Selective binding of nucleotide probes by eosinophilic cationic protein during in situ hybridisation

During in situ hybridisation on frozen and paraffin-embedded sections of bowel for IB, oligodeoxyribonucleotide probes were found to bind more avidly to eosinophils than target mRNA. This binding could not be obviated using strategies previously employed to block either binding of long DNA probes (200-mers) to eosinophils in bone marrow smears, or of riboprobes to eosinophils in sections of bowel, without removing specific hybridisation of probes. That this binding could arise through interaction of anionic oligodeoxyribonucleotides with eosinophil cationic protein, which has an unusually high pI, and is abundant in cytoplasmic granules of eosinophils, was demonstrated in vitro using real-time biomolecular interaction analysis with a BiacoreX instrument. Finally, a relationship between probe hydrophobicity, measured by reverse phase ion-pair high performance liquid chromatography, and in situ binding of individual probes to eosinophils was demonstrated. Effective tissue penetration by hydrophobic probes and subsequent strong probe–eosinophilic cationic protein interactions therefore may confound the interpretation of in situ hybridisation performed with oligonucleotide probes in eosinophil-containing tissues, such as bowel and nasal polyps.     

An immunofluorescent method for a specific demonstration of granulocytes and some of their proteins (ECP and CCP)

Using p-benzoquinone as a fixative the non-specific fluorescence of granulocytes and especially the eosinophils is removed for both FITC and TRITC. In this way we have been able to detect the eosinophil cationic protein (ECP) and the chymotrypsin-like cationic protein (CCP) in human lung tissue and by double immunofluorescent labelling shown that these two proteins very likely are related to eosinophils respectively the neutrophils.     

Étude de la distribution des organes sensoriels de l'antenne et de leurs relations possibles avec le comportement chez deux fourmis myrmicines:Myrmica lævinodis Nyl. etAphænogaster gibbosa Latr. récoltées dans la région des Eyzies

Conclusion. Summary Morphological and statistical researches on the sensory equipment of the Ant's antennæ in chapter I of this work have enabled us to interpret some biological phenomena to be enlarged upon in the next chapters.The part played by the champagne cork shaped organs in the trophallaxic behaviour has been shown by ablations ofMyrmica lævinodis Nyl. antennæ.Aphænogaster gibbosa Latr., a species which does not practise the stomodeal exchanges, possesses twice less numerous champagne cork shaped organs on its antennæ.Besides, in those mixed colonies where the two species are bred together,Aphænogaster sometimes receives food fromMyrmica.However the importance of those needful organs should hot be overvalued becauseMyrmica is partly able to practise the trophallaxic behaviour whithout its feelers. Moreover, ifAphænogaster does not exchange food in its nests, this is certainly not due to a defect of its sensory equipment but very likely to the disappearance of the innate behaviour of «agiver», an inductor of the one of a «receiver». ThusAphænogaster's sensory equipment does need to become as evolued as the one ofMyrmica. The importance (very likely of an olfactory nature) of those organs in other behaviours may have caused its persistence onAphænogaster's antennæ. The crop of that ant has grown much less extensible than the one ofMyrmica.It is further to be noticed that the behaviour ofMyrmica as agiver when its feelers have been cut-off proves easier to carry out but more difficult to acquire or to preserve on the evolutive plane, and thus seems to be more basic as regards trophallaxy.Finaly, beyard the knowledge of the sensory qualities, the interest of the myrmecological material lies in that it affords a new method of approach for the study of the determinism of the behaviour.

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Adresse au moment de la parution de l'article: Laboratoire d'Ethologie Expérimentale «Les Sources», 78-Mittainville.     

Deposition of eosinophil cationic protein in granulomas in allergic granulomatosis and vasculitis: the Churg-Strauss syndrome.

Biopsy specimens and tissues obtained at necropsy from two women who died after developing the Churg-Strauss syndrome were analysed to see whether granulomas in these patients contained activated eosinophils or secreted eosinophil cationic proteins, or both. Immunocytochemical studies with monoclonal antibody EG2 showed large amounts of eosinophil cationic protein and eosinophil protein-X (which are toxic for heart cells and other tissues) in the granulomas. Many activated and degranulating eosinophils were seen to be migrating from the blood into these areas. Eosinophils may play a central part in the development of lesions in the heart and other tissues in the Churg-Strauss syndrome.     

The biosynthesis of neutrophil and eosinophil granule proteins

Composition of azurophil and specific granules from human polymorphonuclear neutrophils and granules from eosinophils is presented. Biosynthesis of the granule proteins is discussed in detail with particular emphasis on neutrophil myeloperoxidase (MPO) and eosinophil cationic protein (ECP).     

