparable.

Notes: Human lymphocytes stimulated with phytohaemagglutinin in vitro show maximal 14C-thymidine incorporation when the gas phase of the cultures contains 20% O2. The rate of DNA synthesis is more affected by hyperoxic conditions than by hypoxia. Cell viability, defined by dye exclusion, is considerably less dependent on pO2 than is cellular replication.

Notes: The cytological effects of 2 mM hydroxyurea upon Chinese hamster cells at various phases of the cell cycle were examined. Cells in the G1, G2, or M phases of the generation cycle treated with hydroxyurea showed no chromosomal aberrations. Cell treated in S phase became moribund and eventually lysed. Some of these moribund S cells reached mitosis much later and were found to have chromatid aberrations. Cells in the log phase of growth, surviving exposure to 2 mM hydroxyurea for six hours, also showed no aberrations. Thus, viable (colony-forming) cells, resulting from synchrony procedures with hydroxyurea are free of chromosomal aberrations.

Notes: Carotenoids were introduced into the egg yolk of the medaka by either injection or feeding. Carotenoids for injection were dissolved in either olive oil, a mixture of olive oil and castor oil or Tween 80. Capsanthin, lutein and canthaxanthin were readily transferred from the yolk to the larval xanthophores, but no β-apo-8′-carotenal and little β-carotene were transferred.

Notes: Ouabain induced inhibition of cation transport and cell division in Ehrlich mouse ascites tumor cells is reversible, suggesting that this agent does not bind irreversibly to its site of action.

Notes: Actinomycin D(AM), an inhibitor of DNA-dependent RNA synthesis, produces a reversible cessation of red blood cell production. This study examines the in vivo cellular uptake of 3H-AM in the hematological tissues and livers of B6D2F1 mice. 3H-Am (sp. act. = 2.97 to 4.20 C/mmole) was given IV at a dose of 4.0 to 5.7 μg (14 μc) per mouse. Spleen, bone marrow, blood, and liver samples were taken for autoradiography at post-injection times of five minutes to 67 hours.We have confirmed the rapid in vivo cellular uptake of AM; substantial quantities of the drug were in the nuclei within five minutes of IV administration.Not all cell types became labeled. Erythroid, hepatic, lymphoid, and reticulo-endothelial (RE) cells and monocytes took up the label, whereas labeling of granulocytic elements was doubtful.Most heavily labeled were liver cells (highest mean grain count = 110.1) and splenic RE(19.1) and erythroid (16.1) cells. Erythroid cells in the spleen were more heavily and more rapidly labeled than those in the bone marrow.All nucleated erythroid maturational stages, in both the spleen and the bone marrow, were labeled, even at five minutes.The time course of erythroid and hepatic labeling was quite different. Whereas early erythroid cells required six hours to become 100% labeled, liver cells were 100% labeled at five minutes and loss of hepatic labeling began as early as 15 to 30 minutes.

Notes: Single cell suspensions have been prepared from rabbit cartilage by collagenase digestion in the presence of fetal bovine serum. When inoculated into medium F12 supplemented with 5% fetal bovine serum and 5% rabbit serum, the cartilage cell suspensions yield large healthy colonies of epithelial-like cells with an average plating efficiency of 47%. At the end of a 14 day incubation period, the centers of many of the colonies are thickened into multilayered aggregates. Such colonies exhibit serveral cartilage-like properties, including metachromatic staining with toluidine blue, dense brown staining with bismark brown, bright green staining with alcian green, and a refractile matrix which can be seen with the phase contrast microscope to surround the living cells. Similar properties have also been observed in second and third passage subcultures.

Notes: To determine whether transcription of the genes coding for 5S ribosomal RNA depends upon the concurrent activity of the genes for 28S and 18S ribosomal RNA we studied the synthesis of 5S RNA in cultured L cells when the synthesis of the 45S precursor of 28 and 18S RNA was inhibited with low doses of actinomycin D. Synthesis of 5S RNA was measured by following the incorporation of 3H uridine into components which, upon acrylamide gel electrophoresis, co-migrated with markers of 5S RNA obtained from 32P labeled ribosomes. We observed that the synthesis of 5S RNA persists in the absence of detectable 28S and 18S RNA synthesis, continuing at the normal rate for several hours and at a reduced rate for at least a generation time. This strongly supports the concept that 45S RNA is not a precursor of 5S RNA, and indicates furthermore that neither the 45S component, nor any of the intermediates involved in its transition to 28S and 18S RNA, are involved in controlling the output of the 5S genes The 5S RNA which is made during actinomycin treatment is retained predominantly in the nucleoplasm and undergoes a turnover of about 3.5% per hour. It apparently cannot be utilized when ribosome synthesis resumes following removal of the actinomycin.

