ALBERT

Notes: Abstract Use is made of the argon ion-bombardment technique to reduce the thickness of calcium carbonate shells of the bivalvesMytilus andMercenaria for transmission electron microscopy and electron diffraction, with comparison of single-stage replicas of similar specimens serving as controls. As an additional approach to shell structure studies, it gives results which support earlier published work with both replicas and scanning microscopy. In addition, a “frothy” structure is detected in the shell aragonites, especially inMytilus nacre. As a heat-induced artifact, it is interpreted as representing trapped water and organic material inclusions, an interpretation consistent with several published chemical and electron microscope studies.

Notes: Abstract The ruffled border of osteoclasts in the fracture callus of rat consists of complex infoldings of the plasma membrane forming narrow channels. Absorption of an exogenous, electron-dense compound, thorium dioxide, has been shown to take place via these channels, apparently through “membrane flow”. The contents of the channels are transferred to lysosomes (“specific granules”) located subjacent to the ruffled border. In “transitional regions” adjacent to the ruffled border, uptake of thorium dioxide particles appeared to occur through “membrane vesiculation” (conventional endocytosis).

Notes: Summary The egg-shell of Japanese quail was studied by several techniques. Semithin sections (1μm thick) of non-decalcified shell were observed by normal and polarized light microscopy. Thin sections of non-decalcified shell, examined by transmission electron microscopy, permitted us to observe the forms and dimensions of crystals of calcite within different layers of the shell: mammilary layer, layer of cones, palissade layer and surface crystal layer. There appears to be two distinct zones in the layer of cones as well as in the superficial crystal layer. Electron microdiffraction revealed the orientation of calcite crystals in the columns. Some crystal defects (twins?) were described and the possibility of their artefactual formation during ultramicrotomy is discussed. Localization of Ca, Mg, P and S were made by X-ray microanalysis of semithin sections. This technique shows that shell membranes, and chiefly the true cuticle, are also mineralized but, in these layers, minerals are not crystallized. Otherwise the distribution of Mg is not uniform throughout the shell thickness; it is less concentrated in the external zone of the layer of cones. These results together with observation of developing shells by scanning electron microscopy allowed us to propose a scheme for shell organization of the quail egg. This organization was related with decalcification which occurs during hatching.

Notes: Summary Newly formed osteodentin obtained from the anterior extremities of fetal or young rat incisors was observed by means of electron microscopy and electron probe X-ray microanalysis. Cells related to osteodentin formation frequently showed membrane bound intracellular bodies containing varying amounts of fine, needle-shaped crystals, which were identified as apatite. The intracellular clusters of apatite crystals were extruded from the cells through membrane fusion or cellular degeneration. These extracellular clusters seemed to be gradually incorporated into the mineralizing collagenous matrix, which developed around them. Frequent occurrence of dense, dotshaped or filamentous profiles suggested that the dense bodies seen in the perinuclear regions or in the Golgi area were the sites of crystal formation. Energy dispersive X-ray point analysis showed that the intracellular or extracellular apatite clusters contained sulfur in a concentration higher than was present in the mineralizing collagenous matrix. Furthermore, wave dispersive X-ray line analysis showed that the concentration of sulfur was higher in the osteodentin matrix than in the dentin matrix. The sulfur detected is presumed to be contained in acid mucopolysaccharides, which were distributed more heavily in the osteodentin matrix than in the dentin matrix. On the basis of these data, it was concluded that the unique chemical and structural characteristics of the osteodentin result primarily from the incorporation of apatite clusters of intracellular origin and associated acid mucopolysaccharides.

Notes: Summary The comparative ultrastructure of fish bone osteogenesis and resorption induced by scale removal was described in the osteocytic (cellular-boned)Carassius auratus and the anosteocytic (acellular-boned)Tilapia macrocephala. Osteocytes, present in osteocytic bone, were lacking in anosteocytic bone. In osteocytic bone the osteoblast secreted a collagenous preosseous matrix in which it became enmeshed and then was termed a preosteocyte. When the preosseous matrix mineralized, the preosteocyte was termed an osteocyte and was completely surrounded by bone. In anosteocytic bone the osteoblasts receded from the mineralizing front and never became trapped as osteocytes. During resorption, types A and B resorptive cells, present in both bone types, invaded the matrix and demineralized the osseous zone. These cells were characterized by large amounts of granular endoplasmic reticulum and intracellular inclusions containing crystal-like material. Although functionally similar to mammalian osteoclasts, these cells lacked a characteristic ruffled border and were not multinucleated. The osteocytes of cellular bone did not appear to be involved during demineralization.

Notes: Abstract A new method is described by which osteoclasts can be isolated for biochemical and electron microscopic analyses. As a source of cells for isolation by microdissection, imprints of metaphyseal bone were used. The use of imprints provides important advantages over bone sections, including a higher degree of cytologic and enzymatic preservation, and the delivery of whole cells which are more readily manipulated and which yield data that are more readily reproduced. By planimetric analysis of the histochemically-stained isolated cell samples, it was shown that osteoclasts represent over 90% of the sample mass. The levels of several of the pyridine nucleotide-linked enzymes involved in citric acid metabolism, as well as acid phosphatase, were determined in osteoclast samples weighing 0.2 to 2.0 μg isolated from normal and parathyroid-treated rats. Aconitase activity measured in the direction of citrate to isocitrate was 0.5–0.8 M/Kd/H, the lowest of the activities studied. The activities of GDH and NADP-ICDH were 5 to 10 times higher than that of aconitase but only a tenth to a third that of acid phosphatase, lactic or malic dehydrogenase.

Notes: Abstract A small patch of cells in the epithelium of the anterior surface of the collar of the serpulidPomatoceros caeruleus contains membrane-bound vacuoles filled with crystalline material. The crystals have rhomboidal or rectangular profiles and have been shown by electron diffraction analysis to be composed of hydroxyapatite and calcium magnesium phosphate. The apices of the cells are bordered by microvilli. Some cells also have apical cilia. The cells contain Golgi complexes in the apical cytoplasm. Large numbers of mitochondria are distributed thoughout the cytoplasm. The possible function of these mineral-containing cells as sites for storage of calcium and/or phosphorus is discussed.

Notes: Summary The nucleotide sequence ofDrosophila melanogaster 5S RNA has been determined and appears to be homogeneous both in the KC cell line and in the insect at different developmental stages. Experimental evidence on the conformation of this molecule is in agreement with a general class of 5S RNA models.

Notes: Abstract The ultrastructural organization of the spores of the sporocarp of Endogone flammicorona was studied. Two types of organization are described. Initially the spore possessed a vacuolate protoplasm and was bound by two cell wall layers. The spore was surrounded by a hyphal mantle formed of a sheet of vacuolized hyphae with uniformly thin walls. Secondly, although the ultrastructural features of the spore appeared the same, it was now surrounded by a hyphal mantle with unevenly thickened walls (i. e., the so-called flaming crown) due to the gradual and irregular deposition of granules and lamellae. This crown gives the spore its most commonly observed morphological feature and is the preminent character employed taxonomically to speciate Endogone flammicorona Trappe & Gerdemann.

Notes: Summary Injection of α-ecdysone into the larval haemolymph of late third instar larvae ofD. virilis induces both the extrusion of secretory proteins and the inactivation of the enzyme glutamine-fructose-6-phosphate-aminotransferase (E.C. 2.6.16) in the salivary glands. In the presence of actinomycin D or cycloheximide the hormone is ineffective. If before adding these inhibitors RNA synthesis is allowed to proceed for 1.5h, or protein synthesis for 2h after ecdysone injection, however, the protein extrusion and the enzyme inactivation do occur. It is proposed that ecdysone controls these two cytoplasmic events at the transcriptional level by the activation of specific Correlations with puff activities are discussed.

Notes: Summary 1. The developmental potentials of dissociated cells of the different regions of the male foreleg disc ofDrosophila melanogaster were analysed. To this end, various amounts of foreleg disc material were dissociated together with an excess of heavily irradiated wing discs (“feeding layer”), and the reaggregates were cultured for 10 days in the abdomens of adult hosts prior to metamorphosis. 2. The foreleg disc cells were in most cases unable to regenerate missing structures in a circular direction within the leg segments. Instead they strongly tended to adopt the specifications of more distal leg segments (distal transformation), irrespective of the region of origin of the ancestor cells within the disc. 3. The distal transformation occurred mainly, if not exclusively, during an early phase (“initial phase”) in the reaggregates. 4. The extent of distal transformation was most pronounced in those series in which the foreleg cells were initially least diluted by the “feeding layer” cells. 5. Cells of the lower lateral quadrant were very poor both in proliferative activity and in the extent of distal transformation, compared to cells of the three remaining quadrants. In the experiments with a low initial dilution of the foreleg cells, cells of the lower medial quadrant underwent distal transformation much more distinctly than cells of the upper medial and the upper lateral quadrants. 6. Allotypic structures occurred exclusively in reaggregates of the upper medial and upper lateral quadrants. In these implants, however, the frequency of transdetermination was extremely high. 7. Two alternative mechanisms are discussed which could have led to the general occurrence of distal transformation. They differ in the basic assumption of whether or not the “feeding layer” cells were able to interact with the leg cells to influence their regulative behaviour. In addition, interactions among the leg cells themselves seemed to stimulate proliferation to varying degrees and may account for the observed differences in the degree of distal transformation.

Notes: Summary Three-hundred and twenty fertile,pal-induced Y-chromosome mosaic males and females were obtained. Fractional analysis of the sons of 55 somatically mosaic flies that were also germinally mosaic tentatively suggests that the number of functional primordial germ cells inDrosophila melanogaster is variable and that it is seldom greater than 24. From the observed 0.17 frequency of germinal mosaicism it was estimated that the average number of pole cells at the end of blastoderm formation is 45. At present, the germ cells afford the only opportunity to compare genetic estimates of the number of blastoderm or primordial cells with available histological counts. The good agreement between them suggests that both the fractional and the mosaic frequency methods for estimating primordial or blastoderm cell numbers of various larval and imaginal anatomical structures provide reasonably close approximations of the actual values.

