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Notes: Pyroantimonate precipitate indicates that the epithelium of the proximal tubule is the only segment of the tubular nephron of the fresh water lamprey where large accumlations of cations are distributed. Unusually large amounts of reaction product are located within the lateral intercellular spaces and within vesicles closely associated with the plasma membrane at the lateral and basal surfaces. This technique suggests the continuity of these vesicles with the plasma membrane and alludes to the possibility of an endomembranous system of vesicles and the intercellular spaces as vehicles for ion transport. Lateral intercellular spaces of proximal tubules of lower vertebrates may play a different role in kidney function that their counterparts in higher vertebrates.Osmium-zinc iodide has a specificity for certain cells within the proximal, intermediate, and distal segments, but no structural differences are noted when these cells are compared to unstained cells. Smooth endoplasmic reticulum remains unstained in the distal segment but the stain has a strong affinity for elements of the Golgi apparatus, lysosomes, and the nuclear envelope of all cell types. This technique does not suggest a structural or functional similarity between cells of the distal segment and the chloride cells of the gills of teleosts.

Notes: Anatomical analysis of the forebrain and midbrain of Anelytropsis, Dibamus and feyliniids reveals structural similarities with those of skinks and snakes. Skinks and feyliniids are probably derived from a common ancestral stock. This is suggested by mutual reduction of several telencephalic nuclei, by similar trends in the development of the dorsal thalamus, and by similarities in the lamination of their optic tecta. Anelytropsis, Dibamus, feyliniids and snakes show interdigitation of the periventricular gray zones of the optic tectum and enlargement of lamina 7 of the posterior colliculus. Of these three taxa, Dibamus is most similar to burrowing snakes and many of its brain characters are intermediate between skinks and burrowing snakes. These similarities may suggest common ancestry between Dibamus and snakes rather than parallelism.

Notes: It is possible to monitor the electrical activity of the motor neurons of Drosophila by recording the electrical activity of the muscle fibers. We have found that it is possible to specify the location of the subcuticular terminations and to describe the orientation within the thorax for the individual muscle fibers, because of the large size of the fibers and because the surface anatomy of Drosophila is known in detail. A map has been made to indicate the location of the muscle fibers with respect to superficial landmarks. The importance of the stereotaxic map for physiological studies is discussed.

Notes: The functional anatomy of pulmonary ventilation in Thamnophis elegans was studied by electromyography. Flow of air into and out of the lungs follows a triphasic pattern and occurs while the glottis is held open by a dilator muscle. Non-ventilatory rest periods characterized by a closed glottis and lack of ventilatory movements occur between breaths. Exhalation is caused by contraction of a pair of dorsolateral sheets of muscle that extends from the ventral surface of the vertebral column to the medial surfaces of the ribs and a pair of ventro-lateral sheets that extends from the medial surfaces of the ribs to the mid-ventral skin. Inspiration is powered by contraction of a series of rib levators and retractors. The last phase of the ventilatory cycle is a passive expiration that may be caused by elastic recoil of the walls of the filled lungs and relaxation of muscles used during inspiration. The site of respiratory pumping is shifted craniad after large prey items have been completely ingested. The significance of this shift and its anatomical basis are considered.

Notes: As seen in transverse section, doublet elements of the axial unit of spermatozoa of Haematolocchus medioplexus, a frog lung-fluke, possess walls made up of protofibrillar subunits 50-60 Å in diameter. The partition between A and B members of a doublet element often show extra protofibrils which may partially occlude the “lumen” of the A tubule. Each A tubule possesses outer and inner lateral arms which repeat at longitudinal intervals of about 215 Å and which appear to be structurally dissimilar; the outer arm is expanded at its free end and the inner arm often connects to the B tubule of the adjacent doublet element. Regularly-spaced radial links connect the central sheath of an inner core complex to the A tubules of the peripheral doublet elements. Tests for magnesium-activated ATPase activity provide evidence that the enzyme is associated with the surfaces of doublet elements and the surface of the central sheath. Finally, study of an axial unit which developed in an abnormal manner suggests that normal differentiation of an axial unit may depend on the elaboration of a core complex and radial links.

Notes: This is the first demonstration of an apparent acrosome reaction by the use of negative staining (phosphotungstic acid) and electron microscopy. It is also the first report of such a reaction in spermatozoa of an oligochaete annelid (Lumbricus terrestris). The morphology of the negatively stained unreacted acrosomal complexes is entirely comparable to that seen in sectioned material. After reaction, a short straight filament is extruded and the compartmentalization seen in unreacted spermatozoa is no longer visible. In the flagellar axoneme there are short cross-bridges linking the two central singlet microtubules at 150-170 Å intervals. Two fibrous elements are also present, forming, with the singlets, a tetragon. All four of these structures apparently change their elastic properties after incubation in distilled water for two hours, so that instead of following the doublet microtubules in gentle coils, the central complex components fall as a unit in jagged configurations.

Notes: Serial sections of embryonic rat eyes were stained with hematoxylin and eosin, quantified (by counting pycnotic and viable nuclei), reproduced by camera lucida on wax plates, and moulded into reconstructions in order to study the normal progression of cellular death during morphogenesis. At least nine distinct necrotic loci (A through I) can be distinguished. Immediately following contact between the retina and surface ectoderm (day 11) degenerating cells were observed in (A) the ventral extent of the optic vesicle, beginning in the mid-retinal primordium and continuing ventrally in the optic stalk, (B) in the rostral optic stalk base, and (C) in the surface ectoderm encircling the early lens placode. No degeneration was observed in the dorsal half of the presumptive retina, in the entire pigment epithelium, or in the lens placode proper. During day 11.5 the lens placode thickens and forms a degenerating locus (D) in its ventral portion opposite the underlying pycnotic zone in the retina (A). During day 12 the ventral pycnotic zone (A) divides into two subunits (A1 and A2). Invagination of the lens displaces its marginal and ventral components (C and D) so that they come to occupy the lens pore area and presumptive corneal epithelium. Simultaneous invagination of the retinal rudiment juxtaposes the pigment epithelium which concurrently forms a necrotic area (E) adjacent ventrally to that in the retina (A1). Degeneration appears in the caudal optic stalk (I). The density of viable cells decreases adjacent to pycnotic areas in the retina and pigment epithelium and increases within these death centers. During day 13 the optic fissure forms within the subunits of the ventral pycnotic zone (A1 and A2). Degenerations are seen in the dorsal optic stalk (F) and in the walls of the optic fissure (G and H). Throughout these stages necrosis appears only in those portions of the eye rudiment where invagination is either retarded or completely absent.In part, these observations suggest that cell death serves (1) to retard or inhibit invagination within death centers, (2) to integrate the series of invaginations which mould the dorsal optic cup and optic fissure, (3) to assist formation of the pigment epithelium monolayer, and (4) to orient the lens vesicle within the eye cup. The spatio-temporal relationship between necrotic loci suggests that pycnotic cells in the retina may influence their production in the lens and pigment epithelium. Preliminary observations on the mouse, pig, and human substantiate those on the rat.

Notes: The ultrastructural features of cardiac muscle cells and their innervation were examined in the tarantula spider Eurypelma marxi Simon. The cells are transversely striated and have an A band length of about three microns. H zones are indistinct and M lines are absent. Thick and thin myofilament diameters are approximately 200 and 70 Å respectively. Eight to 12 thin filaments usually surround each thick one. Accumulations of thick and thin myofilaments occur perpendicular to the bulk of the myofilaments in some cells. The Z line is discontinuous and thick filaments may pass through the spaces in the Z line. Extensive systems of sarcoplasmic reticulum and transverse tubules are present; these form numerous dyadic junctions in both A and I band regions. Sarcolemmal invaginations form Z line tubules; lateral extensions of the plasma membrane portion of these invaginations form dyads. Nerve branches of the cardiac ganglion make multiple neuromuscular synapses with at least some of the cardiac muscle cells. Both large granular and small agranular vesicles are present in the presynaptic terminals. Intercalated discs similar to those present in other arthropod hearts occur between the ends of adjacent cardiac muscle cells.

Notes: A cinematographic analysis of the unrestrained walking, trotting, galloping, jumping and landing movements of 11 adult cats was undertaken to provide previously unavailable information concerning the demands imposed on the nervous system for the control of low and high speed movements and the demands imposed by such natural movements on muscle performance and proprioceptive response.With due regard for the swing (F and E1) and stance (E2 and E3) phases of the step cycle of an individual limb, single frame analysis of the film permitted measurement of instantaneous angles of the lower spine, hip, knee, ankle and metatarsophalangeal joints. Appropriate lever arm measurements were also made on 50 freshly dispatched cats and 25 cadavers such that the Law of Cosines could be used to calculate instantaneous lengths of select hind limb muscles that would apply to the natural movements of adult cats of small (1.5-2.5 Kg), intermediate (2.6-3.5 Kg) and large (3.6-4.5 Kg) size. Muscle displacements were analyzed relative to maximum and minimus in situ lengths and the lengths associated with quiet standing. Use was also made of a previous electromyographic analysis of hind limb muscles during unrestrained locomotion (Engberg and Lundberg, '69).The sequential relations between the four phases of the step cycle are maintained as forward speed increases from walking ( 〈 2 mph) to high speed galloping ( 〉 16 mph). There are significant differences in the time consumed by each phase, however, with a greater reduction in the E3 phase, little reduction in the E2 and E1 phases and virtually no reduction in the F phase. When each phase is expressed as a relative percentage of the duration of the total step cycle, the greatest reduction is again in E3 with little change in the E2 phase. In contrast F and E1 phases increase in the percent of time they occur in each cycle, with the greatest increase in the F phase. For all speeds, analysis of the phase relations between movements of various sections of the hind limb revealed a remarkable unity of knee and ankle joint movement. The hip joint is largely out of phase with the knee and ankle during E1 and E2, all three joints being in phase in F and E3. The digits are essentially out of phase with the other joints except in the stance phase of the gallop.Rates and extents of muscle displacement during natural movements are greater than might be anticipated when expressed in absolute mm's and mm/sec but not when considered in relation to maximum and minimum in situ length and the length associated with quiet standing (Ls). During stepping a progressive increase in forward speed results in: (a) a greater usage of muscles at lengths between Ls and maximum in situ length; (b) for knee and ankle extensors, pronounced increase in the lengthening contraction associated with the E2 (yield) phase of step; and, (c) for both flexor and extensor muscles, an increased active phase of lengthening or near isometric contraction immediately prior to periods of active shortening. In contrast to these changes in active muscle status, the change from walking to galloping has little effect on the extent and rate of passive muscle displacements, particularly the F phase stretch of extensors.For the soleus muscle, calculations were made of the relation between changes in overall muscle length during natural movements and the length of the average muscle fiber and the tendon of insertion. These measurements revealed that the increases in fiber length when passive and decreases in length during active shortening are less than would be anticipated from the extensive liteature on extirpated fibers. In contrast, the increase in fiber length when active is greater than would be expected from the admittedly sparse literature on this subject.The results of this study are discussed largely in relation to two points of neurophysiological interest: the physiological range of muscle stretch as it pertains to the responsiveness of muscle spindles and tendon organs; and those mechanical aspects of lengthening contractions that give insight into the neural control of stepping. For exciting both spindles and tendon organs passive muscle stretch and shortening contractions are shown to be relatively ineffective and lengthening and isometric contractions particularly effective movements. It is suggested that, just as recent literature has emphasized the co-activation of efferent alpha and gamma motoneurons as a muscle becomes active, so too is there a synchronous activation of afferents, particularly the Ia and group II endings of muscle spindles and Ib endings of tendon organs. Finally the thesis is advanced that, while it has been convenient to separate E2 from E3 in the description of the stance phase of the step cycle, extensor muscles are actually undergoing a single mechanical event: an active stretch-shorten cycle for knee and ankle extensors and an active isometric-shorten cycle for hip extensors. This hypothesis has significant implications for the neural control program that regulates the stepping sequence in that it emphasizes the extent to which appropriate changes must be preprogrammed in the mechanical properties of muscles for the smooth execution of stepping.

