Bibliothek

Notizen: Summary An extremely rare case of a conception occurring in a 26-year-old patient with a small virilizing Sertoli-Leydig cell tumor (diameter: 0.6 cm), bilateral polycystic ovaries and non-tumorous adrenal hyperandrogenism is presented. Prepregnancy findings included hirsutism, clitoromegaly, secondary amenorrhea, and elevated peripheral plasma testosterone (T; 5.7 ng/ml). Extensive basal steroid screening, dynamic function tests, conventional radiologic procedures, selective glandular vein catheterization, and laparoscopy failed to localize unequivocally the source of androgen excess, but suggested bilateral adrenal involvement. The patient conceived during the diagnostic work-up; peripheral T levels increased to 12.1 ng/ml within the first trimester. An exploratory laparotomy with left adrenalectomy, right adrenal biopsy and left ovarian wedge resection revealed an incompletely removed Sertoli-Leydig cell tumor, but normal adrenal histology. The pregnancy was terminated, a left oophorectomy and right ovarian wedge resection were performed at 14 weeks' gestation. Subsequently, peripheral androgens returned to normal, regular menses resumed, and hirsutism disappeared. Three years later the patient delivered a healthy female infant.

Notizen: Summary Subcutaneous fat from 3 patients with ischemic necrosis of the legs due to arteriosclerotic obstruction were examined histologically and ultrastructurally. Markedly convoluted membranocystic changes were found in all 3 cases. The light and electron microscopic findings of the membranocystic lesions are very similar to those of fat tissue changes in membranous lipodystrophy. Bone lesions and mental disturbance which suggest membranous lipodystrophy, however, were absent in these cases. It is concluded from these results that the membranocystic changes characteristic of membranous lipodystrophy can be produced by circulatory disturbance and the lesions are one of the non-specific changes of adipose tissue.

Notizen: Summary This report presents the ultrastructural features of a congenital epulis. The granular cells of the epulis were packed with numerous membrane bound cytoplasmic granules containing particles, small vesicles, and electron-dense materials. These granules were negative in immunohistochemical reaction for CEA (DAKO PAP KIT). Cytoplasmic organelles such as mitochondria, rough surfaced endoplasmic reticulum, and Golgi apparatus, were absent. Nuclei were markedly indented. Occasionally, banded intracellular collagen fibrils were observed within the cytoplasm. Some of these fibrils were surrounded by a limiting membrane, whereas others appeared to lie free in the cytoplasm. The collagen fibrils were also seen within a deep invagination of the cell surface. There was no basal lamina around the granular cells. Sporadically, mast cells with many granules containing lamellar formations were found between the granular cells. These observations support the idea that granular cells of the congenital epulis are derived from mesenchymal cells, probably fibroblasts.

Notizen: Summary Nine cases of a hitherto undescribed morphologic entity, termed mammosomatotroph cell adenoma of the human pituitary, are reported. These tumors, occurring mostly in men, are invariably associated with acromegaly (or gigantism) and high-normal or slightly elevated blood prolactin levels, and it cannot be distinguished clinically from well-differentiated growth hormone cell or mixed growth hormone cell-prolactin cell adenomas. They show a slow growth rate and usually exhibit a diffuse pattern and intense cytoplasmic acidophilia by histology. The immunoperoxidase technique detects both growth hormone and prolactin within the same cells. Electron microscopy reveals monomorphous tumors with a fine structure markedly similar to that of well-differentiated, densely granulated growth hormone cell adenomas. An added feature and diagnostic marker of mammosomatotroph cell adenoma is the presence of extracellular deposits of secretory material. One tumor shows a marked abnormality of hormone packaging and storage, resulting in the cytoplasmic accumulation of pleomorphic bodies containing semicrystalline secretory material.

Notizen: Summary Experimental autoallergic sialadenitis was induced in SL/Ni mice by one or two injections of syngeneic submandibular gland homogenate emulsified with adjuvant. Light microscopically, there were marked lymphoid cell infiltration in the submandibular glands with high incidence and proliferation of duct epithelia. Furthermore complete alteration of whole glandular lobules in some cases was observed. Ultrastructurally, small and medium sized lymphocytes and plasma cells constituted a major portion of the infiltrating cells, and lymphocytes were frequently observed inside the basal lamina of ductal and acinar regions, especially observed in the small ductal region. In the aggregates of infiltrating cells, the cell remnants of salivary gland epithelia were scattered. Furthermore some of the epithelial cell remnants in aggregates of infiltrating cells could be recognized as epithelial masses which were composed of proliferated duct epithelial cells, though no typical structure of epimyoepithelial islands seen in Sjögren's syndrome was found. Anti-salivary duct antibody was detected in only one case.

Notizen: Summary In this study the structural changes which occur during human pregnancy were examined by light and electron microscopy. Pregnancy was associated with proliferation and differentiation of the epithelial cells within the lobules. Proliferation was continuous throughout pregnancy with a progressive increase in the size of the lobules. The highest level of mitosis was observed in the first trimester with lower levels in the second and third trimesters. Unexpectedly a number of apoptotic cells were observed during pregnancy. Differentiation was initiated in the second trimester with an increase in the amount of rough endoplasmic reticulum and the appearance of a hypertrophic Golgi body and lipid droplets within a number of epithelial cells. A number of small vacuoles were present close to the apical plasmalemma of a few epithelial cells. As the pregnancy proceeded there was an increase in the number of cells exhibiting these features. There was also an increase in the size of the lipid droplets and the number of apical vacuoles. The apical vacuoles which have not been described previously range in size from 150–600 nm with the contents of the larger vacuoles having a whorled or labyrinth-like appearance.

Notizen: Summary Bladder tumours and non-tumorous bladder mucosa were studied by scanning and transmission electron microscopy in seven patients who had undergone hyperthermic vesical irrigation with bleomycin. The treatment induced sloughing of the outermost tumour cells, an increase of blebs and a decrease of cytoplasmic processes of the deeply located tumour cells as well as cellular degeneration. Although less severe, non-tumorous mucosa showed similar changes. Microvilli also appeared on the superficial cells of non-tumorous mucosa after the treatment. This treatment is effective by inducing cell degeneration and desquamation but not selective to the bladder tumour.

Notizen: Summary The ultrastructure of mouse antimesometrial decidual cells was analyzed during the development of the decidua between days 5 and 8 of pregnancy. The first decidual cells, appearing on the 5th day, are polygonal with rounded nuclei and prominent nucleoli; free ribosomes predominate in the cytoplasm. On the 6th to the 8th days the cytoplasm of these cells is typically that of cells actively engaged in macromolecular synthesis. Large numbers of granular and agranular endoplasmic reticulum cisternae are present in addition to well-developed Golgi complexes, mitochondria and lysosomes. Many bundles of microfilaments and lipid droplets occur during this period. An intense accumulation of autophagosomes and lysosomes with very heterogeneous content was noted on the 7th and especially the 8th days. The presence of these organelles is an indication that involution of this part of the decidua has begun.

Notizen: Summary The ultrastructure of the absorptive cells in the duodenum, jejunum and ileum after 7, 14 and 21 days of starvation was investigated using rats aged from 12 to 18 months weighing about 500 g. In the basal cytoplasm of the absorptive cells (in the duodenum and ileum of 21-day-starved rats and the jejunum of 14- and 21-day-starved rats), the following changes were found: atrophied mitochondrion-like bodies, small vesicles, a short and sparse rough-surfaced endoplasmic reticulum and a lack of density in a portion of the cytoplasm. Moreover, many autolysosomes of various sizes and shapes were encountered in the basal cytoplasm; occasionally these elements accumulated and appeared to fuse to one another. In contrast, in the apical cytoplasm of absorptive cells in the intestine of starved rats, the ultrastructure was similar to that of control rats. It was considered that the apical cytoplasm of the absorptive cells in the starved rat intestine might be preserved as long as possible during starvation in order to absord nutrients when they become available again.

Notizen: Summary Experimental high velocity missile brain injury in the rhesus monkey produces widespread swelling of perivascular astrocytes within 30 min of injury. Possible mechanisms for this lesion include a direct effect of force, chemical mediation secondary to the extravasation of blood, alterations in the permeability of the blood brain barrier and ischaemia. The implications of this finding for the function of the blood brain barrier, for neurotransmission and for neuronal survival are discussed.

Notizen: Summary A brain tumor of a 22-year-old man was composed mostly of round cells with perinuclear halos (clear cells), forming clusters intersected by small blood vessels. In some areas, the tumor cells showed perivascular arrangement and epithelial pattern. Phosphotungstic-acid hematoxylin stain and immunoper-oxidase stain for glial fibrillary acidic protein (GFAP) technique failed to stain the clear cells. Electron microscopy of the clear cells revealed them to be classical ependymoma cells with well developed intercellular junctions, microvilli and cilia. As no reporters in the past showed the evidence to clarify the nature of the clear cells, this case is considered a good example to support the viewpoint that the clear cells (oligodendroglioma-like cells) commonly observed in ependymomas are in reality ependymoma cells. It is stressed that the diagnosis of “mixed glioma” or “oligoependymoma” should be made with sufficient caution despite the recent advances of GFAP technique.

Notizen: Summary A 10-min long ischemic insult followed by up to 60-min survival results in several changes of the synaptic ultrastructure in the hippocampal CA-1 region. The alternations consist of gradual change of synaptic curvature from neutral to positive, cleavage and decrease in thickness of the postsynaptic densities and, in the case of many terminals, wrinkling of their profiles. The most striking form of damage are membrane discontinuities which begin to appear in very small numbers after 20 min of blood reflow and become much more pronounced after 60 min. The development of those modifications seems to be time-related, whereas decrease in the number of synaptic vesicles, as shown by the morphometric analysis, occurs after 10 min and does not progress any further after 20 and 60 min. This decrease is most pronounced in the immediate vicinity of the presynaptic membrane. Although the observed signs of ultrastructural alternations of synapses in the postischemic period appear to conform to the general pattern of synaptic degeneration observed under other conditions, the severity of ischemia is underlined with the rate at which those changes develop, thus pointing toward grossly disturbed metabolism of postischemic neurons. Recently, a number of theories have been advanced, discussing significance of ischemic destruction of membrane phospholipids. These theories are discussed in the context of membrane discontinuities reported in this investigation.

Notizen: Summary An autopsy case of a Japanese male with familial β-galactosidase and neuraminidase deficiency is reported. The clinical picture was characterized by adult onset, a gargoyle-like face, cerebellar ataxia, myoclonus, convulsions, retinal degeneration and cortical blindness. Histopathologically, most neurons seemed to have become degenerated in the whole cerebral cortex. Moreover, the calcarine cortex appeared spongy with depopulation of nerve cells. Stuffed neurons or neuronal storage changes were found throughout the brain, especially in the motor nuclei of the spinal cord and brain stem. The inclusions in the stuffed neurons revealed various profiles on the electron microscope. They were composed of membranous lamellar and/or multilamellar structures, often accompanying vacuoles and reminiscent of lipofuscin-like profiles.