Notes: Summary 1. Histological analyses were made of imaginal discs and histoblasts during the larval development ofDrosophila melanogaster to determine the number of cells, the patterns of cell division and the growth dynamics in these adult primordia. Histological studies were also made of the imaginal rings which are the primordia of the adult salivary gland, fore-and hindgut, the anlage cells of the midgut and several larval and embryonic tissues. 2. In the newly-hatched larva, the immature eye-antenna, wing, haltere, leg and genital discs contain about 70, 38, 20, 36–45 and 64 cells respectively. These numbers include cells destined to form cuticular elements as well as peripodial, tracheal and nerve cells and probably the progenitors of adepithelial cells. The number of cells counted in the various imaginal disc anlagen is 1.5 to 4 times higher than the numbers deduced from genetic mosaic analyses by other investigators and reasons for these differences are given. 3. About 12 h after fertilization, mitosis ceases in all tissues of the embryo except the nervous system. After the larva hatches, mitosis resumes in most of the imaginal anlagen and in some larval tissues. The time of resumption of mitosis in the imaginal anlagen was determined after treating the larvae with colchicine for 2 h. 4. Among the imaginal discs, the eye disc is the first to begin cell division, at about 13–15 h after the hatching of the larva (first instar) followed by the wing (15–17 h), the haltere (18–20 h), the antenna, leg, and genitalia (24–26 h, early second instar), and finally the labial and dorsal prothoracic discs (52–54 h, early third instar). The cell doubling time for various discs was calculated from cell counts and the times agree closely with the doubling times deduced from clonal analyses by other workers: e.g., 7.5 h for the cells of the wing disc. 5. The imaginal ring of the hindgut first shows cell division early in the second instar. The imaginal rings of the foregut and salivary glands, the anlage cells of the midgut and the cells of the segmental lateral tracheal branches begin to divide early in the third instar. 6. The histoblasts which are the anlagen of the integument of the adult abdomen do not increase in number from the time of larval hatching until about 5 h after pupation when they begin to divide. Their behaviour contrasts with that of the histoblasts of the other dipterans such asCalliphora, Musca andDacus, which begin to divide during the second instar. 7. The histoblasts are an integral part of the larval abdominal epidermis and, unlike imaginal disc cells, secrete cuticle during larval life. Each hemisegment consists of an anterior dorsal, a posterior dorsal, and a ventral histoblast nest containing about 13, 6 and 12 cells respectively. The 62 histoblasts in each larval segment represent about 7–8% of the total number of cells that form the integument of that segment. 8. The number of cells in a particular type of histoblast nest was constant for both male and female larvae and among the different abdominal segments, except that the anterior dorsal group of the first and the seventh segments contains fewer cells than those of the other segments. Although the male and female adultDrosophila lack the first abdominal sternite and the male lacks the seventh abdominal tergite and sternite, the ventral histoblast nests of the first and the dorsal and ventral nests of the seventh abdominal segments are present in the larval stages as well as in the prepupa and have the same morphology and cell number as similar nests in the rest of the abdominal segments. 9. The cells of the imaginal discs increase in volume about six-fold and their nuclei increase in volume three-fold between the time of hatching and the initiation of mitosis. The histoblasts increase in volume about 60-fold and their nuclei increase in volume about 25-fold between larval hatching and pupariation. 10. Prior to each cell division, the nuclei of the columnar cells of the disc epithelium and of the histoblasts appear to migrate toward the apical surface of the epithelium. The cells round up and shift toward the apical region where mitosis occurs. After cytokinesis, the daughter cells move back to deeper positions in the epithelium. Because the nuclei of the non-dividing cells continue to lie deep in the epithelium, this intermitotic migration of nuclei gives these epithelia a pseudostratified appearance. 11. Analyses of the growth of larval cells and of organs confirmed the observations of earlier investigators that cell division occurs only in a few larval tissues, whereas growth in the rest of the larval tissues is by cell enlargement and polyteny. During larval life, cell division was detected only in the central nervous system, gonads, prothoracic glands, lymph glands and haemocytes. Each tissue began mitosis at a characteristic stage in larval life. The larval cells that did not divide, grew enormously, e.g., epidermal cells increased in volume 150-fold and their nuclei increased in volume 80-fold. 12. The adepithelial cells, which give rise to some of the imaginal muscles, were first identified between the thick side of the imaginal dise epithelium and the basement membrane at the beginning of the third larval instar (50–52 h). The origin of these precursors of mesodermal structures was analysed and evidence is presented that the adepithelial cells come from the disc epithelium. The question of the origin of the mesoderm of cyclorrhaphan Diptera is reviewed and it is suggested that the imaginal disc ectoderm may become segregated from the rest of the embryo before gastrulation has occurred, that is before the mesoderm has been established.

Notes: Summary The present investigation analyzes intercellular junctions in tissues with different developmental capacities. The distribution of junctions was studied inDrosophila embryos, in imaginal disks, and in cultures of disk cells that were no longer able to differentiate any specific pattern of the adult epidermis. The first junctions —primitive desmosomes andclose membrane appositions — already appear in blastoderm.Gap junctions are first detected in early gastrulae and later become more and more frequent.Zonulae adhaerentes are formed around 6 h after fertilization, whileseptate junctions appear in the ectoderm of 10-h-old embryos. Inwing disks of all stages studied (22–120 h), three types of junctions are found: zonulae adhaereentes, gap junctions, and septate junctions. Gap junctions, which are rare and small at 22 h, increase in number and size during larval development. The other types of junctions are found between all cells of a wing disk throughout development. All types of junctions that are found in normal wing disks are also present in theimaginal disk tissues cultured in vivo for some 15 years and in thevesicles of imaginal disk cells grown in embryonic primary cultures in vitro. However, gap junctions are smaller and in the vesicles less frequent than in wing disks of mature larvae. Thus gap junctions, which allow small molecules to pass between the cells they connect, are present in the early embryo, when the first developmental decisions take place, and in all imaginal disk tissues studied, irrespective of whether or not these are capable of forming normal patterns.

Notes: Summary The lineages of cells on the second-leg basitarsus ofDrosophila melanogaster were analyzed by examining gynandromorphs andMinute mosaics. Bracts lie proximal to bristles on the adult basitarsus, yet bract precursor cells were found to originate lateral to bristle precursor cells. In 6 of the 8 longitudinal rows of bristles on this segment, the bract cells arise ventral to the bristle cells; in the others they arise dorsally. The lateral cell origins are interpreted as reflecting a pattern of lateral cell movements associated with evagination of the leg disc. An unusual discrepancy was observed in the relative frequencies of male vs. female bracts and bristles in gynandromorphs. The discrepancy suggests that there is a cell-autonomous sexual difference in either the time at which cells begin moving during evagination or the speed with which they move. On the basis of the results, it is reasoned that the bristle pattern of the basitarsus does not originate in its final form. Prior to evagination, the bristle cells of each row are apparently closer together than in the final pattern, and the rows are farther apart. Evidence is presented which suggests that the bristle cells of each row may originally be arranged in a jagged line which is later straightened by cell movements. The two locations where the anterior/posterior compartment boundary of the second leg passes through the basitarsus were found to vary relative to the bristle pattern. If this boundary is assumed to be a fixed line of positional values, then the extent of the observed variability — which is estimated to be ± 1 or 2 cell diameters — provides a measure of the precision of patterning around the circumference.

Notes: Summary The differentiation of muscles in primary cultures of cells fromDrosophila melanogaster embryos was investigated. In early cultures, and in the absence of exogenous ecdysone, two main classes of muscle were found. Comparison, by light and electron microscopy, of one of these classes (the “myotube” class) with muscles from third instar larvae shows that this class corresponds to the muscles of the body wall of the larva. When α- or β-ecdysone is added to the cultures, these undergo a number of metamorphic changes. Most of the larval muscles disappear, and two new types of muscle form. Ultrastructural and light microscopic examination of these two types indicates that they correspond to the two classes of skeletal muscle (fibrillar and tubular) found in adult flies.

Notes: Summary Cartilaginous cells of aged newts (Triturus cristatus) were studied during hind limb regeneration. The electron microscope was used to study the structure and distribution of chromatin in the cell nuclei, while the DNA content of the chromatin was measured by means of a scanning cytophotometer. Changes in the ultrastructure of the cytoplasm during regeneration were also studied. It was observed that the structure and distribution of chromatin in the activated cell is greatly modified. In the non-activated cell of the aged newt, the chromatin is found highly condensed and distributed peripherally close to the nuclear membrane. In contrast, in the activated cells, the chromatin is much less condensed and is distributed throughout the nucleus. Moreover, cytoplasmic vacuoles, found only in the non-activated aged cells, disappear and an increase in the mitochondria and rough endoplasmic reticulum is also observed. Changes in the nuclear structure are observed prior to the cytoplasmic modifications. It is interesting to note that the process of activation induces structural changes in the aged cells which make these cells appear to be structurally identical to the young cells. This process of rejuvenation takes 3–5 days in the newt. We suggest that these structural changes of the chromatin and cytoplasm in the aged cells are necessary to increase the metabolic activity which precedes cell division. It may also explain why regeneration takes a longer time in the aged animals than in the young ones.

Notes: Summary The pathway of adult sensory nerves has been analysed in three experimental situations: (i) in flies with grossly abnormal thoracic morphology resulting from X-irradiation early during development, (ii) in flies which had been subjected to surgical operations late in the larval period, (iii) in homoeotic mutants. The results provide experimental support for a simple mechanism in which developing adult axons join the nearest larval nerve and are guided by it up to the central nervous system. In particular, experimental interference with normal development can result in nerves from different segments, or from dorsal and ventral appendages, joining each other and entering the central nervous system together.

Notes: Summary The mature labial disc, when implanted into a larva of the same age, undergoes metamorphosis along with the host and produces one lateral half of the medi- and distiproboscis. On the basis of results obtained from transplanted disc halves (including the separate peripodial membrane) a tentative fate map of the labial disc was constructed, which shows most of the presumptive mediproboscis to be located in the dorsal, and most of the presumptive distiproboscis in the ventral part of the disc. The distal protion of the peripodial membrane also contains imaginal anlagen, viz. part of the mediproboscis, prementum, and labellar cap anlagen. The involvement of this part of the peripodial membrane was checked by a careful histological analysis of labial disc development during the first ten hours after prepupation. The results were compared with the situation described forCalliphora imaginal discs. In addition, a detailed morphological analysis was made of the proboscis of the homoeotic mutantproboscipedia (pb). At 27°C,pb changes the distiproboscis into a “telopodite” (leg segments distal to the coxa); the (unchanged) prementum may therefore correspond to the coxa. At 15° C, the tarsus of this homoeotic “telopodite” is replaced to a greater or lesser extent by an arista. The present analysis thus confirms (a) the fundamental morphological correspondence of the medi- and distiproboscis with the labium of other insects, and (b) the fundamental developmental correspondence of the labial, antennal, and leg discs.

Notes: Summary A temperature-sensitive period during early embryogenesis for three stocks carrying thetuh-3 gene suggests that it is a homoeotic mutation involved in the initial determination of the eye-antennal disc rather in maintenance of the determination.