Notes: Cytological and cytochemical methods were used to identify and characterzie six distinct regions of the crayfish kidney: coelomosac, labyrinth I and II, and nephridial canal I, II, and III. Cells of the coelomosac possess cytoplasm which is strongly PAS-positive, but poor in RNA and protein. Their nuclei possess unusual projections which extend to the basal plasmalemma. Labyrinth I contains columnar cells rich in glycogen. Labyrinth II is characterized by a distended lumen and by shorter cells with high cytoplasmic RNA, many possessing a large intracellular vacuole. A PAS-positive brush border is unique to the two portions of the labyrinth. Cells in the nephridial canal show strong reactions for RNA and Mg++-dependent ATPase. In nephridial canal I and II, cells are flattened to cuboidal with the lumen being more distended in nephridial canal I than anywhere else in the tubule. In nephridial canal III, the cells are large and columnar, and the cytoplasmic RNA concentration is greatest apically. Nuclei in all regions of the tubule epithelium, except coelomosac, are large and react strongly for protein. Coelomosac nuclei and those in blood cells are condensed and contain little protein. The cytoplasm of blood cells displays a significant amount of RNA, and traces of polysaccharide material.These observations demonstrate the presence of highly specialized morphological and histochemical alterations along the length of the kidney tubule and indicate sequential modification of urine in the lumen. Evident morphological and cytochemical likenesses between analogous regions of the mammlian nephron and the crayfish kidney tubule suggest that basic physiological similarities may also exist.

Notes: In the tiger salamnder the distribution of optic fibers, as revealed by the Fink-Heimer method, is compared with the localization of acetylcholinesterase, as revealed by histochemical methods. AChE activity coincides with optic nerve axons in the optic fiber layer of the retina, in the optic nerve, in the optic tracts and in every optic projection. Reginons of optic fiber terminals show heavier activity than optic fibers of passage. Comparison with other vertebrates is also made.

Notes: The spleen of the adult gerbil (M. unguiculatus) is characterized by the absence of venous sinuses and ellipsoid sheaths. The follicle (white pulp) is separated from the surrounding red pulp by a distinct marginal zone. The cell types in the marginal zone are common to both the follicle and red pulp. Separating the marginal zone from the follicle is a vascular channel of capillary dimension, the marginal sinus. A number of terminal segments of the arterial vessels within the follicle were observed to form a direct connection with the marginal sinus. Ultrastructurally, discontinuities were evident within the walls of the marginal sinus that would permit passage of both cellular and plasma components from the marginal sinus to either marginal zone or the follicle.

Notes: The innervation of each of the muscles involved in mediating head movement in the desert locust Schistocerca gregaria is described in detail. The number of motor neurones to each muscle and the neutral pathway and ganglion of origin of each are deduced from both histological and electrophysiological evidence. Only two of the muscles are, on histological evidence, innervated by as few as four different neurones, while several receive more than ten, and one at least 13. Individual muscles are shown physiologically to receive, in a few cases, as many as six different motor neurones. At least six muscles are innervated by motor neurones originating in more than one ganglion. One group of four muscles consisting in total of less than 100 muscle fibres receives more than 20 different motor neurones from three different ganglia through three or four different nerve roots. In these muscles, many single muscle fibres receive innervation from at least two different ganglia. It is concluded that the segmental nature of an insect muscle can not be deduced solely from a knowledge of the ganglion of origin of the motor innervation to that muscle. The innervation patterns that exist today must reflect past evolutionary development, but changes in the peripheral distribution of motor neurones, or migration of motor neurone cell bodies from one ganglion to another, or the development of additional motor neurones, or several of these factors together, must have formed a part of that development.

Notes: 1. Forty-eight species of oribatids in 37 families representing most of the superfamilies were collected from various environments (littoral, salt marsh, litter, sod, and freshwater) and sectioned.2. The coxal gland is composed of a sacculus and a labyrinth in all stages of all oribatid species. Muscles, originating on the body wall, insert at several points on the thin-walled sacculus which opens into the labyrinth. The labyrinth has an internal, chitinous supporting skeleton. The type A labyrinth has 3-180° bends, producing four parallel regions, and occurs in all inferior oribatids. The type B labyrinth has 1-180° bend, producing two parallel regions, and occurs in all superior oribatids. The coxal gland duct and the lateral gland duct join, penetrate the body wall, and empty into the posterior end of the podocephalic canal. All oribatids have lateral accessory glands, but only inferior oribatids have rostral and medial glands. Three ductless coxendral bodies are always present.3. The labyrinth length in oribatids is correlated with body size and the environment of the species. Oribatids from sod, leaflitter, or moss show a simple correlation of labyrinth length (X) to total body length (Y) where Y = 4.64X. Freshwater species have a labyrinth length greater than that of comparably sized terrestrial species and salt water (littoral) species have a labyrinth length less than that of comparably sized terrestrial species. There is a greater reduction in labyrinth length in species restricted to salt marshes than in species not restricted to salt marshes.4. The probable function of oribatid coxal glands is osmoregulation. Hemolymph filtration would occur across the sacculus by positive hemolymph pressure and contraction of the sacculus muscles. Resorption of ions would occur in the labyrinth, which is noncollapsible due to the internal skeleton. The hypothesis is that in freshwater species the rate of filtration is high and resorption of ions would have to be very efficient, therefore they have an elongated labyrinth; but in salt water species water loss must be minimized and preservation of ions would be a disadvantage, therefore they have a shortened labyrinth. Excre ion may also be a function of the coxal glands. The lateral gland may possibly function as an endocrine gland involved with production of a molting hormone. The rostral glands in inferior oribatids may have a salivary function.5. The coxal glands of Peripatus, some millipedes, apterygote insects, decapod crustaceans, and all arachnid orders are homologous. The Tetrastigmata, Notostigmata, Cryptostigmata, and soft ticks have typical arachnid coxal glands. The coxal glands of higher Prostigmata may be modified into salivary, silk, or venom glands. The coxal glands in Mesostigmata, Astigmata, and hard ticks are lacking or highly modified.

Notes: The blood system of the flabelligerid polychaete, Flabelliderma commensalis has been explored by dissection, light and electron microscopy and absorption spectrophotometry. The main longitudinal vessels are the dorsal, ventral, perineural, sub-oesophageal, supra-oesophageal and heart. Each segment has a segmental vessel which communicates with the dorsal vessel in thoracic setigers and the gut sinus in abdominal setigers. Branches of the segmental vessels in setigers 2-9 supply the gonads. A blood sinus envelopes most of the gut. Circulation is maintained by the pumping of the heart which immediately supplies blood to the supra-oesophageal ganglion, the branchiae and the palps. These are paralleled by a system of collecting vessels. The sinus of the supra-oesophageal ganglion receives a number of different axonal endings, some of which may be neurosecretory. The retroperitoneal vessels in their most developed form are composed of an intima, longitudinal and circular muscles and a peritoneum. The heart vessel contains a cardiac body whose cells appear to contain vacuoles of blood pigment. The blood pigment exhibits the absorption characteristics of a chlorocruorin with maxima at 438, 558 and 606 nm.

Notes: Attachment of muscle of the integument was studied in two species of Balanidae, Balanus balanoides and B. improvisus with electron microscopy. Specilalized epithelial cells (tendon cells) intervene between the muscle and overlying cuticle. An extensive desmosome-like junction exists between these two tissues. The tendon cell contains huge amounts of microtubules aligned parallel to the long axis of the inserting muscle. Tension exerted by contracting muscles is conveyed to the chitinous exoskeleton via the specially organized tendon cells and the extensive myo-tendinal cell junctions.

Notes: Among ants, Cataglyphis bicolor shows the best performance in optical orientation. Its eye is of the apposition type with a fused rhabdom. Morphological studies on the general struture of the eye as well as the effect of light have been carried out with transmission and scanning electron microscopy.An ommatidium is composed of a dioptric apparatus, consisting of a cornea, corneal process and a crystalline cone, the sensory retinula, which is made up of eight retinula cells in the distal half and of an additional ninth one in the proximal half. The ommatidia are separated from each other by two primary pigment cells, which surround the crystalline cone and an average of 12 secondary pigment cells, which reach from cornea to the basement membrane.The eye of Cataglyphis bicolor possesses a light intensity dependent adaptation mechanism, which causes a radial and distal movement of the pigment granules within the retinula cells and a dilatation of cisternae of the ER along the rhabdom.Until now, no overall order in arrangement of retinula cells or direction of microvilli has been found from ommatidium to ommatidium. Such an order, however, must exist, either on the retina or the lamina level, since we have proven the ant's capacity for polarized light analysis.

