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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Notes: The foam separation technique of microflotation was studied using A. aerogenes as the test organisms. Gas flow rate, collector and alum (when used) concentrations, and frother dose were held constant. In contrast to E. coli, previously reported, A. aerogenes are removed using both lauric acid and laurylamine as the collectors without prior coagulation with alum. The removals are improved after coagulation, with laurylamine being the more efficient collector. In all cases the removals decrease upon increasing the pH above about 8.

Notes: The rate of dissolution and dehydration of CO2 in a liquid model system was investigated. Components in the model system established the main conditions which may exist, in the extracellular space of a microbiological culture liquid. The charge in voltage of a glass electrode was measured which indicated the formation of H+ ions in the H2CO3 ⇌ HCO3- H+ reaction. The rate of CO2 hydration increased with the increase of temperature from 0 to 40°C. Likewise the equilibrium of the reaction was shifted towards the forward reaction. Similar results were observed when the tip velocity of the impeller was increased. Data suggest that agitation promotes the dissolution of CO2 in the culture liquid through the reduction of gas-liquid film resistance in the diffusion of this gas. The rate of hydration of CO2 into the bulk of the liquid was independent of pCO2 above the surface of the liquid but depended on pCO2 in the gas bubble within the liquid. The concentration of HCO3- was, furthermore, influenced by the buffer components, buffer capacity, and the viscosity of the system. Since pCO2 and the HCO3- concentration in the extracellular space depend on both physical and chemical factors, the ventilation of a culture liquid necessitates both exhaust of CO2 from the gas bubbles of the culture broth and shift of the H2CO3 ⇌ HCO3- + H+ reaction towards the backward direction.

Notes: A method for the determination of total heat generation in a fermentation tank and overall heat-transfer coefficient at the cooling surface is outlined. These data, which are essential for the design of cooling systems, are measured during the actual fermentation by dynamic analysis of controlled temperature variations. Each experiment consists of two stages: one in which cooling is cut off, and one in which cooling is constant. The necessary temperature variation is so limited that, the course of the fermentation is not affected.

Notes: A generalized logistic equation is proposed for the mathematical representation of batch culture kinetic data. Properties of the equation are discussed. A computer program is used to fit the generalized equation to both artificial and actual batch culture data. The equation is shown to be capable of fitting data exhibiting lag, exponential, deceleration, stationary, and death phases, as well as diauxic growth. The fitted equation is useful for differentiation, interpolation, and other manipulations of the data, and it is a convenient means of data storage.

Notes: The growth of batch-cultivated yeast Candida lipolytica on three kinds of gas oil using mineral medium was studied. A linear dependence was found between the production of yeast biomass and the consumption of n-alkanes, while the decrease of freezing point of gas oil during cultivation had a distinct course. This disproportion was explained by different degradation of individual n-alkanes contained in gas oil. The rate of degradation of pentadecane, hexadecane, and heptadecane was the same during the entire cultivation. On the contrary, in the first phase the utilization of shorter chain n-alkanes, nonane to tetradecane, was more rapid while that of longer chain homologs, octadecane to pentacosane, lagged. Rapid utilization of longer chain n-alkanes did not occur before the concentration of the other n-alkanes decreased. Only then the rapid decrease of freezing point appeared.

Notes: The influence of agitation and aeration on growth and on production of glucose oxidase of Asp. niger has been studied. It was found that both rate of growth and glucose oxidase production was higher at an agitation speed of 700 rpm than at 460 rpm. Further increase in speed of agitation resulted in neither a higher rate of growth nor a higher glucose oxidase activity. Total glucose oxidase activity was highest in a medium containing 5% sugar (at an agitation speed of 700 rpm) and did not get higher when the sugar concentration of the medium was increased to 7%. When pure oxygen was bubbled through the culture the rate of growth of the culture (in the linear phase) was 95 mg. mycelial dry wt./100 ml./hr., and only 61 mg. when air was applied. The glucose oxidase activity of oxygenated culture was double the activity of aerated culture. Viscosity of the homogenized culture became higher with higher concentration of mycelia. The viscosity of oxygenated culture was found to be lower than that of aerated culture.

