l-Alanine uptake in brush border membrane vesicles from the gill of a marine bivalve

Summary Brush border membrane vesicles were prepared from mussel gills using differential and sucrose density gradient centrifugation. These vesicles contained both the maximal Na⁺-dependent alanine transport activity found in the gradient and the maximal activities of -glutamyl transpeptidase and alkaline phosphatase. Electron micrographs showed closed vesicles of approximately 0.1–0.5 m diameter. Transport experiments using these vesicles demonstrated a transient 18-fold overshoot in intravesicular alanine concentration in the presence of an inwardly directed Na⁺ gradient, but not under Na⁺ equilibrium conditions. A reduced overshoot (10-fold) was seen with an inwardly directed K⁺ gradient. Further studies revealed a broad cation selectivity, with preference for Na⁺, which was characteristic of alanine transport but not glucose transport in these membranes. The apparent amino acid specificity of the uptake pathway(s) was similar to that of intact gills and supported the idea of at least four separate pathways for amino acid transport in mussel gill brush border membranes. The apparent Michaelis constant for alanine uptake was approximately 7m, consistent with values forK _t determined with intact tissue.     

Opportunistic zygomycotic infections

This is a literature review of 361 opportunistic fungal infections caused by the Zygomycetes. The clinical and laboratory diagnosis, pathogenesis, management, treatment, and outcome of infection are discussed. The Zygomycetes are a group of opportunistic fungi (orders Mucorales and Entomophthorales) which cause severe infections which may be fatal. Early clinical recognition, prompt diagnostic procedures, control of underlying disease and treatment with high doses of amphotericin B and aggressive surgery increases survival in an otherwise lethal infection.     

Performance of nymphal and adult Nezara viridula on selected alternate host plants

Laboratory studies with the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), feeding on seeds or fruits of some alternate host plants, demonstrated differences in nymphal survival and development time, and body weight at adult emergence. Lowest mortalities (25%) were observed for nymphs on fruits of dakota mustard [Brassica kaber (DC.)] and siberian motherwort (Leonurus sibiricus L.). On fruits of castor bean (Ricinus communis L.) nymph mortality was ca. 60%, and on mature seeds of L. sibiricus ca. 75%. Total development time from second instar to adult ranged from ca. 26 days on B. kaber to ca. 42 days on R. communis. Fresh body weight at adult emergence was greatest for nymphs on L. sibiricus and lowest on R. communis. Adult survival after 30 days and mean longevity was greater on L. sibiricus than on the remaining foods, being drastically reduced on R. communis. In general, reproductive performance on all foods was similar, except R. communis, on which no female reproduced. Female weight gain of N. viridula was generally greater on L. sibiricus On R. communis, adults lost weight during their first week and showed the lowest weight gain at the end of day 29. Field observations coupled with these laboratory studies suggest that in the Warta area (northern Paraná state, Brazil, 23° 11 S), L. sibiricus and B. kaber are important alternate food plants in the seasonal phenology of N. viridula. R. communis, however, is of poor nutritional quality, serving primarily as a refuge.

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Résumé L'alimentation au laboratoire de N. viridula avec les graines et les fruits de diverses plantes a entraîné différentes mortalités larvaires, différentes vitesses de développement et des différences dans les poids des adultes à l'émergence. Les mortalités les plus faibles (25%) on été observées avec les fruits de Brassica kaber DC. et de Leonurus sibiricus L.Avec les fruits de Ricinus communis L., la mortalité larvaire était de 60%; sur graines mûres de L. sibiricus, elle était de 75%. La durée du développement du second stade à l'adulte a varié de 26 jours sur B. kaber à 42 jours sur R. communis. Le poids frais des adultes à l'émergence était le plus élevé avec L. sibiricus et le plus faible avec de R. communis. La longévité des adultes au-delà de 30 jours était la plus élevée avec L. sibiricus, et était fortement réduite avec R. communis. Dans l'ensemble, la reproduction était de même importance avec les différents aliments, à l'exception de R. communis avec lequel les femelles ne se reproduisaient pas. Le gain de poids des femelles était généralement supérieur avec L. sibiricus; avec R. communis les adultes ont perdu du poids pendant la première semaine et ont présenté le plus faible gain à la fin du 29ème jour.Des observations dans la nature, associées à ces expériences de laboratoire, laissent supposer que dans la zone de Warta (au Nord de l'état de Paraná, 23° 11 S, au Bresil), L. sibiricus et B. kaber sont des plantes hôtes alternatives importantes dans la phénologie saisonnière de N. viridula. Cependant, R. communis, de faible valeur alimentaire, sert avant tout de refuge.

