The glucose levels and carbohydrate autolysis inVespa orientalis hemolymph

Summary The hemolymph ofVespa orientalis is rich in saccharolytic enzymes which convert poly- and disaccharides into monosaccharides.During the life of the insect the enzymatic activity is probably controlled by a hormonal feedback reaction which is analogous for most living organisms. After removal of hemolymph from the insect, the enzymes start uncontrolled activity, which is limited only by the amount of available substrate. Thus the liberated monosaccharides express the total available energy reserve and the initial levels, which represent the immediate requirement of the insect.Both factors are governed by the seasonal, environmental, physical and physiological requirements of the individual insect and are widely variable.The enzyme levels and the substrate reserves generally tend to increase with maturation, but functional and caste differences are significant.The enzyme assays of the fresh and of the incubated hemolymph provides valuable information on the developmental and the biological status of the insects.

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Résumé L'hémolymphe duVespa orientalis est riche en enzymes saccharolytiques, ceux qui transforment les poly- et disaccharides en monosaccharides.Durant la vie de l'insecte, l'activité enzymatique est contrôlée par la réaction hormonale du retour des aliments, qui est analogue chez la plupart des organismes vivants. Après l'extraction de l'hémolymphe de l'insecte, les enzymes mettent en train une activité incontrôlable, qui ne cesse que par l'épuisement du substrat disponible. Les monosaccharides ainsi libérés font connaître la réserve totale de l'énergie valable et les quantités initiales qui sont l'expression du besoin immédiat de l'insecte.Cex deux facteurs dépendent de la saison, de l'environnement, des besoins physiques et physiologiques de chaque insecte individuel, et sont extrêmement variables.Les valeurs des enzymes et les réserves du substrat sont d'habitude disposées à augmenter en parallèle à la maturation; mais les divergences qui existent quant à la caste et à la fonction spécifique restent significatives.Les essais enzymatiques de l'hémolymphe fraîche ainsi que de l'hémolymphe incubée offrent des renseignements de valeur sur l'état développemental et biologique de l'insecte.

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This research study was carried out with the aid of Ford Research Grant No. B III/6 accorded to Dr. JacobIshay.     

Characterization of COMMD protein-protein interactions in NF-kappaB signalling

COMMD [copper metabolism gene MURR1 (mouse U2af1-rs1 region 1) domain] proteins constitute a recently identified family of NF-kappaB (nuclear factor kappaB)-inhibiting proteins, characterized by the presence of the COMM domain. In the present paper, we report detailed investigation of the role of this protein family, and specifically the role of the COMM domain, in NF-kappaB signalling through characterization of protein-protein interactions involving COMMD proteins. The small ubiquitously expressed COMMD6 consists primarily of the COMM domain. Therefore COMMD1 and COMMD6 were analysed further as prototype members of the COMMD protein family. Using specific antisera, interaction between endogenous COMMD1 and COMMD6 is described. This interaction was verified by independent techniques, appeared to be direct and could be detected throughout the whole cell, including the nucleus. Both proteins inhibit TNF (tumour necrosis factor)-induced NF-kappaB activation in a non-synergistic manner. Mutation of the amino acid residues Trp24 and Pro41 in the COMM domain of COMMD6 completely abolished the inhibitory effect of COMMD6 on TNF-induced NF-kappaB activation, but this was not accompanied by loss of interaction with COMMD1, COMMD6 or the NF-kappaB subunit RelA. In contrast with COMMD1, COMMD6 does not bind to IkappaBalpha (inhibitory kappaBalpha), indicating that both proteins inhibit NF-kappaB in an overlapping, but not completely similar, manner. Taken together, these data support the significance of COMMD protein-protein interactions and provide new mechanistic insight into the function of this protein family in NF-kappaB signalling.     