Notes: Summary The development of the adult abdomen ofDrosophila melanogaster was analyzed by histology, microcautery, and genetic strategies. Eight nests of diploid histoblasts were identified in the newly hatched larva among the polytene epidermal cells of each abdominal segment: pairs of anterior dorsal, posterior dorsal, and ventral histoblast nests and a pair of spiracular anlagen. The histoblasts do not divide during larval life but begin dividing rapidly 3 h after pupariation, doubling every 3.6 h. Initially they remain confined to their original area, but 15 h after pupariation the nests enlarge, and histoblasts replace adjacent epidermis cell by cell. The histoblasts cover half the abdomen by 28 h after pupariation and the rest by 36 h. Polytene epidermal cells of the intersegmental margin are replaced last. Cautery of the anterior dorsal nest caused deletion of the whole corresponding hemitergite, whereas cautery of the posterior dorsal nest caused the deletion of the macrochaetae of the posterior of the hemitergite. Cautery of the ventral nest deleted the hemisternite and the pleura, whereas cautery of the spiracular anlagen deleted the spiracle. Results of cautery also revealed that no macrochaetae formed on the tergite in the absence of adjacent microchaetae. Clonal analysis revealed that there were no clonal restrictions within a hemitergite at pupariation. Cautery of polytene epidermal cells other than those of the intersegmental margin failed to affect tergite development. However, cautery of polytene epidermal cells of the intersegmental margin adjacent to either dorsal histoblast nest caused mirror-image duplications of the anterior or posterior of the hemitergite in 10% of the hemitergites. Forty percent of the damaged presumptive hemitergites formed complete hemitergites, indicating extensive pattern regulation and regeneration. Pattern duplication and regeneration were accounted for in terms of intercalation and a model of epimorphic pattern regulation (French et al., 1976). Histoblasts in adjacent segments normally develop independently, but if they are enabled to interact by deleting the polytene epidermal cells of the intersegmental margin, they undergo intercalation which results in duplication or regeneration. The possible role of the intersegmental margin cells of insects in development was analyzed.

Notes: Summary The development of the rhabdomeric pattern in the compound eye ofDrosophila has been studied using combined transplantation and electron microscope techniques. In a first series of experiments eye imaginal discs of increasing age were implanted into larvae ready to pupate, thus losing variable amounts of the normal time for development. A sequence of differentiative abilities was found in the metamorphosed test pieces. As far as the photoreceptor cells are concerned, the most prominent steps of this sequence are: ability to form groups with other similar elements, anatomical polarization of microvilli, establishment of the rhabdomeric pattern and formation of an equator line. The stability of determination of the equator line was tested in a second experimental series. Fragment of different topographical origin within the mature eye anlage were brought to metamorphosis by implantation into larvae ready to pupate. It was found that an equator line differentiates only in those pieces which according to the published anlage maps contain the prospective equator region prior to metamorphosis. The mitotic abilities of implanted eye imaginal discs were investigated by means of “in vitro”3H-thymidine pulse-labelling and light microscope autoradiography of the differentiated test pieces. During the third larval stage the eye anlage is traversed by two consecutive mitotic waves, each one of them producing different categories of receptor cells. The first, anterior wave predominantly produces cells oriented toward the poles of the eye within the ommatidia, while the second, posterior wave gives rise to elements exclusively in an equatorial position. The dynamics of this proliferation are discussed in relation to the findings in the implantation experiments. Silver-grain counts support the possibility that at least two successive cell divisions occur in the eye anlage between labeling with tritiated thymidine and beginning of morphological differentiation. The relevance of this finding for the understanding of the concept of acquisition of competence is discussed.

Notes: Summary 1. Eggs of the midgeSmittia were investigated by light microscopy and transmission electron microscopy. This paper describes elements and architecture of periplasm and yolk endoplasm before the formation of pole cells. 2. The periplasm is coated externally by the oolemma and a multilayered egg shell. The periplasm consists of a cytoplasmic matrix rich in ribosomes; it contains mitochondria and ER cisternae, some annulate lamellae and an occasional Golgi complex. Microtubuli were demonstrated only rarely. Accumulations of a dense granulated substance resembling in its structure the oosome material were frequently observed. 3. The yolk endoplasm is a cytoplasmic network embodying proteid yolk particles, lipid droplets and accumulations of glycogen. The endoplasm is continuous with the periplasm and shows the same cell constituents. It may form between 3 and 7 cytoplasmic islands free of yolk particles. Rosette-shaped membranous structures in the yolk endoplasm are interpreted as nuclear envelope organizing centres. 4. Three carefully analysed eggs contained 2 nuclei each. both nuclei were situated in the posterior egg half. 5. Periplasm and yolk endoplasm are characterized by random distribution of cell elements. No zonation or special accumulations could be recognized. 6. The spatial distribution of the egg components studied did not indicate that any of these components could function as a determinant in embryonic pattern formation.

Notes: Summary 1. The egg ofPollicipes polymerus, the common intertidal gooseneck barnacle, has been studied by electron microscopy. Constriction rings, similar to the contractile rings of cleaving cells and polar lobes, move unidirectionally from the animal to the vegetal pole of newly fertilized eggs. This is referred to as peristaltic constriction. The present paper describes the fine structure of the egg during first polar body formation and peristalsis. 2. During formation of the polar body, dense bodies are produced by the Golgi and extracellular plaques are observed. Thin microfilaments (40–60 Å) are in the egg adjacent to the polar body. 3. In eggs undergoing peristalsis, the appearance of extracellular spheres, flocculent material and filaments is observed. Intracellularly large numbers of multivesiculate bodies, glycogen granules, mitochondria and protein-carbohydrate and lipid yolk bodies are seen at the level of constriction. 4. Thin microfilaments are found in the cortical area of newly-fertilized eggs exclusively in peristaltic constriction rings. Filaments are oriented primarily in a meshwork, although circumferentially-oriented filaments are also found in rings near the vegetal pole. Microvilli extend into the space created between a constriction and the elevated egg membrane. 5. A model is proposed to explain the peristalsis in this species. It is suggested that information from a pacemaker region activates peristalsis by affecting filament polymerization and orientation. One function of peristalsis may be elongation of the egg from a sphere to an ovoid, although other possibilities such as elevation of the egg membrane, segregation of the lipid yolk to the vegetal pole and predetermination of the first cleavage plane are also discussed.

Notes: Summary The derivatives of 110 mosaic genital discs of gynandromorphs have been analysed microscopically. It has been found that theanalia of both sexes are homologous and derive from a single primordium (see Fig. 1a). Whether male or female anal plates are formed depends on the genetic constitution of the cells. This is analogous to the development of male sex combs versus female transversal rows on the forelegs of gynandromorphs. In contrast, the data for thegenitalia (see Fig. 1 b) are best explained if it is assumed that there are two genital primordia in everyDrosophila embryo: a male primordium that will only develop into genitalia if populated by XY (or XO) nuclei, and a female primordium that will only do so if populated by XX nuclei. This model, as depicted in Figure 2, is compatible with all our gynandromorph data and also with observations onMusca andCalliphora where in fact two separate genital primordia are found.

Notes: Summary Five regions of the compound eye have been found to be preferential boundaries for clones of labelledMinute + cells, and to act restrictively on the growth of cell clones after a given developmental stage. One of these regions is topographically related to the line of pattern inversion existing at the level of the equator. The results of experiments showing independency of origin of restriction lines and line of pattern inversion are reported.

Notes: Summary Collagen fibrils with a main period banding of 610 Å and 220 Å in width were observed in the blastocoel of 72-h embryos of the sea urchin,Strongylocentrotus purpuratus. Non-striated fibrils of 50 Å diameter were also observed. The collagen is seen in highest concentration in the vicinity of mesenchyme cells which are richly endowed with endoplasmic reticulum and secretory vesicles. A role for collagen in cell attachment, orientation and spicule formation is discussed.

Notes: Summary sev LY3,the only existing allele at thesev locus (1–33,2±0,2), behaves as strongly hypomorph or even as amorph. Ommatidia in asev compound eye have only seven receptor cells, the position of the R7 pattern element being vacant. Various criteria showing that the missing cell is R7 have been verified. These include (i) anatomical characteristics ofsev ommatidia; (ii) behaviour of central R cells insev rdgB double mutants; (iii) medullary projection of central R cell axons; and (iv) mitotic pattern ofsev imaginal discs. The analysis of morphogeneticsev-sev + mosaics has shown thatsev is expressed autonomously by R7 cells, indicating that thesev phenotype is not due to asev genotype of ommatidial pattern elements other than R7. The study of third instarsev imaginal discs has not brought any direct evidence for death of clustered presumptive R7 cells; however, clonal analysis of the developingsev compound eye has given evidence of developmental parameters comparable to those ofsev +, therefore favouring the hypothesis that R7 cells die insev mutants. On the other hand,sev + seems to be required for the determination of the R7 cells, since thesev phenotype cannot be uncovered during the last mitoses of heterozygous mutant cells.

Notes: Summary These experiments examined whether inDrosophila immature imaginal disc tissue and tissues from embryonic stages can influence pattern regulation in a disc fragment in the same way as can mature imaginal discs. Immature imaginal discs, or the cells of whole embryos, were mixed with a test fragment (presumptive notum) from a mature wing disc. The immature tissues in each mixture were genetically marked and had been heavily irradiated (25 Kr gamma) prior to mixing to prevent growth and maturation during subsequent culture in vivo. Alteration of the regulative behavior of the test fragment (that is, regeneration of wing) thus provided an assay for the communication of positional information by the immature tissues. The results suggest that this capacity arises well before competence to metamorphose, as early as the 16th hour of embryonic development, whereas prior to 16 h, essentially no stimulation of regeneration occurred. It is suggested that the imaginal disc (or presumptive disc) cells of the embryo may have been responsible for this early stimulatory capacity.

Notes: Summary The ontogeny of allozyme patterns has been studied in embryos ofDrosophilamelanogaster, which are doubly heterozygous for alleles specifying the slow and fast forms of alcohol dehydrogenase (ADH) and α-glycerophosphate dehydrogenase (GPDH). The ontogeny of esterase-2 was studied in embryos and young larvae of the flour mothEphestia kühniella, which are heterozygous for two of the three existing esterase-2 alleles. In freshly laidDrosophila eggs only the maternal enzyme forms are present and during the first 15 hours of development the staining of these forms becomes progressively fainter. After 16 and 17 h, the paternal and hybrid bands of ADH and GPDH respectively become obvious. Before hatching, the intensity distribution in the three-banded pattern of reciprocal hybrids is asymmetric in favour of the persisting maternal enzyme form. InEphestia embryos, however, there is no persistence of the maternal esterases. In all reciprocal heterozygotes a three-banded pattern suddenly appears 96 h after egg deposition, indicating synchronous activation of both parental alleles. The relative intensity distribution in the hybrid patterns approaches that of the mature larvae stepwise and in an allele-specific manner. This result and the fact that the various heterozygous types exhibit unequal total activities suggest that the Esterase-2 alleles have different activities, which are fixed late in embryogenesis.

