ALBERT

Notes: The blood of Perophora viridis is found to contain six types of cells: (1) Green cells, which have green-colored fatty bodies embedded in clear cytoplasm. (2) Orange cells, with orange-colored bodies of unknown composition in the cytoplasm. (3) Colorless berry-like cells, with fluid-filled vesicles in the cytoplasm. (4) Granular amoeboid cells. (5) Compartmental amoeboid cells, which have box-like vacuoles containing brownian granules of a fatty substance. (6) Vesicular, signet-ring type of cell having a single large vacuole. The cytological structure of these cells and their reaction to various dyes are described.An effort has been made to homologize the types of cells found in the blood of other ascidians with those found in Perophora.It is concluded that the variety of colors found in the cells of ascidian blood is due to the varying chemical states of the vanadium-containing chromogen present in the cells.

Notes: Embryological study of bullfrog tadpoles collected from various parts of the United States has shown the existence of local races which differ markedly in regard to the time of occurrence and character of the developmental processes involved in the formation of the definitive testis of the male individuals. Those races in which the gonads of the two sexes are easily distinguished in early larval stages are called differentiated. Other local races show a peculiar gonadic development chiefly affecting the males, the definitive testis sometimes not appearing until near the end of the second year of larval life. Such races are called undifferentiated, because the morphological features of the definitive testes are not established until late. The larvae first develop a peculiar gonad (progonad) which later degenerates and is replaced by the definitive testis. All male animals of the undifferentiated strains exhibit the gonad cycle.The progonad varies among the local frog races in regard to the length of persistence and degree of differentiation attained before undergoing degeneration. Its germ cells may exhibit a typical male maturation cycle ending in degeneration, or the cells may differentiate along both male and female lines or remain sexually neutral.The development and differentiation of the progonad in the various races are described and a detailed account given of the origin of the definitive testis. The problems of sex differentiation and continuity of the Keimbahn in anurans are discussed.

Notes: Disintegration in killing agents was studied throughout development. In the unfertilized egg and cleavage stages the death gradient runs from animal to vegetal pole. In the late blastula stage the future dorsal surface and future point of gastrulation show heightened susceptibility. The gastrula has a gradient from anterior to posterior end along its dorsal surface, with a slight reverse gradient around the blastopore; lateral and ventral regions are least susceptible.Before and after the appearance of the neural groove, the dorsal surface shows increased susceptibility with gradient in it from anterior to posterior end. The neural tube is highly susceptible, with a death gradient from anterior to posterior end and a slight reverse gradient at its posterior end.During late stages and in the larva the double gradient is present; death begins at the two ends and progresses backward from head, forward from anus; from the former most rapidly. The least susceptible place is near the posterior end. The posterior reverse gradient is less developed in the lamprey than in other vertebrate embryos, due, probably, to its lack of a tail bud.Assuming that death differences indicate differences in rate of activity, it appears that such differences in activity may be causes and not results of developmental processes, for the development of certain parts (dorsal surface, blastopore, central nervous system) is indicated by heightened activity before it is evident morphologically.

Notes: The prochordal plate in the chick is a thickening of the entoderm in the anterior part of the area pellucida. It consists at first of several layers of yolk-charged cells, which finally lose their yolk and form a thin layer of entoderm.The prechordal plate (an area of mesodermal proliferation immediately anterior to the notochord) arises immediately anterior to the primitive streak (and head process when the latter appears) in a part of the region once occupied by the prochordal plate. Later the prechordal plate comes to lie upon the dorsal wall of the foregut.The premandibular cavities arise in condensations of the prechordal mesoderm. The cavities seem to appear during the rapid expansion of the prechordal mesoderm which occurs in response to a release from constraint imposed by surrounding structures. They are connected with one another across the midline by a usually solid bridge of mesoderm.Later, the cavities are replaced by a mesodermal condensation which serves as a mold, on the surface of which the oculomotor muscles appear, arising before the disappearance of the premandibular head cavities.Differences in the relations of notochord and hypophysis in the chick and robin may be explained by an analysis of the growth of the two forms. The analysis furnishes a basis for understanding the relations of the anterior end of the notochord in the mammal.

