Lipoxin A-induced inhibition of human natural killer cell cytotoxicity: studies on stereospecificity of inhibition and mode of action

Human leukocyte-derived lipoxin A (LXA; 5S,-6R,15S-trihydroxy-7,9,13-trans-11-cis-eicosatetraenoic acid) inhibits the cytotoxic activity of human natural killer (NK) cells. LXA and three of its isomers were prepared by total organic synthesis and assayed for activity with human NK cells. Dose-response studies showed that biologically derived LXA and synthetic LXA were equally effective in inhibiting NK cell cytotoxicity. 6S-LXA, with its 6S-OH group in an (S) configuration, proved to be approximately half as potent as LXA. In contrast, 6S-11-trans-LXA and 11-trans-LXA displayed virtually no inhibitory activities. The methyl esters of both LXA and 6S-LXA proved to be more potent than their corresponding free acids. Thus, LXA inhibition of NK cells displays clear-cut stereochemistry. In the absence of putative inhibitors, NK cells bind to their targets to form conjugates. This event is followed by polarization of the NK Golgi apparatus, which moves towards the plasma membrane that is in contact with the target cell. However, in the presence of either the methyl ester or free acid of LXA, the Golgi apparati of NK cells bound to their targets were randomly oriented. In contrast, neither 6S-11-trans-LXA nor the potent NK inhibitor prostaglandin E2 affected the polarization. Furthermore, although prostaglandin E2 resulted in a decrease in NK-target cell binding efficiency, LXA and its isomers failed to affect conjugate formation. Together these results indicate that LXA-induced inhibition of NK cytotoxicity does not act on NK cell binding but may block cytotoxicity by disrupting "signals" involved in the specific orientation of the Golgi. Thus, this latter event may appear to be important in cytotoxicity.     

Carbon and nitrogen metabolism in ectomycorrhizal fungi and ectomycorrhizas

The literature concerning the metabolism of carbon and nitrogen compounds in ectomycorrhizal associations of trees is reviewed. The absorption and translocation of mineral ions by the mycelia require an energy source and a reductant which are both supplied by respiratory catabolism of carbohydrates produced by the host plant. Photosynthates are also required to generate the carbon skeletons for amino acid and carbohydrate syntheses during the growth of the mycelia. Competition for photosynthates occurs between the fungal cells and the various vegetative sinks in the host tree. The nature of carbon compounds involved in these processes, their routes of metabolism, the mechanisms of control and the partitioning of metabolites between the various sites of utilization are only poorly understood. Both ascomycetous and basidiomycetous ectomycorrhizal fungi synthesize and some, if not all, accumulate mannitol, trehalose and triglycerides. The fungal strains employ the Embden--Meyerhof pathway of glucose catabolism and the key enzymes of the pentose phosphate pathway (6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, transaldolase and transketolase). Anaplerotic CO2 fixation, via pyruvate carboxylase and/or phosphoenolpyruvate carboxykinase, provides high pools of amino acids. This process could be important in the recapture and assimilation of respired CO2 in the rhizosphere. The ectomycorrhizas are thought to contain the Embden--Meyerhof pathway, the pentose phosphate pathway and the tricarboxylic acid cycle, which provide the carbon skeletons for the assimilation of ammonia into amino acids. The main route of assimilation of ammonia appears to be through the glutamine synthetase-glutamate synthase cycle in the ectomycorrhizas. Glutamate dehydrogenase plays a minor role in this process. Glutamate dehydrogenase and glutamine synthetase are present in free-living ectomycorrhizal fungi and they participate in the assimilation of ammonia and the synthesis of amino acids through the glutamate dehydrogenase/glutamine synthetase sequence. In both in vitro cultures of fungi and ectomycorrhizas, the assimilated nitrogen accumulates in glutamine. Glutamine, but also ammonia, are thought to be exported from the fungal tissues to the host cells. Studies on the metabolism of ectomycorrhizas and ectomycorrhizal fungi have focused on the metabolic pathways and compounds which accumulate in the symbiotic tissues. Studies on regulation of the overall process, and the control of enzyme activity in particular, are still fragmentary.(ABSTRACT TRUNCATED AT 400 WORDS)     

Study of plasmids isolated from saccharolytic Clostridia: Construction of hybrid plasmids and transfer toEscherichia coli andBacillus subtilis

Summary Small plasmids ofClostridium acetobutylicum and related strains were isolated and studied. Their restriction maps were established and different hybrid plasmids were constructed by ligation with plasmid pHV33.     