Notizen: Summary The nervous system of a 22-week-old fetus with GM1-gangliosidosis type 1 was studied by electron microscopy. The tissues thus examined were the cerebral cortex at the parietal region, the cerebellum, the thoracic spinal cord, the Auerbach's myenteric plexus in the large intestine and the radial nerve fibers. In the cerebral cortex, membrane-bound vacuoles, which occasionally contained stacks of fine fibrils, were observed in the large young neurons in the deeper part of the cortical plate. The neurons in the other part of the cerebral cortex carried no storage materials. In the cerebellum, the membrane-bound vacuoles with stacks of fine fibrils were seen only in the Purkinje cells. The neurons in the spinal cord also contained several zebra-like bodies and the above membrane-bound vacuoles. As for the peripheral nervous system (PNS), neurons in the Auerbach's myenteric plexus carried membranous cytoplasmic bodies and zebra-like bodies. Some of the axons in the radial nerve fibers also contained a lot of pleomorphic electron-dense bodies and a few membranous cytoplasmic ones. These results show that the accumulation of storage materials is started in the large neurons which are produced in the early stage of neurogenesis in the central nervous system (CNS). Additionally, the observed membrane-bound vacuoles are considered to be structures which occur before the membranous cytoplasmic bodies and/or the zebra-like bodies. It is also elucidated that the PNS is affected earlier than the cerebral and cerebellar cortices and thoracic spinal cord.

Notizen: Summary An ultrastructural study of four pinealomas was carried out to precise eventual specific markers. Dark and clear cells joined with zonulae adherents, extensive and pleiomorphous processes, a complex vacuolar system, and characteristic organelles (lysosome-like structures, clear and dense-core vesicles, vesicle-crowned rodlets and related structures, microtubular sheaves and centriolar derivatives, membranous whorls, fibrous bodies, microtubules, heterogeneous cytoplasmic inclusions) offered a typical pattern. No correlation could be made between the histological and ultrastructural features. The authors stress the ultrastructural similarities between the human tumor cells and the mammalian pineal cells. Pinealomas appeared as a morphological entity distinct from neuronal and astrocytic tumors.

Notizen: Summary The ultrastructure of the porcine uterine glands is described from material taken from 11 pregnant pigs at exactly known stages of gestation (day 30; 58; 80; 100; 110). Fixation was performed by perfusion via a branch of the uterine artery and the tissue was routinely processed for electrom microscopy. Additionally, cytochemical studies (phosphotungstic acid reaction for glycoproteins, according to Rambourg 1967; acid phosphatase reaction; ultrastructural localization of cellular iron, according to Parmley et al. 1978) were performed. On day 30 of pregnancy the uterine glands are coiled, simple tubular glands with a narrow lumen. The epithelial lining is simple columnar and consists basically of two cell types, ciliated cells and secretory cells. The secretory activity of the glandular epithelium is low; only a few secretory granules are present in the supranuclear cytoplasm. At midpregnancy the ultrastructure of the glands has significantly changed and the cells now show all the characteristics of high secretory activity: numerous parallel cisternae of rough endoplasmic reticulum, an extensively developed Golgi apparatus and many secretory granules which give a positive reaction for acid phosphatase and glycoproteins. The lumina of the glands are significantly enlarged and filled with a great amount of a granular, acid phosphatase-positive material. In the last third of pregnancy, only minor changes in the ultrastructure of the uterine glands are observed. The secretory activity is still high. The amount of rough endoplasmic reticulum has further increased and parallel arrays of cisternae occupy a considerable part of the supranuclear cytoplasm. The importance of the uterine secretion for embryonic nutrition and development is only partly understood. One of the secreted glycoproteins, uteroferrin, is believed to play an important role in the iron transfer from mother to fetus. From midpregnancy onward, a special cell type, the “granule laden cell” is found scattered between normal secretory cells of the uterine glands. Contrary to the opinion of Perry and Cromby (1982), we could demonstrate that these cells frequently extend to the lumen of the gland; hence the term “basal cell” seems inappropriate for this cell type.

Notizen: Summary Tritiated thymidine autoradiography has established that after interrupting the commissural afferents to the dentate gyrus a number of non-neuronal cells proliferate in the molecular layer. In the present study the fine structure of the proliferating cells was analyzed by reembedding the 2-μm thick plastic sections of the dentate gyrus which had been previously coated with a nuclear emulsion and processed for light microscopic autoradiography. The location of the labeled cells was plotted with a camera lucida and a few ultrathin sections were taken from the re-embedded sections. In these the labeled cells were re-identified and photographed in an electron microscope. Most of the identified proliferating cells exhibited the following morphological features: The nuclei were irregularly oval, sometimes with deep indentations and contained dense clumps of chromatin; their diameters ranged between 4.5 and 6.5 μm. The cytoplasm was generally disposed to one side of the nucleus and often extended into a few broad processes. The Golgi apparatus was well developed. Many rosettes of free ribosomes were scattered throughout the cytoplasm, and the rough endoplasmic reticulum usually consisted of a few short cisternae. Small multilamellated bodies were common, but dense inclusion bodies were infrequent. The observations reported in this paper suggest: 1. that the nonneuronal cells which proliferate in a neuropil undergoing a mild denervation are morphologically closely related to microglia; 2. that in young adult animals these cells do not seem to have been previously involved in intense phagocytic activity; and 3. that the proliferating cells are present in the neuropil at the time of the denervation.

Notizen: Summary The histochemical profile and ultrastructural properties of the bulbospongiosus muscle (BSM) fibers from 5–6 month old boars and barrows (castrated at 7 days of age), and intact week old piglets were compared. Based on myosin ATPase, preincubated at pH 4.2, BSM of boars contained predominately intermediately staining fibers, whereas BSM of barrows and piglets had a mixture of staining intensities. Fibers from boar BSM stained intensely for SDH, with subsarcolemmal and diffuse location of reaction product. Staining intensity for SDH was variable in BSM from barrows and piglets, with diffuse location of reaction product. The BSM of boars and barrows contained predominately dark fibers when stained for glycogen and phosphorylase, and the fibers were low in stored lipids. While the fibers were smaller in barrow as compared to boar BSM, ultrastructural differences between boar and barrow BSM were not detectable.

Notizen: Summary The junctional complexes of the principal cells in the guinea pig epididymis were analyzed using freeze fracture and ultrathin section goniometric techniques. Replicas of the seven regions (I to VII) investigated reveal a continuous decrease in the number of tight junctional strands, ranging from 15.73±3.54 in zone I (proximal) to 4.39±0.78 in zone VII (distal tubule). The distance from the adluminal to the basolateral strand also diminishes from proximal, 0.73±0.02 μm to distal, 0.19±0.03 μm. The junctional strands appear on the P-face and anastomose forming compartments which are larger in the basolateral areas than those in the apical. The network of strands frequently form terminal loops and blind endings towards the more basal parts of the lateral membrane. Freeze fracture images also exhibit randomly distributed particulate aggregations which correspond to maculae adhaerentes, the highest number of which are found in zone IV, V, and VII. Desmosomal figures are found not only below, but also adjacent and intermingled among the tight junctional strands. This special junctional arrangement is confirmed upon goniometric analysis of ultrathin sections from zones IV, V, and VII. Electron dense desmosomal plaques are seen parallel and directly subjacent to the membranes of the tight junctions, following the strands in both directions to finally converge on the punctiform connections. Goniometry also reveals a dense feltwork of material closely applied along the lateral cell border. These zonulae adhaerentes are seen to be of greatest length and density in zones I, VI, and VII.

Notizen: Summary In this study the morphological features of lactation in the human breast were examined by scanning and transmission electron microscopy. The lactating lobules comprised large numbers of interconnecting acini which were lined by a single layer of epithelial cells with underlying myoepithelial cells. Marked variations were noted in the shape of the epithelial cells. The myoepithelial cells formed an open meshwork of interconnecting cytoplasmic processes packed with myofibrils. The basal cytoplasm of the epithelial cells was packed with rough endoplasmic reticulum while the apical cytoplasm contained a hypertrophic Golgi body, numerous vacuoles (a few of which contained casein micelles), a number of lipid droplets and small coated and uncoated vesicles. The lipid droplets were released by progressive protrusion from the apical surface. They remained covered by the plasmalemma and were finally budded off into the lumen. In certain cases a portion of cytoplasm was released with the lipid droplet. The vacuoles and small vesicles fused with the plasmalemma and released their contents by exocytosis. Within the samples the majority of epithelial cells were actively lactating although examples of undifferentiated “resting” and dead (lysed) cells were also identified.

Notizen: Summary The guinea pig epididymis is subdivided into seven zones. The ultrastructure and morphology of the principal cells in these zones is analyzed. The position, shape and content of the nuclei are variable along the length of the epididymal duct. Features characteristic of absorptive activity, such as micropinocytotic caveolae, vacuoles, and multivesicular bodies are of high concentration in zone IV and VI. The Golgi apparatus, rough endoplasmic reticulum and secretion granules are organelles and inclusions implicated to secretory functions and in this study are not found in the following concurring amounts within the principal cells of the seven zones: the Golgi apparatus exhibits a trend of increase from zone II to zone VII while the rough endoplasmic reticulum decreases. Secretion granules, though, are detected only in zones II and III, not only in the supra-, but also in the peri-and infranuclear regions. This possibly implies an exocrine secretory functions. Lamellar whorls and profiles of tubular smooth endoplasmic reticulum are concentrated in the supranuclear and adluminal regions of zones I, II and VI. A high concentration of large lipid droplets is a consistent feature of the perinuclear region of zone II. Mitochondia and lysosomes are detected in relatively large amounts along the epididymal duct. The correlations of these morphological characteristics with respect to their possible functional role are discussed.

Notizen: Summary Skin fibroblasts and tumor cells were cultured from four patients with peripheral von Recklinghausen's neurofibromatosis (NF). The cell type enriched in culture from the tumors carried the fibroblastic Thy 1.1. cell surface antigen and produced fibronectin, like fibroblasts from skin of NF-patients or from control persons. In electron micrographs the NF tumor and NF skin cells were similar to the control skin fibroblasts; elongated in shape, contained tubular mitochondria, variable amounts of granular endoplasmic reticulum, numerous lysosomal inclusion bodies and collections of 5 nm filaments. Trypsinized cells were fractionated with centrifugation in a Percoll density gradient. All cell lines produced only one sharp band of viable cells at the buoyant density of 1.03. Compared with the NF skin or control skin fibroblasts the NF tumor cells, however, produced a less well organized peri-and extracellular matrix estimated from fibronectin fluorescence. The nuclear sizes were measured from photographs of the cultures. The nuclei of all four tumor cell lines were larger than those of the skin fibroblasts of the corresponding patients. Neurofibromatosis tumor cells thus resemble skin fibroblasts in their density and in some ultrastructural properties but are different in their growth pattern and synthetic functions.