Notes: Paramecium trichium varies from 50 μ to 105 μ long with most individuals between 80 μ and 90 μ. The width varies from one-third to one-half the length. It is somewhat depressed dorsoventrally. The broad buccal groove extends from the anterior left border diagonally across the ventral surface to the mouth, which is usually slightly anterior to the middle and to the right side of the median line. The mouth leads into a relatively long cytopharynx containing an undulating membrane. The cytopyge is subterminal and the small caudal tuft of longer cilia is subapical. The plastic ectosarc contains numerous trichocysts. A contractile vacuole apparatus occurs near either end. Each is deeply located and stains more intensely than surrounding protoplasm. In life there appear to be two alternately contracting vacuoles with smaller ones adjacent. Contractions are from fifteen to twenty-five per minute. The macronucleus is medium in size and the single micronucleus is of the ‘caudatum’ type.Binary fission appears to be initiated by a metaphase-like condition of the micronucleus. This is followed by great enlargement and the eventual separation of the chromatin threads into two anaphase groups. The metaphase thus appears to precede the changes which correspond to a prophase in other cases. During division of the body, the two old contractile vacuoles persist as the posterior ones for the daughters, new anterior ones being developed.

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: In a leech infesting the Alaskan codfish germinal masses in the ovary proliferate secondary groups, comprising about forty cells in which a follicle and central supporting cell early differentiate. Active division results in approximately 500 cells which apparently develop ductules extending to a point on the surface of the egg. Granules of unknown origin then appear in each nurse cell, and are drawn down the ductules into the egg which can now be distinguished. Reasons are given for the belief that the nutritive material is drained from the nurse cells by amoeboid activity of the egg. In early stages the nutritive mateiral forms a loose reticulum which gradually becomes transformed into a more extensive network, persisting until the maturation divisions. In this latest period the follicle and nurse cells, which become shrunken as the ovum enlarges, usually are stripped off and soon disintegrate.

Notes: The pelvic fins of the ancestors of the Chondrosteoidei possessed a metameric musculature and their skeleton consisted of a large number of metamerically arranged cartilaginous fin-rays, to which were attached osseous lepidotrichia. Evolution has involved the concrescence of separate elements to form the basal cartilage, the proximal end of which forms the girdle of the fin; the loss of a number of the fin-rays, and the atrophy of distal elements of the rays. The adult Chondrosteoidei have retained the primitive fin structure which characterized the elasmobranchs of the Palaeozoic period and which has disappeared in recent forms.

Notes: Three groups of pure lines of diverse clones of Didinium nasutum were maintained in isolation cultures for 778, 786, and 457 generations, respectively, without exhibiting any decrease in fission rate or any increase in encystment rate or death rate. These lines were supplied with Paramecium caudatum in such numbers that a surplus of food was present at all times.Three other groups of lines of the same clones were established simultaneously and cultured in parallel with the preceding groups, but the food of each of these lines was limited to nine paramecia per line daily. The fission rate of these lines fell to zero and the encystment rate increased to 100 per cent after 155, 165, and 113 generations of culture, respectively. The death rate increased appreciably in these lines prior to encystment.Other groups of pure lines were cultured on a diet limited to six paramecia per line daily. These lines encysted after approximately fifteen generations of culture.This evidence indicates that there is nothing in the nature of a definite life-cycle in Didinium and that diminished vitality and encystment do not result from the passage of generations, but from inadequate and unfavorable cultural conditions-specifically, from in-sufficient food. It shows further that it is possible to induce cycles with reference to encystment in Didinium by limiting the food supply and to vary the length of the cycles by varying the quantity of food.

Notes: Chlamydoselachus resembles the Holocephali in possessing no siphon, but a cavity in the proximal portion of the clasper. The cartilages, musculature, and venous sinuses of the clasper are considered. An older specimen of Echinorhinus is proved to have a spine on a soft papilla; Scymnus has no spine; both have the general features of the Spinacidae. Two species of Cestracion are compared with C. philippi. Mustelus lunulatus lacks the pera and pseudosiphon of M. vulgaris. Dicerobatis has a peculiar scaphus, and the claspers end in a fimbriated manner. Pteroplatea resembles Trygon and Benthobatis and Astrape are both like Torpedo.The species are listed below.

