The tumor suppressor Scrib selectively interacts with specific members of the zyxin family of proteins

The zyxin family of proteins consists of five members, ajuba, LIMD1, LPP, TRIP6 and zyxin, which localize at cell adhesion sites and shuttle to the nucleus. Previously, we established that LPP interacts with the tumor suppressor Scrib, a member of the leucine-rich repeat and PDZ (LAP) family of proteins. Here, we demonstrate that Scrib also interacts with TRIP6, but not with zyxin, ajuba, or LIMD1. We show that TRIP6 directly binds to the third PDZ domain of Scrib via its carboxy-terminus. Both proteins localize in cell-cell contacts but are not responsible to target each other to these structures. In the course of our experiments, we also characterized the nuclear export signal of human TRIP6, and show that LIMD1 is localized in focal adhesions. The binding between two of zyxin's family members and Scrib links Scrib to a communication pathway between cell-cell contacts and the nucleus, and implicates these zyxin family members in Scrib-associated functions.     

Endobiotic bacteria and their pathogenic potential in cnidarian tentacles

Endobiotic bacteria colonize the tentacles of cnidaria. This paper provides first insight into the bacterial spectrum and its potential of pathogenic activities inside four cnidarian species. Sample material originating from Scottish waters comprises the jellyfish species Cyanea capillata and C. lamarckii, hydrozoa Tubularia indivisa and sea anemone Sagartia elegans. Mixed cultures of endobiotic bacteria, pure cultures selected on basis of haemolysis, but also lyophilized samples were prepared from tentacles and used for DGGE-profiling with subsequent phylogenetic analysis of 16S rDNA fragments. Bacteria were detected in each of the cnidarian species tested. Twenty-one bacterial species including four groups of closely related organisms were found in culture material. The species within these groups could not be differentiated from each other (one group of Pseudoalteromonas spp., two groups of Shewanella spp., one group of Vibrio spp.). Each of the hosts exhibits a specific endobacterial spectrum. Solely Cyanea lamarckii harboured Moritella viscosa. Only in Cyanea capillata, members of the Shewanella group #2 and the species Pseudoalteromonas arctica, Shewanella violacea, Sulfitobacter pontiacus and Arcobacter butzleri were detected. Hydrozoa Tubularia indivisa provided an amazingly wide spectrum of nine bacterial species. Exclusively, in the sea anemone Sagartia elegans, the bacterial species P. aliena was found. Overall eleven bacterial species detected were described recently as novel species. Four 16S rDNA fragments generated from lyophilized material displayed extremely low relationship to their next neighbours. These organisms are regarded as members of the endobiotic “terra incognita”. Since the origin of cnidarian toxins is unclear, the possible pathogenic activity of endobiotic bacteria has to be taken into account. Literature data show that their next neighbours display an interesting diversity of haemolytic, septicaemic and necrotic actions including the production of cytotoxins, tetrodotoxin and R-toxin. Findings of haemolysis tests support the literature data. The potential producers are Endozoicimonas elysicola, Moritella viscosa, Photobacterium profundum, P. aliena, P. tetraodonis, Shewanella waksmanii, Vibrio splendidus, V. aestuarius, Arcobacter butzleri.     

Increase of fast nutrient cycling in grassland microcosms through insect herbivory depends on plant functional composition and species diversity

Nutrient cycling in terrestrial ecosystems is affected by various factors such as plant diversity and insect herbivory. While several studies suggest insect herbivory to depend on plant diversity, their interacting effect on nutrient cycling is unclear. In a greenhouse experiment with grassland microcosms of one to six plant species of two functional groups (grasses and legumes), we tested the influence of plant species richness (diversity) and functional composition on plant community biomass production, insect foliar herbivory, soil microbial biomass, and nutrient concentrations in throughfall. To manipulate herbivory, zero, three or six generalist grasshoppers (Chorthippus parallelus) were added to the plant communities. Increasing plant species richness increased shoot biomass and grasshopper performance, without significantly affecting root biomass or insect herbivory. Plant functional composition affected all of these parameters, e.g. legume communities showed the highest shoot biomass, the lowest grasshopper performance and suffered the least herbivory. Nutrient concentrations (dissolved mineral N, PO₄‐P, SO₄‐S) and pH in throughfall increased with herbivory. PO₄‐P and pH increases were positively affected by plant diversity, especially under high herbivore pressure. Plant functional composition affected several throughfall variables, sometimes fully explaining diversity effects. Increasing plant diversity tended to increase soil microbial biomass, but only under high herbivore pressure. Faeces quantities strongly correlated with changes in pH and PO₄‐P; frass may therefore be an important driver of throughfall pH and a main source of PO₄‐P released from living plants. Our results indicate that insect herbivory may significantly influence fast nutrient cycling processes in natural communities, particularly so in managed grasslands.     