Notizen: Summary An analysis is presented of the immunohistological and ultrastructural features in a series of 118 surgically removed pituitary adenomas all of which were studied immunohistologically using antisera to growth hormone (GH), prolactin (PRL) ACTH, βFSH, βLH and βTSH, and 75 of which were studied ultrastructurally. Results were analysed according to the mode of presentation of patients. Forty-one (35%) of the tumours were from patients with acromegaly or gigantism, ten (9%) from patients with Cushing's syndrome or Nelson's syndrome, 19 (16%) from patients with clinical features associated with hyperprolactinaemia and 48 (40%) from patients with space occupying lesions which appeared clinically to be overtly endocrinologically functionless. By light microscopy, using the immunoperoxidase (PAP) technique, immunoreactive GH was demonstrated in all the tumours from patients with acromegaly or gigantism, immunoreactive ACTH in all tumours from patients with Cushing's syndrome or Nelson's syndrome and immunoreactive PRL in 95% of tumours associated with effects of hyperprolactinaemia. Forty-five percent of the tumours from acromegalic patients contained some PRL-positive cells as well as GH-positive cells. Among the tumours which appeared clinically to be endocrinologically functionless were three tumours (from males) uniformly stained for immunoreactive PRL. Of the remainder, 60% were negative for immunoreactive hormones and 40% contained small numbers of cells which were positive for a variety of immunoreactive hormones. ACTH-cell and PRL-cell tumours had ultrastructural features as described in previous studies. Fifty percent of GH-cell tumours examined at the EM level contained fibrous bodies, while in the remainder these structures were not identified. Tumours with fibrous bodies were more likely to contain PRL as well as GH with immunoperoxidase. All tumours that were endocrinologically functionless and which were examined at the EM level contained secretory granules. Oncocytic change was common in these tumours. No ultrastructural differences were observed between those which contained immunoreactive hormones by light microscopy and those which did not.

Notizen: Summary A leukoplakic lesion of the lip showing the histologic features of epidermolytic hyperkeratosis above solar elastosis was investigated by electron microscopy. The ultrastructural alterations observed in the upper epidermal layers corresponded in the main with those of epidermolytic hyperkeratosis in other skin diseases. The keratinocytes showed irregularly formed tonofilaments, a marked intracellular edema, and premature cornification. In addition, there were discrete subcellular signs of premalignancy in the cells of the basal and suprabasal layers. These alterations suggest that the epidermolytic leukoplakia represents a rare histopathologic variant of actinic cheilitis.

Notizen: Summary The ultrastructural localization of adenylate cyclase (E.C. 4.6.1.1.) and cAMP phosphodiesterase (PDE) (E.C. 3.1.4.17.) in the ectoderm of the developmental stage 4 chick embryo was studied. Adenylate cyclase was localized in the lateral surfaces of the ectodermal cells. In the primitive streak cells the enzymatic activity was observed on all the lateral surfaces, whereas in the periphery of the blastoderm the reaction product was localized in the apical parts of the lateral plasma membranes only. cAMP PDE localized in the apical cytoplasm of the ectodermal cells, with highest activity in the globular projections.

Notizen: Summary The self-differentiation potency of the endoderm of the chick embryo was investigated mainly by transmission electron microscopy. Endodermal fragments isolated from 4- to 6-day stomach or small intestine were cultured in the absence of mesenchyme and were able to differentiate in vitro into organ-specific epithelia. Endodermal fragments isolated from the stomach region differentiated into a pseudo-stratified epithelium with periodic acid Schiff-positive mucous granules in the apical cytoplasm, while those from the small intestinal region differentiated into a simple columnar epithelium with a striated border which was positive in alkaline phosphatase activity. These features are comparable with those of the mucous secretory epithelium of the normal embryonic stomach and the absorptive epithelium of normal embryonic small intestine, respectively. Next, the self-differentiation potencies were investigated of the upper and lower layers of the blastoderms, at stages 1–5 of Hamburger and Hamilton (H. and H.). Both stomach-type and small-intestine-type epithelia developed only when fragments of the lower layer isolated from the blastoderms older than stage 3 of H. and H. were cultured, suggesting that cells possessing the potency to differentiate into the stomach- and small-intestine-type epithelia exist in the definitive endoderm at the beginning of its formation.

Notizen: Summary A T-cell line, MT-2, derived from human cord blood lymphocytes by cocultivation with adult T-cell leukemia (ATL) cells is a continuous producer of type-C virus particles. Electron microscopy of MT-2 cells cultured for 1–3 weeks in medium containing 10% ATL patients' sera revealed agglutination of type-C virus particles within the electron-dense deposits in the extracellular spaces. No such agglutination occurred in control cultures supplemented with normal human or fetal calf serum. These results provide direct evidence for the specific reactivity of ATL patients' sera with type-C virus particles in the MT-2 cell line at the ultrastructural level.

Notizen: Summary Small electrolytic lesions were made in the flocculus of two adult cats by means of a stereotactic approach avoiding any damage to the cerebellar nuclei. After a survival time of 3 days the animals were killed and the brains fixed and prepared according to standard procedures for ultrastructural studies. The brains of two unoperated cats were similarly treated and served as normal controls. In the experimental animals a large number of boutons in the rostral part of the nucleus prepositus hypoglossi (Ph) ipsilateral to the floccular lesion showed degenerative changes. These were characterized by hypertrophy, a prominent aggregation of densely packed parallel tubules or concentric arrays of cisternae and a filamentous hyperplasia. Only very rarely were such abnormal boutons seen in the caudal half of the ipsilateral Ph, or on the contralateral side or in the unoperated animals. The degenerating boutons contain clusters of pleomorphic vesicles and they establish symmetrical synaptic contacts with somata, dendritic shafts and dendritic spines. Some of the degenerating boutons appear to be of the en passant type. These findings thus affirm the existence of a direct flocculo-prepositus projection in the cat. It is suggested that this pathway could be responsible for mediating information about eye position and velocity to Ph neurons.

Notizen: Abstract The sporogenesis of aerial spores in Streptomyces thermoviolaceus corresponded to a common sporulation type in the genus. The sporulation septum was composed of an outer ring-shaped constriction wall and an inner interspace septum arising by the inwards growth of a double annulus. In mature spores the wall was composed of two layers, the outer one was part of the parent hyphal wall and septum material, the inner one was formed de novo. The spore chains were enclosed by the thin breakable sheath containing small rod-like elements. The ornamentation in the form of knobs, which were a characteristic feature of the species originated from the sheath. The knobs were hemispherical particles with an inner electron dense core and an outer electron transparent shell. The term “cupular knobs” was suggested for this type of tuberculate ornamentation. Frequently, the knobs became detached from the surface in which case the inner core separated easily from the shell.

Notizen: Abstract Two strains of desiccation-tolerant coccoid cyanobacteria, Chroococcus S24, a marine form, and Chroococcus N41, a cryptoendolith isolated from a hot-desert rock, have been characterized. The mol % DNA base compositions of the strains are 47.1 and 48.9% respectively. Plasmid DNA was not detected in either strain. The pigment contents and nutritional characteristics of the strains are identical. Both lack phycoerythrinoid pigments and, in culture, behave as slow-growing halotolerant marine forms with elevated requirements for Na+, Cl−, Mg2+ and Ca2+. Sucrose was the only carbon source of those tested that supported photoheterotrophic growth. Each strain synthesizes nitrogenase under anaerobic conditions but not in air. Morphologically the two strains are indistinguishable. They are considered to be independent isolates of the same cyanobacterial species. Chroococcus N41 was studied in detail with the electron microscope. When brought to equilibrium at matric water potentials of-168 MPa and lower (to-673 MPa=c0.12a w) the protoplast shrinks, but the cells maintain the same size and diameter as those at-2,156 kPa (MN medium; control); the sheath expands and remains attached to the cell wall outer membrane by fibrils. The cell wall, cell membrane, thylakoid membranes, cyanophycin granules and carboxysomes appeared intact in desiccated cells.

Notizen: Summary Unilateral tenotomy of the Achilles tendon was carried out in 60 rabbits. The limb was then either mobilized directly or immobilized for 10 to 35 days using a plaster usually after tendon suture. In certain cases the plaster was removed early (on the 7th or 16th day) and the animals were than allowed to use this leg. Separation between tendon ends was apparent from steel markers, placed close to each cut end of the tendon and examined by X-ray. The separation curve was biphasic and both the first and the inactive phase reflected the degree of tension over the tendon suture. However, during the second separation phase, which began between the 17th and 21st day, the separation gradually reached the same level in all groups. Enzyme histochemistry and electron microscopy revealed severe degenerative changes in immobilized and in shortened muscles. Furthermore, a gradual shift in fibre type characteristics from type 1 slow-twitch fibres towards type 2 fast-twitch fibres occurred. Rapid recovery followed removal of the plaster. The findings indicated that both degenerative and regenerative processes and adaptive processes had been initiated in all experimental muscles when the tendon continuity was broken. The adaptive processes progressed gradually during the five-week post-operative period and might have been responsible for the second phase of the tendon end separation. The fibre adaptation, i.e. the transformation, may be accounted for by changes in structure of the myofibrils and composition of the myosin molecules.

Notizen: Summary Surgical specimens of 15 normal and 106 para-adenomous anterior pituitaries were studied immunocytochemically and in part electron microscopically for the presence of hyperplasia. GH cell hyperplasia was found in 13% of all normal pituitaries, in 6% of the cases with Prolactin secreting adenomas and in 9% of the cases with ACTH secreting adenomas. Prolactin cell hyperplasia occured in nearly equal percentages (17–23%) in normal pituitaries and in areas adjacent to GH-, Prolactin-or ACTH-secreting adenomas or adjacent to inactive adenomas. Previous findings of relatively more frequent Prolactin cell hyperplasia occuring together with Prolactin producing adenomas have to be revised. Prolactin cell hyperplasia as a primary source of hyperprolactinemia is very rare and almost always occurs in conjunction with oncocytic adenomas. ACTH cell hyperplasia was found in 13% of the normal pituitaries, in 14% of the cases with Prolactin secreting adenomas, in 58% of the cases with ACTH producing adenomas and in 40% of the pituitaries with GH secreting adenomas. We have no explanation for the latter result. ACTH cell hyperplasia may be the primary cause of Cushing's disease (18% of all Cushing cases). Hyperplasia of TSH cells in normal pituitaries was rare (7%) and with the exception of Prolactin producing adenomas (22%) was not found near adenomas. Clinical-pathological correlations are discussed.

Notizen: Summary Two cases of undifferentiated carcinoma of the parotid gland were studied by light and electron microscopy. Light microscopy showed nests of ovoid cells with scanty cytoplasm and pyknotic nuclei in two cases. One case was the small-cell type, and another one was the large-cell type histopathologically. Electron microscopy showed two distinct cell types in each tumor: Case 1 (small-cell type). — An epithelial-like cell, and an irregular-shaped cell containing bundles of filaments suggesting myoepithelial differentiation. Case 2 (large-cell type). — An epithelial-like cell, and a large cell containing secretory-like granules. These findings support a salivary duct epithelial origin for these tumors.

Notizen: Summary Axons in the duodenum, ileum and rectum of the domestic fowl were identified as catecholamine-containing (CA) on the basis of positive reactivity following chromaffin fixation for electron microscopy. CA-axons in association with blood vessels in all regions of the intestine and in non-vascular sites in the small intestine had a ‘ typical’ adrenergic appearance, in that they contained many small granular vesicles (SGV) and variable numbers of large granular vesicles (LGV). In the rectum the non-vascular CA-axon profiles were atypical, in that there were many elongated LGV and few SGV, and the chromaffin reactivity was weak. The nerve profiles in the rectum were dramatically reduced following 6-hydroxydopamine and reserpine treatment and were absent in rectum cultured in the absence of extrinsic ganglia. It was concluded that the profiles, in spite of their low chromaffin reactivity, truely represent CA-axons. The possibility was raised that the atypical morphology and reduced chromaffin reactivity is due to the presence of adrenaline.