Notes: The morphology of the rectal pads of two cockroaches, Blattella germanica and Blaberus giganteus is described and contrasted with that of Periplaneta americana, studied previously. The columnar absorptive cells of the three species are similar in structure. However, Blattella and Blaberus rectal pads have a second type of cell, termed the secondary cell, that is not present in Periplaneta. The secondary cells are embedded in the pad epithelium and have crypts of cavities opening into the subepithelial sinus. In addition, a multilayered sheath is present between the pad cells and the sinus. The sheath acts as a barrier, perhaps of low permeability, between the subepithelial sinus and the pads. It is interrupted only at the borders of the secondary cells. A mechanism for fluid absorption is presented and the possible role of secondary cells is discussed. It is proposed that the secondary cells reabsorb solute from the sinus and that this solute can then be recycled through the pad cells. Structures resembling neurosecretory terminals within the cavities of the secondary cells may be involved in regulation of recycling. The arrangement of the rectal pads in these insects has structural and perhaps functional similarities with other transporting systems, particularly the cryptonephric system.

Notes: Light and electron microscope studies indicate that the old ependyma just proximal to the plane of amputation in early lizard tail regenerates shows a sequence of morphological changes which suggests that it as well as the new ependyma growing into the regenerate may play an active role in the initiation and maintenance of early tail regeneration.The old ependyma close to the plane of amputation undergoes hypertrophy and/or hyperplasia causing a partial closure of the central canal and pseudostratification. Its nuclei shift from an original apical position to a basal one. The ependymal processes become more prominent and extend to the pia, a condition not found more rostrally. There is also a significant increase in the amount of Golgi substance and a moderate increase in the rough endoplasmic reticulum. These observations lead to the thought that these cellular changes may be an expression of enhanced secretion and other activities in the old and new ependyma just proximal or distal to the plane of amputation.

Notes: Statistical analyses of chromatophore densities and morphological states in wild type, burnsi, and kandiyohi: adult Rana pipiens integument yielded the conclusion that disruptive pigmentary pattern is permanently defined by regional chromatophore densities. Spatial chromatophore patterning is enhanced and rendered more visible by the morphological and physiological differences among chromatophores within the various pattern regions. Specifically, (1) chromatophores were differentialy distributed among pattern regions, (2) greater complexity of gross disruptive patterning had greater underlying melanophores densities, (3) there were significant density differences among non-sib individual animals, among fertilization, and among shipment batches, but not among full-sibs, and (4)kandiyohi individuals had more while burnsi individuals had fewer total melanophores than wild type. A stellate morphology was significantly correlated with high melanophore density, although the relationship of melanophore density to melanophore morphology differed among pattern regions and among genotypes. The functional interrelationships of density and morphology, and their role in disruptive patterning and coloration, are discussed.

Notes: The male rotifer copulatory organ is composed of a urethral canal extending from the tip of the copulatory organ internally to a layer of microvilli. The microvilli project from two different cell types, referred to as the internal and peripheral microvillar cells according to their location. At this microvillar junction a second canal, the vas deferens, continues posteriorly and enters the sperm duct region of the testis. The channel of the vas deferens is formed from the inner wall of three separate cells; the cap, intermediate and basal cells. Peripheral to these cells and parallel to them for their entire length, cross sections of seven prostate gland cells can be observed. Anteriorly, these gland cells are connected to the basal end of the microvillar layer via a short neck region, through which glandular secretion occurs only during copulation. The mechanism of secretion appears to be a form of exocytosis whereby the secretory granule membrane fuses with the cell plasmalemma so that rupturing at the point of fusion will release the granule content into the neck region.The prostate gland cells contain an abundance of autophagic vacuoles while most of the other cells of the copulatory organ contain primary lysosomes and cytolosomes. These organelles may be associated with the aging process in rotifers, or, as in the case of the prostate gland-autophagic vacuoles, with a fast organelle turnover during secretion.

Notes: Spermatogonia of the nine-banded armadillo, Dasypus novemcinctus mexicanus, were studied morphologically using light and electron microscopy and examined histochemically using light microscopy. Immature flat type spermatogonia have ovoid or irregular nuclei with loosely condensed chromatin. Free ribosomes are abundant while profiles of rough endoplasmic reticulum are scarce. Smooth endoplasmic reticulum is a prominent feature occasionally taking an unusual cylindrical form. Mature spermatogonia exhibit rounder nuclei with greater degrees of chromatin clumping. Smooth endoplasmic reticulum is no longer prominent whereas profiles of rough endoplasmic reticulum are quite common. Occasional lysosomal configurations are found in mature spermatogonia.The majority of spermatogonial cells exhibit weak to moderate reactivity when stained with the periodic acid-Schiff (PAS) reaction. Certain cells in each tubular cross section stain vividly with this reaction and the PAS positivity is removable with salivary amylase. Because of nuclear characteristics, position of the cell immediately upon the basal lamina, intensity of the PAS reaction and the relative paucity of the vividly staining cells, it is suggested that they are members of the immature spermatogonial cell line, perhaps acting as stem cells. None of the several other histochemical procedures employed was capable of selectively demonstrating these cells.

Notes: The duodenal glands of four marsupial species, the kangaroo, native cat, marsupial mouse, and bandicoot, form a narrow lobular collar immediately distal to the gastrointestinal junction. In two species, the koala and wombat, the duodenal glands are diffuse and scattered along the intestinal tract distal to the gastrointestinal junction for considerable distances. The glands of all species examined empty independently into the intestinal lumen. Histochemical studies indicate that the latter two species, the koala and wombat, elaborate an acid mucin whereas the former species produce a neutral mucosubstance. Ultrastructural examination shows that generally the duodenal glands are comprised of large pyramidal cells that show a definite polarity, with basally-placed nuclei and apically-situated secretory granules. Species differences, with regard both to the morphology and nature of the secretory granules and to the proportions of cell organelles, exist. The diversity of diet between species is discussed.

Notes: Cytodifferentiation of the myoepithelial cells (MEC) of the rat submandibular gland (SMG) was observed by studying the prenatal and postnatal development of the gland in vivo and in vitro by light and electron microscopy. The anlage of the SMG first appeared on the fourteenth day of gestation and, from its earliest inception, was surrounded by an intact basal lamina. Presumptive myoepithelial cells were first seen at 18 days of gestation coinciding with the onset of secretion in the rudiment. These cells were flattened, peripherally located and subjacent to the epithelial basal lamina. Initial deposition of cytofilaments in the MEC's was observed during the first three days following birth and fully matured cells were seen as early as one week after birth. Presumptive and immature MEC's were observed undergoing mitosis, but once cytofilament deposition had begun in the cells they did not divide. Myoepithelium developed in relation to embryonic secretory structures and were only observed surounding acini and intercalated ducts in the adult gland. New myoepithelial cells were formed as long as new acinar-intercalated duct units were formed. Myoepithelial cells did not produce secretory type granules at any time during their development or in their mature state.Development of the MEC's in vitro paralleled that in vivo and supported the above observations.

Notes: Larval compound (jointed) setae of the polychaete Nereis vexillosa Grube were examined by scanning and transmission electron microscopy and by polarization microscopy. Long-bladed spinigers and short-bladed falcigers are described. The proximal shaft of each of these types of setae flares distally into a serrated collar and encloses the proximal end of a toothed blade. The collar projects on one side as a boss. The blade and the cortex of the shaft have longitudinal channels. A large excentric cavity in the shaft (the medullary channel) contains a loose meshwork of trabeculae. In the distal part of the shaft these trabeculae are aggregated into diaphragms. The seta is invested with an electron dense layer of enamel. Juvenile setae contain both chitin and protein. With respect to the long axis of the seta, the blade and the cortex of the shaft are positively birefringent and the medullary diaphragms are negatively birefringent. KOH extraction renders the setae negative to a test for protein and reverses the sign of birefringence of the cortical material of the shaft.

Notes: The marine priapulid Priapulus caudatus has a voluminous body cavity filled with a blood-like fluid containing erythrocytes and leucocytes (amoebocytes). The hematocrit of animals weighing 0.5-14 gm was 2-10%. The erythrocytes contain a hemerythrin blood pigment. The structure of the coelomocytes was studied by light and electron microscopy. The erythrocytes are nucleated and contain marginal bands, vacuoles and occasionally crystals. The cytoplasm has few organelles. The leucocytes are amoeboid motile cells, the cytoplasm of which contains numerous organelles. The most conspicuous of these are oval particles, probably representing developmental stages of lysosomes. Most of these organelles contain tubules stretching from one pole to another. In the hind part of the animal, certain tissues, primarily the posterior warts contain large numbers of coelomocytes. The histological picture is complicated, showing some resemblance to the lymphoepithelial tissues of vertebrates.

Notes: The dissected tooth bearing bones of 20 specimens of Amia calva (Pisces:Holostei) ranging from the third to tenth season have been examined radiographically and in alizarin red S stained and cleared specimens. Although forms of alternate (1:1) tooth series replacement (sensu Edmund, '60) were frequently observed, even in the youngest, immature specimens, many examples of irregular replacement were recorded. In several bones, the maxillae in particular, series with every third (2:1) or fourth (3:1) tooth replacing were seen and possible patterns of 2:2, were recorded. It is concluded that these data are not consistent with the Zahnreihen concept but support a morphogenic field concept of tooth development.