Variants of the anti-Müllerian hormone gene in a compound heterozygote with the persistent Müllerian duct syndrome and his family

Summary The human genome contains a large number of interspersed simple repeat sequences that are variable in length and can therefore serve as highly informative, polymorphic markers. Typing procedures include conventional multilocus and single locus probing, and polymerase chain reaction aided analysis. We have identified simple sequences in a cosmid clone stemming from the human Y chromosome and consisting of (gata)_n repeats. We have compared these with two equivalent simple repeat loci from chromosome 12. After amplifying the tandemly repeated motifs, we detected between four and eight different alleles at each of the three loci. Codominant inheritance of the alleles was established in family studies and the informativity of the simple repeat loci was determined by typing unrelated individuals. The polymorphisms are suitable for application in linkage studies, practical forensic case work, deficiency cases in paternity determination, and for studying ethnological questions. The mutational mechanisms that bring about changes in simple repeats located both on the autosomes and on the sex chromosomes, are discussed.Professor Dr. Otto Prokop (Humboldt-Universität Berlin) on the occasion of his 70th birthday     

Réactions observées chez l'Anguille maintenue dans un milieu privé d'électrolytes,en particulier au niveau du système hypothalamo-hypophysaire

Résumé 45 Anguilles sont maintenues (32 h. à 46 jours) en eau déminéralisée (désionisée ou distillée). On note un assombrissement de la pigmentation, une hypertrophie importante de la vésicule biliaire et des modifications du système hypothalamo-hypophysaire.Dans la pars distalis rostrale, les cellules à prolactine subissent une régression, ne semblant plus élaborer de granulations. Les cellules corticotropes sont faiblement stimulées, partiellement dégranulées, en accord avec la légère activation de l'interrénal. Les cellules thyréotropes apparaissent souvent peu actives après 46 jours; cependant, l'examen préliminaire des thyroïdes ne montre pas d'hypoactivité. Dans la pars distalis proximale, les cellules gonadotropes ne sont pas modifiées; les cellules somatotropes sont stimulées: hypertrophie nucléaire et nucléolaire, développement de l'ergastoplasme, sans dégranulation complète. Leur rôle dans l'osmorégulation est discuté. Dans le lobe intermédiaire, les cellules du type 1, hématoxyline au plomb positives, prédominent chez les témoins; elles subissent une stimulation avec dégranulation partielle correspondant peut-être à l'assombrissement de la pigmentation. Le type 2, PAS positif, s'hypertrophie et s'hyperplasie rapidement en eau déminéralisée. Cette hyperactivité indiquerait un rôle dans l'osmorégulation. Le neuroséorétat s'accumule, puis tend à diminuer, la neurohypophyse étant de volume réduit. Le noyau préoptique paraît peu actif. L'hypothèse d'une faible utilisation des produits élaborés par l'axe hypothalamo-neurohypophysaire est envisagée. En plus de l'interrénal et des corpuscules de Stannius, l'hypothalamus et l'hypophyse interviennent donc dans la réponse de l'Anguille à une déminéralisation du milieu ambiant.

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Summary 45 male silver eels were kept from 32 hours to 46 days in demineralized (distilled or deionized) water. They show a darkening of the skin, a hypertrophy of the gall-bladder and some modifications of the hypothalamo-neuro-adenohypophysial system.In the rostral pars distalis, the prolactin cells regressed and appeared chromophobic. The corticotrophic cells are slightly stimulated and partly degranulated, in correlation with some activation of the interrenal, previously described. The thyrotrophic cells do not appear active at the end of the experiment, however the thyroid glands are not often inactive.In the proximal pars distalis, the gonadotrophic cells remain undifferentiated. The somatotrophic cells are strongly stimulated: a cellular, nuclear and nucleolar hyperactivity, without degranulation and a development of the ergastoplasm are observed. Their role in osmoregulation and metabolism of potassium is discussed.The pars intermedia is composed of two cell types: one of which is predominant, leadhematoxylin positive. It could elaborate the MSH (intermedin). In demineralized water, it is partly degranulated and progressively stimulated, perhaps in correlation with the darkening of the pigmentation. The other cell type is PAS positive, and not abundant in the control fish. In the treated fish, rapid hyperplasia and hypertrophy occur, with cytoplasmic degranulation and mitotic activity, so that it becomes predominant in this lobe after a month or more in demineralized water. Cytologic criteria indicate a great hyperactivity which seems to play a role in the processes of the osmoregulation; this hypothesis is discussed.The neurosecretory material tends to increase, as a storage, then to decrease in the neurohypophysis which is of a reduced volume. The preoptic nucleus does not appear active. In addition to the modifications of the interrenal and the corpuscles of Stannius previously described, the changes in the hypothalamus and the pituitary constitute evidence of their important role in the reaction of the eel to a demineralized environment.