Notizen: Summary Corticotropes of rat fetuses aged 16, 18 and 21 days were localized by the indirect antibody-enzyme method on semithin sections of the pituitary. The development of the ultrastructure of these cells was observed on consecutive ultrathin sections. In comparison with previous data our present results show that identification of a fetal cell type cannot be based entirely on morphological criteria. The structural peculiarities of corticotropes obtained from studies in vivo are compared with those observed in cells maintained in vitro.

Notizen: Summary Cyclophosphamide-induced changes in rodent odontogenesis were investigated by light and electron microscopy in four-day-old Sprague Dawley rats given one injection of 40 mg/kg of body weight of cyclophosphamide and killed at intervals of one hour, one day, one week and two weeks. Incisor and molar teeth were dissected from the animals, fixed in 2.0% glutaraldehyde in 0.1 M sodium cacodylate with 3.4% sucrose, and subsequently some were incubated for alkaline phosphatase reaction, and embedded in Spurr's medium for sectioning at light- and electron-microscopic levels. From three days a cell-sparse zone was created in the pulp in the growing end of the tooth and progressive cellular changes were observed which became more severe in the one-week and two-week specimens. Subodontoblast and adjacent pulpal cells were the most affected showing nuclear changes, damage to, or loss of, organelles, and inclusion bodies. Odontogenic epithelium was less affected and odontoblasts appeared to be unaffected by the drug. A new irregular matrix was laid down in the defect area and seemed to be the product of depolarized odontoblasts. This new matrix showed alkaline phosphatase activity, as did the cells embedded in it, and later it became mineralized. It is speculated that the polarity of odontoblasts might be maintained by an intact subodontoblastic layer; when this is lost the odontoblasts become depolarized and capable of secreting matrix from both ends.

Notizen: Summary Work on endothelial cells has been limited by the availability of procedures for obtaining such cells in quantities adequate for direct in vitro analysis. The present paper describes a method for the isolation of endothelial cells from bovine cavernous bodies. A number of cells ranging from 2.5 to 4 × 108 per animal has been obtained. The cells were identified as follows 1) presence of the “Weibel and Palade” bodies in the isolated cells, 2) “cobblestone” appearance of cell cultures, and 3) presence of factor VIII, as demonstrated by immunofluorescence assays. The cell viability at the end of the purification procedure was tested 1) by dye-exclusion tests and 2) by metabolic assays. Features of this preparation are 1) the very high yield of viable endothelial cells, 2) the absence of contamination by fibroblasts and smooth muscle cells and a very low contamination by erythrocytes and 3) the fine dispersion of the isolated cells. These properties allow functional and subcellular fractionation studies on freshly isolated endothelial cells of microvascular origin.

Notizen: Summary The common ventricle in the heart of the Thunnus alalunga was studied. The ventricular myocardium consists of an outer compact layer and a thick inner spongy layer. The compact layer has slightly larger cells (4–6 μm diameter) than the spongy layer (2.5–5 μm diameter). Ultrastructurally the myocardium displays normal arrangements of myofibrils and mitochondria. The sarcoplasmic reticulum is poorly developed. The intercalated discs are simple with the fascia adherens being the most frequent junctional type observed; occasionally a desmosome was seen. Nexus type junctions are present but are unassociated with the intercalated discs. There are no t-tubules evident but the plasmalemma exhibits numerous caveolae which rarely form couplings with the sarcoplasmic reticulum. A morphometric analysis of the volume percent of mitochondria and myofibrils showed that the myocardial cells in the spongy layer of the heart have a significantly greater volume percentage of mitochondria than the compact layer. No significant differences were found between myocardial regions when the volume percentages of myofibrils were compared. The physiological studies revealed that the albacore tuna has heart rates (120 bpm) and ventricular blood pressures (100 mmHg) that are among the highest reported for fish.

Notizen: Summary The anatomical distribution of smooth muscle actin, myosin, fibronectin and basement membrane has been investigated immunohistochemically, using the indirect immunofluorescence technique, in the rat epididymis. The findings were correlated with the ultrastructural organization of the organ. Actin was found to be distributed in the stereociliary region of the epithelial principal cells and in the terminal web region. Actin was also visible along the base of the epithelium. Myosin was detected in the terminal web and in the terminal bar regions of the epithelium. The contractile cells showed a strong stain for both proteins. Basement membrane immunoreactivity was distributed along the epithelial basement membrane and around the contractile cells of the wall. In the cauda, between the epithelium and the contractile cell layers, the lamina propria, containing blood vessels and a thin layer of cells, was negative for all antigens investigated. Fibronectin showed a granular distribution around the contractile cells, mainly in the cauda. The ultrastructural study showed only thin (5–6 nm in diameter) filaments in the stereocilia and terminal web region. Thin filaments were also visible in the cytoplasm of the basal cells, thus suggesting a contractile function of this cell type. The heterogeneous appearance of the contractile cells of the wall seemed to support the different contractile pattern of the epididymal regions: caput, corpus and cauda. The cells present in the lamina propria showed cytoplasmic vesicles with dark granules resembling the “A” cell granules of the endocrine pancreas and gut mucosa cells.

Notizen: Summary Microbasic p-mastigophores, euryteles of two size groups, holotrichous isorhizas and atrichous isorhizas, comprise the cnidom of Chironex fleckeri, a cubozoan that has been responsible for several human fatalities. In its undischarged state each microbasic mastigophore of C. fleckeri consists of a capsule containing matrix and an inverted tube possessing a smooth-walled butt which is loosely coiled helically and which narrows to form a thread that is tightly coiled helically and markedly pleated. Both butt and thread carry three helices of spines and contain a granular matrix. During discharge, the proximal butt spines form initially a piercing stylet. Granular material from the butt and thread is released prior to the release of capsular material. Each eurytele possesses a tube with a butt composed of three bulbs, the middle bulb bearing long spines. Each holotrichous isorhiza possesses a coiled tube bearing small spines along its length. Each atrichous isorhiza exhibits a tube that is devoid of spines and loosely folded in the undischarged condition. The probable role of each type of nematocyst is inferred from its structure and features that enable the ready separation of the nematocysts of C. fleckeri from those of scyphozoan jellyfish are discussed.

Notizen: Summary The ultrastructural characteristics of ciliary epithelium from bovine, pigmented rabbit, and fetal albino rabbit were studied in cultured explants. The tips of ciliary processes were cultured in plastic dishes with Dulbecco Modified Eagle Medium (DMEM) containing 5% fetal bovine serum. More than half of the expiants adhered to the plastic culture dish, and epithelial cells spread as monolayers within a few days. Initially the explant contains two layers, the outer (nonpigmented cells) and the inner (pigmented cells). Later the explant exhibits three layers: 1) outermost lightly pigmented flattened cells, 2) an outer layer of nonpigmented cells, and 3) an inner layer of densely pigmented cuboidal cells. The cells of the outermost layer are continuous with the cells of the inner layer. A narrow space lies between the outermost layer and the outer layer. The columnar cells in the outer layer contain well developed organelles but no pigment granules; they possess a basement membrane, lateral interdigitations, and junctional complexes near their apices. Numerous focal junctions and some ciliary channel-like structures were detected between the columnar cells of the outer layer and the cuboidal cells of the inner layer. The cuboidal cells of the inner layer are filled with pigment granules. These observations suggest that the columnar cells of the outer layer are nonpigmented epithelium, the cuboidal cells of the inner layer are pigmented epithelium, and the flatened cells in the outermost layer are derived from pigmented epithelium.

Notizen: Summary Thyroid glands from 40 human fetuses ranging from 30 to 175 mm in length were examined by electron microscopy. At the 30 mm stage, the cells are arranged in cords or clusters and contain numerous free ribosomes. The rough-surfaced endoplasmic reticulum (RER) is poorly developed and the Golgi complex consists of saccules, smooth and coated vesicles. Mitochondria, microfilaments, glycogen particles, occasional lysosome-like structures, and single cilia are observed. At sites of incipient follicle formation, intercellular clefts containing interdigitating microvilli, delimited by junctional complexes, are evident. In addition, the apical region of some cells contains vacuoles. At the 50-mm stage, intercellular clefts are more apparent, some of them organized into small follicles through the enlargement of the lumen, the incorporation of additional cells, and the formation of a basal lamina. With increasing gestational age, more follicles are established. The lumina vary from round to clover-leaf shaped and have an electron-dense content. Cisternae of RER are dilated and contain flocculent material. The Golgi complex is more extensive, and lysosome-like structures are more numerous. By the 100-mm stage, the follicular cells appear morphologically well differentiated. The results demonstrate an extracellular origin of the follicular lumen in the human fetal thyroid.

Notizen: Summary In a previous study changes in the macrodistribution of fibronectin during rat-lung development were examined. Using the peroxidase-antiperoxidase immunocytochemical technique, we have demonstrated the presence of fibronectin in embryonic, neonatal, and adult rat lung at the ultrastructural level. In the embryo, fibronectin is found both in an intra-and extracellular association with isolated pneumoblasts, and in a periodic distribution along the basal lamina. The neonate displays fibronectin in an intracellular association with early type-I cells and on their basal and luminal surfaces, but not in association with type-II cells. Neonatal basal lamina is diffusely labeled by anti-fibronectin antiserum. Fibronectin in adult tissue is found both intracellularly and on the basal and luminal surfaces of type-I cells but not in type-II cells. The basal lamina and interstitial connective tissue are slightly or non-reactive. These observations confirm and extend our initial suggestion that fibronectin is involved in rat-lung development.

Notizen: Summary The fine structure of conspicuous extracellular materials during the life history of a sea star (Patiria miniata) is described. The outer surface of the developing sea star is covered by two morphologically different cuticles that appear sequentially during ontogeny. The primary cuticle, which is about 120 nm thick and two-layered, is present from mid-blastula through the end of the larval stage. The secondary cuticle, which is about 1 μm thick and three-layered, first appears on the epidermis of the rudiment region of the larva and, after metamorphosis, covers the entire epidermis of the juvenile and adult stages. During ontogeny, there are only two conspicuous gut cuticles: the first lines the newly invaginated archenteron at the start of the gastrula stage, and the second lines the esophagus during the larval stage. A blastocoelic basal lamina first appears at mid-blastula and persists as subectodermal and subendodermal basal laminae. Ruthenium red-positive granules are detectable between the lateral surfaces of adjacent ectodermal cells during part of the gastrula stage; this transient intercellular material may possibly aid in lateral adhesion between cells.

Notizen: Summary When grown in iron-free media, the youngest leaves of healthy green sugar beet plants became completely yellow after 6 to 8 days. This chlorosis was quickly reversed by resupplying iron. A study of the ultrastructure of the iron -stressed leaves revealed apparently normal subcellular organization except for the plastids which were small and undeveloped, contained a rudimentary, disorganized grana-fretwork and clusters of vesicles in the periphery. Twelve to 16 hours after resupply of iron, aggregates of phytoferritin were observed in the stroma, and the granal fretwork underwent further development. There was an increased orientation of the membranes along the long axis of the plastids and an increase in the length of the individual grana stacks. By 48 hours, leaf chlorophyll content was about 40% of the control. At the ultrastructural level, parallel alignment of membrane orientation was complete and the grana stacks began to increase in the number of thylakoids per stack.

