Characteristics of bacterium GB-2, a presumptiveCytophaga species with novel immunoregulatory properties

A Gram-negative, psychrophilic bacterium, designated GB-2, was isolated from fetal calf serum and analyzed for its morphological, physiological, and biochemical characteristics. Gliding motility, sensitivity to actinomycin D, the presence of the pigment flexirubin and cytochrome c, growth on a variety of carbohydrates, the production of acid fermentation products, and a 34.9 mol% guanine+cytosine (G+C) content of bacterial DNA indicate GB-2 to be a member either of the generaCytophaga orFlexibacter. Growth of GB-2 was optimized in a simple defined medium to facilitate isolation and characterization of bacterial products. Liquid growth of GB-2 resulted in the release of significant quantities of a macromolecule free of both endotoxin and protein into the growth supernatant, which activated the proliferation of murine lymphocytes. The relationship between this bacterium and its end-products to other species of theCytophaga/Flexibacter group is discussed.     

Repair inhibition of potential mutations of ultraviolet-irradiatedEscherichia coli by chloroquine and quinacrine

The frequency of ultraviolet(UV)-induced mutations drops rapidly whenEscherichia coli Hcr⁺ cells (strains WP-2 Hcr⁺; B/r) are incubated on phosphate-buffered agar (PBA), but is reduced only slightly if chloroquine or quinacrine are incorporated into the medium. The excision-deficient WP-2 Hcr^– strain shows little reduction in the number of mutants when incubated on PBA. During postirradiation incubation on PBA, cell viability was relatively unaffected by the presence of the chemicals in the PBA (25 g/ml quinacrine; 50 g/ml chloroquine). When cells were given optimal doses of photoreactivating light, no further decline in mutations was obtained during subsequent incubation on PBA. Approximately 64% of the mutants seen when cells are treated with UV-PBA-chloroquine and 90% seen with UV-PBA-quinacrine can be repaired if cells are incubated on PBA. When these chemicals were added to the PBA, both excision-proficient strains (WP-2 Hcr⁺; B/r) demonstrated a marked reduction in the repair of UV-induced mutations to streptomycin resistance. Our results indicate that these chemicals interfere with the excision of UV-induced pyrimidine dimers, a process that normally occurs during postirradiation incubation on PBA.     

Epithelial differentiation at the edentulous alveolar ridge in man

Summary The epithelial lining of the mucosa of the edentulous, maxillary alveolar ridge was subjected to an ultrastructural and stereological analysis. Four biopsies collected from the non-inflamed crest, i.e., the center over former tooth sockets, in non-denture-wearing female patients 30 to 55 years of age were processed for light and electron microscopy. At the light-microscopic level, epithelial thickness was determined histometrically. Electron micrographs were sampled at two levels of magnification, from five strata in regions of epithelial ridges and from three strata over connective tissue papillae. Standardized stereological pointcounting techniques were employed to analyze a total of 990 electron micrographs. Observations and data revealed that at the alveolar ridge the oral epithelium is truly keratinizing and comprises four strata including a 40±5 m-thick stratum corneum, which displays the oral keratin pattern. The histoand cytodifferentiation were peculiar: (1) Compared to the neighbouring gingival and hard palate epithelium, that of the alveolar crest was markedly thicker, with elongated rete ridges indicating acanthosis. (2) The cytoarchitecture was identical neither to the gingival nor to the hard palate epithelium but revealed a mixture of features typical for either of these two epithelia. Reasons for this are explained on the basis of factors, possible genetic, inherent in epithelial cells that are possibly derived from both the gingival and the palatal environment.     

The early postnatal development of the primary immune response in TNP-KLH-stimulated popliteal lymph node in the rat

Summary The popliteal lymph nodes were removed from young rats of various ages five days after a single immunization with TNP-KLH in the hind footpads. Cryostat sections of the lymph nodes were investigated by means of enzyme and immunohistochemical techniques at the light-microscopical level.The presence and localization of anti-TNP antibody-containing cells were examined using a new technique to visualize specific antibodies. Moreover, the development of the lymph nodes following exogenous antigenic stimulation was compared with that of unstimulated lymph nodes.Specific antibody-containing cells could not be found before day 15 after birth, in rats immunized at day 10. From that time these lymphoid cells were located primarily at the border between cortex and medulla. Younger popliteal lymph nodes showed only aspecific immunoglobulin-containing lymphoid cells. With age, the number of specific antibody-containing cells tended to increase. These cells were more mature, according to morphological criteria and were located nearer the medulla.The first primary follicles were seen at day 19, as was the case in unstimulated animals. The first secondary follicles, containing germinal centers, were detected at day 23, whereas in unstimulated popliteal lymph nodes they were never found.Trapping of immune complexes could not be demonstrated before day 33 after birth. The later appearance of this phenomenon might be a consequence of the techniques applied to demonstrate specific antibody-containing cells.Abbreviations PLN popliteal lymph node - FDC follicular dendritic cell - IDC interdigitating cell - HEV high endothelial venule - TNP trinitrophenyl - KLH keyhole limpet hemocyanin - PBS phosphate-buffered saline - GCPC germinal center precursor cell - sIg surface immunoglobulin - cIg cytoplasmic immunoglobulin     

Immunocytochemistry of magnocellular neurons of supraoptic and paraventricular nuclei of normal and Brattleboro rats with vasopressin anti-idiotype antibody