Notes: The first description of vertebral development in a plethodontid salamander is presented. Eurycea bislineata has larvae that hatch at a rather early stage of development. Somites and the notochord appear early. Somitic differentiation is slight, and no distinct sclerotome can be found. As a result, there is no clear primary segmentation of the skeletogenous tissue. No evidence of a sclerocoele can be found. The amount of sclerotomal cells surrounding the notochord is very low, relative to other tetrapods. Yet discrete perichordal rings of cells do form, in nearly midsegmental positions, and these give rise to the intervertebral cartilages. Osteogenesis of the centra is initiated prior to hatching and is coincidental with ossification of the neural arch. There is no sign of a neurocentral suture. The centrum forms as a thin shell of bone directly from sclerotomal cells. The notochord is a prominent feature of the vertebral column throughout life, retaining its integrity until late in life when some disintegration occurs locally. The notochord is filled with cartilage midvertebrally in late larval stages, and some additional cartilage forms later in life. The intervertebral cartilage enlarges greatly in late larval life. An opisthocoelous joint forms in this cartilage, apparently as a result of differential changes in the cells of the perichordal ring rather than by an invasion of cells from an external source. The intervertebral cartilage is a dominant structural and highly important functional feature of the adult vertebra. In metamorphosed individuals it may become extensively mineralized, and it consists of many different structural kinds of cartilage.The cranio-vertebral joint seems to form in a single segment, contrary to the condition reported by some early investigators. It is complex, and consists of articulations between the odontoid process of the atlas and the occipital arch, as well as between the occipital condyles and atlantal cotyles. The notochord plays a dominant role in the early development of the odontoid, but then changes radically and is absent in the adult process.The anterior trunk region seems to be much more conservative than posterior parts of the column. The patterns of nerve routes and nature of development of the ribs and rib bearers differs greatly from conditions elsewhere in the column. The rib patterns are similar to presumed ancestral conditions. Rib development on the sacral and caudosacral vertebrae is in some ways more similar to that of the anterior vertebrae than of the central trunk vertebrae.Quantitative aspects of variation in the vertebrae of adult salamanders are presented. There is more regional variation and less site variation than would be expected from literature reports.Evolutionary aspects of the origin of the cranio-vertebral joint, transverse process and ribs, patterns of segmentation, and centrum development are considered in the light of the new information on Eurycea. There is no evidence that more than one vertebra is involved developmentally or evolutionarily in the cranio-vertebral joint. The most generalized condition of rib bearers in living salamanders is one in which the dorsal and ventral bearers are in cartilaginous continuity during development. There are many variations on this theme in living species. It is inappropriate to speak of a resegmentation of the sclerotome in Eurycca, even though the adult vertebra is a transsegmental structure, because there is no primary segmentation of the scanty sclerotome. The important feature found in vertebral development in all tetrapods is the perichordal tube and its subsequent differentiation. Questinos concerning precise homologies of the salamander vertebral centrum with those of other vertebrates cannot be answered by data from development sequences with currently used criteria of homology. On the other hand, it appears that all centra, regardless of subdivision, are homologous in all tetrapods.

Notes: The Oberhautchen of scales from the dorsal, parietal, and ventral regions of Sceloporus occidentalis (Iguanidae), Gerrhonotus multicarinatus (Anguinidae), and Anniella pulchra (Anniellidae) were examined with a scanning electron microscope. At low magnification, all scales of S. occidentalis exhibit well-defined outlines of cells belonging to the Oberhautchen layer and the previously overlying clear layer. The dorsal and parietal cells of this species exhibit a minutely dentate Oberhautchen that forms tooth-like spinules 0.2 to 0.5 μ long and arranged in irregular rows. Minute pits 0.1 to 0.3 μ in diameter characterize the Oberhautchen of a ventral scale.Cell outlines are not evident on the scales of G. multicarinatus. The Oberhautchen of dorsal and parietal scales of this species is prominently laminated. Laminae are less prominent on scales of the lateral fold, and no intrinsic surface structure is evident on a ventral scale. In contrast, the fossorial anguinomorph Anniella pulchra exhibits Oberhautchen surfaces with practically no intrinsic microornamentation. However, what appear to be outlines of Oberhautchen cells are visible on the dorsal and ventral scales.These observations suggest that modifications of Oberhautchen microornamentation may have evolved to reduce friction with the substrate or other scales. The lack of pronounced microornamentation of the Oberhautchen on some body scales may indicate that a complex interdigitation between clear layer and Oberhautchen cells is not essential to the sloughing process.

Notes: Stolonic growth has been compared among several genera of calyptoblastic (Campanularia flexuosa, C. calceolifera, Gonothyraea and Sertularia) and gymnoblastic (Pennaria, Bougainvillia, Eudendrium and Cordylophora) hydroids with respect to such features as the (1) complexity and uniformity of the tip movements, (2) constancy of growth cycle duration, (3) variability in growth among cycles, (4) growth rate, and (5) variability of retractions among cycles.The “growth cycle,” previously described in C. flexuosa, is the basis for elongation in all species observed. Its pattern is indistinguishable between the Campanularia species; however, at the generic level the tip movements show peculiarities which delineate each genus from all others. In addition, the movements in all calyptoblasts are uniform from cycle to cycle and comparatively simple, whereas those of the gymnoblasts are complicated by the variable appearance of one or more secondary forward thrusts between crests.A dichotomy is seen between the calyptoblasts and most gymnoblasts in other respects as well. Cycle time is very predictable and cycle-to-cycle variability in growth is relatively narrow in all calyptoblasts. However, in most gymnoblasts both features are much more loosely regulated: only Pennaria shows some degree of control to its timing mechanism, and only in Bougainvillia and Eudendrium does the uniformity of growth per cycle reach the calyptoblast level.On the premise that calyptoblasts are evolutionarily the more advanced group, simplicity and regularity of growth movements are concluded to be the evolved, and complexity and variability the primitive, conditions. On this basis the evolutionary relations among the four gymnoblasts, as deduced from their stolonic growth behavior alone, are consistent in many but not all respects with those interpreted from morphological considerations.

Notes: In order to clarify the morphology of the circulatory system of amphioxus the blood vessels were investigated using modern techniques of light and electron microscopy.The pattern of circulation in amphioxus is forward ventrally and backwards dorsally. In addition, circulating corpuscles, usually associated with the blood of higher chordates, are absent.The circulatory system of amphioxus consists of well defined contractile vessels and vascular spaces or sinuses within a connective tissue matrix. The contractile vessels have a discontinuous endothelial lining resting on a basal lamina and are enclosed by a simple layer of contractile myoepithelial cells. Discontinuous endothelial linings occur throughout the vascular tree, including major and minor afferent and efferent vessels and blood sinuses. This is in contrast to higher animals where the endothelium forms a more or less continuous lining along the inner surface of the boundary layer.It is suggested that the endothelial cells of amphioxus, like the endothelial cells in capillaries of higher chordates, most likely play a role in the physiology of the circulatory system by removing residues of filtration from the basal lamina, thereby facilitating an exchange of materials to and from the surrounding tissues.

Notes: The development of the polarity and bilateral asymmetry of the future adult zooid has been traced to their earliest morphological expression in the palleal bud of Botryllus. The account is based upon continued observation of living buds. The polarized antero-posterior and dorso-ventral axes are first expressed by the skewing of a symmetrical, hemispherical bud towards the anterior end of the parental bud. Identification of these axes is reinforced by the development of a loop-like blood circulation, the primary circulation, in the horizontal plane during the enlargement of the skewed hemisphere to form a stalked vesicle. Bilateral asymmetry is first expressed by the asymmetrical expansion of the vesicle stage. The right posterior corner of the vesicle expands further posteriorly and becomes more acute than the left posterior corner. This larger expansion persists throughout the development of the right atrial cavity, which finally expands across the mid-line to partially surround the gut. The bilateral asymmetry, expressed in the expanded vesicle, is reinforced by the development in sequence of a rounded gut rudiment and a pericardial rudiment in positions that would be expected from the asymmetry of the expanded vesicle. The first appearance of the gut rudiment occurs earlier than had been recognized previously. Conflicting accounts of the time and mode of formation of the pericardial rudiment have been clarified. The results of this study are discussed in the context of determination of bud territory, polarity and bilateral asymmetry.

Notes: Four glands of the house sparrow, chicken and turkey were examined histologically and for their content of amylase. These were the external and intermediate mandibular glands, the maxillary gland and glandula anguli oris of the sparrow and the anterior and posterior mandibular, maxillary and anguli oris glands of the chicken and turkey. Amylase was determined by a starch substrate slide method and by biochemical assay. General morphology and mucopolysaccharide staining are described.All four glands of the sparrow demonstrated significant amylolytic activity by the assay. In the external mandibular and anguli oris glands this activity could be traced to mucous and seromucous cells of origin by means of the starch substrate slide procedure. None of the glands of the chicken or turkey displayed significant amylolytic activity.

Notes: The distribution of monoamine (catecholamine and 5-hydroxytryptamine)-containing nerve cell bodies in the brain stem and hypothalmus of the frog (Rana temporaria) was investigated with the help of the histofluorescence technique of Falck and Hillarp ('62). At the level of the hypothalmus of this amphibian brain, catecholamine-containing nerve cell bodies are found mainly within three areas of the periventricular gray substance, namely the peroptic recess organ, the paraventricular organ and the lateral recess region. On the other hand, most of the 5-hydroxytryptamine (serotonin)-containing nerve cell bodies of the brain stem of Rana temporaria appear to be concentrated within the midbrain tegmentum. This huge mesencephalic nerve cell collection can be subdivided into medial and lateral groups. More caudally, at the level of the isthmic tegmentum, another group of 5-hydroxytryptamine-containing perikarya located close to the midline, within the so-called raphae region, is clearly outlined. The latter group of neurons extends caudally as far as the level of the medulla oblongata. In addition, a small group of catecholamine-containing nerve cell bodies is also found in the ventromedial portion of the rostral midbrain tegmentum, whereas a few other catecholamine type neurons are scattered throughout the lower brain stem of the frog and more especially near the ependymal wall of the fourth ventricle. As a whole, the 5-hydroxytryptamine-containing neuronal systems of the brain stem of Rana temporaria are much more elaborated than the catecholamine neuronal systems of the same structure.

