Temporal target restriction of olfactory receptor neurons by Semaphorin-1a/PlexinA-mediated axon-axon interactions

Axon-axon interactions have been implicated in neural circuit assembly, but the underlying mechanisms are poorly understood. Here, we show that in the Drosophila antennal lobe, early-arriving axons of olfactory receptor neurons (ORNs) from the antenna are required for the proper targeting of late-arriving ORN axons from the maxillary palp (MP). Semaphorin-1a is required for targeting of all MP but only half of the antennal ORN classes examined. Sema-1a acts nonautonomously to control ORN axon-axon interactions, in contrast to its cell-autonomous function in olfactory projection neurons. Phenotypic and genetic interaction analyses implicate PlexinA as the Sema-1a receptor in ORN targeting. Sema-1a on antennal ORN axons is required for correct targeting of MP axons within the antennal lobe, while interactions amongst MP axons facilitate their entry into the antennal lobe. We propose that Sema-1a/PlexinA-mediated repulsion provides a mechanism by which early-arriving ORN axons constrain the target choices of late-arriving axons.     

Stainless steel sponge: a novel carrier for the immobilisation of the white-rot fungus Trametes hirsuta for decolourization of textile dyes

Alginate beads, polyurethane foam, nylon sponge and stainless steel sponge were tested as carrier materials for the white-rot fungus Trametes hirsuta for laccase production under submerged fermentation conditions. Stainless steel sponge was the best carrier material leading to the highest laccase activities of up to 800 U/l after 8 days of cultivation. These values are higher than those reported to date operating with inert supports and without inducer addition. In a 1-l bioreactor containing T. hirsuta immobilised on stainless steel sponge laccase activities of about 2200 U/l were obtained when the culture medium was supplemented with 1 mM copper sulphate. There were no operational problems with this system during culturing time. The textile dye Indigo Carmine was almost totally degraded in 3 days by T. hirsuta grown in this bioreactor, while Lanaset Marine was degraded in two successive batches, reaching in the first batch a decolourization percentage of about 82% in 15 h and in the second one by 71% in 28 h. Results obtained after inhibition of growth of T. hirsuta by antibiotics indicated that dye decolourization could not exclusively be attributed to laccase activity.     

Multiplexed homogeneous proximity ligation assays for high-throughput protein biomarker research in serological material

A high throughput protein biomarker discovery tool has been developed based on multiplexed proximity ligation assays in a homogeneous format in the sense of no washing steps. The platform consists of four 24-plex panels profiling 74 putative biomarkers with sub-pm sensitivity each consuming only 1 μl of human plasma sample. The system uses either matched monoclonal antibody pairs or the more readily available single batches of affinity purified polyclonal antibodies to generate the target specific reagents by covalently linking with unique nucleic acid sequences. These paired sequences are united by DNA ligation upon simultaneous target binding forming a PCR amplicon. Multiplex proximity ligation assays thereby converts multiple target analytes into real-time PCR amplicons that are individually quantified using microfluidic high capacity qPCR in nano liter volumes. The assay shows excellent specificity, even in multiplex, by its dual recognition feature, its proximity requirement, and most importantly by using unique sequence specific reporter fragments on both antibody-based probes. To illustrate the potential of this protein detection technology, a pilot biomarker research project was performed using biobanked plasma samples for the detection of colorectal cancer using a multivariate signature.     

Intertidal rocky shore seaweed communities subject to the influence of shallow water hydrothermal activity in São Miguel (Azores, Portugal)

The volcanic origin of the Azores archipelago (Portugal) gives rise to active deep sea and shallow water hydrothermal activity that affects benthic communities. Intertidal seaweed surveys were conducted at two shores affected by intense shallow water hydrothermal vents. Water temperature, acidity and salinity were monitored. Seaweed communities were found to be species poor and have a disproportionally larger number of filamentous early successional species on shores that are subject to the effect of hot and acidic freshwater of volcanic origin. There is an ecological resemblance between hydrothermally affected seaweed communities in the Azores and those affected by acid mine drainage in the UK, thus indicating that hydrothermalism can be a useful scenario for pollution studies under conditions of ocean warming and acidification.     

Flower color change accelerated by bee pollination in Tibouchina (Melastomataceae)

Floral color changes are common among Melastomataceae and have been interpreted as a warning mechanism for bees to avoid old flowers, albeit increasing long-distance flower display. Here the reproductive systems of Tibouchina pulchra and T. sellowiana were investigated by controlled pollinations. Their pollinators were identified, and experiments on floral color and fragrance changes were conduced to verify if those changes affect the floral visitation. Both Tibouchina species are self compatible. The flowers lasted three days or more, and the floral color changed from white in the 1st day to pink in the following days. Pollen deposition on stigma induced floral color change. The effectiveness of the pollination is dependent on bees’ size; only large bees were regarded as effective pollinators. In experimental tests, the bees in T. pulchra preferred the natural white flowers while the visitors of T. sellowiana were attracted by both natural and mimetic 1st-day flowers (2nd-day flowers with experimentally attached 1st-day flower petals). During the experiments on floral fragrance, the bees visited both natural and mimetic 1st-day flowers (2nd-day flowers with 1st-day flower scents). In both experiments, the bees avoided natural 2nd-day flowers, but seldom visited modified 2nd-day flowers. The attractiveness of T. pulchra and T. sellowiana flowers cannot be attributed exclusively to the color or the fragrance separately, both factors seemingly act together.     