Notes: This is the first part of a systematic study on the mechanisms of the Waldhof fermentor. When an agitator is rotating in a liquid, a vortex will develop on the surface. Air is dispersed into the liquid when the vortex is deep enough to reach the agitator. This is the basic mechanism of air dispersion in a Waldhof fermentor. In this work, experimental results, empirical correlations, and theoretical equations were obtained to relate the vortex depth to a number of physical factors including agitator diameter, agitator speed, tank diameter, liquid depth, liquid viscosity, and so on. The vortex depth was found a function of both Reynolds number and Froude Number.

Notes: A mathematical model is presented for both batch and continuous cultures of microorganisms utilizing inhibitory substrates. The key feature of the model is the use of a inhibition function to relate substrate concentration and specific growth rate. Simulation studies show that the primary result of inhibition by substrate in a batch culture is an increase in the lag time whereas in continuous culture inhibition by substrate may result in process instability. The model should be of value in investigations of the stability of biological processes used for the treatment of certain industrial wastes such as those containing phenols, thiocyanates, nitrates, ammonia, volatile acids, etc., which are known to be inhibitory to many of the organisms metabolizing them.

Notes: Growth of Saccharomyces cerevisiae LBG H 1022 on ethanol under transient-state conditions was studied. As a cultivation device, an aerated Chemap fermentor combined with continuously working gas analyzers for oxygen and carbon dioxide was used. Yeast cell dry matter, substrate concentration, specific oxygen uptake, specific carbon dioxide release, and respiration quotient were measured during the different transient states. Depending on which range of the dilution rate the initial steady state was found, we obtain different responses to the shift experiment. For the lower range, up to D = 0.07, we deal with damped oscillations ranging above and below the steady-state values. For the higher specific growth rates, the rate of damping is strongly enhanced and the shape of the curves becomes an asymptotic approach to the final steady states.

Notes: A relatively simple, autoclavable medium was developed that would support the growth of the mouse fibroblast (MFL) cell line in suspension culture. This medium was prepared in three stages with decreasing quantities of serum. As the serum was reduced from 5% to none, the amount of Bacto-Peptone was increased from none to 0.5%. The reduction and finally elimination of serum did not affect proliferation adversely, but actually seemed to produce more rapid growth and higher levels of cell population.

Notes: A mathematical model based on the Einstein law of photochemical equivalence is proposed to describe the batch growth of unicellular algae. The model was applied in an integrated form to cell concentration versus growth time data taken over an extended range of cell concentrations which include both the regions of “exponential” and “linear” growth. It is shown that a certain function of cell concentration contained in the integrated form of the model is linearly dependent on the growth time over both the “exponential” and “linear” growth regions.

Notes: Toxicological investigations were conducted on two new antibiotics, Muconomycin A and Muconomycin B. These non-nitrogenous antibiotics were found to be highly toxic and capable of inducing profound inflammation in the peritoneal cavity of male albino rats. Either antibiotic produced large volumes (10-20 ml) of inflammatory exudate even when injected intraperitoneally in quantities of 1.6 × 10-10 moles. An extensive profile of the electrolytes and proteins found in inflammatory exudates was developed. Simultaneous assays of the blood serum of treated rats provided a basis for comparing the concentrations of constituents of serum with those of the exudate. The results of these assays showed that the exudate contained lower concentrations of sodium and proteins, and greater amounts of potassium, calcium, and phosphorus than the serum. Chloride ion concentrations were variable. Since previous work showed that one of the manifestations of toxicity of these substances was the production of creatinuria, further studies were carried out with ATP/Creatine Phosphotransferase. These studies show that these antibiotics are potent in vitro inhibitors of the enzyme ATP/Creatine Phosphotransferase.