Notes: It is becoming increasingly evident that the in vitro induction of vertebral chondrogenesis may not be due to the acquisition of a new metabolic pattern, but to the stabilization of one that is already existing. The sclerotome region of embryonic chick somites will undergo chondrogenesis in vitro as a response to various stimuli, the most effective being the embryonic notochord. Before stimulation, the somites have all of the metabolic machinery necessary for chondrogenesis, as evidenced by the fact that they can synthesize chondromucoprotein. They cannot, however, accumulate matrix except under special circumstances, e.g., when they are stimulated by an “inducer” such as the notochord (induced cartilage) or when they are permitted to express their chondrogenic bias under suitable culture conditions, such as in enriched nutrient medium (spontaneous cartilage).A study of chondroitin sulfate synthesis in these tissues has shown that the embryonic somites utilize glucosamine differently when compared to cartilage tissue. Analysis of the metabolic steps between glucosamine and UDP-N-acetylgalactosamine indicates the possibility of a metabolic “block,” which would prevent the efficient transformation of N-acetylglucosamine-1-P to UDP-N-acetylglucosamine.

Notes: Since Euglena gracilis Klebs var. bacillaris Pringsheim contains a species of DNA unique to the chloroplast, an important question concerns the extent to which light unblocks the reading of the organelle's template to provide the informational RNA's necessary to construct the plastid proteins. Experiments with 32Pi labeling of chloroplast and nonchloroplast RNA's during light-induced chloroplast development show that both the RNA of the chloroplast and of the rest of the cell become labeled during this process, with the chloroplast RNA's displaying the higher specific activity. The fact that chloroplast RNA is not uniquely labeled indicates that process other than a simple reading of the chloroplast DNA are involved. If we are to preserve the concept of a reasonable degree of chloroplast informational autonomy, we may assume, from this and other data, that the light induction of chloroplast development involves not only the unblocking of chloroplast DNA to make information available, but also a concomitant unblocking of other sites of informational RNA synthesis (e.g., nuclear and mitochondrial DNA's). Such sites external to the developing chloroplast may be concerned with making available the building blocks and energy necessary for the synthesis of chloroplast constituents coded for by the chloroplast DNA. This model leads to the prediction that photosynthesis could be gratuitous for chloroplast development if these nonchloroplast sites were providing most of the building blocks and energy. Experiments are reported which show that chloroplast formation and the acquisition of photosynthetic competence can be achieved under conditions where photosynthesis is completely inhibited for the entire span of development by using the highly selective inhibitor 3, (3,4-dichlorophenyl) 1, 1-dimethyl urea (DCMU), in agreement with the proposed model. The fact that more than just the chloroplast responds to the inducing signals for chloroplast differentiation raises the problem of experimental measurement of interaction among cellular organelles. Since chloroplast development is usually carried out in resting cells to avoid complications due to cell division, we discuss the limitations imposed by turnover in such nondividing systems and present evidence that most of the RNA labeling observed, although actinomycin-D-sensitive, is due to turnover and/or the utilization of preexisting pools. Evidence obtained with mutants of Euglena that form only partial chloroplasts or that lack plastid DNA and plastid-related structures is reported. Such evidence indicates that the functional proplastid restrains overall RNA labeling in the uninduced cells and suggests that the proplastid might be the source of regulatory metabolic signals in the normal plastid-containing cells.

Notes: The biochemical information now available on the cellular slime mold is sufficient to attempt a kinetic model of a portion of the metabolism essential to differentiation. A computer model has been constructed which is consistent with the in vivo and in vitro data obtained for this system, and which makes specific predictions now being tested in the laboratory.Uridine diphosphoglucose (UDPG) is a precursor of soluble glycogen as well as of the saccharide end products of differentiation, such as cell wall material. The rate of turnover of UDPG in vivo is known, and is sufficient to account for the observed accumulation of end products over the period of time normally required during differentiation. The accumulation patterns during development of the various polysaccharides and of UDPG, glucose-1-phosphate (G-1-P), and uridine triphosphate (UTP) are known. Km values of UDPG synthetase for G-1-P and UTP have been determined, as have changes in specific enzyme activity during development. A series of differential equations describing the synthesis and utilization of UDPG have been used to construct a computer model for the conversion of glycogen through G-1-P and UDPG to the end products of differentiation. An analysis of this model demonstrates that an increase in UDPG pyrophosphorylase concentration in vivo cannot account for the enhanced rate of synthesis of UDPG nor for the accumulation patterns observed. The major controlling factor in this system is shown to be the availability of G-1-P. This analysis reflects upon: (1) the general significance of alterations in the concentration of enzymes during differentiation, and (2) the importance of understanding control mechanisms responsible for the availability in vivo of precursors for synthetic pathways necessary to morphogenesis.