Notizen: Summary The karyosome is a spherical body up to 1 μm in diameter that lies on the nucleolus of certain plant species, particularly those with a relatively low nuclear DNA content and an areticulate nuclear structure. It can be seen in the light microscope after impregnation with silver; in the electron microscope its structure consists of fibrillo-granular material. Nucleoli of cells in root apices may bear 0, 1, or 2 karyosomes. The frequency with which these numbers of karyosomes are observed depends on the location of the cells within the apex. In roots ofPisum sativum andZea mays the nucleoli of both slowly-dividing and young differentiating cells bear karyosomes more frequently than the nucleoli of rapidly-dividing cells. The karyosome seems to adopt a preferred location on the nucleolus, lying most frequently on the nucleolar surfaces directed towards the apex or base of the root. The origin and functional significance of the karyosome are discussed. Morphological evidence suggests that it may be material that formerly was part of a fibrillar centre.

Notizen: Summary Cuticle and pore development in the guard cells ofFunaria were investigated with the electron microscope. Pore cuticle formation is simultaneous with the creation of the pore itself. The morphology of the pore cuticle is unlike that of any cuticle described in the literature. It has many lamellae which are penetrated by electron dense fibrils. Three different cuticular morphologies exist from the pore to the subsidiary cell walls. The cuticles on the pore and outer walls contain fibrils that sometimes reach to the surface. The subsidiary cell cuticle lacks fibrils altogether. It is hypothesized that (1) cuticularization of the middle lamella contributes to ventral wall separation and (2) differences in extent of cuticular fibrils are related to greater water loss from stomata than from subsidiary cells (peristomatal transpiration).

Notizen: Summary Microtubule systems appear sequentially at the distal and proximal poles of tetrad members during mid-sporogenesis in the mossTetraphis pellucida Hedw. The distal microtubule system emanates from a microtubule organizing center (MTOC) located between the single plastid and the nucleus. The distal MTOC and associated microtubules, which appear immediately after cytokinesis, are ephemeral and do not appear to be associated with the deposition of exine occuring at the same time. The proximal microtubule system, which appears slightly later than the distal system, is a more stable component of mid-sporogenesis. The proximal MTOC is an irregularly lobed, patelliform aggregation of electron-dense granules located beneath the plasma membrane at the proximal spore pole. Several bundles of microtubules radiate from the proximal MTOC and traverse the cell, enclosing the nucleus in an cone of microtubules. The proximal microtubule system is thought to function in aperture development and organelle migration. The relatively large nucleus migrates a short distance in the small spore early in the tetrad stage and maintains its acentric position at the proximal pole throughout later stages of sporogenesis. The plastid migrates later in the tetrad stage from its meiotic position parallel to the distal surface to a position perpendicular to the distal surface with one tip in close proximity to the proximal MTOC. The proximal microtubule system reaches its maximum development by the end of the tetrad stage and all micrographic evidence of it is lost in the maturation stages of late sporogenesis.

Notizen: Summary The differentiating stages of coremia and rhizomorphs inSphaerostilbe repens were studied by transmission electron microscopy. Vegetative mycelium is characterized by highly cytoplasmic cells rich in ribosomes and mitochondria and with few vacuoles as well as endoplasmic reticulum. Cell walls are thin attaining a maximum thickness of 0.10 μm. During the aggregating phase a prosenchymatous mass of randomly oriented cells is produced by localized elongation and branching of the filaments. The hyphae in this region have the appearance of actively metabolising cells. In the course of the differentiating phase, numerous hyphae of the median zone of the aggregate grow upward and downward to give rise to coremium and rhizomorph primordia respectively. The individual hyphal tips lay parallel to each other and cells of the growing apices retain their meristematic characteristics. At the periphery of the aggregate and to a lesser extent in the subapical rhizomorphic zone, cells reduce their cytoplasmic density as a consequence of a decrease in the number of ribosomes. These cells also increase in size and become isodiametric and vacuolated. During cellular differentiation walls increase steadily in thickness and at the elongating phase they reach 0.30 μm in the rhizomorphic cortex. Mucilaginous material is progressively deposited around hyphae and in the most differentiated zones, coalesce to fill interhyphal spaces. This extracellular matrix seems to play a role in maintaining cohesiveness of the aggregated organs. The tissue in the process of differentiation is scattered with cells highly enriched in mitochondria and with cells virtually undifferentiated. Accumulation of microfilaments takes place in the differentiating zone localized behind the immersed meristematic apex. These structures might be involved in wall synthesis. Glycogen rosettes accumulate in the vegetative mycelium surrounding the aggregating centers, suggesting the possibility of supplying energy during the differentiating processes. The vacuolar system, represented by autophagic vacuoles which are present until the differentiation phase, presumably may also participate in the biochemical changes that occur during aggregation. Coremial cells are characterized by an increase in wall thickness, a highly sinuous plasma-membrane as well as large amounts of mucilaginous compounds accumulated between hyphae, but in all other respects they resemble the cells of actively growing vegetative hyphae.

Notizen: Summary Sorogena stoianovitchae Bradbury andOlive is an unusual kinetophragminophoran ciliate characterized by the aggregation of individual trophic cells to form a multicellular sorogen that rises aerially from the medium surface to produce a sorocarp. A light and electron microscope study of sorogenesis revealed how the sorogenic cells produce the stalk. The feeding ciliates contain numerous subpellicular expansions of rough endoplasmic reticulum cisternae that are only one element of the secretory system necessary for stalk formation. Upon aggregation, the stalk secretory system develops further, initially with the proliferation of the subpellicular expansions. Then, during later aggregation and early sorogenesis, other secretory elements appear: stalk material vesicles, dense granule vesicles, and electron lucent vacuoles. These are all derived from the rough endoplasmic reticulum. Only the subpellicular expansions and stalk material vesicles appear to add to the stalk matrix. The dense granule vesicles could be precursors of the stalk material vesicles; the electron lucent vacuoles, which can be demonstrated to contain fibrillar material, are perhaps by-products of the secretory process. Sorogenesis inSorogena occurs by a mechanism unique among the stalk-producing protists that have been studied.

Notizen: Summary The sorogenic cells of the haptorid ciliateSorogena stoianovitchae Bradbury andOlive secrete a hydrated, fibrillar matrix that becomes organized to form a stalk. This light and electron microscope study examines the role of the secreted stalk matrix in the development of the sorocarp. The stalk material is secreted as a compact clump or clumps of material that expands as it hydrates. The expanded stalk material is initially viscous, but soon, through dehydration or cross-linking, solidifies to form a sheath. The sheath is the outside layer of the stalk, provides structural support, and controls the expansion of the newly secreted stalk material. The sorogenic cells continue to secrete stalk material that expands once it is outside of the cell. The resulting expansion force is directed upward by the sheath, thereby moving the cells further above the surface. The continued dehydration of the sheath material provides an inwardly directed force that also contributes to the upward movement of the sorogen.

Notizen: Summary Cytochrome oxidase (EC 1.9.3.1) activity of conidia of wild-typeNeurospora crassa was detected cytochemically at the ultrastructural level by 3,′-diaminobenzidine (DAB) reaction during heat treatment at 46 °C and after shift down to 25 °C. At 46 °C the decrease in the enzyme activity was shown to be time-dependent (0–10 hours) in the progressively overswelling conidia. Gradual recovery of the DAB reaction occurred in mitochondria of conidia outgrowing conidiogenous germ tubes (microcycle conidiation) when shifted down to 25 °C. Inactivation of cytochrome oxidase was efficiently prevented during heat treatment in the presence of Tween 20. Such conidia no longer overswelled but still required shift down to 25 °C to produce germ tubes with delayed conidiogenous ability.

Notizen: Summary The effect of UV-C (254 nm) and UV-B (290–320 nm) radiation on leaves ofBeta vulgaris L. at the ultrastructural level was investigated. Although the damage caused by UV-C radiation was more striking than that resulting from UV-B radiation, several structural changes were seen in the UV-B treated material. Generally the effects of UV-B and UV-C radiation were different, suggesting different mechanisms of action, discernible even at the ultrastructural level.

Notizen: Summary A tapetum is found around all higher plant meiocytes and is thought to nourish them. It may, in turn, be influenced by their development. The mature tapetal membrane in amoeboid (or periplasmodial) tapeta, of whichArum italicum is an example, fits closely around the developing meiocyte. This description of tapetal ultrastructure starts from the meiotic prophase when the tapetum is still cellular and comprises two rows of cells on the inside of the tetrasporangiate anther. The radial walls of the still cellular tapetum start to dissolve during leptotene of the first meiosis. The lysis begins in the middle lamella in those areas penetrated by the most plasmodesmata. The walls in contact with the meiocytes on the other hand do not disappear until after the first meiosis. At telophase the now fused cytoplasmic tapetal mass begins to extend its joint plasmamembrane, amoeboid fashion, into the loculus. A cluster of microtubules can be seen at the apex of this intrusion apparently initiating or maintaining the shape of the invading plasmamembrane front. The tapetum now adheres closely to the microspores. The tapetal zone closest to the spores has a prominent population of microtubules and just a little ER, whereas the outer zone away from the spores contains all the other organelles. The inner zone, in a squash preparation, is not readily separated from the spores. The microtubules, at the middle microspore stage, are no longer to be seen in a circle around the microspores, but spread out with some lying orthogonal to the now-forming exine surface. In places the tapetal plasmamembrane begins to retract from the exine leaving roughly cone-shaped zones (spines) which become filled with fibrillar material. This material begins to be deposited on the exine surface. These “spines” are both PAS. and Coomassie-blue positive and susceptible to acetolysis.

Notizen: Summary Cytoplasmic reserves and extracellular substances were progressively broken down and utilized during carpogenic germination of sclerotia ofSclerotinia minor. Glycogen, wall polysaccharides and polyphosphate granules were removed first from regions of the sclerotium distant from developing apothecia, while protein bodies near the base of apothecial stipes were hydrolysed before those further away. The number of profiles of mitochondria and endoplasmic reticulum in cortical and medullary hyphae increased at the onset of germination, indicating increased metabolism in the hyphae. In contrast to developing sclerotia, simple pores with Woronin bodies were frequent in walls and septa during germination. Hyphae that appeared to converge towards the base of apothecial initials retained their cytoplasm and organelles until late in germination and hydrolysis of their reserves was delayed; these are interpreted as translocatory hyphae, although further work is required to determine their role unequivocally. When apothecia were fully developed, hyphae throughout the sclerotium were empty and the walls and extracellular matrix of cortical and medullary hyphae had almost completely broken down.