     

Susceptibility of autologous target cells to lysis by lymphokine-activated effectors from interferon-α-treated chronic myelogenous leukaemia patients

Summary Chronic myelogenous leukemia (CML) patients in chronic phase display compromised lymphokine-activated killer (LAK) cell induction, which is partly restored after therapy with interferon . However, the relative resistance of the leukemic cells from these patients to autologous or allogeneic LAK lysis is not affected by this treatment. In an attempt to render CML cells more susceptible to lysis or cytostasis, they were precultured in serum-free medium with or without recombinant growth factors. In eight patients studied, interleukin-3 (IL-3) significantly enhanced the spontaneous short-term (6-day) proliferation of CML cells, with retention of ability to form colonies in methylcellulose. Culture in either medium alone or IL-3 led to a significant enrichment of CD14⁺ and CD33⁺ cells but to a reduction in CD34⁺ cells. In contrast, culture of the same cells in IL-2 (to generate autologous LAK activity) resulted in a loss of CD14⁺ and CD33⁺ as well as CD34⁺ cells but in a significant increase in CD3⁺ and CD56⁺ cells. Despite similarities in their phenotypes, IL-3 cultured cells but not those cultured in medium alone acquired susceptibility to lysis by the IL-2-cultured autologous LAK cells. These results may have significance for the design of novel combination immunotherapy in CML.This work was supported in part by the Deutsche Forschungsgemeinschaft (SFB 120)     

Contrasting breeding systems in twoEriotheca (Bombacaceae) species of the Brazilian cerrados

The pollination biology and breeding systems ofEriotheca pubescens andE. gracilipes have been studied. These two species occur as trees in cerrado vegetation, the neotropical savannas of Central Brazil, with partially sympatric distributions. They have similar phenology and floral structure, although the flowers ofE. pubescens are larger. Both species have nectar flowers pollinated by largeAnthophoridae bees but the main pollinators of each species differ in size. The species have markedly different breeding systems: late-acting self-incompatibility inE. gracilipes and apomixis stimulated by pollination inE. pubescens.     

Glutamic acid decarboxylase of embryonic avian retina cells in culture: Regulation byγ-aminobutyric acid (GABA)

1. Retina-cell aggregate cultures expressed glutamate decarboxylase activity (L-glutamate 1-carboxylase; EC 4.1.1.15) as a function of culture differentiation. 2. Glutamic acid decarboxylase (GAD) activity was low in the initial phases of culture and increased eight-fold until culture day 7, remaining high up to day 13 (last stage studied). 3. The addition of GABA to the culture medium 24 h after cell seeding almost totally prevented the expression of GAD activity. 4. In association with decreased enzyme activity, aggregates exposed to GABA did not display immunoreactivity for GAD, suggesting that GAD molecules were either lost from GABAergic neurons or significantly altered with GABA treatment. 5. Control, untreated aggregates showed intense GAD immunoreactivity in neurons. Positive cell bodies were characterized by a thin rim of labeled cytoplasm with thickest labeling at the emergence of the main neurite. 6. Heavily labeled patches were also observed throughout the aggregates, possibly reflecting regions enriched in neurites. 7. The GABA-mediated reduction of GAD immunoreactivity was a reversible phenomenon and could be prevented by picrotoxin.     