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Ce travail a été réalisé avec la collaboration technique de Mademoiselle Jacqueline Olivereau, du C.N.R.S., que nous remercions bien vivement: elle nous a beaucoup aidée au cours de ces expériences, en particulier lors des changements quotidiens de l'eau distillée ou désionisée, et a réalisé toutes les préparations histologiques et les microphotographies illustrant cet article.     

Interaction between cells and elastin fibers: An ultrastructural and immunocytochemical study

Mesenchymal cells (fibroblasts, smooth muscle cells) and endothelial cells were shown to interact with elastin fibers. The strong adhesion of elastin fibers to these cells is mediated by a cell membrane complex with a major glycoprotein component of 120 kDa designated as elastonectin. This interaction was studied by transmission electron microscopy (TEM) and immunocytochemical techniques using antibodies raised against the elastin adhesive proteins. When fibroblasts and smooth muscle cells were cultured in presence of elastin fibers, TEM showed an adhesion mechanism that takes place over several sites along the plasma membrane of these cells. Endothelial cells showed a very close association with elastin, emitting “pseudopodia” that embody the fibers. TEM, indirect immunofluorescence, immunoperoxidase, and confocal microscopy showed the presence and localization of cell membrane components synthesized in large quantities when cells were incubated in presence of elastin. Cells without elastin fibers barely revealed the adhesive membrane complex. These results confirm and extend previous findings concerning the presence of an inducible cell membrane complex that mediates the adhesion of elastin fibers to these cell types. © 1994 Wiley-Liss, Inc.     

Ozone-induced microscopical changes and quantitative carbohydrate contents of hybrid poplar (Populus × euramericana)

Summary Cuttings of hybrid poplar (Populus × euramericana var. Dorskamp) were exposed to ozone (80 g/m³ from 2100 hours to 0700 hours, 180 g/m³ from 0700 hours to 2100 hours) for 3 months. Ozone reduced the starch content in leaves and stem bark, whereas starch granules accumulated in bundle sheath cells along small leaf veins. At the same time, sucrose and inositol content increased in the leaves. Mesophyll cells in the vicinity of the stomata were injured first, and droplet-like material appeared on their walls. In the sieve plates of fumigated trees, the pores showed a higher degree of narrowing than those of the control treatment. Cell collapse in the leaves was accompanied by water loss and an increase in air space. In the stems, the ozone treatment led to a reduced radial width, particularly in the xylem tissue. These results are discussed in relation to reduced or inhibited phloem loading and ozone-induced drought stress. The plants injured by ozone showed quite distinct patterns of metabolite responses as well as enzyme activities (PEP- and RubP-carboxylase) in the leaves from the top to the bottom. There were also remarkable differences in the reaction of sucrose and inositol between leaves and stem bark. Future research should therefore increasingly follow a whole-plant approach for a better understanding of complex plant reactions.     

Two genes present on a transposon-like structure in Lactococcus lactis are involved in a Clp-family proteolytic activity

The lactose-protease plasmid pUCL22 of Lactococcus lactis subsp. lactis strain CNRZ270 contained two inverted copies of IS 1076 flanking a region of 3.7 kb. This internal region was sequenced and found to contain two large open reading frames, ORF1 and ORFP in opposite orientations. ORF1 consists of 2289 bp; the deduced 763-amino-acid sequence is similar to the ATPases of the ClpA family. It contains two well-conserved consensus ATP-binding sites. It was named ClpL. ORFP consists of 930 bp encoding a protein of 310 amino acids. No similarity with any known protein was found in GenBank data for ORFP. Increased ATP-dependent proteolytic activity was detected in extracts from Escherichia coli cells expressing the clpL and ORFP genes.