Notes: A novel and convenient method for the synthesis of guanosine is described. The reaction of AICA-riboside with sodium methylxanthate gave 2-mercaptoinosine in almost quantitative yield. The latter was oxidized with hydrogen peroxide to afford inosine-2-sulfonic acids, which was readily animated to give guanosine in excellent yield. Similarly, the preparation of N2-methylguanosine and N2,N2-dimethylguanosine, minor constituents of transfer RNA, was also accomplished. Furthermore, this procedure was extended to the synthesis of 2′,3′-O-isopropylideneguanosine and the isopropylidene derivatives of various N2-substituted guanosines from 2′,3′-O-isopropylidene-AICA-riboside. Guanosine via 2′,3′-O-isopropylideneguanosine was successfully phosphorylated to give 5′-guanylic acid.

Notes: A HeLa cell line was propagated in semicontinuous suspension culture, 85 liters final volume, and in continuous flow culture with a volume of 300 ml. or 5 liters in an autoclavable medium to which 8% calf serum had been added. A medium containing 0.1% Methocel and 2% calf serum was also tested. Maximum productivity was obtained at a dilution rate of 0.33 day-1 with a cell density of about 1.0 × 106 cells/ml. The same cell line was also infected with Rubella virus and the production of virus was followed at the 5-liter cultivation level.

Notes: The purpose of this experimental study was to examine the transient response of a chemostat-type continuous culture of Escherichia coli B to step changes in temperature by following transient limiting substrate concentration and calculating from it the transient growth rate. The transient response to step changes of temperature was tested for four different situations. In the first two cases, temperature was shifted down from 37 to 27°C., and 37 to 32°C. In the last two, it was shifted up from 32 to 37°C., and 27 to 37°C. When the temperature was shifted up, the growth rate increased rather rapidly to its transient maximum value and then decreased slowly until it, settled back into the steady-state value. On the other hand, when the temperature was shifted down, the growth rate decreased relatively rapidly to its transient minimum and then it slowly increased and returned gradually to the steady-state value. The magnitude of the transients was less than would be expected if the transient growth rates followed an Arrhenius function.

Notes: Equipment has been developed for the recovery of precipitates from rotors of industrial tubular centrifuges. A double piston is described for nonaseptic discharge of precipitates through the outlet holes of the clarifier rotor. For the aseptic resuspension or dissolving of valuable precipitates a closed-circulation system has been developed, which is applied without opening of the clarifier rotor.

Notes: The inhibitory effect of ethanol concentration p in a medium on the specific rates of growth μ and ethanol production ν of a specific strain of baker's yeast was studied in a chemostat, where except for ethanol as the product, only the concentration of glucose S was controlled to limit the metabolic activity of the yeast. This was designed to supplement the previous findings from the batch experiment, in which ethanol was added artificially and no substrate components were limiting the metabolism of the same yeast, that μ = μ0e-k1p and ν = ν0e-k2p, where k1 and k2 are empirical constants and subscript the 0 denotes respective values at p = 0. The effects of p on the values of μ and ν were confirmed by the Lineweaver-Burk plot to belong to noncompetitive inhibition. The formulas here for μ and ν as affected by p, if extrapolated to the case of no limiting substrates, were in good agreement in respective forms with those derived previously from the batch experiment, though the values of corresponding coefficients in these formulas were different. The differential equations for μ and ν as functions of both p and S and, in addition for the rate of glucose consumption as correlated by the yield factors either with the cell growth rate or the rate of ethanol production, were solved properly with a digital computer. A kinetic, pattern calculated so far was discussed with reference to the data obtained in the batch experiment and those relevant to actual “sake” brewing.

Notes: As a rate equation of microbial cell growth, the Monod equation is widely used. However, this equation cannot fully correspond to real courses of microbial cell growth in many batch cultivations. Especially, predicted values based on this equation do not agree with observed values in many continuous cultivations. In this paper, which introduces new concepts of critical concentration and coefficient of consumption activity, the growth rate equation which corresponds to the whole period including lag period is newly derived and characteristics of microbial cell growth in batch cultivation are clarified. Further, applying the new rate equation to continuous cultivation, a general equation with which to calculate cell concentration is derived and characteristics of microbial cell growth in continuous cultivation are clarified. The calculated values of cell concentration based on the new theory showed quite good agreement with the observed values in both batch and continuous cultivation.