Notes: In the golden-mantled ground-squirrel, Callospermophilus, a spatulate glandular area has been noted in the skin of the back. It has been found in the following species: C. l. lateralis, C. l. arizonensis, C. l. caryi, C. l. saturatus, C. l. tescorum, C. c. chrysodeirus, and C. bernardinus. Probably it is common to the genus.The individual glands making up this area are modified and enlarged sudoriparous glands. They are divided into a tightly coiled and branched fundus, a large sinus, and a duct which passes caudad and outward to its exit at the surface.The glands secrete a strongly smelling oil, which is probably left on vegetation and other objects in the animal's environment and serves as a source of information to other members of the species. The glands are more active in spring and summer than in winter. They are stimulated by excitement. While present in both sexes, both adult and juvenile, they are best developed in adult males.Callospermophilus has three anal glands. These have flat-topped, straight-sided nipples which are protruded from the anus if the animal is frightened. A milky substance with a very weak odor can be extruded.

Notes: The study is divided into three parts. Part I deals with the chromosome number and morphology in the amniotic cells of rabbit embryos. The number of chromosomes has been found essentially constant in amniotic cells of young, but more variable in older embryos. The somatic number is 44. Part II deals with the chromosomes of race crosses (Flemmish Giant X Polish) in which the homologous chromosomes were found to be alike. Part III deals with spermatogenesis. There are forty-four chromosomes in spermatogonia, and twenty-two in primary spermatocytes. The sex chromosomes are of the usual X-Y type.

Notes: 1Monovalent cation salts induce reversal in the direction of the stroke of the cilia; bivalent and trivalent cation salts with a few exceptions do not. Some acids induce reversal, others do not.2The duration of reversed action varies with the kind of salt and with the concentration. As the concentration increases, the duration of reversed action increases to a maximum and then decreases to zero.3Bivalent and trivalent cation salts neutralize the effect of monovalent cation salts. The relative amount required varies with the kind of salt used and with the concentration.4The amount of a given salt required to neutralize another salt is not proportional to the concentration of the salt neutralized. Weber's law does not hold.5The results seem to indicate that ciliary reversal is associated with differential adsorption and consequent changes in electric potential, but that there are also other factors involved.

Notes: Webbing of toes or fingers in man is produced by a local arrest of development, causing retention of the normal embryonic webbing. This type of digital fusion involves only the skin, the skeleton being unaffected. The extensor tendons of the toes may sometimes be fused.Webbed digits occur normally in some marsupials, rodents, and insectivores, in a number of lemurs and catarrhines, and in the siamang and gorilla. They also may occur in varying degree in other Primates, notably Hylobates. An analysis of five new pedigrees together with those already published demonstrates that webbing of toes in man may be inherited in either a mendelian or sex-linked manner. In one case this character follows the course of the Y-chromosome.

Notes: The flagellate, Tetramitus rostratus Perty, appeared in cultures of certain amoebae obtained from the coecum of rats. In a typical life-cycle a cyst, planted in an appropriate medium, gives rise to an amoeba which may divide a number of times, but eventually some of the amoebae transform into flagellates identical with Tetramitus rostratus. These divide frequently through several days, sometimes for weeks or months, and then transform back to amoebae which become encysted.During excystment the smooth cyst wall dissolves. Usually both the amoeboid and flagellate phases pass into a “gel” state during division. A “gel” state sometimes occurs during transformation. The time required for transformations varies from a few minutes to several hours.Many culture media and methods have been tested. In certain cultures the flagellate phase was prolonged for weeks or months. These cultures were characterized by: 1) great variation in size, from minute “dwarfs” to oversized “monsters”; 2) frequent multiple fission; 3) pairing and fusion, and, 4) some evidence for the origin of secondary nuclei from chromidia. In cases of pairing and fusion, the process of maturation and union of nuclei could not be definitely proved, although suggested by the observations.The flagellate phase is more probably the “adult” phase because of its complex organization and possible sexual phenomena. This case is considered an extreme for this type amoeba-flagellate transformation.