A comparative study of mechanisms underlying digenean-snail specificity: in vitro interactions between hemocytes and digenean larvae

A panel of 4 digenetic trematode species (Echinostoma paraensei, E. trivolvis, Schistosoma mansoni, and Schistosomatium douthitti) and 5 snail species (Biomphalaria glabrata, Helisoma trivolvis, Lymnaea stagnalis, Stagnicola elodes, and Helix aspersa) was examined to determine if known patterns of host specificity could be explained by the tendency of digenean larvae to be bound by snail hemocytes, or by the ability of larvae to influence the spreading behavior of hemocytes. In short-term (1 hr) in vitro adherence assays, there was no overall pattern to suggest that sporocysts were more likely to be bound by hemocytes from incompatible than compatible snails. Compared with the other parasites, sporocysts of E. paraensei were less likely to be bound by hemocytes from any of the snail species tested. All rediae examined, including those of another species Echinoparyphium sp., were also remarkably refractory to binding by hemocytes from any of the snails. Of all the larvae examined, only sporocysts and young daughter rediae of E. paraensei caused hemocytes to round up in their presence. This was true for hemocytes from the compatible species B. glabrata and the incompatible lymnaeid species S. elodes and L. stagnalis. The patterns of host specificity shown by this particular panel of parasites and snails were not predicted by either the extent of hemocyte adherence to digenean larvae or by the ability of larvae to affect hemocyte spreading behavior. The results of this study suggest that a role for hemocytes, although likely, may require different assays, possibly of a more prolonged nature, for its detection. Also, different parasite species (notably E. paraensei) and intramolluscan stages have distinctive interactions with host hemocytes, suggesting that the determinants of specificity vary with the host-parasite combination, and with the parasite life cycle stage.     

Collagen-platelet interactions: recognition and signalling

The collagen-platelet interaction is central to haemostasis and may be a critical determinant of arterial thrombosis, where subendothelium is exposed after rupture of atherosclerotic plaque. Recent research has capitalized on the cloning of an important signalling receptor for collagen, glycoprotein VI, which is expressed only on platelets, and on the use of collagen-mimetic peptides as specific tools for both glycoprotein VI and integrin alpha 2 beta 1. We have identified sequences, GPO and GFOGER (where O denotes hydroxyproline), within collagen that are recognized by the collagen receptors glycoprotein VI and integrin alpha 2 beta 1 respectively, allowing their signalling properties and specific functional roles to be examined. Triple-helical peptides containing these sequences were used to show the signalling potential of integrin alpha 2 beta 1, and to confirm its important contribution to platelet adhesion. Glycoprotein VI appears to operate functionally on the platelet surface as a dimer, which recognizes GPO motifs that are separated by four triplets of collagen sequence. These advances will allow the relationship between the structure of collagen and its haemostatic activity to be established.     

寄生蜂畸形细胞特性及与蜂幼虫生长发育的关系

本文综述寄生蜂畸形细胞特性及其功能研究的最新进展。一些进化高等的膜翅目内寄生蜂,在胚胎发育过程中,包裹在胚胎周围的浆膜会随着蜂卵的孵化,以游离的细胞团或单个细胞的形式释放到寄主昆虫的血淋巴中。这些源自浆膜被称作“畸形细胞（Teratocyte）”的特殊类群,在伴随蜂幼虫的生长发育过程中,自身也经历一系列结构和功能的改变。它们主要通过分泌蛋白质或酶类,改变寄主的物质代谢途径等方式为发育中的蜂幼虫提供营养和能量。对寄生蜂重要的调控因子一畸形细胞的特性及与蜂幼虫相互关系的揭示,可以阐明寄生蜂一寄主昆虫协同进化的内在机理,也可为开发害虫生物防治新策略提供新的思路和途径。     

Siglecs: sialic-acid-binding immunoglobulin-like lectins in cell-cell interactions and signalling

Siglecs are sialic-acid-binding immunoglobulin-like lectins involved in cell-cell interactions and signalling functions in the haemopoietic, immune and nervous systems. Significant advances have been made in our understanding of the link between carbohydrate recognition and signalling for two well-characterised siglecs, CD22 and myelin-associated glycoprotein. Over the past few years, several novel siglecs have been discovered through genomics and functional screens. These "CD33-related" siglecs have molecular features of inhibitory receptors and may be important in regulating leucocyte activation during immune and inflammatory responses.     