Summary A vasopressin anti-idiotype antibody was generated by immunization with purified IgG of a primary vasopressin antiserum. The anti-idiotype antibody immunostained neurons in the supraoptic and paraventricular nuclei of the hypothalamus of normal and Brattleboro rats. The distribution of immunostained perikarya in these hypothalamic nuclei together with the staining of fibers in median eminence and neural lobe was similar to that observed in normal rats with anti-vasopressin and suggests strongly that vasopressinergic neurons are being stained. Absorption studies with vasopressin and a vasopressin-binding receptor protein further indicate that a receptor associated with vasopressinergic neurons is recognized by the anti-idiotype antibody.Supported by NIH grants ES03239, NS18626 and NSF grant BNS-8310914. D.T.P. is the receipient of RCDA award NS00869     

Lectin cytochemical evaluation of somatosensory neurons and their peripheral and central processes in rat and man

Summary Paraffin sections of the trigeminal nerve root of the rat, and human spinal nerve root and trigeminal ganglion were stained with a battery of lectin-horseradish peroxidase conjugates to localize and characterize glycoconjugate (GC) in situ. In the rat the myelin sheath of the peripheral segment contained GC with sialic acid most probably linked to the penultinate disaccharide galactose(1 4)-N-acetylglucosamine (Gal(1 )-GlcNAc), and complex type N-glycosidic side chains. The myelin sheath in the central segment differed in containing little if any of the GC named above and in containing GC with terminal -Gal linked to N-acetylgalactosamine (GalNAc), terminal GalNAc and fucose. Schwann cells stained for GC with GlcNAc or mannose whereas oligodendroglia stained for GC with the terminal disaccharide Gal-(1 3)-GalNAc and N-glycosidic side chains, especially in presumed Golgi zones, but also in processes continued as the outer myelin sheath. The human myelin sheath in the central segment differed from that of the rat in not staining with lectins specific for fucose and terminal GalNAc. Sialic acid and terminal -Gal were seen in the human central segment but these sugars appeared to bind to astroglial structures rather than to the myelin sheath as in the rat. Astrocytes in both rat and man were stained by two fucose-binding lectins. Several lectins revealed affinity for GC in the neurilemmal sheath, and staining of this structure was stronger in the human specimens. Neurons in the human trigeminal ganglion ranged from unstained to strongly positive for fucoconjugate in cytoplasmic bodies and plasmalemma. Positive ganglion cells gave rise to unmyelinated fibers which also stained for fucoconjugate. Remak fibers and their extensions into the substantia gelatinosa of the human spinal cord stained strongly for content of fucose.The stronger lectin affinity for N-glycosidic core sugars in the peripheral as compared with the central segment suggests that lectins localize P_o protein in peripheral myelin. The reactivity for several sugars in the central segment can possibly be attributed to myelin-associated glycoprotein (MAG) of central myelin, but lectin staining for GalNAc shows in addition a biochemically unrecognized GC with O-glycosidic linked oligosaccharides in myelin. The lectin cytochemistry indicates that the 170 K Dalton glycoprotein with PNA affinity obtained from rat sciatic nerves occurs in nodes of Ranvier.This research was supported by NIH Grants AM-10956, HL-29775 and United Health and Medical Research Foundation of South Carolina, Inc. Grant No. 79     

Dinitrogen fixation by obligate and facultative anaerobic bacteria in association with cellulolytic fungi

The anaerobic bacteriumClostridium butyricum is the major contributor to nitrogen gains by a cellulolytic/nitrogen-fixing population isolated from straw. Growth of the anaerobe is supported by the products of fungal cellulases. The facultative anaerobeEnterobacter cloacae does not make a significant direct contribution to nitrogen fixation but in association withC. butyricum allows the anaerobe to grow under aerobic conditions. The major function ofE. cloacae is though to be provision of oxygen-depleted microsites.     

The in vitro screening of methylated 4-oxypiperidine compounds for inhibition of protein synthesis in a rabbit reticulocyte cell-free translation system

A variety of methylated 4-oxypiperidine derivatives were tested for their ability to inhibit protein synthesis in vitro. A direct correlation was found between the extent of methylation of these compounds and their inhibitory activity in a rabbit reticulocyte lysate cell-free translation system.Abbreviation IC₅₀ 50% inhibitory concentration     

Ontogeny of the photoreceptors in the embryonic retina of the viviparous guppy,Poecilia reticulata P. (Teleostei)

Summary Ultrastructural analyses of retinal development in the guppy embryo show that at midgestation all types of photoreceptors are differntiated in the fundus, and at birth differentiation extends over the whole retina. Formation of discs of outer segments is more rapid in rods than in cones. Double cones differentiate simultaneously with long single cones and are formed by the adhesion of two primordial inner segments; short single cones develop last. Wherever cones are differentiated, they are arranged in an adult-type square mosaic. The rods in the embryo, as opposed to the adult, are likewise regularly arranged within the mosaic unit.These results are at variance with the generally held opinion that adult teleosts which possess duplex retinae have larvae with pure cone retinae, and that rods, double cones and mosaics appear in late larval life or only at metamorphosis.In the double cones of the guppy embryo subsurface cisternae develop along the adjoining primordial inner segments. Additionally, regularly distributed subsurface cisternae are formed in the regions of intimate contact of long single cones with double cones and rods.We suggest that the early development of rods and double cones, and a square-mosaic with regular distribution of rods and subsurface cisternae, provide the newly born with a fully functional optical apparatus, especially suited to perception of movements. This is necessary for its survival against predatory, especially maternal, attacks.This work was in part subsidized by a grant from the Medical Research Council of Ireland to Y.W.K.