IgE-Independent Activation of Human Mast Cells Indicates their Role in the Late Phase Reaction of Allergic Inflammation

Mast cells are tissue dwelling cells that have a clear-cut pathologic role in allergy. Besides that, they participate in several chronic inflammatory conditions, helminitic parasitosis, and in some solid tumor reactions, but also in physiological situations, such as wound healing and innate immunity. Mast cells produce and release various mediators after activation induced by either IgE-dependent or IgE-independent mechanisms. Although much information has been gathered on the immunological (IgE-dependent) mast cell activation both in vivo and in vitro, not much is known about the non-immunological (IgE-independent) activation particularly in human mast cells. Mast cell IgE-independent activation is mediated through Gi3alpha which has been identified in rat mast cells as the pertussis toxin (Ptx)-sensitive heterotrimeric G protein that interacts with cationic secretagogues inducing PLC-independent mast cell exocytosis. Mast cell IgE-independent activation in allergy most likely occurs when mast cells encounter eosinophils, the main inflammatory cells that persist throughout the late and chronic phases of the allergic reaction. This review summarizes the influence of eosinophils on mast cell activation demonstrating that IgE-independent activation has a significant role in pathophysiological processes.     

Über sekundäre Geschlechtsmerkmale bei einigen Ameisenlarven

Summarium In larvis, etiam juvenilibus, et præpupis generumSolenopsis etTetramorium (Subfam.Myrmicinæ) genus ex signis secundariis cognosci potest.Solenopsis fugax: Mas: Pili e vertice et fronte provenientes (sicut pili per corpus sparsi) in duos ramos divergentes dividuntur; truncus pili brevior ramis vel etiam æqualis; pili genæ similes, sed eorum truncus multum largior; pili clypei ambo cum acumine quadripartito. Mandibula cum brevi acumine; dentes molares desunt, eorumque locum denticulus acutus tenet. Femina: Pili corporis ut in masculo; pili capitis omnes longiores, in apice tantum bi-vel. tripartiti; duo pili clypei simplices. Mandibula cum dente incisivo robusto et duobus distinctis molaribus.Tetramorium cæspitum: Mas: Pili corporis discissi in multos ramos conflexos; pili capitis penicillo non dissimiles. Palpus maxillaris crassus et tuberosus. Galea lata, paulo minor quam palpus. Femina: Pili corporis cum ramis expansis; pili capitis dupli quam in masculo, in apice tantum bi- ad quadripartiti; pili clypei breves ut in masculo, in apice bifurci. Palpus maxillaris minus crassus. Galea procera, altior quam palpus.     

La glande sternale dans l'évolution des Termites

Résumé Des différenciations glandulaires de l'épiderme au niveau de certains sternites abdominaux (glandes sternales) existent chez tous les Termites.Mastotermes darwiniensis possède une glande sur chacun des sternites 3, 4 et 5; dans les sous-familles des Stolotermitinæ, Porotermitinæ, Hodotermitinæ, la glande sternale, unique, est située sur le 4^e sternite; tous les autres Termites examinés ont une glande sternale sur le 5^e sternite. Ces glandes existent dans toutes les castes, mais subissent une régression chez les sexués fonctionnels, qu'ils soient imaginaux ou néoténiques.

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Summary Glandular swellings of the epidermis in the region of some abdominal sternites (sternal glands) are always present in Termites.Mastotermes darwiniensis shows 3 glands, one on each of the 3rd, 4th and 5th sternites; in the sub-families Stolotermitinæ, Porotermitinæ and Hodotermitinæ, there is a single sternal gland on the 4th sternite; in all other Termites examined, one finds this sternal gland on the 5th sternite. These glands have been observed in every caste, but they regress in functional reproductives, both imaginal and neoteinic.

     

CXCR3 expression and activation of eosinophils: role of IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma

CXC chemokine receptor 3 (CXCR3), predominately expressed on memory/activated T lymphocytes, is a receptor for both IFN-gamma-inducible protein-10 (gamma IP-10) and monokine induced by IFN-gamma (Mig). We report a novel finding that CXCR3 is also expressed on eosinophils. gamma IP-10 and Mig induce eosinophil chemotaxis via CXCR3, as documented by the fact that anti-CXCR3 mAb blocks gamma IP-10- and Mig-induced eosinophil chemotaxis. gamma IP-10- and Mig-induced eosinophil chemotaxis are up- and down-regulated by IL-2 and IL-10, respectively. Correspondingly, CXCR3 protein and mRNA expressions in eosinophils are up- and down-regulated by IL-2 and IL-10, respectively, as detected using flow cytometry, immunocytochemical assay, and a real-time quantitative RT-PCR technique. gamma IP-10 and Mig act eosinophils to induce chemotaxis via the cAMP-dependent protein kinase A signaling pathways. The fact that gamma IP-10 and Mig induce an increase in intracellular calcium in eosinophils confirms that CXCR3 exists on eosinophils. Besides induction to chemotaxis, gamma IP-10 and Mig also activate eosinophils to eosinophil cationic protein release. These results indicate that CXCR3-gamma IP-10 and -Mig receptor-ligand pairs as well as the effects of IL-2 and IL-10 on them may be especially important in the cytokine/chemokine environment for the pathophysiologic events of allergic inflammation, including initiation, progression, and termination in the processes.     