Notes: The guinea pig soleus muscle is a convenient model for the study of slow-twitch intermediate (STI) fiber ultrastructure because it is composed entirely of fibers of this class. Such fibers were compared with fast-twitch red (FTR) and fast-twitch white (FTW) fibers from the vastus lateralis muscle.FTW fibers are characterized by small, sparse mitochondria, a narrow Z line and, an extensive sarcoplasmic reticulum arranged primarily in longitudinal profiles at the A band and with numerous expansions at the I band. Abundant mitochondria with a dense matrix and subsarcolemmal and perinuclear aggregations are typical of FTR fibers. These fibers contain a plexus of sarcoplasmic reticulum at the A band and a less extensive network at the I band. The Z lines are wider (890 ± 74 Å) than those of FTW fibers (582 ± 62 Å). STI intermediate fibers are distinguished from other types by wide Z lines (1205 ± 58 Å), a faint M band, and a less extensive sarcoplasmic reticulum. Compared to FTR fibers, STI fiber mitochondria are usually smaller with less notable subsarcolemmal accumulations.FTW fibers have a more limited capillary supply, rarely contain lipid inclusions, and thus may be restricted to phasic activity. Extensive capillarity, mitochondrial and lipid context, and fast contraction times indicate possible phasic and tonic roles for FTR fibers. STI fibers, characterized by numerous lipid inclusions, extensive capillarity, relatively numerous mitochondria, but slow contraction-relaxation cycles, are morphologically suited for tonic muscle activity.

Notes: The haemocytes of Calpodes ethlius are described with the light and electron microscopes. Five fine structurally distinct types are distinguishable. However only three of these, the granular haemocytes, sphaerule cells and oenocytoids can be positively identified using a series of histochemical stains on smears and on thick sections of araldite-embedded material. The classification is based entirely on the structural features even though several suggestions concerning their function can be made from their fine structure. Intermediates having features of more than one cell type suggest developmental relationships.

Notes: Four hemocyte types have been identified in the late last larval instar of Galleria mellonella. Plasmatocytoids are round to spindle shaped cells, 10-20 μ long and 5-10 μ wide. The cytoplasm contains no distinguishing inclusions. Golgi complexes, rough endoplasmic reticulum and free ribosomes are abundant. Granular hemocytes are oval shaped cells, 10-20 μ long and 5-10 μ wide. The granules, their most characteristic feature, have a diameter of 0.2 μ, a microtubular sub-structure, and are made up of acidic mucosubstances. Lipid droplets may be present in these cells at some stage of development. These cells appear to be phagocytic. Spherule cells are oval shaped, 15-20 μ long and 5-10 μ wide. The spherules, approximately 2 μ in diameter, have a highly ordered substructure and are made up of acidic mucosubstances. Oenocytoids are the largest cells, 20 by 40 μ. The cytoplasm contains mostly free ribosomes and microtubules.

Notes: The epineurium of the esophageal complex of the gastropod pulmonate Triodopsis divesta was examined by electron microscopy. The epineurium consists of two main regions: an inner dense fibrous region adjacent to the avascular neural tissue of the ganglion and an outer cellular region comprised of a variety of cell types embedded in a connective tissue matrix. The dense fibrous region contains smooth muscle cells and associated nerve processes and is invested on the neural side by thin processes of glial cells. The outer highly cellular region contains smooth muscle cells, nerve processes, wandering cells (amebocytes), globular cells, and myoepithelial cells comprising the walls of the vascular system. In addition, a cell type not previously identified in other gastropod epineuria is present. These cells resemble neurosecretory cells. The morphology and structural interrelationships of these various constituents are presented and the possible functions of individual cell types and the epineurium in general are discussed in relation to information available on other molluscs.

Notes: The muscles of the head, neck and thorax of the adult cabbage looper Trichoplusia ni are described for the first time. They are compared to those of Choristoneura fumiferana, Manduca sexta, Smerinthus geminatus, Antheraea polyphemus, Sphinx convolvuli, Crymodes devastator, and Danaus plexippus.

Notes: A precise sequence of four morphological stages of head regeneration in the planarian Dugesia tigrina has been determined by light and electron microscopy. Each stage is identified by a particular morphogenetic process: I, wound healing; II, blastema development; III, growth; IV, differentiation. A wound epidermis consisting of a thin, sheet-like layer of cells, rapidly forms from undamaged epidermal cells at the wound margin. The early blastema is comprised of neoblasts which mature into regeneration cells. The maturational changes include the appearance of a nucleolus, nuclear pores, and perinuclear dense aggregates of granulofibrillar material in these cells. These elements are not evident in the neoblasts of the younger blastema. No mitotic cells are encountered in the blastema or wound epidermis. Cytoplasmic expansion of the regeneration cells is correlated with the formation of numerous microtubules radiating from a juxtanuclear centrosphere. During differentiation of muscle cells, distended, granule-studded cisternae, having moderately fibrillar contents, are regularly disposed adjacent to small patches of myofilaments. Presumptive epidermal cells are recognized by prominent “islands” of finely fibrillar cytoplasm. These cytoplasmic zones persist for a time during definitive differentiation when Golgi bodies, vacuoles, mucous droplets, and rhabdites become evident. The newly formed epidermal cells become inserted among the cells of the wound epidermis. Thus, cells of both the blastema and of the wound epidermis contribute to the reconstituted epidermis.

Notes: Five different kinds of sense organs have been identified on the antennal flagellum of males of Ptilocerembia sp: (1) tactile hairs, (2) thick-walled chemoreceptors, (3) chemoreceptors with thick walls and many pores, (4) thin-walled chemoreceptors and (5) campaniform sense organs. The third type has not been reported previously for other species of insects and combines the characteristics of typical thick-walled and thin-walled chemoreceptors. The campaniform sense organs are situated in sets of three, 120° apart, on the periphery of the distal end of nearly every subsegment. They lie at the transition zone where the thicker cuticle of the subsegment changes to the thin membrane between subsegments.

Notes: The paddlefish (Polyodon spathula) is one of the most primitive and unique freshwater fishes of North America. It is adapted as a plankton filter-feeder. The wide mouth and greatly expansible pharyngeal cavity allows it to process a large quantity of water. Numerous setiform gillrakers, which operate against the flattened surfaces of the gill arches and opercula, are capable of filtering out small food particles. The abundance of taste buds and mucus secreting cells in the buccal cavity and pharynx probably aids in selecting and entangling the food. The first segment of the stomach is provided with peculiar, large longitudinal rods of fat which may help in selecting food from water by causing occlusion of the small lumen when the striated fibers of the muscularis contract. Teeth, unnecessary for microphagous fishes, are absent in adults.The digestive tube is one of the most complex present in fishes. It has two distinct divisions of the stomach and four distinct divisions of the intestine besides possessing a large caecum. Ciliated epithelial cells occur in many parts of the gut and may be an aid in moving or processing the small items of food.Besides the presence of cilia, other primitive features of the digestive tract include the lack of distinction between the esophagus and stomach, the presence of a spiral valve, and the presence of some of the diffuse pancreatic tissue within the wall of the gut.The numerous Peyer's patches in the wall of much of the intestine may be an immunological response to the heavy parasitic infestation.

Notes: The cranial glands of ten species of turtles were studied by the use of histochemistry applied to serial sections of whole heads. The majority were stenohaline species, but one brackish water form, Malaclemys, was included. The results show that all species have two major orbital glands, an anterior Harderian gland, and a posterior lachrymal gland. The latter is seromucous in all species except Malaclemys terrapin in which the gland shows little evidence or organic secretion. External and medial nasal glands are found in all species studied, and also are seromucous glands. With these reslts, combined with a review of the literature the following conclusions are made. The Harderian gland is by definition the orbital gland opening through the medial surface of the nictitating membrane at or near the anterior canthus. It is of constant occurrence, and histological appearance, probably serving the same function. However, despite much recent study this function remains unknown. The lachrymal gland is defined as the orbital gland which opens through the lateral surface of the nictitating membrane, or medial surface of the lower eyelid, at or near the posterior canthus. It is of variable occurrence, absent in many reptiles, and has a histological structure which is also variable. In the stenohaline species it is apparently involved in organic secretion, while in the brackish water Malaclemys it may be involved in salt secretion, as it is in Cheloniidae. The nasal glands in turtles are probably homologous with the nasal salt glands of lizards and birds, but they do not appear to subserve the same function. In all species of turtles studied the nasal glands are seromucous. They are perhaps involved in the maintenance of the epithelium of the olfactory cavity.

Notes: The dorsal proctodeal wall of the Coturnix contains a gland which is especially well-developed in the sexually active male and which secretes a foamy exudate. This gland has not been recorded in any other genus of birds and appears to be unique to members of the genus Coturnix. On the basis of its location in the adult we have named this gland the proctodeal gland (glandula proctodealis). Structure of the gland does not conform to any curently defined category of glands because it is an aggregate of individual glandular units each of which opens independently into the proctodeal cavity but with the entire structure enclosed by a connective tissue capsule. We have thus characterized it as an aggregate gland. Observations on the arrangement and composition of the fibrous capsule and its intimate relationship to the perimysium of M. sphincter cloacae indicate that release of the secretion probably results from contraction of the cloacal musculature. Preliminary data suggest that development of glandular tissue in the female may be a function of age rather than of androgen sensitivity which characterizes the male gland. It is concluded that the chemical nature of the glandular secretion and its function in the sexually active bird, as based on presently available information, remains unknown.

Notes: Wound healing in the integument of the sea cucumber, Thyone briareus, was studied for up to 50 days after inflicting wide excisional wounds and for 14 days after producing incisional wounds. Rapid re-epithelialization of the wound was effected by the migration of epidermal cells and pigment cells from the periphery of the wound margin. This occurred without apparent evidence of concomitant mitotic activity. Dermal wound healing was completed by the fourteenth day in the incision wounds but occurred very slowly in the broad excision wounds. Morula cells seem to be involved in both epidermal and dermal wound healing, although their precise role is unknown. In excisional wounds the integument was never completely restored to its normal appearance during 50 days of observation.

