Macroevolution of perfume signalling in orchid bees

Theory predicts that both stabilising selection and diversifying selection jointly contribute to the evolution of sexual signalling traits by (1) maintaining the integrity of communication signals within species and (2) promoting the diversification of traits among lineages. However, for many important signalling traits, little is known about whether these dynamics translate into predictable macroevolutionary signatures. Here, we test for macroevolutionary patterns consistent with sexual signalling theory in the perfume signals of neotropical orchid bees, a group well studied for their chemical sexual communication. Our results revealed both high species‐specificity and elevated rates of evolution in perfume signals compared to nonsignalling traits. Perfume complexity was correlated with the number of congeners in a species’ range, suggesting that perfume evolution may be tied to the remarkably high number of orchid bee species coexisting together in some neotropical communities. Finally, sister‐pair comparisons were consistent with both rapid divergence at speciation and character displacement upon secondary contact. Together, our results provide new insight into the macroevolution of sexual signalling in insects.     

Antioxidant and anti‐inflammatory effects of the monocarbonyl curcumin analogs B2BRBC and C66 in monocrotaline‐induced right ventricular hypertrophy

For 22 days after monocrotaline injection two groups of rats received either of the monocarbonyl curcumin analogs (2E,6E)‐2,6‐bis(2‐bromobenzylidene)cycloxehanone (B2BrBC) and (2E,6E)‐2,6‐bis([2‐tri?uoromethyl]benzylidene)cyclohexanone (C66), and their right ventricle parameters were compared to those from the control and the monocrotaline injected animals. B2BrBC and C66 treatments did not prevent the monocrotaline‐induced right ventricular hypertrophy but attenuated the changes in antioxidant enzyme activities and reduced inflammation. The level of thiol‐based nonenzymatic antioxidants did not change in the function of monocrotaline or curcumin analogs treatment. However, due to its stronger antioxidant properties, only B2BrBC treatment was effective in the reduction of monocrotaline‐associated lipid peroxidation. The obtained results suggest that increasing the levels of antioxidant enzymes may not be sufficient to reduce oxidative stress and chronic inflammation optimally and our current study supports the potential of compounds with more than one beneficial biological activity as a promising treatment against the progression of cardiac hypertrophy.     

Grandisol as an attractant for the sugar beet pest Bothynoderes affinis and data on other Lixinae species

As a result of field tests in Bulgaria and Hungary, cis‐2‐isopropenyl‐l‐methylcyclobutane ethanol (racemic grandisol) is reported for the first time as an attractant for Bothynoderes affinis (Schrank) (Coleoptera: Curculionidae, Lixinae), a member of the pest weevil complex of sugar beet. Dose–response experiments in the field using Csalomon TAL (modified pitfall) traps (Plant Protection Institute, CAR HAS, Budapest, Hungary) showed that catches of B. affinis adults increased with increasing attractant dose. In a subsequent experiment studying the effect of trap color (white, blue, yellow, fluorescent yellow, and transparent) all traps with the lure caught more than non‐baited control traps, and the highest number of adults was recorded in transparent and yellow baited traps. Trap color had a significant effect on the number of B. affinis females captured. Transparent TAL traps baited with 1–10 mg grandisol applied on rubber dispensers are recommended for the detection and monitoring of B. affinis. In addition to the target species, 17 other Lixinae species were captured during the field experiments, demonstrating for the first time the possible role of grandisol in the chemical communication systems of some of these species. A second locality of Lixus punctiventris Boheman (Lixinae, Lixini) in Bulgaria is reported. TAL traps baited with grandisol might be a useful tool for surveying Lixinae diversity in different biotopes.     

Female sex pheromone of Cameraria ohridella Desch. and Dim. (Lepidoptera: Gracillariidae): structure confirmation,synthesis and biological activity of (8E,10Z)-8,10-tetradecadienal and some analogues

Mass spectrometric investigations confirmed the structure of the female produced sex pheromone of the horse-chestnut leafminer Cameraria ohridella Desch. and Dim. to be (8E,IOZ)-8,10-tetradecadienal. Pure samples, prepared in a straightforward synthesis, were highly attractive in field tests and proved to be suitable for monitoring of flight activities and population dynamics. In mixtures with the synthetic pheromone, analogues like 9-tridecynal and 7-dodecynyl formate were shown to reduce trap catches. In electroantennographic experiments, pheromone analogues were less active than the pheromone. 9-Tridecynal was the most EAG active analogue tested, followed by 7-dodecyn-1-yl formate and 7-undecyn-1-yl formate.     

Correlation between nerve terminal size and muscle fibers diameter in urodela extrinsic eye muscle.

Extrinsic eye muscles of newts and salamanders were investigated by means of electron microscope. It was possible to distinguish two types of muscle fiber tonic-slow and twitch-fast acting ones. It was shown that mioneural junctions in both types of fibers differ in their ultrastructural organization because of lack of post-synaptic infoldings on the surface of slow tonic fibers. After "cholinesterase" "staining" it was possible to measure the surface of junctional area and correlate it with the diameter of particular muscle fiber. The results show a positive correlation.     