Notes: Summary The females produced in the crossesD. melanogaster×D. simulans andD. melanogaster×D. mauritiana are sterile and have reduced ovaries. Normal and fertile ovaries were produced when genetically marked pole cells ofD. melanogaster were transplanted into eggs which gave rise to the hybrid females. These results eliminate the possibility that the sterility of these hybrids is due to the somatic component, i.e. the follicular cells of the ovaries, or to other physiological causes. The results also suggest that the control of gonadal morphogenesis is dependent mainly on the germ line.

Notes: Summary Gynandromorphs with female XX-and male XO-areas result from the loss of an unstable ring-X-chromosome in the early cleavage mitoses of ring/rod-X-chromosome heterozygotes. The phenotypes of the recessive alleles on the rod-X-chromosome are expressed in the XO-areas. 377 larval gynandromorphs of the genotypeR(1)2, In(1)w vC /y w sn3Iz50e mal were examined and scored for the phenotypes of 13 paired and 10 unpaired structures (Table 2, Fig. 2). This was possible mainly by the cell-autonomous expression of aldehyde oxidase activity in soft tissues and by the comparison of the distribution of enzyme activity in wildtype and gynander larvae. The distances between pairs of structures were calculated in sturt-units (Tables 3 and 4). A morphogenetic fate map with the presumptive areas of larval structures was constructed (Fig. 3). The relative positions of the structures agree well with Poulson's fate map (Fig. 4). In addition, the distribution of phenotypes was scored in 380 adult gynandromorphs Table (5). The fate map (Fig. 5) which was constructed from these data is very similar to the fate map of larval structures. This similarity becomes even more pronounced if fate maps are constructed which contain only structures analogous in larva and imago (Table 6, Fig. 6). Therefore an attempt was made to set up an integrated morphogenetic fate map containing the presumptive areas of both larval and imaginal structures (Fig. 7). The possibilities of further blastoderm mapping are discussed.

Notes: Summary Females homozygous for a newly isolated mutation induced by ethyl methane sulphonate,fs(1)K10, lay abnormally shaped eggs in which the dorsal appendages of the chorion are enlarged and fused ventrally. The eggs are usually not fertilized and development is never normal beyond the blastoderm stage. The mutant was mapped to the tip of the X-chromosome with a meiotic position of 1–0.5 and a cytological location between 2B17 and 3A3. Using germ line mosaics constructed by transplantation of pole cells, it was shown that the abnormal morphology and the sterility are obtained only when the germ line is homozygous for the mutant.

Notes: Summary TheDrosophila chorion is produced normally in isolated follicles in Robb's chemically defined culture medium. The complex architecture of the shell developed in vitro from follicles as young as early stage 10 is completely normal morphologically. In addition, the time required for in vitro development closely approximates that observed for in vivo development. Comparisons of insect culture media developed by Robb, Grace, Schneider, and Echalier show large variations in their ability to supportDrosophila chorion development.

Notes: Summary The bristle pattern of the second-leg basitarsus inDrosophila melanogaster was studied as a function of the number and size of the cells on this segment in well-fed and starved wild-type flies, in triploid flies, and in two mutants (dachs andfour-jointed) that have abnormally short basitarsi. The second-leg basitarsi of well-fed, wild-type flies from 22 otherDrosophila species were studied in a similar manner. There are typically 8 longitudinal rows of evenly-spaced bristles on the second-leg basitarsus, and in each row the number of bristles was consistently found to vary in proportion to the estimated number of cells along the segment, and the interval between bristles was found to vary in proportion to the average cell diameter on the segment. These correlations are interpreted to mean that the spacing of the bristles within each row is controlled developmentally, whereas the number of bristles is not. The interval between bristles is evidently measured either as a fixed number of cells or as a distance which indirectly depends upon cell diameter.

Notes: Summary Electron microscopic analysis of primary cultures derived from larvalXenopus liver has shown that these cells, although they form only two-dimensional aggregates, retain and presumably also develop structural characteristics typical of liver parenchyma cells, such as bile canaliculi with microvilli and epithelial junctional complexes. As judged from structural criteria, primary cultures contain 80–90% hepatocytes. In contrast to the intact tissue, primary cultures showed excessive development of microfilaments, however. Incorporation of labeled amino acids has revealed further that the capacity for protein synthesis is maintained in culture and that synthesis of liverspecific protein albumin is maintained in vitro, even in liver cultures derived from thyrostatic tadpoles. This latter result suggests that initiation of albumin synthesis in the larval liver is probably not dependent upon thyroid hormones but rather reflects the protodifferentiated state of this tissue.

Notes: Summary Salivary gland cells of members of theDrosophila melanogaster group (from four different subgroups) were examined electron microscopically and histochemically during the late larval period of development. The secretory product, which is supposed to be utilized as ‘glue’ at the time of puparium formation, appears, by analogy to Palade and Jamieson's results, to be synthesized partially in the rough endoplasmic reticulum (RER) and partially in the Golgi complex. The latter is also the usual site of the packaging of the product into secretory granules, except in the case of one of the secretory granule components ofD. lucipennis. The phylogenetic relationships among the subgroups, implied by the morphological appearance of the secretory granules, fit well with the existing phylogenetic relationships within the group. The secretory granules of each species have their own morphological features; granules of species of the same subgroup share some of these features. Secretion occurs from the cells via exocytosis during which the morphology of the secretory granules changes. Light microscope examination of PAS (Periodic Acid-Schiff reaction) stained glands shows a strong positive reaction in most species, with the exception of the species of thesuzukii subgroup which show a weak, or a negative reaction (D. rajasekari). Electron histochemical localization of polysaccharides in the secretory granules was possible inD. melanogaster and the species of theananassae subgroup.

Notes: Summary The generalogical relationships of photoreceptor cells within the compound eye ofDrosophila have been studied using cell labelling, with either3H-thymidine or recessive mutations, during the third larval stage. It has been found that photoreceptor and secondary pigment cells arise from different precursor cells. Under the present experimental conditions, precursors of receptor cells give rise to about 8 elements which differentiate as R cells of two different groups. One of the cells differentiates as R7 and the remaining as any one of the R1 to R6. The last cells behave initially as equivalent, and can differentiate within the same or within different, but neighbouring, ommatidia. The class of R1 to R6 cell in which each one of these elements differentiates, seems to depend on the time of its origin. The implications of these findings for the formation of the ommatidial pattern are discussed.

Notes: Summary We report on the size distribution of clones marked by mitotic recombination induced by several different doses of X-rays applied to 72 h oldDrosophila larvae. The results indicate that the radiation significantly reduces the number of cells which undergo normal proliferation in the imaginal wing disc. We estimate that 1000 r reduces by 40–60% the number of cells capable of making a normal contribution to the development of the adult wing. Part of this reduction is due to severe curtailment in the proliferative ability of cells which nevertheless remain capable of adult differentiation; this effect is possibly due to radiation-induced aneuploidy. Cytological evidence suggests that immediate cell death also occurs as a result of radiation doses as low as 100 r. The surviving cells are stimulated to undergo additional proliferation in response to the X-ray damage so that the result is the differentiation of a normal wing.

Notes: Summary Immature ovaries ofDrosophila mercatorum were injected into young larvae and into adult males ofD. mercatorum, D. melanogaster, D. hydei, D. virilis, andZaprionius vittiger. These homo- and heteroplastic transplantations allow normal vitellogenesis to occur in the donor ovary. By SDS gel electrophoresis, we identified the major species-specific yolk proteins of mature eggs (stage 14) which were exclusively of donor-specific origin. Other experiments withD. hydei andZ. vittiger showed that, when females were used as hosts, the host-specific yolk proteins became incorporated into the donor eggs. When two immature ovaries, one ofD. mercatorum and one ofD. hydei, were co-cultured in males, again only the donor-specific yolk proteins were found in the mature eggs implying that these yolk proteins were not released into the host hemolymph. A parthenogenetic strain ofD. mercatorum was used to demonstrate the ability of transplanted immature ovaries to produce viable eggs which can give rise to fertile adults. The role of the species-specific yolk proteins is discussed with respect to the dual origin of these proteins during normal vitellogenesis, i.e., an autonomous synthesis within the ovary itself in addition to the well-known production by the fat body. Further experiments with pupae as hosts indicate that even in the absence of juvenile hormone and in the presence of high doses of ecdysone, vitellogenesis can proceed within the donor ovary. Based on these experiments, a new hyopthesis on the hormonal control of vitellogenesis inDrosophila is presented. We propose that yolk proteins derived from the fat body are controlled by juvenile hormone, whereas the independent and autonomous vitellogenesis within the ovary itself is controlled by endogenously synthesized ecdysone.

Notes: Summary We estimate the number of blastoderm cells which generate the thoracic imaginal discs ofDrosophila. At hatching the wing disc is twice the size of the haltere disc, but the results suggest that both discs develop from a similar number of blastoderm cells. Two homeotic mutations, which transform the haltere into wing, affect embryonic growth but not the primordial number. All the segmental primordia may be of similar size and each may be similarly subdivided into a larger anterior, and a smaller posterior polyclone.

Notes: Summary The embryonic organization of the sexually dimorphic genital disc was studied in genetic mosaics resulting (a) from early loss of a chromosome or (b) from mitotic recombination. (a) Early Loss of a Chromosome. Three types of mosaics were produced — purely female mosaics, purely male mosaics, and gynandromorphs. They show that the genital disc arises from a group of cells in the ventral region of the embryo somewhat larger than that giving rise to a single foreleg (Table 2). Within this group of cells three regions can be distinguished that are present in both sexes: an anterior, a medial, and a posterior one, with distances of only 3–4 sturts between adjacent regions. The anterior region gives rise to the female genitalia, the medial region to the male genitalia, and the posterior region forms the analia of both sexes and the parovaria of the female (Figs. 2 and 3). The relative positions of the three regions were deduced from sturt distances (Tables 1 and 5), and from frequencies of mosaicism (Table 2). (b) Mitotic recombination was induced at the blastoderm stage in order to produce twin spots in the external genitalia and analia of purely male and female flies. Clone sizes indicate that these structures arise from a small number of precursor cells (Table 4). Clones overlapped right and left sides, but no clones were found extending over analia and genitalia. However, within either the analia or the genitalia of each sex, no clonal restrictions could be observed, and the clones comprised structures that were up to 12 sturts apart. A comparison of clone sizes and sturt distances in the foreleg and in the genital disc indicates that equal gynandromorph distances involve equal numbers of cells in different regions on the ellipsoid egg (Fig. 5). The results obtained from all mosaics provide a consistent picture of the embryonic organization of the genital disc. This becomes apparent in the summarized fate maps (Fig. 4), where the map derived from normal gynandromorphs can be produced by a simple superposition of the male and the female maps. The data are also discussed with respect to mechanisms of sexual differentiation in the genital disc.