Rhythmic expression, light entrainment and alpha-MSH modulation of rhodopsin mRNA in a teleost pigment cell line

To investigate whether teleost fish GEM-81 erythrophoroma cells were photosensitive, the cells were submitted to constant darkness (DD), 14 h of light and 10 h of darkness (14L:10D), and 10 h of light and 14 h of darkness (10L:14L). The doubling times (hours) were: DD 35.33+/-0.05; 14L:10D 67.85+/-0.04; and 10L:14D 49.60+/-0.08. In order to verify whether proliferation was dependent on light phase length, GEM-81 cells were submitted to 7L: 5D. The proliferation curves and doubling times were similar in 14L:10D and 7L:5D (respectively 69.44+/-0.03 and 67.85+/-0.04), suggesting that the cell cycle was regulated by the length of the light phase within 24 h, or by the light/dark ratio. We have also demonstrated the expression of Carassius retinal rhodopsin mRNA in GEM-81 cells, which cycles in a circadian rhythm, entrained by light. In addition, we showed that alpha-melanocyte stimulating hormone (alpha-MSH, 10(-10) to 10(-8) M), a conspicuous hormone that exerts mitogenic and melanogenic activity in most vertebrates, decreased rhodopsin mRNA in the first 3 days; after 4 days the inhibition was reversed, and after 5 days an increase in rhodopsin mRNA level was elicited. This is the first report of rhythmic expression of extra-ocular rhodopsin and its modulation by light and hormones.     

Modeling interactions between photic and nonphotic entrainment mechanisms in transmeridian flights

In transmeridian flights, photic and nonphotic entrainment mechanisms are expected to interact dynamically in the human circadian system. In order to simulate the reentrainment process of the circadian rhythms, the photic entrainment mechanism was introduced to our previous model, which consisted of three coupled oscillators. Regardless of flight direction, a large time difference beyond 10 h tended to induce the antidromic reentrainment. The partition between the oscillators resulted for the eastward flight over a 10-h or longer time difference and the westward over 6 h or longer. The simulated reentrainment processes almost coincided with empirical knowledge. Simulated effects of physical exercise showed that some antidromic reentrainments were switched to the orthodromic ones for the eastward flight and most of the partitions between the oscillators were prevented in the westward flight. These results are due to an augmentation of the entrainment pressure of the rest–activity cycle on the oscillators. The mechanisms underlying these various reentrainment patterns were explained based on the photic response, the interactions between the oscillators, and their adaptive modification. The simulation results suggest that an appropriate selection of departure time and physical exercise could ease the jet lag caused by transmeridian flight.This research was supported by a Grant-In-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (Nos. 14015205, 15014204, and 15560372).     

Characterization of some membrane-active components of Vespa orientalis venom

The molecular weight distribution of the components of giant hornet (Vespa orientalis) venom was studied, using gel-filtration on a column with Sephadex G-50. The effects of the venom and its constituent fractions on the permeability and stability of artificial bilayer phospholipid membranes, potassium ions release from the erythrocytes and mitochondrial oxidative phosphorylation parameters, as well as on the activity and stability of polyenzymic systems of the mitochondrial respiratroy chain, were studied. The data obtained suggest that the high molecular weight fractions contain phospholipases, whose activities are much higher than those of presently known venoms. Despite the fact that the hemolytic effect is typical for two low molecular weight fractions, no fractions possessing high activity of bee venom of the melitin type were found.     

Protein-protein interactions in receptor activation and intracellular signalling

We review here signalling complexes that we have defined using X-ray analysis in our laboratory. They include growth factors and their receptors: nerve growth factor (NGF) and its hetero-hexameric 7S NGF storage complex, hepatocyte growth factor/scatter factor (HGF/SF) NK1 dimers and fibroblast growth factor (FGF1) in complex with its receptor (FGFR2) ectodomain and heparin. We also review our recent structural studies on intracellular signalling complexes, focusing on phosducin transducin GPry, CK2 protein kinase and its complexes, and the cyclin D-dependent kinase, Cdk6, bound to the cell cycle inhibitor p19INK4d. Comparing the structures of these complexes with others we show that the surface area buried in signalling interactions does not always give a good indication of the strength of the interactions. We show that conformational changes are often important in complexes with intermediate buried surface areas of 1500 to 2000 A2, such as Cdk6INK4 interactions. Some interactions involve recognition of continuous epitopes, where there is no necessity for a tertiary structure and very often the binding conformation is induced during the process of interaction, for example phosducin binding to the betagamma subunits (Gtbetagamma) of the heterotrimeric G protein transducin.     