Notes: The electron microscopy of the epiphysis cerebri of the antarctic seal Leptonychotes weddelli revealed a highly organized organ. The general cytological characteristics of the pinealocytes and the glial cells are described. The capillary blood vessels are the nonfenestrated type. The organ is richly innervated by mainly unmyelinated nerve fibers. Most of the axons end in the inner part of the organ, around vessels, some of them in relation with pinealocytes. The significance of the findings is discussed.

Notes: Cleavage and blastoderm formation in Coelopa frigida are extremely rapid developmental processes. In short (6-7 minutes) successive cell cycles, nuclei multiply and spread out through the egg. The movement seems to be aided by endoplasmic vesicles and cisternae which are in direct contact with the nuclear membrane. The first cells to separate from the egg plasmodium in early superficial cleavage stages are the pole cells.Precursor material from multivesicular bodies forms the pole cell membranes. The primary nuclei from the posterior pole region are removed from the blastoderm by the pole cell segregation. Blastoderm nuclei from the regions adjacent to the posterior pole migrate into the residual periplasm after pole cell segregation has been completed and constitute the blastoderm nuclei in that region of the egg. Nucleoli are not revealed during internal cleavage. They appear in pole cells shortly after their segregation. The generation time of the blastoderm nuclei increases after the twelfth cleavage. Concurrently, nucleoli form in the blastoderm nuclei and permanent cell membranes separate individual blastoderm cells. After blastoderm cells have been separated from each other, they remain in contact with the interior yolk sac by means of cytoplasmic canals. This contact is maintained at least during the early phases of blastokinesis. Observations on nuclear migration and rapid membrane formation are discussed as examples of protein assembly from subunits as an alternative to de novo protein synthesis in early stages of development.

Notes: Ovarian follicular histology of the lizard Anolis carolinensis was studied in relation to follicular size and rank. The structure of the Anolis follicular granulosa is similar to that described in other reptiles. Changes in the granulosa during follicular hypertrophy are also similar to those described in other species. Follicles of low rank in the linear size-gradation exhibited atresia.Counts were made of erythrocytes per millimeter of thecal tissue in crosssections of follicles of different size and rank. Follicles greater than 2.6 mm in diameter exhibited a marked increase in the number of thecal erythrocytes. Also, follicles of a similar diameter tended to possess a greater number of thecal erythrocytes the highter the rank in the linerar size-gradation. These observations support the hypothesis that follicular selection involves appropriation of blood and blood-borne hormones by larger follicles.

Notes: Some of the cytological and histochemical characteristics of hemocytes of Thyropygus poseidon were studied. Jone's system of classification of hemocytes was extended to millipede hemocytes. Seven classes were identified as prohemocytes, plasmatocytes, granular hemocytes, cytocytes, oenocytoids, spherule cells and adipohemocytes. Oenocytoids were rarely found. All cells except prohemocytes and oenocytoids sequestered oxidized products of catechol. Results of the histochemical tests for proteins also suggest that these cells sequester plasma proteins. Cytocytes do not bring about any visible coagulation similar to the hemocytes of some hemipteran insects. Granular hemocytes contain phenol-oxidase capable of oxidising tyrosine as well as catechol, similar to the hemocytes of crustaceans. In this respect millipede hemocytes differ from insect hemocytes. It is suggested that these cell types may represent distinct cell lines and may not represent transformation of one cell type to another.

Notes: Twenty-three stages in the embryonic development of the western painted turtle, Chrysemys picta bellii are described. The staging is based primarily on morphological changes, as well as age and size of the embryo.

Notes: Scanning and transmission electron microscopy were used to investigate the surface area and number per unit area of microvilli from jejunal villus epithelial cells in the rat, hamster and dog.The calculated mean microvillus surface area was 0.419 μ2, 0.573 μ2, 0.751 μ2 for the rat, hamster and dog respectively.The largest number of microvilli per square micron freeze dried villus surface was measured in the rat with a mean value of 65. Hamster and dog freeze dried specimens had lower mean values of 54 and 34 microvilli respectively.The total microvilli surface area in square micron per square micron villus surface was more closely related for the three species with values of 27.23 for the rat, 30.94 for the hamster and 25.53 for the dog.These data indicate an inverse relationship between the mean microvillus surface area and population density in the species studied. However, the total microvilli surface area per unit villus surface is relatively similar for the three species.The observed number of microvilli per unit villus surface was shown to vary depending upon the dehydration technique employed for preparation of scanning electron microscopic specimens. This variation probably reflects shrinkage artifact and should be considered in soft tissue studies involving the scanning electron microscope.

Notes: In Esox americanus and Esox niger (Esocidae) there are two types of surface pits, sensory and nonsensory. Both types of pits are morphologically very similar and are associated with modified (cardioid) scales. The pits are distinguished by the presence and absence respectively of neuromasts. Ultrastructure of the neuromasts indicates that they are basically similar to previously described neuromasts. However, the hair cells have crystalline-like deposits in the nuclear region. The nonsensory pit contains a small group of distinct epidermal cells, called central cells. These cells occupy a central position in the pit, similar to that of the neuromasts in the sensory pit. The central cells are unlike any cell types described for fishes, and on an ultrastructural basis no function can be ascribed to them and the significance of the nonsensory pit is unknown.

Notes: Esox americanus and Esox niger (esocidae) possess two types of surface pits, sensory and nonsensory. The pits are morphologically very similar and both are associated with modified (cardioid) scales. The pits are distinguished by neuromasts in the sensory and central cells in the nonsensory pit.Neuromasts and central cells are responsible for the formation and maintenance of respectively the pit and cardioid scale. This supports an extension of the epidermal co-participation hypothesis that epidermal cells control the form of dermal structures in addition to contributing to composition.The central cells are similar in structure to induced primordial cells of regenerating neuromasts.The development of primordial cells may be arrested and the cells maintained in a static condition during which time they still influence the dermal tissues and maintain a pit and cardioid scale. Essentially this represents the experimental conversion of a sensory pit into a nonsensory pit. This evidence is taken in support of the hypothesis that the nonsensory pit and central cells represent an aplasic condition of the lateral line system.

Notes: The gular gland of the bat Tadarida brasiliensis is a specialized sebaceous gland located in the skin of the suprasternal region of adult males. It consists of an aggregation of simple branched tubulo-acinar gland units, the number of which varies seasonally. Each acinus is composed of densely packed sebaceous cells at various stages of differentiation. Acinar basal cells and cells of the epithelium of the ducts can differentiate into sebaceous cells. Two main changes appear in the cytoplasm concurrent with the sebaceous transformation: the differentiation of cytoplasmic organelles and the deposition of lipid material. The appearance of a different type of mitochondrion and the development of large numbers of ribosomes and polyribosomes can be recognized in the cytoplasm at an early stage of differentiation. Concomitant with the deposition of significant numbers of lipid droplets, the cells develop abundant agranular endoplasmic reticulum occurring mainly as scattered tubular cisternae. These at times form whorls surrounding lipid droplets. At later stages, the cisternae of the agranular endoplasmic reticulum often occur in crystalline arrays between secretory oil droplets. The roles of the different cytoplasmic organelles, especially in relation to the production of sebum, are discussed.

Notes: The mutant gene srn produces sireniform abnormalities in the mouse. The effects of this gene on normal prenatal development were examined histologically by retrograde analysis from day 10 through day 18 of gestation.The sireniformphenotype is characterized by the fusion of the hindlimbs and accompanying defects of the posterior appendicular skeleton. Visceral defects involve the urogenital system and the lower intestinal tract. A single umbilical artery arises from the dorsal aorta and controls the fetal circulatory pathway normally mediated by the lateral umbilical arteries. This artery is believed to have resulted from the fusion of the paired primitive umbilical arteries.It is suggested that the srn mutation causes a narrowing of the posterior amnion early in development. The resulting abnormal pressure causes a dorsal displacement of the hindgut, allowing fusion of the hindlimb buds. Abnormal structural relationships and pressure in the narrowed posteriior region disrupt normal urogenital and intestinal development and increase the proximity of the primitive umbilical arteries.

Notes: The retinofugal projections of the snake Thamnophis sirtalis were studied by the method of experimentally induced Wallerian degeneration stained by the Fink-Heimer method. The retinal ganglion cells project to all parts of the contralateral lateral geniculate complex, nucleus lentiformis mesencephali, nucleus geniculatus pretectalis, nucleus posterodorsalis, basal optic nucleus and superficial layers of the optic tectum. In addition, the retinofugal projections were observed terminating in portions of the ipsilateral lateral geniculate complex and nucleus posterodorsalis.Examination of the morphology of the retinal terminal areas stained for Nissl substance with cresyl violet led to the conclusion that these regions are well differentiated and should not be considered poorly developed when compared with other reptilian forms such as turtles.

Notes: The course of the statoblast formation in Pectinatella gelatinosa was divided into four stages and studied histologically and histochemically. The bottom of the cystigenous cup is a center of cystigenous cell differentiation and the peripheral zone of the inner cystigenous layer turns to the outer cystigenous layer as the cystigenous cup grows. The annulus is formed by migration and transformation of the outer cystigenous cells.During early stages, the yolk cells have an intensely pyroninophilic or RNA-rich cytoplasm. The cytoplasmic pyroninophilia then diminishes as the amount of yolk granules increases.Several kinds of yolk substances occur in the mature statoblast. During statoblast formation glycogen appears first, then glycoprotein and finally neutral unsaturated lipid.Acid phosphatase activity is associated with granular structures in the cytoplasm. In the cystigenous vesicle, acid phosphatase activity is low and confined to the apical extremity of the cell. Histochemically detectable alkaline phosphatase activity is not involved in the formation of the statoblast.

Notes: The anatomy and physiology of feeding in miniature swine were studied, using gross dissection, electromyography, cinematography, and cineradiography.Small particles are preferred for ingestion, and large items are usually broken down outside of the oral cavity. The particles are initially picked up with the lower lip and then retrieved by the tongue; the tongue very rarely leaves the oral cavity. Geniohyoid, mylohyoid and digastric are the most active muscles during food collection.Mastication is fairly rapid (3 cycles/sec) and involves a transverse component which may be either medially or laterally directed. The direction of motion is generally reversed with every chew. Electromyography indicates that the transverse rotation is caused by a force couple consisting of protrusors (masseter, medial and lateral pterygoids) on one side and retrusors (zygomaticomandibularis and temporalis) on the other. The direction of the rotation is not necessarily related to the side containing the bolus. Mandibular depression is electromyographically biphasic. Concurrent tongue and hyoid movements complicate the interpretation of activity patterns in the oral floor.Both masticated food and liquid are stored between the tongue and the soft palate and epiglottis before being swallowed. Swallowing involves very strong activity in all of the hyoid and tongue muscles and very low activity in the adductors. Liquid is taken in by suction rather than lapping.Comparisons with other mammals are presented.