Notes: New trends and developments in vaccines emphasized the growing need for safety tests of increased complexity, rather than a need for ultrasophisticated and complex devices. The Vaccine Development Branch of the National Institute of Allergy and Infectious Diseases, operating with an annual budget of $6,000,000 has the task of expediting the collaborative efforts of 50 laboratories for developing, producing, and testing experimental vaccine lots. Rubella has a major priority; other priorities are assigned to respiratory syncytial, parainfluenza, adenoviruses, influenza, and rhinoviruses, and Mycoplasma pneumoniae. To accomplish this requires a very expensive and complex program involving rapid information exchange and increased emphasis on safety testing with regard to extraneous viruses and the oncogenic potential of the vaccines. The latter need resulted from such experience as that with the Salk vaccine and the tumorproducing potential of some adenoviruses. Electron microscopy has been useful in discovering possible viral contaminants. Of all material produced for experimental work, from 65 to 70% is used for safety tests.

Notes: It has been demonstrated experimentally that the thickness of fluid overlay in conventional tissue culture systems limits the oxygen available to mammalian cells growing as a submerged monolayer. A rocker culture system is described which circumvents critical problems associated with thin film culture while permitting nearly unlimited access of oxygen to the cell monolayer. Good growth of primary hepatic cells as isolated sheets has been obtained.

Notes: A 3-liter culture vessel has been developed for the growth of animal cells in suspension at controlled pH and dissolved oxygen partial pressure (pO2). The culture technique allows metabolically produced CO2 to be measured; provision can be made to control the dissolved CO2 partial pressure. In cultures containing a low serum concentration, gas sparging to control pO2 was found to cause cell damage. This could be prevented by increasing the serum concentration to 10%, or by adding 0.02% of the surface-active polymer Pluronic F68. The growth of mouse LS cells in batch culture without pO2 control was found to be limited by the availability of oxygen. Maximum viable cell populations were obtained when dissolved pO2 was controlled at values within the range 40-100 mm Hg.

Notes: Rat liver was subjected to three different, disruption procedures (homogenization, explosive decompression, and Chaikoff press) and mitochondria were subsequently isolated by conventional differential Centrifugation and by zonal Centrifugation. The properties of these mitochondria were investigated by polarographic measurement of oxygen uptake and they were examined by electron microscopy. All three methods of disruption gave mitochondria which showed respiratory control. Nitrogen cavitation gave the most reproducible conditions for cell breakage and zonal Centrifugation gave good separation of subcellular organdies in extracts produced by this method. Some separation of the heterogenous mitochondrial populations was achieved by zonal Centrifugation.

Notes: An interesting cholinergic compound has been isolated from the fungus Rhizoctonia leguminicola grown on extracts of red clover hay. The compound was characterized as 1-acetoxy-8-aminooctahydroindolizidine and given the name “slaframine.” It has been shown that slaframine is not the active compound but is converted to the active metabolite by liver microsomal enzymes. Physiological studies with slaframine point out that it is a potent stimulator of exocrine glands. In addition, its long duration of action and low toxicity suggest that it may have therapeutic value. Preliminary data suggest that slaframine is a potent stimulator of pancreatic activity, and its long duration of action results in a stimulation of protein synthesis by the gland.