Physiological mechanisms underlying the control of meal size in Manduca sexta larvae

Abstract. Fifth stadium larvae of the tobacco hornworm, Manduca sexta (L.), ate larger meals than usual when they had been deprived of food for periods of time longer than the usual intermeal interval (c. 45 min). Meal size increased with time since the last meal until 180 min, when it was about 3 times normal. There was no evidence of a role for volumetric feedback from the gut in controlling meal size. Injections of a paraffin oil/wax mixture, or of petroleum jelly (Vaseline) into the foregut, midgut or rectum failed to decrease meal size. Cutting the recurrent nerve failed to alter meal size compared to sham-operated controls (although both groups took smaller meals than unoperated controls). By contrast, injections of an extract of soluble nutrients from the diet into the midgut inhibited feeding in some insects and reduced subsequent meal size in others. Appropriate controls showed that these effects were not due to the volumetric or osmotic effects of the injections. These results imply that nutrient feedback plays an important role in controlling meal size in Manduca caterpillars, while volumetric feedback is probably unimportant.     

Cell lineages in the embryonic kidney: their inductive interactions and signalling molecules.

The first signalling genes acting in the inductive interactions in the kidney have now been identified. Differentiation of the permanent kidney or the metanephros is critically dependent on inductive signalling between the nephrogenic mesenchyme and ureteric bud epithelium. Further inductive interactions occur between developing nephrons, interstitial stroma, endothelial cells and neurones. Glial-cell-line-derived neurotrophic factor is a signal for the ureteric bud initiation and branching, and Wnt4 is an autocrine epithelializing signal at the pretubular stage of nephron formation. The signals for renal angiogenesis and innervation are less well defined, but seem to include vascular endothelial growth factor and neurotrophins, at least. The ureteric-bud-derived signal for induction of the nephrogenic mesenchyme (to bring the cells to the condensate stage) is not yet known, but fibroblast growth factor 2 is a good candidate. None of the signalling genes identified from the embryonic kidney is specific to the organ, which raises some general questions. How do the organs develop from similar rudiments to various patterns with different cell types and functions? Does the information for organ-specific differentiation pathways retain in the epithelial or mesenchymal compartment? The present, rather fragmentary molecular data would favour the view that similar molecules acting in different combinations and developmental sequences, rather than few organ-specific master genes, could be responsible for the divergence of patterning.     

Binocular interactions in the entrainment and phase shifting of locomotor activity rhythms in Syrian hamsters

To assess binocular interactions and possible ocular dominance in entrainment of circadian rhythms, Syrian hamsters maintained in LL were subjected for several weeks to schedules of eye occlusion with opaque contact lenses. In separate groups, the opaque lens was inserted into the left or right eye for 12 h at the same clock time each day. The left and right eyes of other groups were alternately occluded for 12 h each day, with initial occlusion of either the left or right eye for different groups. A majority of hamsters entrained their locomotor activity rhythm when 1 eye was occluded for 12 h. The modified visual input imposed by covering 1 eye is sufficient to induce entrainment. Locomotor rhythms of most animals in which the 2 eyes were alternately occluded for 12 h each day phasedelayed onset of activity during the 1st few days of the lensing procedure; activity onset then free ran with tau < 24 h for several weeks until entraining with tau of 24 h regardless of whether the left or right eye was initially occluded. Entrainment eventually occurred when activity onset coincided with occlusion of the eye contralateral to the one that was first lensed. Photic and nonphotic explanations for eventual entrainment of locomotor rhythms are discussed, and evidence for asymmetrical photic input from the 2 eyes to the SCN is considered.