Notes: Summary Mutations of the bithorax complex result in segmental transformations in the thorax and abdomen ofDrosophila. The haltere discs from larvae homozygous forbx 3 orpbx are transformed so that the discs contain cells that will produce wing cuticle as well as cells that produce haltere cuticle. The pattern regulation behavior of these discs has been examined. The fate maps of the two discs were established, and then the regulative behavior of a number of fragments from both types of mutant discs was established by culturing the fragments in vivo prior to metamorphosis. The most important conclusion from this work is that the cells producing, haltere cuticle and wing cuticle within the same disc share the same positional information and that they communicate during pattern regulation.

Notes: Summary The ultrastructural changes in the wing bud afterapical ectodermal ridge (A.E.R.) removal was studied to re-examine the issue of distal mesenchymal cell death. The A.E.R. of the right wing bud was removed microsurgically from chick embryos of stages 18 to 22 (HH 1951). The wing buds were examined at three hour intervals up to twelve hours after the operation with light, transmission and scanning electron microscopy. The main findings were: (1) Immediate and temporary shrinkage of the mesenchymal extracellular space 100 to 150 μm and chromatin condensation in the cells 50 to 75 μm from the wound. (2) Death of ectodermal and mesenchymal cells in the immediate vicinity of the wound. (3) Formation of a single squamous-like layer of mesenchymal cells to cover the wound. (4) Occasional evidence of cell death in the distal mesenchyme at later times after the operation. The pattern of cell death observed suggests only a traumatic etiology, and gives little evidence for the postulated developmental significance of cell death following A.E.R. removal.

Notes: Summary Pairs of eye-antennal discs, attached to the cephalic ganglia, were cultured in vitro with a concentration of β-ecdysone optimal for imaginal differentiation. The eye-antennal discs fused to form a vesicle inside which the antennae were partially everted, and on the inner surface of which imaginal structures differentiated. The epithelium of the discs was continuous, and an integrated pattern of bristles and hairs differentiated in vitro. In particular, the median ocellus, a unified structure derived partially from each disc, differentiated normally.

Notes: Summary l(1)su(f)mad-ts (mad) is a new temperature-sensitive (ts) lethal mutant ofDrosophila melanogaster which produces duplicated legs after temperature pulse treatment during larval development. The ts-lethality was studied in temperature experiments and genetic mosaics. Temperature pulses given during two distinct TSPs of larval development result in two different types of leg pattern duplication. “Total” differ from “partial” duplications with respect to the affected leg compartments and the orientation of the planes of symmetry which are perpendicular to the dorso-ventral and the proximo-distal leg axes in total and partial duplications, respectively. Genetic mosaic studies indicate (i) disc autonomy of leg pattern duplication, (ii) clonal separation of the anlagen of the two pattern copies, and (iii) clonal restriction along the antero-posterior compartment border in the two pattern copies of totally duplicated legs. The results suggest thatmad leg pattern duplication is caused by a change in positional information rather than by cell death and subsequent regeneration. Our data are compatible with the assumption that during normal development the leg disc cells acquire information about their position within the disc with respect to the different leg axes independently and at different times.

Notes: Summary Two possible mechanisms are considered for the occurrence of experimentally or genetically induced duplications of bristles: extra cell division of a bristle mother cell versus determination of more than one mother cell. From a clonal analysis it appears that duplications induced by actinomycin-D arise by the latter mechanism, whereas those found in the mutantspl seem to arise by the former mechanism.

Notes: Abstract Removal of the soluble proteoglycans from slices of bovine costal cartilage by extraction in 4 M guanidinium hydrochloride permitted the visualization of abundant amounts of dispersed and disaggregated collagen in the matrix. Proteoglycans which are resistant to extraction are seen as small granules which are concentrated in the perilacunar regions. Large proteoglycan granules appear to originate in the chondrocyte. As they come to occupy positions in the matrix distant from the chondrocyte, the granules become smaller. A non-granular, “amorphous” component masks the collagen fibers so that they cannot be readily seen in the intact cartilage.

Notes: Summary The mantle edge of the freshwater pulmonate snailsLymnaea stagnalis andBiomphalaria pfeifferi was investigated with histochemical and ultrastructural methods. The mantle edge gland, which is involved in shell formation, consists of the periostracal groove and the belt. This belt appears to be composed of various regions. In the area of the periostracal groove a number of subepithelial gland cell types occur; these release their products into the groove. Between the groove cells ciliated free nerve endings terminate; the corresponding perikarya occur in the subepidermal connective tissue. Also in the posterior belt region free nerve endings were observed between the epithelial cells; in addition, a particular type of subepithelial gland cell was found in this area. The epithelial cells of this part of the belt have the ultrastructural characteristics of ion and water transporting cells; they are probably involved in calcium deposition and resorption. The possible role of the free nerve endings and of the subepithelial gland cells is discussed.

Notes: Abstract Ten male weanling Holtzman rats, injected intraperitoneally with aqueous estradiol (Progynon, Schering), in daily doses of 1 μg. per g body weight, were sacrificed, simultaneously with controls receiving an equivalent amount of diluent, at intervals ranging from one hour to six days. Upper tibial epiphyseal cartilage plates (ECP), procesed for electron microscopy, revealed, as early as three hours after injection, appreciable enhancement of secretory activity, evidenced, in the zone of matrix secretion, by the abundance in Golgi cisternae of stippled material representing proteinpolysaccharide complexes. Disintegration of the lining membrane of individual Golgi vesicles was advanced after twenty-four hours; following three days of treatment, few vesicles remained intact, and pools of initially intravacuolar material were observable in the gound plasm. Long filaments, suggestive of primary or precursor collagen fibrils were apparent in this secretion. After six days, virtual lakes of this substance filled cells in the zone of prehypertophy, with consequent displacement of the rough endoplasmic reticulum against the cell periphery. Cytoplasmic vacuoles, containing mateerial similar to that found in the lacunar moat, and displaying finely beaded, radially arrayed filaments on the lining membrane were frequently encountered. Our observations suggest an initial acclleration of chondrocytic secretory activity, with subsequent retardation of transport. The resultant retention and intracellular polymerization of precollagenous products accelerates hypertrophy, thereby promoting early degeneration of chondrocytes. These ultrastructural alterations are apparently estrogen-specific.

Notes: Abstract A dorsal and ventral gland composed of large, flask-shaped cells located in the margin of the mantle ofLittorina give rise to the inner and outer layers of the periostracum respectively. The material comprising the periostracum is derived from secretory granules elaborated by the Golgi apparatus. When the Golgi granules of the ventral gland which consist of a substance exhibiting a definite periodicity, are discharged at the surface in contact with sea water, they are widely dispersed and consist of particles embedded in a substrate. Formation of the outer periostracum involves the re-aggregation of the Golgi secretory particles into a dense sheet which exhibits a periodicity of 300 Å. Viewed in transverse section the periostracum exhibits a lattice pattern suggestive of a crystalline substance. The outer layer eventually reaches a thickness of 4–5 μ. The inner layer is derived from the secretory granules of the dorsal gland. The formation of the inner layer occurs in a manner similar to that of the outer layer. It does not, however, exhibit a periodicity. In the mature state it attains a thickness of 0.4–0.5 μ. In addition to a protective function the periostracum provides a barrier between the sea water and the pallial space and also regulates the site and arrangement of mineral crystal formation at the growing margin of the shell.

Notes: Abstract A solution of chromium sulphate was used as a combined fixative, stain and demineralizing agent for the ultrastructural study of mineralizing cartilage. This technique revealed the presence of an organic ‘crystal ghost’ associated with each crystallite. The effectiveness of chromium sulphate as a demineralizing agent is discussed.

Notes: Abstract Although experimental and clinical experience indicates that large doses of testosterone lead to premature cessation of growth, the exact mechanism and precise site of action of this hormone on the growth apparatus of long bones remain unknown. In this study, plateaued male rats were injected with supraphysiologic doses of testosterone to observe the submicroscopic effects on the various zones of the epiphyseal cartilage. In the zone of cell division there were increased numbers of dividing cells. The maturing cells accumulated larger amounts of secretory products at earlier stages of their life cycle, and appeared to undergo a more abrupt hypertrophy. In the zone of prehypertrophy, the interterritorial matrix contained foci of early and premature calcification and thicker and longer collagen fibers than at comparable levels in controls. It is concluded that in intact animals, even large doses of testosterone initially cause a stimulation of chondrocyte proliferation, prior to promoting maturation processes.

Notes: Abstract The initiation of prisms occurs in the proximal region of the outer surface of the outer mantle fold in the pallial space bounded externally by the periostracum. The first step in the formation of a prism is similar to that observed in the formation of nacre, namely, the elaboration of an electron-dense lamella that serves as the internal boundary of the future prism. Fragments of the lamella become detached and migrate to a chamber bounded externally by the periostracum. These lamellar fragments form envelopes within which crystal initiation and growth oocur. At the same time stout interprismatic walls appear. They are also derived from the lamellae. Growth consisting of the formation of additional crystals and the organic components finally give rise to the mature elongated prism. Growth of the shell occurs at the margin chiefly by formation of new prisms in this area. In addition a modified environment consisting of duplicature of the periostracum on the distal surface results in the formation of thin spurs containing prisms that occur within the confines of the space created by the periostracal loop.

Notes: Abstract Nine day old experimental calluses in radii of young rats were treated with Gomori's (1939) calcium-cobalt method to demonstrate ultrastructurally the presence of alkaline phosphatase in a search for its possible role in the desposition of calcium. Enzyme activity first appeared as globule-like precipitates outside the cell membrane of early hypertrophic cartilage cells. This precipitate layer then seemed to give rise to spherical bodies of alkaline phosphatase which occur at a slight distance from the cell. The spherical bodies were also observed further away from the cell in an intermediate zone between neighboring cells. Needle-like crystal formation, apparently calcification, occurred inside single or aggregated bodies, leaving their peripheral rim clearly visible, even when calcification had increased to such an extent that individual crystals could no longer be recognised. In controls, treated in the same way but without substrate, or with EDTA, no enzyme precipitate or spherical bodies were seen. The appearance of crystalline deposits in bodies which contain alkaline phosphatase seems difficult to explain on any other basis than that there is a close functional association between the enzyme and calcification.