Release of granule proteins from eosinophils cultured with IL-5.

Eosinophils isolated from normal individuals were cultured in the presence of human rIL-5 (hrIL-5) for up to 14 days, and the effects of this exposure were determined. First, the hrIL-5-cultured eosinophils were activated and degranulated more readily than freshly isolated eosinophils. For example, eosinophils cultured for 7 days with hrIL-5 released 30 and 10% of granule eosinophil-derived neurotoxin (EDN) when exposed to Sepharose 4B beads coupled to secretory IgA and IgG, respectively, whereas freshly isolated eosinophils released only 19 and 4%, respectively, of their EDN in response to the same stimuli. Degranulation of hrIL-5-cultured eosinophils was not augmented by further exposure to hrIL-5, whereas degranulation of freshly isolated cells to secretory IgA and IgG beads was increased by exposure to hrIL-5. Second, eosinophils cultured with hrIL-5 had prolonged viability in vitro. For example, after four days of culture with 50 U/ml of hrIL-5, 86% of eosinophils were viable compared to 12% in medium alone. Third, hrIL-5-cultured eosinophils became hypodense, and electron microscopy showed that they contained granules with core and matrix lucency and with evidence of granule fusion. Fourth, hrIL-5-cultured eosinophils spontaneously lost 30 to 60% of their EDN, eosinophil cationic protein, and eosinophil peroxidase and about 50% of their eosinophil granule major basic protein content compared to freshly isolated eosinophils, and all four of the granule proteins were released into the culture medium. Fifth, detailed studies of eosinophils cultured in hrIL-5 showed that 89 +/- 10% of the starting quantity of EDN could be recovered at 7 days. Whereas 99 +/- 1% of the EDN at day 0 was cell associated, by 7 days 60 +/- 9% was in the cell supernatants. Thus, hrIL-5 activates eosinophils, increases their viability, decreases their density, and their content of granule proteins and causes release of the granule proteins into culture fluids. The striking loss of granule proteins during culture with hrIL-5 may be an important mechanism for deposition of these cationic toxins in various diseases where IL-5 plays a role.     

Usurpation de nids étrangers et parasitisme facultatif chezHalictus scabiosæ (Rossi) (Insecte hyménoptère)

Résumé Les auxiliaires deHalictus scabiosæ (Rossi), expulsées de leurs nids lors de la dissolution des sociétés polygynes dont elles sont membres, sont contraintes de se reloger: ou bien elles creusent de nouveaux terriers, ou bien elles utilisent des nids préalablement édifiés par des femelles d'une espèce différente. Le parasitisme facultatif, décrit dans cette note, atteintEvylæus nigripes (Lep.) et va de la simple occupation d'un nid étranger au massacre de sa propriétaire avec ponte dans des cellules déjà pourvues de pains d'abeille.

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Summary The break-up of pleometrotic spring associations ofHalictus scabiosæ (Rossi) causes the expelled auxiliaries to establish their own nests. They either dig their own burrows or utilise those of other species already present. The facultative parasitism described in this note was mainly directed againstEvylæus nigripes (Lep.) and ranged from mere nest occupation to the killing of the original owner and ovopósition in already provisioned cells.

     

The role of zoobenthos in energy flow in two shallow lakes

Net production of zoobenthos in two shallow and eutrophic lakes, i.e. the S-basin of Mývatn, Iceland (maximum depth 4.2 m, mean depth 2.3 m) and Hjarbæk Fjord, Denmark (maximum depth 6.5 m, mean depth 1.9 m) were calculated as 878 and 1093 kJ m^-2 yr^-1, respectively. The zoobenthos in both lakes was dominated by Chironomidae (Diptera) living partly as filtrators feeding on suspended particles (phytoplankton) and partly as surface feeders foraging on benthic algae and/or seston. Respiration and consumption were estimated from the literature. Net production efficiency averaged 0.41 and 0.48 in Hjarbæk Fjord and Mývatn, respectively. Ingestion was dominated by herbivorous chironomids, while detritivorous tubificids were insignificant. Zoobenthic production made up 86% of total secondary production (zooplankton plus zoobenthos) in both lakes. The trophic efficiency between net primary production and benthic net secondary production was 8% and 11% in Hjarbæk Fjord and Lake Mývatn S-basin, respectively.     