Notes: In the blastula the region of accelerated cell division is on the ventral side of the egg, nearly opposite the site of the future beginning blastopore. The ventral wall of the blastocoele is thicker than the dorsal. In the very early gastrula a new region of accelerated cell division appears in the vicinity of the dorsal lip of the blastopore.A downward movement of material comprising the marginal zone of micromeres and the portion of the wall of the blastocoele immediately above it occurs during all the later blastula stages and continues until this material is carried below the level of the equator and involved in the process of gastrulation. On the dorsal side of the egg, this movement is more rapid than on the ventral side.In the late blastula stage there are evidences of growth in the region of smaller micromeres. In the very late blastula, a vertical groove appears at the dorsal margin of the floor of the blastocoele; this groove is believed to indicate the operation of factors concerned with gastrulation.In connection with the first nuclear division, evidences of cytoplasmic activity leading to the formation of the first cleavage furrow are described. As the blastomeres become smaller, progressive changes take place in the distribution of their cytoplasm.

Notes: A summary is given of the previous memoirs, and an attempt has been made, upon the three characters of this investigation, to arrange the genera into families, which agree, with two exceptions (Mitsukurina and Dicerobatis), with preexisting families. Some deductions are drawn as to relationships and attention is drawn to Cestracion galeatus and Triakis as connecting links. Separate families are erected for Dicerobatis and Mitsukurina, and the latter is placed near to Notidanus. Resemblances are shown between Chlamydoselachus and the Holocephali. The Trygonidae, Torpedinidae, and Rhinobatidae are singularly uniform families. An index is added both general and specific.

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 testis is differentiated from the indifferent gonad in embryos of fourteen days. The indifferent gonad and the sex cords contain large and small cells, both of which are derived from the coelomic epithelium. The large cells have generally been called primordial germ cells, they increase by mitosis and reach their maximum in embryos of seventeen days. But all the large cells degenerate and have completely disappeared by nine days after birth. The large cells, therefore, do not contribute to the formation of cells in the tubules of the adult testis. After the disappearance of the large cells, the solid seminiferous tubules are composed of indifferent epithelial cells of a uniform type, which are descendants of the small cells of the genital ridge, and which later form both the germ cells and the Sertoli cells of the seminiferous tubules of the adult testis.Definitive spermatogonia begin to form fifteen days after birth, when the solid cords are becoming hollow tubules; spermatocytes occur at twenty-three days, when the testes are entering the inguinal canals; spermatids are first observed about thirty-three days, when the testes enter the scrotum, and perfect spermatozoa at forty-eight days. Sertoli cells cannot be clearly distinguished until almost puberty; these cells may be only indifferent epithelial cells. During the development of the seminiferous tubules, a good many cells of all stages of spermatogenesis degenerate.

Notes: The claspers of Centrina are adnate with the pelvic fin and bear a spine as in other Spinacidae. Mustelus canis resembles M. lunulatus rather than M. vulgaris. The claspers of Chiloscyllium end in a pointed spike. Pseudotriakis resembles the Carchariidae. The three North American Atlantic species of the genus Raia are considered, and R. laevis and R. erinacea are placed in the pseudogenus containing R. batis, and a new pseudogenus erected for R. ocellata. A gross and histological account is given of the Cowper's glands of Homo, and they are shown to be homoplastic with clasper glands, similar in structure, arrangement, development, and function.