Zygoparity and sex steroid hormone profiles in bluemouth Helicolenus dactylopterus

Two hundred and seven individuals (103 females and 104 males) of bluemouth Helicolenus dactylopterus (Scorpaeniformes, Sebastidae), a commercially important deep‐water species with an unusual reproductive strategy, from the eastern Atlantic Ocean ranging from 13·9 to 37·5 cm total length (L_T) were analysed from September 2011 to October 2012. The analysis included gonad maturity phases and blood‐plasma levels of oestradiol‐17β (E₂), 11‐ketotestosterone (11‐KT) and 17,20β‐dihydroxypregn‐4‐en‐3‐one (17,20β‐P). Results confirmed the existence of an annual reproductive cycle with asynchrony between females and males and a spawning season from January to May. A pronounced peak in 17,20β‐P in October for both sexes was associated with possible mating behaviour and recent copula. Levels of E₂ increased preceding the elevation of the gonado‐somatic index during ovarian growth and were lower during regression and regeneration. The frequency distribution of oocyte–embryonic stages and variation of hormone levels suggest the existence of daily rhythms. Fertilization was detected between 2000–0000 and 0800–1200 h and spawning took place throughout the day peaking between 2000 and 0000 h. The cyclic pattern of sex steroids and ovarian recruitment provides a new insight into the reproductive strategy of this species.     

Hydrolysis of the brain dipeptide N-acetyl-L-aspartyl-L-glutamate. Identification and characterization of a novel N-acetylated alpha-linked acidic dipeptidase activity from rat brain

High performance liquid chromatography studies documented the presence of an enzyme activity, N-acetylated alpha-linked acidic dipeptidase (NAALA dipeptidase), in rat brain membranes that cleaves the endogenous brain dipeptide, N-acetyl-L-aspartyl-L-glutamate to N-acetyl-aspartate and glutamate. With ion exchange chromatography, which quantitatively separated [3,4-3H]glutamate from N-acetyl-L-aspartyl-L-[3,4-3H]glutamate, we found that NAALA dipeptidase activity was essentially restricted to nervous tissue and kidney. We characterized NAALA dipeptidase activity in lysed synaptosomal membranes obtained from rat forebrain. Membrane-bound NAALA dipeptidase activity was optimal between pH 6.0 and 7.4 at 37 degrees C. Eadie-Hofstee analysis of kinetic data revealed a rather high apparent affinity for N-acetyl-L-aspartyl-L-glutamate with a Km = 540 nM and a Vmax = 180 nM/mg of protein/min. While NAALA dipeptidase showed a requirement for monovalent anions such as Cl-, the polyvalent anions phosphate and sulfate inhibited enzyme activity 50% at 100 microM and 1 mM, respectively. The divalent metal ion chelators EGTA, EDTA, and o-phenanthroline completely abolished activity, which was partially restored by manganese. Treatment of membranes with 1 mM dithiothreitol abolished NAALA dipeptidase activity. NAALA dipeptidase activity was also sensitive to the aminopeptidase inhibitors bestatin and puromycin, although not to the selective aminopeptidase A inhibitor amastatin. Structure-activity relationships inferred from inhibitor studies suggest that this enzyme shows specificity for N-acetylated alpha-linked acidic dipeptides. NAALA dipeptidase was also potently inhibited by the excitatory amino acid agonist L-quisqualate. Comparison of the properties of NAALA dipeptidase to those of previously characterized enzymes suggests that this is a novel peptidase which may be involved in the synaptic degradation of N-acetyl-L-aspartyl-L-glutamate.     

Dye removal by immobilised fungi

Dyes are widely used within the food, pharmaceutical, cosmetic, printing, textile and leather industries. This has resulted in the discharge of highly coloured effluents that affect water transparency and gas solubility in water bodies. Furthermore, they pose a problem because of their carcinogenicity and toxicity. Therefore, removal of such dyes before discharging them into natural water streams is essential. For this, appropriate treatment technologies are required. The treatment of recalcitrant and toxic dyes with traditional technologies is not always effective or may not be environmentally friendly. This has impelled the search for alternative technologies such as biodegradation with fungi. In particular, ligninolytic fungi and their non-specific oxidative enzymes have been reported to be responsible for the decolouration of different synthetic dyes. Thus, the use of such fungi is becoming a promising alternative to replace or complement the current technologies for dye removal. Processes using immobilised growing cells seem to be more promising than those with free cells, since the immobilisation allows using the microbial cells repeatedly and continuously. This paper reviews the application of fungal immobilisation to dye removal.     

Olfactory Attraction of the Hornet Vespa velutina to Honeybee Colony Odors and Pheromones

Since the beginning of the last century, the number of biological invasions has continuously increased worldwide. Due to their environmental and economical consequences, invasive species are now a major concern. Social wasps are particularly efficient invaders because of their distinctive biology and behavior. Among them, the yellow-legged hornet, Vespa velutina, is a keen hunter of domestic honeybees. Its recent introduction to Europe may induce important beekeeping, pollination, and biodiversity problems. Hornets use olfactory cues for the long-range detection of food sources, in this case the location of honeybee colonies, but the exact nature of these cues remains unknown. Here, we studied the orientation behavior of V. velutina workers towards a range of hive products and protein sources, as well as towards prominent chemical substances emitted by these food sources. In a multiple choice test performed under controlled laboratory conditions, we found that hornets are strongly attracted to the odor of some hive products, especially pollen and honey. When testing specific compounds, the honeybee aggregation pheromone, geraniol, proved highly attractive. Pheromones produced by honeybee larvae or by the queen were also of interest to hornet workers, albeit to a lesser extent. Our results indicate that V. velutina workers are selectively attracted towards olfactory cues from hives (stored food, brood, and queen), which may signal a high prey density. This study opens new perspectives for understanding hornets’ hunting behavior and paves the way for developing efficient trapping strategies against this invasive species.