SNARE proteins are critical for regulated exocytosis of ECP from human eosinophils

The SNARE hypothesis, describing a protein assembly-disassembly pathway, was recently proposed for the sequential steps of synaptic vesicle docking, activation and fusion. To determine if SNARE proteins are involved in regulated exocytosis in eosinophils, the presence and functional role of SNAREs was examined in human blood eosinophils. Immunoblotting, subcellular fractionation, and immunocytochemistry documented that vesicle-associated membrane protein-2 (VAMP-2), a vesicle-SNARE, was expressed in human eosinophils. Syntaxin 4 and SNAP-25 were also detected. Sequencing of cloned RT-PCR products amplified from a domain conserved among VAMP isoforms revealed identity only to VAMP-2 but not to VAMP-1 or cellubrevin. Functional experiments revealed that tetanus toxin pretreatment, which cleaved VAMP-2 in eosinophils, significantly inhibited both IgE receptor- and phorbol ester-mediated exocytosis of eosinophil cationic protein (ECP) from streptolysin-O-permeabilized eosinophils. Thus, these results strongly suggest a critical role of SNAREs in regulated exocytosis in eosinophils.     

Observations on the nesting behavior ofMoniæcera asperata (Fox) (Hymenoptera,sphecidæ, crabroninæ) with comments on communal nesting in solitary wasps

Summary Moniæcera asperata (Fox), a species occuring in southwestern United States and provisioning its nests in the soil with small Diptera and Hemiptera, has been found to exhibit communal nesting, two or three females sharing the same burrow. Similar behavior has been reported for four other Sphecidæ, all five belonging to the tribe Crabronini. Thus both types of social evolution believed to occur in bees, by way of communal nesting and by way matrifilial associations, are exemplified, in their initial stages, in the solitary digger wasps.

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Zusammenfassung Moniæcera asperata (Fox), eine Art von den südwestlichen Vereinigten Staaten, versorgt ihre Erdnesten mit kleinen Diptera und Hemiptera, und hat gemeinschaftliche Nester mit 2–3 Weibchen die denselben Bau teilen. Ähnliches Verhalten ist von vier andere

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Résumé Moniæcera asperata (Fox) est une espèce existant dans le Sud-Ouest des Etats-Unis et qui approvisionne ses nids creusés dans le sol de petits Diptères et Hémiptères. On a découvert chez cette espèce une tendance sociale qui se manifeste par le fait que plusieurs femelles (deux ou trois) partagent le même terrier. Ce comportement a déjà été observé chez quatre autres Sphecidæ, appartenant tous, comme celui-ci, à la tribu des Crabronini. Ainsi, les deux types d'évolution sociale qu'on croit s'être réalisés chez les abeilles, soit par nidification en commun, soit par association des filles avec leur mère, se retrouvent dans leurs phases initiales chez les Hyménoptères fouisseurs.

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Research supported by theNational Science Foundation as part of a comparative study of the behavior of solitary wasps, grant no. G 17497.     

Activation of eosinophils in cancer patients treated with IL-2 and IL-2-generated lymphokine-activated killer cells

Of 16 patients, a total of 13 who received IL-2 and autologous IL-2-generated lymphokine-activated killer LAK cells developed eosinophilia late during the course of treatment. To understand the direct or indirect effects of IL-2 on eosinophils, the physical and functional characteristics of the late-treatment eosinophils were compared to those of early-treatment and control eosinophils. Late-treatment eosinophils differed from early-treatment and control eosinophils in the following respects: they had somewhat reduced density, hypersegmented nuclei, eosinophil cationic protein converted from the storage form to the secretory form, and a greater than 200% increased ability to kill larvae of Schistosoma mansoni by an antibody-dependent mechanism (cytotoxic function). In vitro, IL-2 (1000 U/ml in medium as used to culture LAK cells) did not affect the cytotoxic function of eosinophils from cancer patients or from control subjects. However, LAK cell-conditioned medium enhanced the cytotoxic function of eosinophils from early-treatment cancer patients and from normal subjects by greater than 150%. Thus, eosinophils late in the course of IL-2/LAK cell treatment undergo physical changes and become functionally activated. The involvement of IL-2 in these changes is probably indirect, as an inducer of factors that enhance eosinophil function.     