Notizen: Summary Mitotic ultrastructure was observed in meristematic cells of carposporophyte generations of the freshwater red algaBatrachospermum ectocarpum. Prior to nuclear division, cell elongation occurs resulting in the nucleus being located at the proximal end of the cell and separated by a large central vacuole from a distal “empty region” which the daughter nucleus eventually occupies. In late prophase, nuclear envelope-attached polar rings are situated adjacent to shallow nuclear invaginations. At prometaphase the nuclear invaginations extend deeply into the nucleus forming continuous cytoplasmic channels containing microtubules. Perinuclear ER and a typical equatorial chromosomal plate are formed by metaphase at which time the nuclear envelope lining the cytoplasmic channels has dispersed. Chromosomal and non-chromosomal microtubules converge at the poles where a single, large gap is seen in the otherwise intact nuclear envelope. Polar rings were not detected in the few mitotic cells observed beyond prometaphase but are thought to be present. During anaphase an interzonal midpiece is formed and the distal-most incipient daughter nucleus moves laterally past the central vacuole into the apical “empty region”. Features of mitosis inBatrachospermum are believed to be intermediate between those exhibited by the lower and higher orders of red algae, this being consistent with the taxonomic placement of the genus in theNemaliales, the least advanced order of the classFlorideophyceae.

Notizen: Summary Nuclear bodies are found in interphase nuclei of root apices of a number of plant species. They often show differences in structure and position relative to the nucleolus and this has led to an attempt to define two classes of body. However, in some species their separation into two classes on structural grounds alone breaks down, indicating that although they may occupy different positions within the nucleus they may in these particular cases be only different forms of the same body. The two extremes of the range of bodies examined represent what have been called “nucleolus-associated body” (karyosome) and “dense body”. The nucleolus-associated body is typically attached to, or adjacent to, the nucleolus. It is composed of fibrils 4–8 nm wide and often has an open structure showing compound threads or fibrils separated from each other by electron-lucent spaces. The dense body is more compact in structure and typically lies free in the nucleoplasm. Both types of body have an affinity for silver ions which, together with their staining reaction following treatment with EDTA, indicates that they consist of ribonucleoprotein. The characteristics of nuclear bodies found in different plant species have some relationship with the structure and DNA content of the interphase nucleus. Nucleolus-associated bodies are characteristic of species with an areticulate nuclear structure (2 C DNA content 〈6 pg), while dense bodies are common in species with a reticulate nuclear structure (2 C DNA content 〉6 pg). The possible functions of the two forms of nuclear body are discussed.

Notizen: Summary Emiliania huxleyi is a marine coccolithophorid which produces coccoliths,i.e., particles consisting of calcite and macromolecular organic material. The coccoliths are formed intracellulary in specialized organelles which comprise a coccolith vesicle (CV) and a reticular body (RB), together forming the CV/RB system or calcifying system. After termination of calcification, the coccolith is extruded and incorporated into the coccosphere,i.e., one or several layers of extracellular coccoliths surrounding the cell. Apart from the coccolith-producing cells (C cells) ofE. huxleyi, there are naked cells (N cells) which seem to have lost the capacity to produce coccoliths but are very similar to the C cells in other morphological respects. Biochemical studies have revealed that polysaccharides may play a regulatory role in calcification. The aim of the present study was to determine the localization of polysaccharides in both C and N cells electron microscopically. For this purpose, a cytochemical staining technique according toThiéry (1967) was applied. The CV/RB system of C cells was conspicuously stained. Due to the excellent stainability of this system, a putative succession of morphological stages during coccolithogenesis could be described. The staining pattern of the N cells closely resembled that of the C cells. It was found, however, that the “calcifying” system of N and C cells differed in both morphology and position. It is suggested that the divergent morphology of the “calcifying” system of N cells accounts for its failure to produce coccoliths.

Notizen: Summary The endodermis of bothTilia cordata andPicae citchensis progressess through 3 characteristic phases of development. These developments are delayed somewhat in the xylem pole endodermis ofT. cordata, while inP. sitchensis 3–5 passage cells are found. The cortex ofT. cordata is characterised by very thick walls, while that ofP. sitchensis is characterised by a thick walled layer just outside the endodermis and by 2–3 outer layers of transfer cells.

Notizen: Summary The ultrastructural characteristics of multinuclear foreign-body giant cells (MCG) in a human vocal cord Teflon granuloma are described. The cells were found to contain varying numbers of Teflon particles within their cytoplasm. The particles, rounded or oval in shape, were surrounded by a rather elctron-dense membrane. Numerous lysosomal structures were discerned within the cells. The fine structure of MCG in granulomas induced by foreign materials other than Teflon has already been described in man as well as in several experimental animals, and the Teflon MCG resembled these cells in many respects. No evidence of malignant change was found.

Notizen: 13C-NMR. Spectra of Dicyanohydroquinones and Dicyano-p-benzoquinonesThe 13C-NMR. spectra of 12 dicyanohydroquinones and fully substituted dicyano- and diazido-p-benzoquinones have been measured and assigned by comparison with related benzene and p-benzoquinone derivatives. For p-benzoquinones the influence of the cyano and azido substituents on the chemical shift of the CO-, α- and β-C-atoms has been investigated.

Notizen: Thermal cyclization of the pyrimidine-N-oxide dicarbamate 2 gives a 95:5 mixture of the 2-oxo-2H-[1,2,4]oxadiazolo[2,3-a]- and [2,3-c]pyrimidinecarbamates 3 and 4. An efficient procedure for the conversion of 3 to 4, and vice versa, is described.

Notizen: Polarographic Investigations on Symmetrical Triazacarbocyanine Dyes in MethanolThe reduction mechanism of eleven symmetrical triazacarbocyanine dyes with different heterocycles in methanol (containing lithium chloride or lithium acetate/acetic acid) has been investigated by means of polarography, cyclic oscillopolarography and coulometry.The reduction occurs in a reversible to irreversible two electron transfer step, followed by an irreversible chemical step. Reduction potentials, transfer coefficients and specific currents have been measured and compared with properties measured in acetonitrile.The effect of substituents X has been studied on the compounds 1-ethyl-2-[3-(1-ethyl-6-X-1,2-dihydroquinolin-2-ylidene)-1-triazeno]-6-X-quinolinium tetrafluoroborate 1f(X) and 3-ethyl-2-[3-(3-ethyl-5-X-Δ4-1, 3-thiazolin-2-ylidene)-1-triazeno]-5-X-1, 3-thiazolium tetrafluoroborate 1g(X), and ρ-values of 0.20 and 0.40 V, respectively, have been found.

Notizen: The anion [(OH)3(O)AsCH2As(O)(OH)2] has been trapped in an X-irradiated single crystal of methylene diarsonic acid. The g-tensor and the hyperfine coupling tensors of the two 75As-nuclei have been obtained and the spin densities in the arsenic 4s- and 4p-orbitals determined.

Notizen: Excited singlet states of bicyclobutylene-benzene (2), 1,2:4,5-bis (bicyclo-butylene)benzene (3), 2,3-bicyclobutylene-naphthalene (4), and 1,8-bicyclobutylene-naphthalene (5) are investigated by means of linear dichroic absorption spectroscopy of molecules oriented in stretched polyethylene films and by semiempirical model calculations. The results indicate a strong hyperconjugative impact of the bicyclobutylene group on the aromatic chromophores in these compounds. Valence isomerization to aromatic products is predicted as the preferred photochemical pathway.

Notizen: The hydrogenation of 2′, 3′-O-isopropylidene-5-methyluridine (1) in water over 5% Rh/Al2O3 gave (5 R)- and (5 S)-5-methyl-5, 6-dihydrouridine (2), separated as 5′-O-(p-tolylsulfonyl)- (3) and 5′-O-benzoyl- (5) derivatives by preparative TLC. on silica gel and ether/hexane developments. The diastereoisomeric differentiation at the C(5) chiral centre depends upon the reaction media and the nature of the protecting group attached to the ribosyl moiety.The synthesis of iodo derivatives (5 R)- and (5 S)-4 is also described. The diastereoisomers 4 were converted into (5 R)- and (5 S)-2′, 3′,-O-isopropylidene-5-methyl-2, 5′-anhydro-5, 6-dihydrouridine (7).

Notizen: The Acylation of 5-Amino-1 H-1,2,4-triazoles. A 13C-NMR. StudyThe acylation of 3-substituted-5-amino-1 H-1,2,4-triazoles (1) with methyl chloroformate or dimethylcarbamoyl chloride yielded mainly 1-acyl-5-amino-1,2,4-triazoles (2 and 3). Acylation of 3-methyl-, 3-methoxy- and 3-methylthio-5-amino-1 H-1,2,4-triazole (1b, 1c and 1d) with methyl chloroformate gave up to 10% of the 1-acyl-3-amino-1,2,4-triazoles. For the unsubstituted 5-amino-1,2,4-triazole (1a), a (1:1)-mixture of the 3- and 5-isomers 2a and 4 was obtained in dioxane in the presence of triethylamine. No 4-acylated product was detected in contrast to earlier reports. The structures of the reaction products were determined with the aid of proton coupled 13C-NMR. spectra using the corresponding N-methyl-1,2,4-triazoles as reference compounds.

Notizen: The X-ray crystallographic molecular dimensions and thermal response of the 5, 13-diphenyl-cyclonoa [b]naphthalene 1a, are described.

Notizen: 7,7-Dimethyl-6-methylidenetricyclo[6.2.1.01,5]undec-9-ene (10) has been prepared from allylic alcohol 8 in one step in 16% yield. Selective hydrogenation of 10 with diazene gives the 2-norzizaenes 1a and 11.

Notizen: STO-3G calculations show that the nitrocyclopropyl anion prefers a triplet ground state with a tetrahedral C(α)-atom and a perpendicular orientation of the three-membered ring and the NO2-group, 1-90° (Table 1). This unusual result is in agreement with recent experimental findings of Kai et al. (Helv. Chim. Acta 65, 137 (1982)). Other NO2-anions as well as all CHO-, COOH- and CN-substituted anions 2-12, have singlet ground states 2-0°-12-0° (Table 1), in agreement with copious experimental data (Wagner & Boche, Z. Naturforsch. 37b, 1339 (1982)).

Notizen: A general method for enlargement of carbocyclic rings by the so called zip reaction is given. The Michael adducts of 2-nitrocycloalkanones with 3-oxo-4-pentenoates in the presence of tetrabutylammonium fluoride give in high yield compounds with the ring enlarged by four C-atoms. By this method 7-, 8-, and 12-membered cycloalkanones were converted respectively to 11-, 12-, and 16-membered functionalized carbocycles (see Scheme 2 and 3).

Notizen: Pure alkaline phosphatase (EC 3.1.3.1; 1500 U/mg) was dialyzed at 4° during 168 h against water, 10-4M EDTA or 10-4M o-phenanthroline. During the dialysis, samples were periodically removed and analyzed for metal content and activity. The results indicate that 1 mol of native calf intestinal alkaline phosphatase contains 4 g-atom of zinc and 4 g-atom of magnesium tightly bound, and that both metal ions are necessary for full enzyme activity. The dialyzed, partially demetallized enzyme could be reactivated by the addition of zinc and/or magnesium salts.