Notes: A Candida sp. was grown on a fraction of n-alkanes (dodecane 22%, tridecane 48%, tetradecane 28%) as sole carbon source. The growth rate was increased most markedly by using high concentrations of n-alkanes (16.7% v/v). When grown in a 5 liter fermentor, the yeast reached its highest yield (60 g. of cell dry wt/l) with a concomitant high yield of fatty acids (21 g of fatty acids/l), by using a nitrogen-deficient medium. To achieve good growth, it was essential to use an inoculum (1 part into 10) of rapidly growing cells and beneficial to increase the agitation rate gradually once growth had begun. After 108 hr maximum conversions of substrate to product were: 71.5% (w/w) for alkanes into cells and 24.8% (w/w) for alkanes into fatty acids. Of the, total fatty acids at the end of the fat-accumulating phase of growth 54% were shorter in chain length than palmitic acid (C16H32O2). When grown on glucose, as sole carbon source, less than 2% of the total fatty acids were shorter than palmitic acid. When n-alkanes were added to cells growing on glucose, short-chain fatty acids (C10 to C14) were synthesized immediately, indicating a derepressed enzyme system for hydrocarbon assimilation and the absence of diauxie. The production of these acids was at the apparent sacrifice of linoleic acid synthesis. In spite of the high conversion ratios, it is concluded that it would be uneconomical to produce fatty acids, even expensive ones such as lauric acid, by microbial transformation of n-alkanes.

Notes: Various multisurface mass-scale tissue culture propagators ranging in capacity up to 200 liters have been developed at Abbott Laboratories. These patented units consist, of an enclosed vessel containing a multiplicity of separated glass plates or disks on which cells may attach and proliferate. Means for mixing and aeration of the medium are provided. Sample ports facilitate the addition of cultures and media, the withdrawal of samples, the washing of cell monolayers, and the harvesting of cells and cell products. The large cell growth surface per occupied volume, the provision for separating tissue cells from media simply and easily, and the minimization of the amount of labor required per cell growth area are some of the many advantages of the multisurface tissue propagator that are described.

Notes: The herpes-type virus found in certain cell cultures derived from Burkitt's lymphoma, other human leukemias, and normal human leukocytes, was concentrated and partially purified by large-volume density gradient centrifugation using zonal centrifuge systems. Using the Jiyoye (P-3) cell line as a model, rate-zonal runs on disrupted cell suspensions in sucrose gradients yielded concentrates with high virus particle counts when 10-15 ml of packed cells were processed per liter of gradient. Isolation and removal of cell nuclei or fluorocarbon treatment of cell sonicates permitted virus recovery from larger volumes of cells per experiment. Zonal centrifugation of concentrated cell-free spent media from highly infected cell cultures yielded more purified virus than obtained from cells. Viral concentrates were prepared with particle counts of 1010-1011/ml and total protein concentrations of 0.2-0.5 mg/ml. Subsequent isopyenie-zonal centrifugation of the various high-count virus fractions from the zonal centrifuge showed a heterogeneity in buoyant virus density ranging from 1.18 to 1.27 in potassium tart rate. The spread in virus density was attributed to the different morphological forms of the virus observed by electron microscopy.

Notes: The production of large quantities of microbial mass, or their by-products, frequently requires aeration and mixing of fluid media. This operation often results in copious production of foam which cannot be exhausted with the effluent air or gas. Foam is usually controlled with antifoam agents which may interfere with product purity, oxygen uptake, and with subsequent product, handling. The process herein described obviates the requirement for antifoam agents or other foam-control methods. In essence, the air (or other gases) and foam in the headspace are continuously withdrawn, entrained in the intake side of a self-priming pump, and reintroduced into the bulk of the process liquid medium. The headspace may be enriched with oxygen or other gases.

Notes: Dried vaccinia virus was found to be very stable in aerosol form in our chamber at a temperature of 75°F and a relative humidity (RH) of 85% when the virus was protected with 1.5% lysine, 1.5% sodium glutamate, 0.5% isoniazid, 0.5% thiourea, and either 2.5% heart infusion broth (HIB) or a combination of 3.75% lactose plus 1.25% raffinose prior to freeze-drying. Utilization of the Environmental chamber technique resulted in (a) selection of two diluents which protected vaccinia virus against the effects of high relative humidity, (b) measurement of the effect of various chemical additives in the diluents, and (c) quantitative measurement of the moisture absorbed by the various dried products.