Notes: Abstract An ultrastructural study of peripheral human coronal dentin in patients aged 11 to 75 years, has shown main aspects of the dentinal tubular content. In transverse sections, they consist of dead tracts and annular or solid content to the tubular lumen. No nerve endings were observed in the outer dentin.

Notes: Abstract The organic phase (or crystallite ghost) associated with each crystallite, together with the background material associated with each crystallite cluster, was demonstrated in calcified cartilage using basic chromium sulphate as a combined fixative, stain, and demineralizing agent. Subsequent treatment of the tissue with papain, or with hyaluronidase, suggests that the crystallite ghosts represented a protein-polysaccharide complex and that the background material was principally protein together with some acid polysaccharide. The relationship between inorganic and organic phases is discussed.

Notes: Abstract Tillers of Festuca arundinacea Schreb. were subjected to-8°C in a bath of methylated spirits for three-quarters of an hour. They were thawed at room temperature and some material taken from the shoot apical meristem and leaf blade for electron microscopy. Similar material was taken from control plants for electron microscopy. Nine tillers subjected to-8°C and thawed subsequently failed to regrow. Nine control tillers regrew. All the treated meristem cells and about half the treated leaf mesophyll cells were extensively altered. Their nuclei were contracted, organelles were swollen or partly disrupted, plasmalemma and nuclear membranes were broken or absent and vacuoles were sometimes disrupted. Strongly osmiophilic material accumulated in the vicinity of membranes. About half the leaf mesophyll cells differed from the control mesophyll cells only in having more spherosomes and narrower thylakoids. Parallels with other ultrastructural studies of stress damage and the indications the results give of possible primary damaging events are discussed.

Notes: Abstract Embryogeny in the nasturtium is characterized by the development of a large, tripartite suspensor and storing cotyledons. A light and electron microscopic study revealed an early diversification of the plastids in the various regions of the suspensor and the embryo proper. Amyloplasts are found in the developing cotyledons of the heart-like embryo, while chloroplasts occur within the meristematic part of the embryo and the adjacent portion of the suspensor. The cells between the meristem and the storing cotyledons display undifferentiated leukoplasts, whereas leukoplasts with an electron-dense matrix occur in the basal cell mass of the embryo-suspensor. Etioplasts develop in several cells of the placental haustorium of the suspensor. The carpel haustorium shows rather undifferentiated leukoplasts, which are transformed into electron-dense plastids during autolysis of the suspensor. This early plastidal differentiation in discussed with respect to its control and functional significance.

Notes: Abstract Bristles radiating from openings were detected on colonies and unicells ofScenedesmus culture N 46, when examined with transmission and scanning electron microscopes. Although narrower, they correspond in gross appearance and ultrastructure to previously describedScenedesmus bristles. Openings, bordered by a series of props, are unlike those ofScenedesmus culture 614. Additional props are observed scattered independently on the cell wall; ridges are composed of a linear row of props. Sections of cells, or cell walls, reveal an additional prop, situated inside the openings; these props are composed of several tubules. Possible extrusion of bristles through these tubules, as well as the origin of the bristle from the cavity and vesicles immediately under the opening are discussed.

Notes: Abstract The morphology and ultrastructure of aJaponochytrium sp. has been studied by light, scanning and transmission electron microscopy. The wall has been shown to be multilamellate and persistent. Stages in zoospore cleavage are described and sagenogenetosomes reported in mature sporangia.

Notes: Abstract Ultrastructural observations on encysting haploid zoospores of Allomyces arbuscula are presented with special reference to cell wall deposition. Multivesicular bodies are observed in the cytoplasm of zoospores 15 min after inoculation, lomasomes after 30 min and fine membrane profiles between the plasmalemma and the cyst wall are observed after 4 h indicating a possible system for secretion of cell wall components.

Notes: Abstract Observations of sporophore development in fresh and glutaraldehydeosmium sequentially-fixed material ofProtophysarum phloiogenum show the following sequence. Small plasmodia cease streaming and round up. Food vacuoles collect in the lower center of the cytoplasmic mass. As the cytoplasm rises the food vacuolar contents are excluded from the plasmalemma and become the stalk core. A continuous, fibrous peridium and stalk tube enclose cytoplasm and stalk core respectively. Capillitial formation just precedes spore cleavage. Sporophore development is marked by autophagic activity and calcium deposition. Stalks of dried herbarium specimens of seven additional species have been examined. A mature stalk morphology very similar toProtophysarum with recognizable remnants of microorganismal food material is seen in all of them. It is thought that this marker is indicative of non-stemonitaceous stalk development.

Notes: Abstract A strain ofSchizophyllum commune carrying a mutation in theB-mating factor (B-mut) shows septal dissolution when grown at 30° C for 2 to 3 days. The septa are intact if the organism is grown at 25° C for the same time, but begin to break down within 1 h after transfer to 30° C. At the ultrastructural level the dolipore swelling is the first part of the septal apparatus to be degraded, closely followed by the disorganization of the parenthesomes. A progressive thinning of the septal cross-wall produces an enlargement of the septal aperture sufficient to allow the passage of nuclei. It appears that degradative enzymes are probably carried to the site of septal dissolution in vesicles derived from endoplasmic reticulum in the area of the septal apparatus.

Notes: Abstract Nitrogen starvation, effected by incubating a culture ofAnabaena cylindrica in a medium free from combined nitrogen and under an atmosphere of 1% CO2 in argon, leads to rapid and characteristic changes in the appearance, structure and function of the alga. Change of colour, due apparently to a decrease in the amounts of nitrogenous pigments, is accompanied by a structural transformation of vegetative cells: cyanophycin granules and polyhedral bodies disintegrate, lipid and glycogen accumulate, and large membrane-bound spaces form by means of thylakoid swelling and vesiculation. The rate of heterocyst differentiation and nitrogenase activity is increased. These changes are fully reversed on addition of ammonia to the culture. It appears that thylakoids reform by coalescence of small vesicles assembled in the intrathylakoidal space. Rapid ammonia assimilation is indicated by ample formation of cyanophycin granules in vegetative cells and of “plugs” in the heterocysts.

Notes: Abstract Complexes of twisted ribbons composed of ordered arrays of microtubules are identified in close association with the plasmalemma and the surfaces of some organelles in senescent cells of photoheterotrophically cultured Chlamydomonas dysosmos. The ribbon complexes occur throughout the cytoplasm, and do not appear related to the flagellar insertions. The component microtubules are approximately 26 nm in width, exhibiting a center-to-center spacing of about 44 nm. Additional cytoplasmic microtubules are often closely related to the tubular complexes. A detailed description of their fine structure is presented here which tends to support the ascribed function of microtubules in maintaining the structural integrity of the protoplasm.

Notes: Abstract The structural and functional symptoms of molybdenum deficiency inAnabaena cylindrica grown in a medium without combined nitrogen and thus dependent on fixation of elemental nitrogen, resemble those brought about by nitrogen starvation. However, the substantially increased rate of heterocyst differentiation in this culture is not accompanied by a corresponding increase in nitrogenase activity; on the contrary, enzyme activity is severely impaired in the absence of molybdenum. When the supply of molybdenum, or of ammonia, is restored, the alga recovers rapidly. Vanadium exerts an inhibitory effect upon nitrogen-fixing ability of the alga, and its presence in the molybdenum-deficient culture results in the amplification of the symptoms of mlybdenum deficiency.

Notes: Abstract Microbody-like organelles occur in the cytoplasm of two chloromonadophycean algae,Vacuolaria virescens Cienkowsky andGonyostomum semen Diesing. Microbodies ofVacuolaria andGonyostomum have a granular matrix which lacks a crystalloid core; they are often present in close association with elements of the endoplasmic reticulum. The occurrence of microbodies in other algae is briefly reviewed.

Notes: Abstract This paper deals with electron microscopic observations on cultivated plants of the marine red alga Gracilaria verrucosa which developed simple galls; also sea collected material, without galls, had been studied. The galls showed unusual but characteristic cell structures, caterpillar-like bodies, containing rows of fusiform bodies. These were found mostly in the cytoplasm near the plastids, in one case connected with the endoplasmic reticulum, occasionally even inside the nucleus, and are described here, as far as we know, for the first time. It does not seem probable that the caterpillar-like bodies represent mitochondria or bacteria, but the hypothesis that fusiform bodies are related to virus-like structures is discussed. The normal tissues as well as the gall tissue of the laboratory plants contained, besides plastids typical for the red algae, another type of plastids characterized by tubular thylakoids.

Notes: Abstract The mode of attack and the infection structures of the necrotrophic mycoparasite, Pythium acanthicum, as well as the responses of various fungal hosts to parasitism were studied using both electron and light microscopy. Many taxonomically distinct fungal hosts were used, though Phycomyces blakesleeanus, Pythium aphanidermatum, Rhizoctonia solani and a basidiomycete identified as Corticium sensu lato were studied in greatest detail. Parasitism was by direct penetration of the fungal host without appressorium formation by the parasite. The host's cells responded to contact by P. acanthicum by forming papillae. The morphological features of the papillae varied with the particular host. In P. blakesleeanus they were comprised of vesicles and segments of cytoplasm entrapped in a fibrillo-granular matrix, while in R. solani and the Corticium basidiomycete they contained considerable amounts of electron-opaque and electron-translucent material. Evidence for both mechanical and enzymatic penetration of the host fungi by the parasite are presented. Details of host wall and septum penetration by the parasite are presented using time-lapse light microscopy with in vivo systems. Many of these stages of parasitism were examined ultrastructurally. Some comparisons of these mycoparasitic relationships are discussed in relation to what is known from the literature about phytoparasitic interactions.

Notes: Abstract The ultrastructure of gametangial development in Allomyces macrogynus was determined from longitudinal sections of gametophytic hyphae at stages of differentiation from vegetative apices at time zero to fully cleaved gametangia at about 150 min. Whereas vegetative hyphae show an apical clustering of mitochondria, cytoplasmic vesicles and microtubules, this arrangement was sharply altered in early development. Mitochondria were evenly redistributed, apical vesicles and microtubules disappeared, and autophagic vacuoles became prominent. Subsequently, electron-dense granules and microbody/lipid droplet complexes became evident and later, during gamete cleavage, developed into gamma bodies and side-body complexes respectively. Meanwhile cytoplasmic vesicles were involved in exit papilla formation. The significance of autophagic vacuoles and multivesicular bodies is discussed.