Notizen: Stereochemical Correlations between (2R,4′R,8′R)-α-Tocopherol, (25S,26)-Dihydroxycholecalciferol, (-)-(1S,5R)-Frontalin and (-)-(R)-LinalolThe optically active C5- and C4-building units 1 and 2 with their hydroxy group at a asymmetric C-atom were transformed to (-)-(1S,5R)-Frontalin (7) and (-)-(3R)-Linalol (8) respectively; 1 and 2 had been used earlier in the preparation of the chroman part of (2R,4′R,8′R)-α-Tocopherol (6a, vitamin E), and for introduction of the side chain in (25S,26)-Dihydroxycholecalciferol ((25S)-4), a natural metabolite of Vitamin D3. The stereochemical correlations resulting from these converions fit into a coherent picture with those correlations already known from literature and they confirm our earlier stereochemical assignments. A stereochemical assignment concerning the C(25)-epimers of 25,26-Dihydroxycholecalciferol that was in contrast to our findings and that initiated the conversion of 1 and 2 to 7 resp. 8 for additional stereochemical correlations has been corrected in the meantime by the authors [26].

Notizen: With a modification of the cyclization procedure of Richman & Atkins [8] the two macrocycles 1,4,7-tritosyl-11-benzyl-1,4,7,11-tetraazacyclotetradecane (8) and 1,7,11-tritosyl-4-trityl-1,4,7,11-tetraazacyclotetradecane (15) were prepared. After selective cleavage of the benzyl and trityl group, respectively, one obtains the two key products 1,4,7-tritosyl-11-benzyl-1,4,7,11-tetraazacyclotetradecane (9) and 1,7,11-tritosyl-4-trityl-1,4,7,11-tetraazacyclotetradecane (16) which have three N-atoms protected by tosyl groups and one accessible for further reactions.To test some of the possibilities we have alkylated 9 and 16 with iodoacetamide, 1-tosyl-aziridine and acrylonitrile. After detosylation with HBr in glacial acetic acid in the presence of phenol mono-N-functionalized tetraazamacrocycles were thus obtained. The advantage of this synthesis is that the cyclization which is the most difficult step of the whole procedure, has to be done only once, regardless of the nature of the pendant arm. In addition a large number of derivatives can be prepared by varying the alkylation component.With Ni2+, Cu2+ and Zn2+ metal complexes of these new ligands were prepared and their IR. and VIS. spectra studied. In the case of the carbamoyl derivatives 12, 14 and 18 the Cu2+-complexes exist in two forms. Whereas at low pH the carboxamide group of the pendant arm is probably not bound to the metal ion, at high pH after deprotonation it coordinates in one of the axial positions.

Notizen: Tentative Synthesis of ‘Bis(triasterane)’ and Synthesis of the Heterocyclic (Tricyclo[4.4.1.01,6]undeca-3,8-diene-11,11-dimethyl)sulfiteThe synthesis of the bis(triasterane) (1) has been tried; the reaction of ‘isotetraline’ (1,4,5,8-Tetrahydronaphthalene; 2) with diazomalonate yielded the tricyclic systems 5 and 6, and not 4. Hydrolysis of 5 gave the monocarboxylic acid 7, and not the dicarboxylic acid 9. The latter could be obtained from the dibromoderivative 8, but 9 couldn't be converted to the acyl chloride 10. The reduction of 9 with LiAlH4 yielded the crystalline diol 11, which was cyclized with SOCl2 to the heterocycle 12. The spectral data of the new compounds 5, 6, 7, 11 and 12 are reported and discussed.

Notizen: A detailed study of the biodegradation of one of the fish toxic chlorinated resin acids, 12,14-dichlorodehydroabietic acidThe correct IUPAC-name is 12,14-dichloro-8,9,11,12-tetradehydro-7,8-dihydroabietic acid. (3) is presented. When exposed to the fungus Mortierella isabellina, 3 is converted into a number of hydroxylated and keto derivatives which show low levels of toxicity to fish. These metabolites were isolated and characterized.

Notizen: Zero-current Ion-transport Studies and Potentiometric Measurements on Membranes Containing Chromoionophores for Li+ and UO22+Non-macrocyclic ionophores for Li+ and UO22+ have been substituted with a diphenylmaleimide unit. In membranes the obtained chromoionophores induce the same ion-selectivity and ion-transport behavior as the unsubstituted ligands.

Notizen: Reactivity-structure correlations for anisole and eleven of its substituted derivatives established from bromination rate constants in liquid SO2, unlike observations in water, show the reaction to be highly sensitive to substituent effects, (ρH2O+ = -7.1; ρSO2+ = -10.51). This result is ascribed to the solvation of the methoxy group which decreases the conjugation of para-substituted (ρO+ = -9.70) compared to that of ortho-substituted derivatives (ρP+ = -8.86). The highly solvated transition state lies far from reactants on the reaction coordinate and the positive charge developed in this state is nearly unity.

Notizen: Peptidic model conjugates carrying a single 2-carboxy-4,6-dinitrophenyl haptenic group, and as carbohydrate moieties D-gluconoyl, β-D-mannopyranosyl, 2-deoxy-β-D-glucopyranos-2-yl, or lactobionoyl residues, including the pseudocarbohydrate residue 1,3,4,5-tetrahydroxycyclohexane-1-carbonyl, were synthesized. Conjugates carrying the lactobionoyl of the bis(2-deoxy-β-D-glucopyranos-2-yl) moiety were anaphylactogenic in the guinea pig, passively sensitized against 2-carboxy-4,6-dinitrophenyl antigen.

Notizen: The stereochemical aspects of the intramolecular Ramberg-Bäcklund reaction, i.e. the 1,3-elimination of hydrogen halide followed by sulfur dioxide extrusion, have been studied on the α-bromosulfones of the 1-thiadecalinThroughout this paper ‘1-thiadecalin’ will be used in place of ‘decahydro-1-thianaphthalene’. series. Whereas the cis,exo-bromosulfone 23a containing the ideal W-type arrangement of the reacting atoms undergoes a clean Ramberg-Bäcklund reaction, the trans,exo- and trans,endo-bromosulfones, 24a and 24b, respectively, lead to an α,β-unsaturated sulfone by simple 1,2-elimination of HBr. Application of the Ramberg-Bäcklund reaction to 9-bromo-8-thiabicyclo[5.2.1]decane-8,8-dioxide (17) permits a short synthesis of the Bredt olefin bicyclo[5.1.1]non-1(8)-ene (5), which can be isolated but shows the typical high reactivity of other methylene-bridged (E)-cyclooctenes.

Notizen: Conversion of 2-bromomethylstyrene 22 and benzocyclobutenyl carbamate 28 to the benzophenanthridine alkaloids (±)-chelidonine (1, five steps, 25% from 28) and to (±)-norchelidonine (2, six steps, 24% from 28) are described. The key step 29 → 31 involves a highly regio- and stereocontrolled intramolecular Diels-Alder reaction of the (E)-quinodimethane 30.

Notizen: Dehydrogenation of hydroaromatic compounds with quinones was reinvestigated in the light of recent criticism of the reaction mechanism. Kinetic and spectroscopic evidence shows that disappearance of substrate proceeds at the same rate as the product-forming step. A mechanism consisting in fast formation followed by slow decomposition of an intermediate can be ruled out. The order of reactivities of 1,4-cyclohexadiene (1), 1,4-dihydronaphthalene (8) and 9,10-dihydroanthracene (11) changes in going from benzoquinone to chloranil or 2,3-dichloro-5,6-dicyanobenzoquinone. It is suggested that this behaviour could be due to contribution of CT-complexes or HOMO-LUMO interactions for determining the reactivity of the substrates.

Notizen: Dependence of the Stereoselectivity on the Leaving Group, the Temperature and the Solvent in the Favorskii-Reaction of Enolisable CyclobutanonesThe base-induced contraction of enolisable 2-halocyclobutanones is often not stereoselectiv. The cis/trans-isomer ratio in the cyclopropanecarboxylic acid 9 derived from 2,4-cis-substituted cyclobutanones 7 depends on the nature of the leaving group. In addition, the choice of the base, the temperature and the solvent can profoundly affect the stereoselectivity of the Favorskii-reaction. The use of the p-bromophenylsulphonyloxy group as leaving group and an inhomogenous reaction medium leads exclusively to cis-carboxylic acid 9.

Notizen: Synthesis of Glyceryletherphosphatides, 2nd Communication. Preparation of 2-O-Acetyl-1-O-[(Z)-9-octadecenyl]-sn -glyceryl-3-phosphorylcholin (‘Oleyl-PAF’), of its Enantiomer and Some Analogous, Unsaturated CompoundsSyntheses of the unsaturated glyceroletherphospholipid 1a as an olefin-analog of ‘Platelet Activating Factor’ (PAF) are described together with the methods for the preparation of the enantiomer 1′a, the corresponding ‘lyso compounds’ 1b and 1′b and their positional isomers 21a, 21′a, 26, 26′, 25, 25′ obtained on formally exchanging the attachment site of the functional groups at the glycerol moiety. Structural assignments and optical purity of the compounds are confirmed.

Notizen: Three macrocyclic polyamines [24]ane-N6 1, [32]ane-N8 2 and [27]ane-N6O3 3 of ring size 24, 32 and 27, respectively, have been synthesized. They contain either trimethylenediamine of ethylenediamine units. The acyclic analog 4, as the reference compound, was also prepared. Compounds 1-3 are macrocyclic analogs of natural polyamines and are potential ligands for metal cations as well as, when protonated, for anions. The protonation constants of compounds 1-4 have been determined. They are high enough for compounds 1-4 to be fully protonated in a pH-range close to neutrality, as required for binding of anions of weak acids. The effect of structural features on the protonation constants are briefly discussed in relation to the design of macrocyclic polyamine ligands.

Notizen: Irradiation (λ〉280 nm) of 3-hydroxypyrroles 1 in acetonitrile leads to the formation of 3,3′-bi-3H-pyrroles 2 in reasonable to good yields.

Notizen: On the Course of the Intramolecular Diels-Alder-Reaction of Cyclopentadienes with Olefinic SubstituentsThe 1:3 mixture of 4-bromobicyclo [3.2.0]hept-2-en-6-one and -7-one (1/2), available by N-bromosuccinimide bromination of bicyclo [3.2.0]hept-2-en-6-one, reacted rapidly with the organo-magnesium and -zinc reagents 3, 10a, 10b and 10d by cyclobutanone ring opening and bromide ion expulsion to give the 5-substituted cyclopentadienes 5, 12a, 12b/12c, and 12d as non-isolated intermediates. Further transformation occured in situ either by a direct intramolecular Diels-Alder reaction (path a) or by a [1,5]-H-migration prior to the intramolecular Diels-Alder reaction (path b). The intermediate 5 followed only path a to give the bridged norbornene derivative 7, the intermediates 12a, 12b and 12c followed only path b to give the annellated norbornene derivatives 15a, 15b and 15c, respectively, and the intermediate 12d followed both paths to give the bridged 14d and the annellated norbornene derivative 15d (in the ration of about 1.4:1). These observations are discussed in terms of the relative velocities of [1,5]-H-migrations and intramolecular Diels-Alder reactions. The major conclusions are: (1) bridged norbornene derivatives with a six-membered ring C (such as 14d) can be prepared by an intramolecular Diels-Alder reaction from 5-alkenyl-cyclopentadienes 12, as long as the dienophilic double bond is activated by an appropriate substituent (as in 12d); (2) such 5-alkenyl-cyclopentadienes 12 are available from the reaction of the bromo-bicyclo-heptenones 1/2 with suitable C-nucleophiles 10.