Adiponectin stimulates release of CCL2, -3, -4 and -5 while the surface abundance of CCR2 and -5 is simultaneously reduced in primary human monocytes

The adipokine adiponectin is well known to affect the function of immune cells and upregulation of CCL2 by adiponectin in monocytes/macrophages has already been reported. In the current study the effect of adiponectin on CCL2, -3, -4, and -5 and their corresponding receptors CCR1, CCR2, and CCR5 has been analyzed. Adiponectin elevates mRNA and protein of the CC chemokines in primary human monocytes. Simultaneously the surface abundance of CCR2 and CCR5 is reduced while CCR1 is not affected. Downregulation of CCR2 by adiponectin is blocked by a CCR2 antagonist although expression of the CCL2 regulated genes CCR2 and TGF-beta 1 is not altered in the adiponectin-incubated monocytes. CCL2, -3, and -5 concentrations measured in supernatants of monocytes of normal-weight (NW), overweight (OW), and type 2 diabetic (T2D) patients positively correlate with BMI and are increased in obesity and T2D. In contrast CCL4 is similarly abundant in the supernatants of all of these monocytes. The degree of adiponectin-mediated induction of the chemokines CCL3, -4, and -5 negatively correlates with their basal levels and upregulation of CCL3 and CCL5 is significantly impaired in OW and T2D cells. Serum concentrations of these chemokines are almost equal in the three groups and do not correlate with the levels in monocyte supernatants. In conclusion these data demonstrate that adiponectin stimulates release of CCL2 to CCL5 in primary human monocytes, and induction in cells of overweight probands is partly impaired. Adiponectin also lowers surface abundance of CCR2 and CCR5 and downregulation of CCR2 seems to depend on autocrine/paracrine effects of CCL2.     

Epitope mapping of the neuronal growth inhibitor Nogo-A for the Nogo receptor and the cognate monoclonal antibody IN-1 by means of the SPOT technique

Nogo-A is a potent inhibitor of axonal outgrowth in the central nervous system of adult mammals, where it is expressed as a membrane protein on oligodendrocytes and in myelin. Here we describe an attempt to identify linear peptide epitopes in its sequence that are responsible for the interaction either with the Nogo receptor (NgR) or with the neutralizing monoclonal antibody IN-1. Analysis of an array of immobilized overlapping 15 mer peptides covering the entire amino acid sequence of human Nogo-A (1192 residues) revealed a single epitope with prominent binding activity both towards the recombinant NgR and the IN-1 F(ab) fragment. Further truncation and substitution analysis yielded the minimal epitope sequence 'IKxLRRL' (x not equal to P), which occurs within the so-called Nogo66 region (residues 1054-1120) of Nogo-A. The bacterially produced Nogo66 fragment exhibited binding activity both for the recombinant NgR and for the IN-1 F(ab) fragment on the Western blot as well as in ELISA. Unexpectedly, the synthetic epitope peptide and the recombinant Nogo66 showed cross-reactivity with the 8-18C5 F(ab) fragment, which is directed against myelin oligodendrocyte glycoprotein (MOG) as a structurally unrelated target. On the other hand, the recombinant N-terminal domain of Nogo-A (residues 334-966) was shown to specifically interact on the Western blot and in an ELISA with the IN-1 F(ab) fragment but not with the recombinant NgR, which is in agreement with previous results. Hence, our data suggest that there is a distinct binding site for the Nogo receptor in the Nogo66 region of Nogo-A, whereas its interaction with NgR is less specific than anticipated before. Although there probably exists a non-linear epitope for the neutralizing antibody IN-1 in the N-terminal region of Nogo-A, which is likely to be accessible from outside the cell, a previously postulated second binding site for NgR in this region (called Nogo-A-24) remains elusive.     

Bacterial diversity in the breadcrumb sponge Halichondria panicea (Pallas)

The aim of this study was to investigate the diversity and variability of bacterial communities associated with the marine sponge Halichondria panicea with respect to tissue compartmentalization as well as seasonal and small-scale geographic variation. Diversity of microorganisms in sponges was investigated recently, but work on the variability and succession of associated bacterial communities is rare. Despite some information on Pacific and Mediterranean sponges, it is still uncertain whether bacteria and sponges are specifically associated. In this study, H. panicea specimens were sampled throughout the year at different stations around the island of Helgoland (North Sea) and investigated using molecular tools. The bacterial community associated with H. panicea was diverse, consisting of one denaturing gradient gel electrophoresis (DGGE) band occurring in most 'tissue' samples and additional variable bands. Variability was observed between different sponge fractions (i.e. the aquiferous system and the 'tissue'), sampling locations, and sampling dates. A PCR-DGGE specific for the Roseobacter group of marine Alphaproteobacteria displayed low diversity and a marked similarity between all samples. Phylogenetic analysis also pointed to specific Alphaproteobacteria of the Roseobacter group, which was predominant in most sponge 'tissue' samples. We conclude that H. panicea harbour a specific Roseobacter population with varying bacterial co-populations occurring seasonally or on a small-scale geographically, sometimes even dominating the bacterial community.     