Notes: Abstract The morphology and ultrastructure of the aerobic, Gram-negative multicellular-filamentous bacteria of the genus Simonsiella were investigated by scanning and transmission electron microscopy. The flat, ribbon-shaped, multicellular filaments show dorsal-ventral differentiation with respect to their orientations to solid substrata. The dorsal surface, orientated away from the substrate, is convex and possesses an unstructured capsule. The ventral surface, on which the organisms adhere and glide, is concave and has an extracellular layer with fibrils extending at right angles from the cell wall. The cytoplasm in the ventral region contains a proliferation of intracytoplasmic membranes and few ribosomes in comparison to the cytoplasm in other parts of the cell. Centripetal cell wall formation is asymmetrical and commences preferentially in the ventral region. Quantitative differences in morphology and cytology exist among selected Simonsiella strains. Functional aspects of this dorsalventral differentiation are discussed with respect to the colonization and adherence of Simonsiella to mucosal squamous epithelial cells in its ecological habitat, the oral cavities of warm-blooded vertebrates.

Notes: Abstract Two strains of unicellular cyanobacteria which reproduce exclusively by budding are described and assigned to genus Chamaesiphon.

Notes: Abstract A new genus of marine methanogenic bacteria and two species within this genus are described.Methanogenium is the proposed genus andMethanogenium cariaci the type species. Cells of the type species are Gram-negative, peritrichously flagellated, irregular cocci with a periodic wall surface pattern. Colonies formed by these bacteria are yellow, circular and umbonate with entire edges. The DNA base composition is 52 mol% guanine plus cytosine. Formate or hydrogen and carbon dioxide serve as substrates for growth. Cells ofMethanogenium marisnigri are of similar shape but smaller diameter thanM. cariaci. The colonies ofM. marisnigri are convex, and the DNA base composition is 61 mol % G+C. Formate or hydrogen and carbon dioxide are growth substrates. Sodium chloride is required for growth of both methanogens.

Notes: Abstract The predacious watermold Zoophagus insidians traps loricate rotifers on short, lateral branches of the main, hyphal axis. These branches or “traps” are packed at their distal ends with a number of vesicles filled with an electron-dense matrix. Electron micrographs of the mycelium disclose a two-layered wall; the outer layer is electron dense and the inner, electron transparent. The outer dense layer on the tip of the “trap” is organized into a number of fine ridges and occasional discontinuities. Thin sections through recently trapped rotifers indicate that the cilia of the animals are stuck to the trap by a glue. This adhesive is derived from secretion of the matrix of the vesicles aggregated in the tip of the trap. The secretion mechanism is triggered by the animal and is accompanied by: 1. The separation of the two layers of the wall, 2. fusion of the vesicles with the cell membrane and 3. extrusion of the glue through pits in the tip of the inner wall of the “trap”. After snaring a rotifer, the previously arrested branch grows as a haustorium into the body cavity of the animal. The host tissues disintegrate within a few hours and appear to be the main nutrient source of the fungus.

Notes: Abstract The cell wall of Coelastrum is usually composed of three layers. The outermost layer was studied most extensively. It consists of erect tubules which often bear long bristles whose function may be to stabilize the algae in its environment. The cell wall can modify its morphology according to the environment.

Notes: Abstract Effects of diazepam (Valium) on photosynthesis, chlorophyll/photosynthesis ratios, respiration, uptake of rubidium ions, and ultrastructure of Scenedesmus obliquus synchronized by a light-dark regimen of $$14:\overline {10}$$ hrs were determined. 80 and 160 μM diazepam, added to the nutrient medium at the start of the light-dark change (i.e., start of the cell cycle) gradually reduced rates of photosynthesis below the initial rates from the beginning of the experiment. Contents of chlorophyll, however, remained nearly unaffected. Consequently, the diazepam-treated cells had a higher chlorophyll/photosynthesis ratio—also with regard to respiration in order to calculate the gross photosynthesis. The occurrence of photorespiration cannot be assumed. The net influx or rubidium was slightly reduced by 100 μM diazepam 0.5 and 2.0 hrs after the start of the cell cycle and was strongly inhibited after 5 to 14 hrs. 80 and 160 μM diazepam caused separation of thylakoids, formation of giant mitochondria and enlargement of vacuoles. The results are discussed and it is finally suggested that diazepam acts on different membrane systems. Furthermore an ATP deficiency cannot be excluded.

Notes: Abstract Spores of the thermophilic, acidophilic, Bacillus acidocaldarius were covered by a thick outer coat and a laminated inner coat (5.5 nm periodicity). Small membranous vesicles were present in the spore core and they disappeared as germination proceeded. After depolymerization of the cortex, and a 30% increase in spore diameter, a localized gap appeared in the laminated inner coat only. This inner coat gap was narrow and could be the whole length of the spore. The germ cell appeared to grow, or to be pushed towards the inner coat gap, at which stage the outer coat disappeared in the same localized area. As the vegetative cell grew out the spore coat fell away, with loose cortical material still attached to it. The young germ cell developed a large spherical electron dense inclusion body in the cytoplasm, at the same time as the ribosomal and nuclear areas became distinct.

Notes: Abstract An ammonia-oxidizing, autotroph growing, slender, curved rod was isolated from the soil of Hawaii. It is well distinguishable from any other nitrifying bacteria thus far described by their morphology. The cells are 1.1–3.0 μm long and 0.3–0.4 μm wide. They are motile by means of 1–4 subpolar to lateral flagella. In contrast to most of the ammonia-oxidizing bacteria the isolated vibrio is void of an extensive cytomembrane system. To categorize this not yet described species we propose to create the new genus Nitrosovibrio and to classify the isolated strain as Nitrosovibrio tenuis.

Notes: Abstract The cell morphology, the arrangement and fine structure of flagella and the piliation of the following Gram-negative aerobic hydrogen bacteria have been studied: Alcaligenes eutrophus, Alcaligenes paradoxus, Alcaligenes ruhlandii, Pseudomonas flava, Pseudomonas pseudoflava, Pseudomonas palleronii, Pseudomonas facilis, Aquaspirillum autotrophicum, Paracoccus denitrificans, Corynebacterium autotrophicum, and strains MA 2 and SA 35. The identity of the bacteria was examined by their substrate spectra and type of flagellation. Three types of flagellar fine structure were differentiated. The presence of pili was noted in strains of Alcaligenes paradoxus, Pseudomonas flava, P. pseudoflava, P. palleronii, and P. facilis.

Notes: Abstract In a related brewing study detailed characteristics of fermentations displaying effective yeastaminopterin interaction were presented. Fermentative yeast types (certain Saccharomyces species and Selenotila intestinalis) proved effective aminopterin reactors whereas oxidative yeasts (certain Candida, Cryptococcus, Pichia, Rhodotorula, Saccharomyces, and Trigonopsis species) proved ineffective reactors. In general effective reactors were polyploids characterized by the lack of film or pellicle formation and ineffective reactors the opposite. In stationary fermentations the Fleischmann 139 strain of S. cerevisiae proved a fair reactor. When aerated it proved an ineffective reactor and aminopterin or products there-of stimulated growth. Conversely aeration enhanced aminopterin activity of effective reactor yeasts. The positive effect of biotin on aminopterin activity and the negative effect of yeast extract, L-asparagine, adenine and thymine is shown and compared and contrasted with earlier reported studies. These findings supported by outside data suggest that oxidative yeasts (and bacteria) can readily elicit enzymes capable of inactivating aminopterin whereas fermentative types are lacking in this capability. Finally that past yeast-aminopterin studies were conducted with oxidative yeast types. Advantages of effective aminopterin reactor yeasts to be published elsewhere include improved ultrastructure using KMnO4−OsO4 fixation, a yeast bioassay procedure for detecting aminopterin in plasma and urine, and cell synchronization.

Notes: Abstract Three types of structure of the ascospore wall were found among the haploid Saccharomyces species examined: a warty wall (S. rouxii), a smooth wall with a single electron-light inner layer (S. bailii) and a smooth wall with a double light inner layer (S. montanus, S. florentinus). The latter type also occurred in Kluyveromyces thermotolerans and K. waltii. In K. fragilis spores the wall had a single light inner layer. The taxonomic implications of these findings were discussed.

Notes: Abstract The freeze-fracture technique and electron microscopy have been used to demonstrate that localized damage is inflicted upon the cytoplasmic membrane of Spirillum serpens VHL within 20 to 30 min after the start of its association with Bdellovibrio bacteriovorus 109D. This damage is not observed in uninfected Spirillum cells, nor in infected cells within the first 10 min. This damage takes the form of a “blister” which, when viewed stereoscopically in electron micrographs, is seen to project toward the interior of the Spirillum cell. Shortly after its formation, the blister becomes elaborated into a series of ridges which may assume forms ranging from an elaborate spiral to a series of loops or knots. The formation of a blister is shown to involve both the inner and outer leaves of the membrane bilayer, and evidence is presented to indicate that the blister site corresponds to the site of attachment of the Bdellovibrio cell. The hypothesis is proposed that this ultrastructural damage is the cytological basis for the controlled and localized leakage through the cytoplasmic membrane into the periplasmic space of the Spirillum cell at locations adjacent to the Bdellovibrio cell. It is suggested that this localized membrane damage may be the ultrastructural basis for the high efficiency with which bdellowvibrios are known to incorporate cytoplasmic materials from the other bacteria in whose periplasmic spaces they develop.

Notes: Abstract 1. Tubular-like structures were regularly revealed in the surface sheath of the aerial mycelium of the parent strain of Streptomycetes roseoflavus var. roseofungini. In their shape and dimensions these structures were highly reminiscent of those massive accumulation of which was earlier reported to occur in cultures of dedifferentiated nocardioform “fructose” mutant of the same parent strain. 2. The tubular-like structures of the aerial mycelium sheath were shown to be markedly susceptible to brief acetone washing, undergoing almost complete desintegration. 3. On addition of water to crude acetone extract of the aerial mycelium precipitation and possible selfassembly of a spectrum of various structures occurred (folded scaly, bubble-like, spout-like). Some among structures so produced were reminiscent of those found in spectra of structures observed in reconstruction experiments with tubules from the mutant as well as of structures found in the aerial sheath of the parent strain and some other actinomycetes. Similarity in subunit structure of above materials was also noticeable. The presence, in the sheath of aerial mycelium, of material with a tendency to selforganization is discussed in relation to the possible involvement of selfassembly processes in the formation of the surface sheath.

Notes: Abstract Gloebacter violaceus gen. and sp. n. is a unicellular photosynthetic prokaryote of unusual cellular structure. The only unit membrane in the small, rod-shaped cells is the cytoplasmic membrane, which has a simple contour, without intrusions. Immediately underlying it is an electron-dense layer 80 nm thick. Gloeobacter is an aerobic photoautotroph which contains chlorophyll α, β-carotene and other carotenoids, allophycocyanin, phycocyanin and phycoerythrin. Chlorophyll and carotenoids are associated with the particulate fraction of cell-free extracts, and are thus probably localized in the cytoplasmic membrane. The phycobiliproteins may be associated with the electron-dense 80 nm layer. The DNA contains 64.4 moles percent GC. The cellular lipids have a high content of polyunsaturated fatty acids, largely linoleate and γ-linolenate. Despite its atypical fine structure, Gloeobacter is evidently a cyanobacterium, sufficiently different from other unicellular cyanobacteria to be placed in a new genus.