Notizen: Spiro 3-Oxazolines from the 1,3-Dipolar Cycloaddition of Benzonitrilio-2-propanide and 1,4-QuinonesOn irradiation with light of wavelength 290-350 nm, 2,2-dimethyl-3-phenyl-2H-azirine (1b) reacts with 1,4-naphthoquinone to give the 1H-benzo [f]isoindol-4,9-dione (11) (Scheme 3) via cycloaddition of the benzonitrilio-2-propanide (2b) onto the quinone C, C-double bond. With 2-methyl- and 2,3-dimethyl-1,4-naphthoquinone, the nitrile ylide 2b undergoes cycloaddition preferentially onto the C, O-double bond of the quinone, leading to spiro-oxazolines 12 and 14 (Scheme 4). Steric as well as electronic effects can be discussed to explain the observed site selectivity of the cycloaddition.With the 1,4-benzoquinones 15a, 15b, 15d and 15f, nitrile ylide 2b undergoes the 1,3-dipolar cycloaddition exclusively onto the C, O-double bond. The corresponding spiro-oxazolines have been isolated in 17-32% yield. This contrasts with the previously reported results with benzonitrilipo-phenylmethanide (2a), which undergoes cycloaddition to the C, C-double bond of 1,4-benzoquinones (cf. [1]). This difference in the site selectivity of the 1,3-dipolar cycloaddition can be explained with Houk's concept of LUMO-polarization, that is, the stronger nucleophilic dipol 2b polarizes the LUMO of a α,β-unsaturated carbonyl compound more efficient than the less nucleophilic 2a. This leads to a preference of the cycloaddition to the C, O-double bond in the case of 2b.With 2,3-dimethyl- (15c) and 2,3,5,6-tetramethyl-1,4-benzoquinone (15e), nitrile ylide 2b undergoes C, O- as well as C, C-cycloaddition (Schemes 7 and 8).

Notizen: About the Availability of 13C-NMR Substituent Increments in 1,4-Benzo- and 1,4-NaphthoquinonesThe 13C-NMR spectra of a number of substituted 1,4-benzo- and 1,4-naphthoquinones have been obtained and assigned. Effects of CH3-, O-alkyl-, Cl-, NH2- and N3- substituents have been deduced for the quinoid part of the molecules and used for the calculation of 13C-chemical shifts.

Notizen: Flow thermolysis of 2-propynyl propiolate (5) at 580° afforded butatriene (6) (ca. 50%) and, as by-products, 4-methylene-2-cyclobuten-1-one (7), 2-ethynylpropenal (8), 1-penten-4-yn-3-one (9), 4-penten-2-ynal (10) (total ca. 10%), along with some propynal, acetylene, CO2 and CO. In the same way, propiolic acid (1,1-D2)-2-propynyl propiolate (11) led to (1,1-D2)-butatriene (12) and a little 4-((D2)methylene)-2-cyclobuten-1-one (13). A mechanism is proposed for the transformation of 5 into 6 and of 11 into 12, which also accounts for the formation of 7,8,9 and 10, as well as 13. The position of one of the published 13C-NMR signals of butatriene (6) must be revised. Thermolysis of methyl-(1) and ethyl propiolate (2) resulted in small yields of 2-buten-4-olide (3) and 2-penten-4-olide (4).

Notizen: 13C-NMR. Substituent Increments of the Azido-group in 1,4-BenzoquinonesThe empirical substituent increments of the azido-group in 1,4-benzoquinones have been derived from the 13C-NMR. chemical shifts of 2-azido-5-chloro-1,4-benzoquinone. The validity of the obtained values was tested by comparison of the empirical and computed 13C-NMR. chemical shifts of other azido-chloro-1,4-benzoquinones.

Notizen: The synthesis of a large variety of enantiomerically pure substituted reactive cyclopentenes 16, 23, 24 and 28 have been synthesized from the readily available (-)-quinic acid 1. The straightforward strategy involves a high-yielding intramolecular aldolization-dehydration of acyclic 1,6-dialdehydes 13, 18, 19 and 27 obtained by oxidative cleavage of cyclohexanediols 5, 7, 11 and 12, using either lead tetraacetate or triphenylbismuth carbonate. Neither sulfoxide formation nor racemization of the intermediate dialdehydes at the oxygenated chiral centre was observed. Transformation of the thioacetal 25 to the corresponding ketone 26 using phenylselenic anhydride is also described.

Notizen: Photochromism of spiropyrans. Photodegradation of aza-, oxaza-, and thiaza-cyclopentane- and -cyclohexanespiropyransStudy of the photodegradation of saturated azaheterocyclic spiropyrans showed that electron-donor substituents improve, as in the benzoheterocyclic series, the fatigue properties towards. UV. light. The introduction of oxygen or sulfur in the 5- or 6-membered saturated azacycles or the substitution with an orthofused aromatic nucleus have practically no influence on the results of photodegradation. The indolinospiropyrans allow the greatest number of photochromic cycles before falling to half the initial absorbance on photolysis under accurate conditions.

Notizen: Low-Temperature 13C-NMR. Spectroscopy of Organolithium Derivatives. - 13C, 6Li-Coupling, a Powerful Structural InformationThe 13C-NMR. spectra of thirteen lithiated hydrocarbons (1c-13c. Table 2) and of eighteen a-halo-lithium carbenoids (14c-31c, Table 3) have been recorded in donor solvent (R2O, R3N) mixtures at temperatures down to -150°. The organolithium species were generated from singly or doubly 13C-labelled precursors by H/6Li- or Br/6-exchange. - 13C, 6Li-Coupling was observed of all species but those which supposedly contain contact ion pair C,Li-bonds (benzylic and acetylenic derivatives). The multiplicities of the signals are correlated with the degree of aggregation in solution: the triplets of the halocarbenoids must arise from monomers or heteroatom-bridged oligomers, the quintuplets of butyl-, cyclopropyl-, bycyclo[1.1.0]butyl-, vinyl-, and phenyllithium from dimers with planar arrangement of two Li- and two C-atoms, as known from crystal structures (Scheme 3). All 13C, 6Li-couplings are temperature-dependent, dynamic processes cause them to disappear above ca. -70° (Fig. 1-4). - Types of organolithium compounds are categorized according to the change of chemical shift δΔ (H, Li) upon H/Li-substitution, according to the 13C, 6Li-coupling constants ranging from 0 to 17 Hz, and according to the multiplicities which indicate the aggregation: type A are Li-derivatives of alkanes and cycloalkanes, type B are s̰-bonded vinyl, aryl, and alinyl derivatives, type C are a-heterosubstituted (RS, hetero=halogen) organolithium compounds, and type D are π-bonded allylic and benzylic systems (Table 5). The C,Li-distances in the crystal structures of representatives of all four classes are within the small range of 2.18-2.28 Å (cf. Scheme 3). - Some surprising observations and their interpretations and consequences are: (a) butyllithium solutions in THF, THF/TMEDA, and dimethyl ether contain increasing amounts of dimer upon cooling, the equilibrium (tetramer · 4 THF)+4 THF ⇌ 2 (dimer · 4 THF) being shifted to the right (Fig. 1 and Scheme 4); thus, more of a different species is present at low temperatures, with the accompanying changes in reactivity; (b) mixed higher aggregates are formed upon addition of butyllithium to bicyclobutyllithium; these are broken up to dimers upon addition of TMEDA (Tetramethylethylene-diamine) (Fig. 2 and Scheme 5); (c) the solid state, the calculated gas-phase and the solution species of phenyllithium all have dimeric structures, and so do vinyl and cyclopropyl lithium derivatives; the 13C-deshielding observed upon replacement of H by Li on sp2- and sp-C-atoms is related to a polarization of the π-electrons (Table 3, Fig. 3 and Scheme 6); (d) the spectra of halo-lithium carbenoids show three striking features as compared to the C,H-compound: deshielding of up to 280 ppm (Table 3), strong decrease of the coupling constant with 1H- and 13C-nuclei attached to the carbenoid C-atom (Table 4), and a structure-independant, almost constant, large 13C, 6Li-coupling constant of 17 Hz (Table 3); as shown in Scheme 7, these effects might be the consequence of a reduced degree of hybridization of the carbenoid C-atom. - The preparation of the labelled compounds and the generation of solutions of the organolithium compounds for NMR. measurements are described in full detail.

Notizen: From treatment of D-labelled 1, 2exo-, 1, 2endo- and 2endo, 6endo-trimethylene-norbornane (1, 2 and 3, resp.) with aluminum bromide in carbon disulfide, the evidence is gained that a degenerate rearrangement is involved in the admantane rearrangement of both 1 and 2 to 3.

Notizen: The 1,3-dipolar cycloaddition of the C-t-butyloxycarbonyl-N-mannosyl-nitrone 5, formed in situ from the partially protected D-mannose-oxime 3 and the glyoxylate 4, to ethylene gave preferentially the (3S)-N-glycosyl-isoxazolidine 6 which was transformed into the 3-isoxazolidine-carboxylate (L-5-oxaproline ester) 12 and into some derivatives thereof. The (S)-configuration of 12 was proved by chemical correlation with a derivative of L-asparagine. The D-5-oxaproline ester was obtained from the corresponding N-ribosyl-nitrone 24. Two protected dipeptides containing either C-terminal- (28) or N-terminal-5-oxaproline (= Opro) (30) were synthesized. Starting from 12, the analogue 1 of captopril® (2) was prepared and its activity as an inhibitor of the angiotensin-converting-enzyme (ACE) was examined.

Notizen: (3S, 8E)-1,3-Dihydroxy-8-decen-5-one, a Metabolite of Streptomyces fimbriatus (MILLARD and BURR 1926)From the cultures of Streptomyces fimbriatus, strain Tü 2335, the title compound 1 was isolated by solvent extraction, chromatography, and distillation. Its constitution was determined by spectroscopic investigations of 1 and some of its derivatives, and the chirality by transformation to (R)-1,3-isopropylidenedioxy-decane (12); a reference sample of the latter was prepared from (R)-methyl 3-hydroxydecanoate (10). In the equilibrium mixture the keto form 1 predominates the two hemiacetals 2a and 2b. Compound 1 is inactive against bacteria, yeasts and fungi.

Notizen: (Phenacetin)4·2I4·2H2O is triclinic, a = 13.641 (7), b = 12.807 (6), c = 7.201 (3) Å, α = 99.8 (4), b̃ = 86.5 (4), γ = 104.0 (5)°, P1, Z = 1. The ordered crystal structure has been refined to RF = 0.050, using 4173 independent reflections measured on a four-circle diffractometer with MoKa (graphite monochromator) radiation. The crystals are composed of alternating positively and negatively charged slices; each positive slice contains a double layer of stacks of hemi-protonated phenacetin molecules which are H-bonded through their carbonyl groups (d(O - - - O) = 2.432 (4) Å) while each negative slice contains a single layer of I2-4-ions linked in chains along [100] through H-bonds to pairs of water molecules. The axes of the phenacetin stacks are parallel to the planes of the (I2-4·2H2O)-layers. The I2-4-ion is centro-symmetric and can be approximately represented as I-- - - I-I- - - I- (d(I- - - - I) = 3.404 (1) Å; d(I-I) = 2.774 (1) Å). The compound is a pseudo-type A basic salt.