Notes: It was shown previously that the specific catalase activity of human diploid cell strains falls immediately after subculture and then progressively rises in an exponential fashion. In this paper evidence is presented suggesting that the rise in catalase activity cannot be due to an accumulation within the cell of a small molecule which enhances enzyme activity in cell-free extracts. It is also shown that activity per cell, as well as per unit cell protein, rises as the culture grows.The rate of fall of specific catalase activity immediately after subculture is greater if the cells are at a low population density than if they are at a high one. The rate of fall can be made more sharp by increasing the frequency with which the cultures are fed.It is shown that used medium, which has previously been incubated with cultured cells of the same strain, does not significantly change either the rate of fall of specific catalase activity following subculture, or the rate of its subsequent rise. It is postulated, as one possibility, that the cells liberate into the medium an enhancer of cell catalase activity which is highly labile. The steady state concentration of this enhancer in the medium might be expected to increase as the culture grew, but to decrease when the cells are subcultured into fresh medium or when the frequency of feedings is increased.

Notes: Erythropoietic activity of spleen cell grafts was measured (Fe59 uptake) in X-irradiated recipient mice under conditions in which these grafts were engaged in homograft reactions against allogeneic target cells or in graft-versus-host reactions. Such Fe59 incorporation was greatly reduced at 7 to 10 days after graft implantation relative to that of control grafts. This reduced erythropoiesis did not occur when the spleen cell graft was immunologically incompetent. Transplantation of bone marrow-lymph node cell mixtures also resulted in a relative decline in Fe59 uptake, but only when minimal numbers (105 to 106) of marrow cells were injected. The incorporation of I125 UdR in the spleen of irradiated recipients was used to assess cellular proliferation. Incorporation of this label was reduced when measured 7-10 days after implantation of the lympho-hemopoietic cell graft, but reached a peak at five days - the latter indicating stimulated lymphopoiesis. These data are consistent with the concept of depletion of a pluripotent stem cell pool (limited in size under these experimental conditions) due to excessive and concurrent functional demands for erythropoiesis and lymphopoiesis. An alternative explanation would involve cytotoxic effects on hemopoietic elements present in the milieu of the immunologic reaction.

Notes: The periodic mating behavior of some stocks of Paramecium aurelia, syngen 3, exhibits features typical of circadian rhythms. In the most extensively studied stock (37p), rhythmicity persists at least four days in continuous darkness, but disappears rapidly in continuous illumination (200 foot-candles). The period of the free-running rhythm is 22.2 hours, and relatively insensitive to ambient temperature. Cycles of illumination and temperature can regulate the mating rhythm. Changes in illumination at specified times in the circadian cycle will induce shifts in the phase of the rhythm. Stock differences with respect to the persistence of the rhythm and the environmental control of its phase have been observed.

Notes: Differential equations have been written and solved to describe the concentration gradients for the two hydra growth factors. The equations consider diffusion, catabolism and synthesis of the materials based primarily on the model of hydra growth described by A. L. Burnett. Concentration gradient profiles were obtained which correspond with the concentration gradients deduced from experimental evidence. It has also been possible to show that in areas of high mitotic rate, mitotic rate is related to the calculated concentration ratio of the two growth factors (stimulator/inhibitor). However, the correspondence of mitotic rate to growth factor concentration ratio does not hold for extreme values of the ratio indicating that very high and very low concentration ratios are not conductive to mitotic activity.

Notes: Using the hematopoietic colony technique, we have investigated the repopulating potential of bone marrow cells and leukocytes of blood from normal mice and have demonstrated that the frequency of hematopoietic stem cells in bone marrow is 50 to 150 times that of stem cells in the circulating blood. The differentiation capacity of these stem cells has also been examined. Results of comparative studies of the serial sections of hematopoietic colonies formed from marrow and blood leukocytes indicate that the differentiation capacity of stem cells from marrow and blood is similar, and that at least 80% of these cells differentiate along a single cell line. Thus, peripheral blood stem cells can effect a complete hematopoietic graft, establishing in the host, donor red cells, granulocytes, and platelets.The possibility that blood leukocytes may serve as a potential source of stem cells for hematopoietic transplants has been considered. Although blood contains stem cells, their frequency is so low as to make it unlikely that they would become a useful source of precursor cells for transplantation purposes.

Notes: Visible light of moderate intensity causes two and perhaps three types of division inhibition in Euglena gracilis are related cells. Fluorescent light causes a general inhibition of growth and division which is temperature-dependent. Pigmentation or complex organic media partially lifts this inhibition. A second type of inhibition, which is transient, can be caused by either fluorescent or incandescent light, and is found with an irreversibly bleached strain of Euglena grown on a limiting concentration of acetate; this inhibition could not be demonstrated in cells grown on limiting concentrations of glucose.

Notes: Photoreactivation of growth and DNA synthesis in UV-irradiated cells and photoreactivating-enzyme activity of cell-free extracts can be demonstrated in a cell line derived from liver tissue of the African clawed toad, Xenopus laevis. This cell line, A8W2, is a favorable system for the quantitative study of photoreactivation in vertebrates.