Notes: Abstract During the process of degradation of the cell wall of the yeast form of Pullularia pullulans by the lytic system of Micromonospora chalcea samples were withdrawn at different times and observed under phase contrast and electron microscope. The progressive lysis of the walls reveals a fibrillar component inside the apparently amorphous wall. Freeze etched preparations of cells during the formation and regeneration of protoplasts show that the cellular membrane is split and this method allows the smooth external face of the membrane and other internal face covered by particles to be seen. The fact that the smooth face of the membrane is only visible during the preparation or the regeneration of protoplasts and very rarely when intact cells are fractured, suggests a strong adherence between cell wall and this external layer of the membrane. During the regeneration which takes place as in most of the yeasts and moulds, a special study of the extension of the cell wall is made and a possible mechanism for this extension of the regenerated cell wall is proposed.

Notes: Abstract Infection of the mucoraceous host Cokeromyces recurvatus by Piptocephalis unispora was studied ultrastructurally, using a new technique involving yeast-phase cells of the host to obtain large numbers of infection sites for thin-sectioning. Morphologically, the haustorial apparatus was similar to that of fungi parasitic on higher plants, and comprised an appressorium, a neck region with a collar and a neck ring, and a lobed region surrounded by a sheath matrix enclosed in an extra-haustorial membrane. Penetration of the host by the infection peg probably involved both enzymatic degradation and physical pressure. Reaction of the host to infection is described and the results related to the theory of host infection by haustorial fungal parasites.

Notes: Abstract An ammonia-oxidizing bacterium was isolated from a sample of brackish water (North Sea, Harbour of Husum). It is a motile large coccus 1.5–1.7 μm in diameter. The extensive cytomembrane system occurring as flattened vesicles in the peripheral region of the cytoplasm and as intrusions into the center of the cytoplasm is to be emphasized as a characteristic mark of identification. The lithoauto-trophically growing bacterium turned out to be an obligate halophile. Because of its physiological and morphological properties, we assigned it to the genus Nitrosoccus and propose the name Nitrosococcus mobilis.

Notes: Abstract Ultrathin sectioning of submerged mycelium of Claviceps purpurea Tul. producing clavine alkaloids revealed yeast-like budding resulting in asexual sporesblastospores. These deciduous spores were born by extended hyphal cells and retained the same ultrastructure of cell organelles. Both the extended hyphae and the blastospores resembled the cells of ergot sclerotial tissue. A surface culture of C. purpurea Tul. producing no alkaloids was used as a reference.

Notes: Abstract The fine structure of the cell envelope, of membrane systems and of cytoplasmic inclusions of Gram-negative aerobic hydrogen bacteria has been studied. The results have been tabulated, and three main groups could be recognized: Group 1: Alcaligenes eutrophus, A. paradoxus, A. ruhlandii, Pseudomonas facilis, P. flava, P. pseudoflava, P. palleronii, and Aquaspirillum autotrophicum; Group 2: “Corynebacterium” autotrophicum and strains MA 2 and SA 35; Group 3: Paracoccus denitrificans. Special structures related to the chemoautotrophic way of life of the hydrogen bacteria were not observed.

Notes: Abstract The photosynthetic bacterium,Rhodopseudomonas sphaeroides, can be grown phototrophically (light, anaerobiosis), of chemotrophically (dark, aerobiosis). In the first case, it contains intracytoplasmic membranes with photosynthetic pigments. When shifted from phototrophy to chemotrophy these membranes disappear in an unknown fashion. In the present experiment, samples were taken for electron microscopy, cell density and bacteriochlorophyll determinations after shift from phototrophy to chemotrophy. The density of intracytoplasmic vesicles was measured on micrographs. During the first 2h growth is very slow and the ultrastructure remains unaltered. As growth resumes, the vesicles disappear at a rate which implies that they are not incorportated into the cytoplasmic membrane, nor actively digested, but remain intact and become increasingly diluted in the cytoplasm as the culture grows. The size of the vesicles was estimated to about 500 Å. The number of vesicles in phototrophically grown cells was calculated to about 575 per cell, and after 6h chemotrophic growth to about 100. The areas of the cytoplasmic and intracytoplasmic membranes are roughly calculated.

Notes: Abstract The mode of spore differentiation in a strain of Streptomyces melanochromogenes was followed by analysis of ultrathin sections of sporulating aerial hyphae at various stages of sporogenesis. A special accent was laid on the formation of the sporulation septum and its alterations in the course of spore delimitation and separation. Distinct differences in formation and substructure have been observed between the cross walls of vegetative hyphae and the sporulation septa. Cross walls of vegetative hyphae are formed in a way typical for Gram-positive bacteria by a centripetal annular ingrowth of cytoplasmic membrane, on which wall material immediately is deposited. The development of the sporulation septa is characterized by the accumulation of amorphous material in addition to the newly synthesized wall layer inside the invaginating cytoplasmic membrane. This amorphous septal material will later be decomposed presumably by two lytic systems which cause the separation of the spores. The central region of the finished sporulation septum is perforated by microplasmodesmata. Spores are released by a break down of the surface sheath. The complete spores are enveloped by a twolayered cell wall and the spiny surface sheath.

Notes: Abstract Gram-negative, anaerobic gliding bacteria were isolated from normal supragingival plaque and from periodontal lesions. Isolates could be divided into two size classes: small 2.4–4.2 μm×0.38–0.5 μm and large 4.8–5.8 μm×0.42–0.6 μm cells. The outer membrane was either loose-fitting and wavy, or taut, and of variable thickness. An electron-dense fuzz was discernible on several of the isolates. The periplasmic region was of variable electron-density. The genus Capnocytophaga has been proposed for these organisms based on morphological and cultural characteristics.

Notes: Abstract The colonial sheath of Microcystis marginata has a definite structure as seen by light and electron microscopy, consisting of a relatively smooth inner surface and densely packed, long fibrils on the outer surface. The sheath initially forms around the single cell and expands by continual deposition of sheath material to accomodate the synchronously dividing cells of the colony.

Notes: Abstract 1. After a resting period of up to 6 months cells of Nitrobacter agilis grow with acetate, formate, and pyruvate as carbon and energy source. Yeast extract and peptone were added to supply the organism with nitrogen and to meet possible vitamin requirements. 2. The length of the growth period depends on the substrate; it increases according to the following sequence: pyruvate, formate, acetate. The highest growth yield is observed with pyruvate, the lowest with formate. 3. O2 consumption is increased in the presence of substrates as compared to endogenous respiration. With pyruvate and acetate twice as much O2 is consumed, with formate 7 times, with yeast extractpeptone 10 times as much. 4. The ability of nitrite oxidation is largely preserved, except in cells grown with acetate or pyruvate in the presence of 0.015% yeast extract and peptone. Such cells have nearly no cytochrome a 1. Accordingly, the cytochrome spectra of nitrite oxidizers grown under chemoorganotrophic and lithoautotrophic conditions coincide qualitatively. 5. The nitrite oxidizing system is inducible. It is induced by nitrite but also by substances present in yeast extract and peptone. Cells grown on acetate and yeast extract and peptone (0.015%) require 3–4 weeks before they regain the ability to grow with nitrite. Cells grown chemoorganotrophically with the same substrates and yeast extract and peptone (0.15%) start growing with nitrite as energy source without a lag. 6. Cell size and form, distribution of storage materials, order and fine structure of double membranes are correlated with growth conditions.

Notes: Abstract This is the first report of longitudinal sections of synaptonemal complexes in oömycetous fungi. These indicators of meiosis were observed in antheridial nuclei of Achlya ambisexualis E87. They were attached to a platelike structure at the inner membrane of the nuclear envelope. The lateral elements were separated from each other by an average distance of 160 nm. These results provide new ultrastructural evidence for gametangial meiosis in Oömycetes.

Notes: Abstract Heat-shock of macroconidia of Neurospora crassa at 46°C followed by shift-down to 25°C determines premature conidiogenesis. The nuclei and cytoplasm of heat-treated, swollen conidia contain spots of a dense material especially concentrated around the nucleolus in short time treated ones. In the first proconidium apically budding on the enlarged tip of the premature conidiophore, small vesicles are peripherally spread. A few such vesicles are later seen lining the initially simple septum separating the proconidial units into conidia. The doubling of this interconidial septum is surface viewn as a thick annulus. Disarticulation of the conidial units intervenes along a septal furrow of electroluscent material. Interconidial continuity through the septal pores is transiently insured by a connective which is ruptured for final liberation of the conidia.

Notes: Abstract Polyhedral inclusion bodies were observed in cells of a Nitrosomonas species. They were present in growing cells as well as in resting cells. In thin sections their size was about 130 nm in growing cells and about 185 nm in diameter in resting cells. The bodies were commonly located in the nucleoplasm. They appeared to be bounded by a nonunit membrane and had a granular substructure. In thin sections about 70% of the exponentially grown cells and about 20% of the resting cells of the investigated strain showed 1–7 respectively 1–3 inclusion bodies.

Notes: Abstract Individuals of an alcohol dehydrogenase-negative strain of Drosophila melanogaster designated Adh n5 are resistant to ethanol poisoning at low but not at high temperatures. The basis for this behavior is that Adh n5 flies contain a mutant form of alcohol dehydrogenase which is less heat stable than that of wild-type flies. The mutation in Adh n5 maps at, or very close to, the presumptive structural locus and is not complemented by any of 11 other alcohol dehydrogenase-null mutants.

Notes: Abstract 3-Hydroxykynurenine is virtually absent from st larvae but accumulates during adult development in the puparium. Over the period of adult emergence, the accumulated 3-hydroxykynurenine is excreted so that st adults contain none. Larvae of st fed on tryptophan-C 14 medium produce labeled 3-hydroxykynurenine, at a reduced rate, perhaps, compared to wild type. Xanthurenic acid levels in st pupae are similar to those in wild type. Thus the failure of st larvae to accumulate 3-hydroxykynurenine does not seem to be due either to an inability to synthesize this compound or to an excessive rate of its conversion to xanthurenic acid. Rather, it appears that the mechanism of 3-hydroxykynurenine storage during larval life is defective, so that this compound is excreted at an abnormally high rate. The inability of the pigment cells of the eyes of st to synthesize xanthommatin may result from a similar defect in their ability to take up or store 3-hydroxykynurenine.