Ca2+ calmodulin-dependent protein kinase activity in the ascomycetes Neurospora crassa

DEAE-cellulose column chromatography of Neurospora crassa soluble mycelial extracts leads to the resolution of three major protein kinase activity peaks designated PKI, PKII, and PKIII.PKII activity is stimulated by Ca²⁺ and Neurospora or brain calmodulin. Maximal stimulation was observed at 2 µM-free Ca²⁺ and 1 µg/ml of the modulator. The stimulatory effect of the Ca²⁺-calmodulin complex was blocked by EGTA and by some calmodulin antagonists such as phenothiazine drugs or compound 48/80.PKII phosphorylates different proteins, among which histone II-A at a low concentration and CDPKS, the synthetic peptide specific for Ca²⁺-calmodulin dependent protein kinases, are the best substrates. Some phosphorylation can be detected in the absence of any exogenous acceptor. PKII activity assayed in the presence of histone II-A or in the absence of exogenous phosphate acceptor (autophosphorylation) co-elute in a DEAE-cellulose column at 0.28 M NaCl. As result of the autophosphorylation reaction of the purified enzyme a main phosphorylated component of 70 kDa was resolved by SDS-polyacrylamide gel electrophoresis. It is possible that this component is an active part of this enzyme.     

Segmental release of amino acid neurotransmitters from transcranial stimulation

The present study used microdialysis techniques in an intact rabbit model to measure the release of amino acids within the lumbar spinal cord in response to transcranial electrical stimulation. Dialysis samples from the extracellular space were obtained over a stimulation period of 90 minutes and were examined using high pressure liquid chromatography. Neuronal excitation was verified by recerding corticomotor evoked potentials (CMEPs) from the spinal cord. A significant increase in the release of glycine and taurine compared to sham animals was measured after 90 minutes of transcranial stimulation. Glutamate and aspartate release was not significantly elevated. GABA concentrations were consistently low. CMEP components repeatedly showed adequate activation of descending fiber pathways and segmental interneuron pools during dialysis sampling. Since glycine, and to a lesser extent taurine, have been shown to inhibit motor neuron activity and are closely associated with segmental interneuron pools, suprasegmental modulation of motor activity may be, in part, through these inhibitory amino acid neurotransmitters in the rabbit lumbar spinal cord.     

Potato Lectin: A Cell-Wall Glycoprotein

The activity and the amount of potato lectin were measured inpotato tuber slices (Solanum tuberosum cv. Huinkul) aeratedfor 48 h. Lectin was found in a soluble form, liberated to themedium and associated with insoluble structures. Polyacrylamidegel electrophoresis in denaturating conditions and immunologicaltechniques indicated that the lectins associated to cell wall,soluble or liberated to the medium, were identical. The cell-wallfraction was found to contain 65% of total lectin in the tuber.The possible role of potato lectin in tubers was discussed. (Received June 5, 1985; Accepted September 3, 1985)     

The cytoskeletal architecture of interdigitated microvilli in the photoreceptors of some nocturnal spiders

Summary The photoreceptor microvilli of some nocturnal spiders (Isopeda andOlios in theSparassidae, andClubiona in theClubionidae) are wide (80–140 nm), and microvilli from adjacent receptors are interdigitated. Because microvillar diameters are relatively large in relation to the thicknesses of thin sections, it is possible to examine cytoskeletal structures closely associated with the microvillar plasmalemmae directly.Retinae were treated with a specific inhibitor of cysteine proteases before primary fixation for electron microscopy in a Ca²⁺-chelating medium. Cytoskeletal components were stabilized with tannic acid. A variety of microvillar profiles was obtained, consistent with an assumption that they represent imperfect preservation of anin vivo plasmalemmal undercoat, inferred to consist of longitudinally-disposed microfilaments, presumptively bonded to the microvillar plasmalemma. The microvillar lumen is inferred to be empty of cytoskeletal components in life.This model is discussed in terms of 1. the cytoskeletal organisation of microvilli of the primitive photoreceptors of a leech (Blest et al. 1983), where the arrangement of microfilaments resembles that in the vertebrate intestinal brush-border; 2. the large complement of membrane-associated oligomeric actin in rhabdoms of crayfish, where identifiable microfilaments cannot be resolved within microvilli by transmission electron microscopy (de Couet et al. 1984), and a single visualizable axial filament of uncertain composition is linked to the plasmalemma by side-arms.