Ultrastructural investigations of the female genital system of Epiperipatus biolleyi ( Bouvier 1902) (Onychophora,Peripatidae)

Brockmann, C., Mummert, R., Ruhberg, H. and Storch, V. 1999. Ultrastructural investigations of the female genital system of Epiperipatus biolleyi (Bouvier 1902) (Onychophora, Peripatidae). — Acta Zoologica (Stockholm) 80: 339–349. The female genital system of the neotropical peripatid Epiperipatus biolleyi was examined using transmission and scanning electron microscopy. Special attention is given to the two accessory organs of the paired oviducts: the receptacula seminis and the ovarian funnels (Ovarialtrichter). The latter occur only in the Peripatidae, whereas receptacula seminis may also be present in the Peripatopsidae, the only other family in the Onychophora. The ovarian funnels of E. biolleyi are thin-walled and closed from the haemocoel. This trait has also been reported from Epiperipatus trinidadensis, Macroperipatus torquatus, and Eoperipatus weldoni, whereas other peripatids have ovarian funnels which have been reported to open into the haemocoel. The occurrence of artificially opened ovarian funnels, caused by tissue rupture during specimen preparation, is discussed. The presence of spermatozoa both in the receptaculum seminis and in parts of the uterus of the female examined in this study supports the hypothesis that in E. biolleyi insemination of juvenile females occurs directly via the genital opening. The female contained one unstalked cleavage embryo in each uterus horn. Two features of the uterus were found to be unique to E. biolleyi: (1) a second cell layer overlying the uterine epithelium with a pronounced secretory activity (2) embryos are enclosed in a noncellular coat interspersed with numerous transport vesicles.     

Diurnal Modulation of Arginine Decarboxylase Activity of Chenopodium rubrum L. by Temperature and Light

Arginine decarboxylase (ADC), one of the key enzymes of polyaminemetabolism in plants, was investigated in Chenopodium rubrumL. seedlings under constant and alternating temperature andlighting conditions. With seedlings grown at constant temperature,ADC activity of the whole seedling increased rapidly betweenthe second and the third day after sowing. This effect was alwayshigher under continuous light than in continuous darkness. Fromthe third to the seventh day after sowing, there was a markeddecrease in ADC activity of the whole seedling almost down tothe level of the second day. Under "normal" lighting and temperatureconditions (32.5?C/10?C, light/dark) there was a marked increasein ADC activity when plants were transferred to 10?C and a rapiddecrease when they were transferred to 32.5?C. The same timecourse was observed when an "inverse" light-temperature program(32.5?C/10?C; dark/light) was applied. This means that the timecourse of ADC activity in the seedlings is slightly light-dependent,but strongly temperature-dependent. The data are discussed withrespect to the chronopathological effects of the "inverse" light-temperatureprogram. (Received March 5, 1985; Accepted October 2, 1985)     

Ultrastructural Analysis of the Sensilla of Austroperipatus aequabilis Reid, 1996 (Onychophora,Peripatopsidae)

The head of Austroperipatus aequabilis bears five types of sensilla. which were examined by electron microscopy. They differ from each other in position, shape of outer sensory elements and cuticular socket structures. Thus, we distinguish sensilla with sensory hairs, sensilla with sensory bulbs, cone-shaped sensilla. sensilla with sensory bristles, and sensilla of the lips. They are composed of up to 15 cells, which can he separated into four cell types. The most frequent cell type is the bipolar receptor cell that occurs in all sensilla. The apical surface of this primary receptor cell is characterized by one or two partly branched cilia with a basal 9 × 2 + 0 pattern of microtubules. A modified bipolar receptor cell was found in all sensilla bearing a sensory peg except for the sensilla equipped with sensory bristles. The apical dendrite extends to a long pale process which exclusively contains mitochondria and single microtubules. In all sensilla examined in this study at least one supporting cell occurs which is characterized by parallel microvilli. An additional function of this cell type as a part of the stimulus-conducting system is possible. In the sensillum with a sensory bulb two kinds of supporting cells occur. A unique cell type with an upside down position has regularly been found in all sensilla bearing a sensory peg. Apart from the sensilla they also occur within the labial epidermis. Since most sensilla contain several different receptor cells, they can be considered as complex sense organs. © 1998 The Royal Swedish Academy of Sciences. Published by Elsevier Science Ltd. All rights reserved