Notes: A laboratory process for the isolation of type B acid phosphatase from human erythrocytes has been scaled up 80 times. The enzyme was purified 1300-fold by adsorption on calcium phosphate gel, ammonium sulfate precipitation, and gel filtration. The yield was 27%. Results for the type A enzyme are also presented.

Notes: A survey of interactions of membrane filters with viruses has included 28 types of membranes, 4 types of enteroviruses, and 1 reovirus. Losses of these viruses in filtration, due to adsorption to the filter membranes, appear to be governed by three factors: the chemical composition of the filter membrane, the ratio of pore diameter to the diameter of the virus particle, and the presence of substances, such as those occurring in serum, which interfere with adsorption. Membranes of cellulose triacetate and of certain other materials have a very low affinity for these viruses. Cellulose triacetate filters adsorb virtually none when the pore size exceeds the virus diameter by a factor of more than 3. At porosities nearer the virus diameter, even low-affinity membranes adsorb large quantities of virus unless serum or some other additive interferes. Cellulose nitrate membranes, in the absence of interfering substances, adsorb enterovirus significantly at a pore size 285 times the virus diameter.

Notes: The intracellular levels of seven enzymes in mouse LS cells growing in suspension culture at controlled dissolved oxygen partial pressures (pO2) have been measured. During the growth of each culture large fluctuations were observed in the levels of some enzymes, particularly aldolase and cytochrome oxidase. Mean values for the concentration of each enzyme during the growth phase have been calculated. These results are discussed in relation to previous observations made on the growth of mouse LS cells.

Notes: Continuous cultivation of the yeast Candida lipolytica on gas oil was studied from the viewpoint of biomass production and oil deparaffination. Optimum conditions wore found at the dilution rate D = 0.16-0.19 when biomass productivity 1.7 g/l/hr and yield coefficient. y = 0.92 were achieved. At deparaffination to the same freezing point, more than double the production of biomass and deparaffined oil during a given time unit was achieved in a continuous process than in batch cultivation. Consumption of substrate was followed in both cultivation processes and it was confirmed that individual n-alkanes of gas oil were degraded at various rates and yields. Results proved optimum cultivation conditions to depend on concentration and composition of the paraffinic fraction of gas oil used. To achieve these conditions the continuous process may be controlled by choice; of suitable dilution rate and concentration of gas oil.

Notes: The biochemical oxygen demand (BOD) of a complex, partially colloidal substrate is divided into oxidation (fo) and synthesis (fs) fractions. The substrate being investigated is a suspension containing 10 mg./l. of dry skim milk. The oxidized and synthesized fractions of the BOD are shown to be 38.4% and 61.6%, respectively. The division of BOD is related to the time at which maximum cell growth appears. The time required to obtain maximum cell production is shown to be at least 10 hr. The relationship between the time to maximum cell production and the apparent initial activity of the seed organisms is discussed.

Notes: This study reports some findings on the effects of centrifugation on the viability of mammalian cells. The authors used Burkitt lymphoma cells cultivated in a synthetic medium containing 10% fetal calf serum for all experiments. Batch centrifugations were conducted in a RC2-B centrifuge (Ivan Sorvall, Incorporated, Norwalk, Connecticut USA) operated at 0 and 25°C. During centrifugation we exposed the cells to gravitational fields ranging from 24,800 to 42.200g. The results showed that at, 0°C and 25,800 or 42,000g no loss in cell viability occurred for up to 90 min exposures in the centrifugal field. However, at 25°C and for gravitational fields of 24,800 and 42,000g, there were appreciable losses in cell viability. Continuous centrifugation studies in the Sharples supercentrifuge (Division of Penn Salt Corporation, Warminister, Pennsylvania USA) were also conducted with bowl speeds up to 28,000 rpm (19,000g) and flow rates ranging from 1.4 to 20 1, hr. Slight, losses in cell viability were noted and postulated as caused by the shear stresses encountered by the cells. Some pumping studies using the lymphoma cells substantiate this conclusion.