Notes: The wall of the gametolytic gland in Aplysia and Phyllaplysia consists of a muscular layer lined with an epithelium incorporating four distinct cell types, some of which show intense secretory activity. The cytoplasm of these cells, rich in RNA, has a peripheral layer of densely-packed mitochondria in association with a smooth endoplasmic reticulum. Lipid globules, perhaps resulting from absorption of lumen contents by microvilli, appear in the cytoplasm in the course of growth and reproductive activity. A basal band of fibrillar material appears to serve as a cytoplasmic skeleton. The elaboration of cell types, extensive areas of cytoplasmic RNA, and vigorous secretory activity suggest important roles for this organ such as development of food reserves, recycling of sexual materials of high physiological value, and interaction with the bag cells of the nervous system in coordinating ovulation with the availability of capacitated, exogenous sperm.

Notes: The predaceous neotropical characoid fish Ctenolucius has an essentially homodont dentition, the number of teeth increasing linearly with age. The basic manner of tooth replacement suggests that Ctenolucius is a primitive characoid. Tooth replacement continues throughout life and is similar to that of tetrapods, involving replacement waves which pass from the back to the front of the jaws. The waves containing the greatest number of teeth are found just anterior to the middle of the jaws. In the upper jaw the increase in the number of teeth is restricted to the anterior portion (premaxillary) whereas the number on the posterior part (maxillary) remains constant. In specimens measuring from 68-230 mm in standard length the posterior portion of the upper jaw doubles in length whereas the anterior portion triples. It is suggested that the area immediately anterior to the middle of the jaw, where replacement waves are longest, is where most of the increase in tooth numbers occurs.During growth of the teeth the absolute height is always greater than the absolute width as the shape changes. The final shape of the recurved conical teeth is determined only in the last stages of tooth formation when the main axis of growth abruptly changes.

Notes: Improved histological procedures have revealed in some detail the form and structure of auditory receptor organs in the frog. These organs, the papilla amphibiorum and papilla basilaris, are located in extensions of the saccular cavity, and lie in a path of vibratory fluid motion beginning at the columellar footplate and extending in a continuous circuit through the perilymph spaces and the round window back to the middle ear region.These organs exhibit great uniformity, and for the amphibian papilla there is a high degree of complexity in the tectorial structures that overlay the ciliated ends of the hair cells. In both papillae a “sensing membrane,” which is a derivative of the tectorial body, lies in the path of fluid flow and aids in the stimulation process.The mode of stimulation of the hair cells in the amphibian ear is peculiar, and in principle is the reverse of the familiar one of reptiles, birds, and mammals.

Notes: To determine the prism sheath configurations in human cuspal enamel 80 teeth were initially ground to produce flat surfaces through the following planes: a horizontal series at successively greater distances from the dentinoenamel junction and longitudinally through the center of the cusps. Individual teeth were suspended in an acid-alcohol solution (1 cm3 conc. HCl in 100 cm3 95% ethanol) at 37°C for seven to ten days. The treatment “softened” the enamel to a depth of approximately 1 mm. The teeth were embedded in Epon and sectioned at 0.5 to 10 μm with a diamond knife. Thick and thin ground sections for phase contrast microscopy and acid-etched ground sections for Nomarski differential interference microscopy were prepared through the same regions. In thicker longitudinal sections, the prisms in gnarled enamel formed a zig-zag pattern which was unlike the twisting pattern generally observed in ground sections. The thinnest transverse sections showed the sheath outlines to be dramatically different from those seen elsewhere in the enamel. Some prism sheaths were circular, others were in the form of spirals. What could be described as sheaths within sheaths were also seen. In the thinnest longitudinal sections the prisms were seen to be elongated and discontinuous. Sheath outlines in enamel adjacent to the central core of gnarled enamel were similar to those described elsewhere in the body of the enamel. Keyhole, modified keyhole patterns and arcade forms were the dominant sheath patterns. Other atypical sheath configurations were seen scattered throughout this region.

Notes: The differentiation and degeneration of the cement gland in Xenopus laevis is described. The gland is first observed histologically at stage 19 (neural tube stage) as a packed group of apical ectoderm cells heavily laden with oocyte pigment granules, lying ventral to the cranial neural fold. By tailbud stage 35/36, the gland cells have increased in height and are approximately ten times taller than nonglandular apical ectoderm cells. The nuclei divide the gland cells into an apical region that is eosinophilic and contains oocyte pigment granules, and a basal region that contains clear droplets. The cells are decreasing in height by stage 40 (early tadpole) and begin to lose their pigment granules. Between stages 45 and 48, the pigment is extruded and the clear basal droplets diminish in number. From stage 48 to 49 the cells become vacuolated and the histotypic characteristics of the functional gland are lost. The gland is not vascularized, nor do phagocytic cells appear in its vicinity during any stage of its development. It remains bordered at its base by subjacent basal ectoderm during its entire life cycle of 10 to 12 days at room temperature.

Notes: Methods for the production of protein concentrates, with a low content of nucleic acid, in kilogram quantities from yeast have been studied with the aid of equipment designed for operation on pilot-plant scale. The influence of drum drying and mechanical disintegration on the nutritive value of the yeast was also investigated. Drum drying and mechanical disintegration improved the nutritive value of the yeast but high extractability of protein and nucleic acid was only obtained after mechanical disintegration.Protein concentrates without and with cell walls were produced from mechanically disintegrated yeast. The different fractions which were obtained when separating cell walls and precipitating protein by heating at alkaline pH, were analyzed. After protein precipitation, about 90% of the RNA could be precipitated from the supernatant by addition of acid, giving a product containing 50% RNA of the dry weight. The protein precipitate obtained after cell wall separation had an RNA content of less than 2% and contained 70-l75% of the amino acids in the starting yeast material. Protein concentrates containing cell walls were produced by precipitating protein by heating at alkaline pH directly after mechanical disintegration. The content of RNA was about 2% and the yield of amino acids was 70-80%. It was found that the nutritive value of the protein concentrate was higher than that of the starting yeast material. To produce such a protein concentrate on a large scale, the process described can probably be employed.

Notes: Xanthan gum, a heteropolysaccharide with unusual and useful properties, is now produced commercially by fermentation with Xanthomonas compestris NRRL B-1459 in a medium containing glucose, minerals, and a complex nitrogen source - distillers' dried solubles (DDS). Understanding the kinetics of the fermentation should contribute to process improvements and increase the market potential for the gum. Earlier studies showed that although DDS determined initial growth rate, growth was stopped by some mechanism other than substrate exhaustion, probably an effect related to product formation.Product formation did not require active growth, but its rate increased with cell concentration. Specific product formation rate declined at high viscosities. Varying glucose concentration from 0.5 to 5.0% and dissolved O2 tension between 20 and 90% air saturated had no effect on the rates, but pH had to be maintained near 7 and temperature near 28°C to permit continued product formation. Xanthan yield could be explained by the energy required for growth and polymerization, that energy coming from dissimilation of the part of the carbohydrate substrate not converted to polymer.

Notes: Barely β-amylase (α-1,4-glucan maltohydrolase, EC 3.2.1.2) has been immobilized by covalent fixation to amino derivatives of epichlorohydrin crosslinked Sepharose mediated by cyclohexyl isocyanide and acetaldehyde. The enzyme conjugates contain up to 35% of the total activity of the β-amylase added to the coupling mixture. The profiles of activity versus pH and ionic strength are essentially the same for free and immobilized β-amylase, whereas the resistance to inactivation during storage and use is considerably enhanced by immobilization. Columns with immobilized β-amylase have been used for continuous degradation of starch. At 45°C, half of the initial activity remains after seven weeks, and the corresponding figure at 23°C is 85 percent.

Notes: Of 24 different polymer and surfactant materials examined, a carboxypolymethylene (“Carbopol”) was found to cause enhancement of respiration rates in an Aspergillus niger culture by as much as 200%. Enhancement of other fermentation parameters, such as cellular growth and amylase production, was also observed.The enhancement effects of Carbopol were examined with clusters of spores and mold pellets. In the first case, it appears that the ionized carboxyl groups of Carbopol induced electrostatic repulsion among the spores thus initiating pulp growth with increased interfacial area of contact between the mold and the nutrient medium. In the second case, the Carbopol additive formed a thin film attached to the surface of the pellets which seemed to be responsible for an increased rate of potassium transport and, hence, fermentation yields. Additive utilization as substrate and physiological changes in the culture were not observed in these cases.It was also found that the probability of pellet formation, the size of pellets formed, and the number of spores per pellet can be correlated to the energy input to the fermentation system.

Notes: The steady state, nonlinear diffusion equations which describe reactions in constrained enzyme solutions are of great interest in many biological and engineering applications. As in other types of nonlinear differential equations, exact analytical solutions do not exist except in some simplified cases. In this paper, a general procedure is presented for solving numerically for the substrate concentration profile and effectiveness factor utilizing the transformation method suggested by Na and Na. Design correlations for enzyme solutions constrained within spherical membranes are included. The use of a unique definition of the Thiele Modulus in these charts permits the clear illustration of the effects of substrate concentration and external mass transfer resistances on the overall effectiveness factor for the catalyst particle.