Inhibition of eosinophil activation in bronchoalveolar lavage fluid from atopic asthmatics by Y-24180, an antagonist to platelet-activating factor

We examined the effects of Y-24180, a potent and long-acting antagonist to platelet-activating factor (PAF) receptor, on the expression of adhesion molecules in peripheral blood and bronchoalveolar lavage fluid (BALF) eosinophils from atopic asthmatics. Y-24180 (20 mg/day) was administered to 4 atopic asthmatics for 8 weeks. The number of eosinophils, the level of eosinophil cationic protein (ECP), the bindings of soluble intercellular adhesion molecule-1 (sICAM-1) and fibronectin (FN), and the expressions of CD11b (alpha chain of Mac-1) and CD49d (alpha chain of VLA-4) on eosinophils were evaluated in peripheral blood (n=4) and BALF (n=3) before and after the administration of Y-24180. The infiltration of eosinophils into the bronchial wall was also examined by taking biopsies. Eosinophil count, sICAM-1 and FN binding to eosinophils in BALF significantly decreased after the administration of Y-24180 (p<0.05). The level of CD11b expression also decreased remarkably after the administration (n=2). In peripheral blood, eosinophil count and ECP level did not change. The binding of sICAM-1 and FN, and expression of CD11b on eosinophils in peripheral blood showed a tendency to decrease after the administration. The level of CD49d expression on eosinophils changed neither in BALF nor in blood. Eosinophil infiltration into the bronchial wall markedly decreased in one out of 3 cases after the administration. These results suggest that Y-24180 inhibits the activation of eosinophils by antagonizing the actions of PAF in atopic asthmatics.     

Endosymbionten bei Ameisen

Summary Some ants of the generaCamponotus andFormica have been known to carry endosymbionts. The author has found such ones additional in the following ant species:Plagiolepis pygmæa, Camponotus æthiops, C. alii, C. cruentatus, C. foreli, C. lateralis, Formica (Serviformica) lemani, F. (S.) cinerea, andF. (S.) pilosissima.InPlagiolepis pygmæa the shape of the bacteria and the position of the mycetome fit well with the figures known fromFormica species. It is argued thatPlagiolepis andFormica got their endosymbionts from a common ancester.TheServiformica species show a sort of facultative symbiosis: In a number of species some colonies happened to be strongly infected, some were weakly or not infected. It must be questioned whether the infected ants have any essential benefit from their symbionts.

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Mit Unterstützung derKarl Hescheler-Stiftung.     

Responses to food web manipulation in a shallow waterfowl lake

The effects of fish stock reduction have been studies in 3 Dutch lakes (Lake Zwemlust, Lake Bleiswijkse Zoom and Lake Noorddiep) and 1 Danish lake (Lake Væng) during 4–5 years. A general response id described. The fish stock reduction led in general to a low fish stock, low chlorophyii-a, high transparency and high abunuance of macrophytes. Large Daphnia became abundant, but their density decreased, due to food limitation and predation by fish. The total nitrogen concentration became low due to N-uptake by macrophytes and enhanced denitrification. In Lake Bleiswijkse Zoom the water transparency deteriorated and the clear water state was not stable. The fish stock increased and the production of young fish in summer was high. lear water occurred only in spring. Large daphnids were absent in summer and the macrophytes decreased.In Lake Zwemlust, Lake Væng and Lake Noorddiep the water remained clear during the first five years. In summer of the sixth year (1992) transparency decreased in Lake Zwemlust (with high P-concentration of 1.0 mg P l^-1). Also in Lake Væng (with a low nutrient concentration of 0.15 mg P.^-1) a short term turbid stage (1.5 month) occurred in summer 1992 after a sudden collapse of the macrophytes. Deterioration of the water quality seems to start in summer and seems related to a collapse in macrophytes. At a low planktivorous fishstock (e.g. Lake Væng)thhe duration of the turbid state is shorter. than in presence of a high planktivorous fish biomass (e.g. Lake Zwemlust, and later years of Lake Bleiswijkse Zoom).     

Exploring the Architecture of Potassium Channels Using Chimæras to Reveal Signal Transduction

The chimæric channel, 4N/1, generated from two outwardly rectifying K⁺ channels by linking the N-terminal cytoplasmic domain of hKv1.4 with the transmembrane body of hKv1.1, functions as an inward rectifier. The operating range of the channel is shifted to hyperpolarizing potentials and it is inactivated at resting membrane potentials. Co-expression of a truncated form of hKv1.1 with the N-terminal domain of hKv1.4 results in the same physiology as the chimæra implying specific interactions between the two segments.