Notes: The activities of the larva and the responses and orientations it makes to external stimulation, and the structural organization by which these activities are produced, are described and figured.Existing systematic confusion of M. citrina with other species (M. manhattensis, M. nana, and M. microsiphonica) is cleared. The duration of the period of larval life is found to vary between 5 and 170 minutes. A proportionally small number retain the larval form a longer time. During larval life, periods of swimming movements alternate with periods of inactivity, the latter, at first of momentary duration, becoming longer and longer until activity ceases.No light receptor exists and no response to light is made. A statolith is found in the sensory vesicle, and frequent geonegative orientations are made during the free-swimming period. These are of short duration and tend to occur, 1) when it emerges from the parent; 2) at the beginning of each of the frequently recurring periods of swimming activity; 3) immediately upon making contacts. Unoriented movements follow each orientation and constitute much the greater part of behavior.The larva lacks definite organs of attachment. The entire surface of the tunic becomes adhesive at the time of metamorphosis. Only those structures that make up the action systems of the larva are fully differentiated, all other parts are embryonic in condition.

Notes: The integument of the vermilion-spotted newt contains three kinds of pigment--yellow, red, and black. The yellow is uniform and continuous over the whole body; the red and black are discrete, and are chiefly in large spots. The pattern does not change measurably in an adult individual.Functional changes in contraction of the scattered dorsal melanophores were uncommon. The only efficient single factors found were a long subjection to low temperature (expansion) and injection of pituitrin (contraction).Removal of skin was followed by a rapid mass migration of dermal elements into the wound area. Melanophores were thus furnished to a dorsal wound, and after several months black spots formed from these. Red pigment never regenerated nor migrated into a wound. Yellow pigment was formed in situ after the wound had completely healed.Auto-, homo-, and heterotransplants lost their pigment patterns and were gradually reorganized so as to conform in gross appearance to the surrounding pattern of the individual host's skin. The transformation was never complete in eight months, but new black spots and new yellow pigment eventually formed.The behavior of melanophores during morphogenesis differed for specific areas of the integument. Their movements and aggregation were highly coordinated among themselves.

Notes: In the epidermal cells of green-frog tadpoles (Rana clamitans) there are present coarse, conspicuous mitochondrial threads. In the dorsal body regions, epidermal cells of the middle layers also contain pigment granules, grouped in crescentic masses in the distal portions of the cells.Administration of thyroid extract results in permanent disappearance of the mitochondrial threads and disappearance to a large extent of the epidermal pigment granules. Processes of cellular dedifferentiation and proliferation occur rapidly over widespread areas, the mitochondria undergoing intracellular resorption. A new type of epidermis is developed containing many cutaneous glands, adapted for the approaching terrestrial life.Wound infliction induces similar processes of cellular dedifferentiation and proliferation in the epidermis in the immediate vicinity of the wound. Cells in this region lose their mitochondrial threads by intracellular resorption, and there is also some disappearance of pigment granules. The new epidermal cells in the early stages of regeneration produce neither mitochondrial threads nor pigment. This condition is not permanent, however, and in the later stages the larval characteristics appear again, both mitochondrial threads and pigment being redifferentiated.In the hyperthyroid animals there is also a significant mobilization of mesenchymal chromatophores, correlated to some extent with the loss of epidermal pigment.The significance of epidermal changes is discussed with reference to cutaneous abnormalities associated with hyperthyroid and hypothyroid conditions.

Notes: Animal posture cannot be regarded as an actual inheritance, for it constitutes a current physiological interaction between gravity and an organism, according to the structure and physical powers of the latter. Hence, ‘inherited posture’ merely implies that in a given species physical qualifications are congenitally transmitted, which facilitate a certain characteristic position of the body. Evolution of posture, therefore, has inevitably been associated with corresponding evolutionary changes in organic structures.Evidence furnished by the application of biomechanics to studies of ancient and modern primate structures indicates that man's erectly supported body posture could only have originated from a vertically suspended posture (arboreal).The fact that the prehuman stem passed through an earlier arboreal and brachiating period is attested to by the grasping character of his hands, the ratio of arm-body length, the extreme mobility of the shoulder-joints, as well as the extension of his legs on the body.The semierect posture of the great apes is not an advance toward human bipedism, but a modern reversion toward quadrupedism.Postural evidence conforms with the many other lines of testimony which maintain the close relationship of the human and anthropoid stems, and signifies that man has been an erect terrestrial biped since the time of his physical origin.

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.