Notes: Studies are reviewed on the composition of hemolymph from Japanese beetle larvae, Popillia japonica. This research was conducted to determine the chemical environment required for sporulation of the milky disease organism, Bacillus popilliae. Amino acids, proteins, organic acids, lipids, carbohydrates, and oxygen content were determined in hemolymph from healthy and diseased larvae. Oxygen content of hemolymph was measured by microelectrodes implanted in the hemocoel of living larvae. Vegetative growth of the pathogen reduces the dissolved oxygen; however, as sporulation occurs, oxygen concentration approaches normal values. Trehalose, the major hemolymph sugar, is reduced by ca. 50% during the course of milky disease. Neutral lipids, phospholipids, and hydrocarbons decrease markedly with infection. Both branched-chain and normal alkanes occur in hemolymph. Milky disease causes an increase in concentration of pyruvic, malic, glycolic, tartaric, and glyoxylic acids. The major hemolymph protein, a lipoglycoprotein, has a mol. wt. of ca. 500,000. Reduced concentration of this protein is accompanied by the appearance of a smaller protein. Amino acids that increase significantly as a result of infection are: Glutamic acid, aspartic acid, β-alanine, phenylalanine, threonine, lysine, and serine. Hemolymph contains few peptides of low molecular weight.

Notes: Cellulase production by strains of Myrothecium verrucaria, Stachybotrys atra and Trichoderma viride was examined. Myrothecium verrucaria was found to give the greatest yields. A variety of media were examined as potential substrates for the industrial production of cellulase. The salts content of the medium was varied and was found to affect cellulase production. Glucose, carboxymethylcellulose (CMC), filter paper and three industrial wastes were examined as possible cellulase inducers. Filter paper was found to be the most effective, followed by sugar cane bagasse and CMC.

Notes: Flavor enhancing nucleotides may be produced by streptomycetes. Mutants of streptomyces 772 were found to produce, under ordinary fermentation conditions, considerable amounts of 5′-IMP and 5′-XMP, but only traces of 5′-GMP. This was probably due to the feedback inhibition of the end product GMP. It was shown that through suitable fermentation techniques it was possible to overcome this internal control mechanism, leading to increased yields of 5′-GMP.

Notes: Quantitative studies are reported of external film and internal pore diffusion effects for glucose oxidase immobilized on porous glass in a continuous, tubular, packed bed reactor. The McCune and Wilhelm model for external film diffusion is shown to describe these data over a particle Reynolds number range from 0.2 to 25. The Thiele analysis provides a good description of internal pore diffusion resistances. It appears that immobilized enzymes free of internal diffusional restrictions generally require the use of supports with pore sizes larger than those currently available in porous glass.

Notes: The response of aerobically growing heterogeneous microbial populations of sewage origin to step increases and decreases in pH were studied in both once-through and cell recycle systems. The pH range studied was 2.7 to 8.0. All studies were conducted at a dilution rate of 0.125 hr-1, and all shocks were administered from a base or preshock pH level of 6.4 to 6.7. In each experiment, the preshock or initial “steady state” was assessed, the pH of the feed changed, and the resulting transient behavior of the system examined until attainment of the new or final “steady state” was approached. The major objectives of the work were to characterize the nature of the response with respect to biomass and effluent substrate concentrations, types of microbial populations present and chemical composition of the biomass, and to obtain guidelines as to allowable change in pH in waste streams. It was found in once-through systems that substrate removal efficiency recovered from pH levels as low as 3.0 after rather long periods of transient leakage of substrate. Cell recycle attenuated the severity of substrate leakage. In all cases of severe acid shock, the microbial population changed from predominantly bacterial-protozoan to one consisting predominantly of filamentous fungi. Changes in chemical composition of the sludge (protein and carbohydrate content) were consistent with the population changes. Based upon the results, it can be conservatively estimated that changes in pH of no more than one unit from the neutral preshock range can be tolerated without possible disruption of biochemical efficiency of substrate removal.

Notes: Wheat germ agglutinin protein, which is able to agglutinate tumor cells better than normal cells, was covalently bound to polyacrylamide gel beads. The specific binding activity of the protein was preserved on these beads and was expressed heterogeneously by the binding of mouse leukemia cells (L1210) to the protein coupled gels. The selective activity of the immobilized protein was maximal when the number of sites available to covalently couple the protein was lowest. The application of this observation to the general field of covalent immobilization of proteins and enzymes may be of considerable utility.

Notes: It is sometimes easier and cheaper to extract a drug's target sites from cells than it is to purify, concentrate or manufacture the drug itself. This is especially true in the case of DNA and some of the exotic antibiotics that react with it. Phleomycin, an inhibitor of replication, was considered as a specific case in point. Biologically active components of this antibiotic mixture bind nondestructively to DNA, in vitro and in vivo, and thereafter may be processed by simple, economical methods usually reserved to macromolecules of much greater size.

Notes: A procedure for the preparation of about 50 g batches of tRNA from 25 kg E. coli W is described. The method involves phenolic extraction of the cells, batch absorption of the tRNA on DEAE-cellulose, washing the DEAE-cellulose and packing it into a column, elution of the tRNA from the column and precipitation of the tRNA with ethanol. The method is less time and labor consuming than the methods described in the literature and can be carried out with relatively simple equipment.

Notes: Several techniques for investigating the multiplicity and stability of open isothermal enzymatic reactors are discussed and some of the pitfalls in previous thinking pointed out. The example which is used to illustrate these methods exhibits several interesting features. Among these is the existence of a stable oscillatory state which surrounds a unique steady state which is asymptotically stable to certain finite disturbances.

Notes: A simplified model of cell metabolism, consisting of a series of linked reversible enzymatic reactions dependent on the concentration of a single external substrate has been developed. The general mathematical solution for this system of reactions is presented. This general solution confirms the concept of a rate-limiting step, or “master reaction”, in biological systems as first proposed by Blackman. The maximum rate of such a process is determined by, and equal to, the maximum rate of the slowest forward reaction in the series.Of practical interest in modeling the growth rate of cells are three cases developed from the general model. The simplest special case results in the Monod equation when the maximum forward rate of one enzymatic reaction in the cell is much less than the maximum forward rate of any other enzymatic reactions.More realistic is the case where the maximum forward rates of more than one enzymatic reaction are slow. When two slow enzymatic reactions are separated from each other by any number of fast reactions that overall can be described by a large equilibrium constant, the Blackman form results: \documentclass{article}\pagestyle{empty}\begin{document}$$\mu = [S]/A, \rm{when} [S] 〈 A\mu_{\rm{max}}$$\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$$\mu = \mu_{\rm{max}}, \rm{when} [S] \rm{〉} A\mu _{\rm{max}}$$\end{document}A third case is that in which two slow enzymatic steps are separated by an equilibrium constant that is not large. Unlike the Monod and Blackman forms, which contain only two arbitrary constants, this model contains three arbitrary constants: \documentclass{article}\pagestyle{empty}\begin{document}$$[S] = \mu A + \frac{{\mu B}} {{(\mu_{\rm{max}} - \mu)}}$$\end{document}The Monod and Blackman forms are special cases of this three constant form.In comparing equations with two arbitrary constants the Monod equation gave poorer fit of the data in most cases than the Blackman form. It is concluded that workers modeling the growth of microorganisms should give a t least as much consideration to the Blackman form as is given to the Monod equation.

Notes: Immobilized invertase was prepared by binding native invertase to a polyamine type ion-exchange resin. Kinetic behavior of the immobilized invertase on these small pellets was investigated in a packed-bed reactor. For low flow rates, an effect of interparticle diffusion on the Machaelis constant was observed and a correlation was proposed to evaluate the effect. At high flow rates, Michaelis and inhibition constants were determined and compared with those for native invertase. With large pellets, intraparticle diffusion was found to be important and at high substrate concentrations, the effectiveness factor exceeded unity. Good agreement was found between the theoretical analyses and the experimental data.

Notes: A process for the large-scale isolation of choline acetyltransferase from ox brain has been developed, by scaling-up an existing laboratory method. 20% of the enzyme was obtained from 17 kg of brain tissue with a 50-fold purification.

Notes: The effect of potassium sorbate addition to a continuous culture of Saccharomyces cerevisiae was observed. The influence on the yeast performance depended on the way the inhibitor was added. The unsteady state continuous method seems to be the best one to obtain microorganisms' adaptation to inhibitors, and for the experimental study of microbial inhibition.

Notes: New methods of crosslinking enzyme molecules inside a matrix with or without an inactive protein are described. Enzyme activity yields range between 30 and 80% of the activity of the untreated preparations. Even fragile enzyme systems, for instance those using mobile cofactors, can be efficiently immobilized. Increased resistance towards heat denaturation and proteolysis results.

Notes: Penicillin amidase, an enzyme which hydrolyzes benzylpenicillin to 6-aminopenicillanic acid and phenylacetic acid, is produced by Bacillus megaterium ATCC 14945 as an extracellular enzyme. We used this system as a model to examine the effects of nitrogen, sulfur, and phosphorous limitation on enzyme production in continuous culture. For these studies, we developed a minimal medium for B. megaterium which contained histidine as the sole nitrogen source. Batch experiments showed that this enzyme is produced as a growth-associated metabolite. Enzyme production was shown to be a function of the growth-limiting conditions and the concentration of the inducer, phenylacetic acid. Sulfur limitation in continuous culture yielded enzyme activities approximately three to five times those observed in nitrogen- and phosphorous-limited chemostats. These results are discussed in terms of the environment's influence on enzyme production in continuous culture.

Notes: A periodic countercurrent whole beer process was developed for the recovery of novobiocin to eliminate the high cost of mycelium filtration and accompanying antibiotic losses in the filter cake. In such a process screened, but unfiltered, novobiocin fermentation beer is contacted with a special grade of an anionic-exchange resin in a series of specially designed, well mixed columns. Each column is fitted with a screen sized to retain resin within the column but allow the cells to press. Periodically the lead column is isolated from the column train, washed free of beer solids, and eluted. A freshly eluted column is placed in the trail position to allow countercurrent operation. The eluate is then processed to crystalline product.A mathematical model for the sorption of novobiocin was developed based on a suitable continuity equation and mass transfer and equilibrium relationships determined in the laboratory. Digital computations of this model correlated well with laboratory and pilot plant data, and predicted well the performance of the production units. This simulation has been in continued use to predict and reoptimize plant operation as process changes (such as improved beer titers, increased production rate, and the evaluation of superior ion-exchange resins) occurred or were anticipated.