Sex pheromone components ofMamestra suasa: chemical analysis, electrophysiological activity, wind tunnel activity and field tests in two European countries

(Z)-11-hexadecenyl acetate (Z-11-16:Ac), (Z)-11-hexadecenal (Z-11-16:Ald), (Z)-11-hexadecenol (Z-11-16:OH) and hexadecanyl acetate (16:Ac) were found in pheromone gland extracts of femaleMamestra suasa (Den. et Schiff.) in the relative amounts 100/2/10/5. All four compounds were also present in collections of aiiborne volatiles from calling females in a 100/7/5/5 ratio. No traces of 14 carbon aldehydes or acetates were detected. In gland extracts the presence of methyl hexadecanoate methyl (Z)-9-hexadecenoate and methyl (Z)-11-hexadecenoate was demonstrated by base methanolysis. No methyl tetradecenoates were detected. In EAG tests Z-11-16:Ac gave the best responses, followed by (Z)-9-tetradecenyl acetate (Z-9-14:Ac) Z-11-16:Ald and Z-11-16:OH. In single sensillum recordings large spike amplitude cells in sensilla responded to Z-11-16:Ac, while small spike amplitude cells to both Z-11-16:OH and Z-9-14:Ac. Cells responding to Z-11-16:Ald were found in one out of 60 sensilla tested. In wind tunnel tests 0.1 g of a 10:1 blend of Z-11-16:Ac/Z-11-16:Ald evoked the same responses and at a similar intensity as 3 isolated female pheromone glands did. In field tests a 10:1 blend of Z-11-16:Ac/Z-11-16:Ald caught significant numbers of males in both Bulgaria and Hungary. The addition of 16:Ac to the binary blend did not have any effect, while more than 1% of Z-11-16:OH or 0.1% of Z-9-14:Ac dramatically decreased captures. In comparing different ratios of the, acetate/aldehyde blend at different dose levels, best catches were recorded at the 10:1 ratio and at the highest (1000 g) dose level.

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La composition de la phéromone sexuelle deMamestra suasa: analyse chimique, étude de l'effet par éléctrophysiologie et à la chambre de vol, et piégeages dans deux pays de l'Europe

Résumé On a trouvé l'acetoxy-1 hexadécene-11 Z (Z-11-16:Ac), le hexadécene-11 Z al-1 (Z-11-16:Ald), le hexadécene-11 Z ol-1 (Z-11-16:OH) et l'acetoxy-1 hexadécene (16:Ac) dans des extraits de glandes phéromona les des femelles deMamestra suasa. La proportion relative des composés était 100/2/10/5. Tous les quatre composés ont été présents aussi dans les collections d'émanations des femelles en stade d'appel, dans la proportion un peu différente de 100/7/5/5. On n'a détecté aucune trace des tétradécenes al-1 ou d'acetoxy-1 tétradécenes. On a démontré la présence de hexadécenoate-1 methyl, hexadécene-9 Z oate-1 methyl et héxadécene-11 Z oate-1 methyl dans des extraits des glandes, par la méthode de base methanolysis. On n'a trouvé pas des tétradéceneoates methyl.En éléctroantennographie, Z-11-16:Ac a donné les meilleurs réponses, suivis par l'acetoxy-1 tétradécene-9 Z (Z-9-14:Ac), Z-11-16:Ald et Z-11-16:OH. Dans des études de single sensillum les cellules à amplitude grande ont répondu à la stimulation avec de Z-11-16:Ac, cependant les cellules à amplitude petite ont répondu à la stimulation avec des deux composés Z-9-14:Ac et Z-11-16:OH. On a trouvé des cellules sensitives à Z-11-16:Ald dans 1 entre 60 sensilla étudiés.À la chambre de vol, le dose de 0.1 g d'un mélange de 10:1 de Z-11-16:Ac/Z-11-16:Ald a provoqué les mêmes réponses et à l'intensité pareille comme 3 glandes phéromonales isolées des femelles.En piégeages sur le champs des males en quantité importante ont été capturé par un mélange de 10:1 de Z-11-16:Ac/Z-11-15:Ald en Bulgarie et Hongrie. L'addition de 16:Ac au mélange binaire n'avait aucun effet, cependant l'addition de plus de 1% de Z-11-16:OH ou 0.1% de Z-9-14:Ac a sérieusement diminué les captûres. En comparant des proportions différentes du mélange de l'acetoxy/aldéhyde dans des doses différentes, on a observé les meilleurs captûres avec de la proportion 10:1 et à la dose la plus haute (1 000 g).

     

Novel characteristics of cassava,Manihot esculenta Crantz,a reputed C3-C4 intermediate photosynthesis species

The cassava plant, Manihot esculenta, grows exceptionally well in low fertility and drought prone environments, but the mechanisms that allow this growth are unknown. Earlier, and sometimes contradictory, work speculated about the presence of a C₄-type photosynthesis in cassava leaves. In the present work we found no evidence for a C₄ metabolism in mature attached cassava leaves as indicated i) by the low, 2 to 8%, incorporation of ¹⁴CO₂ into C₄ organic acids in short time periods, 10 s, and the lack of ¹⁴C transfer from C₄ acids to other compounds in ¹²CO₂, ii) by the lack of C₄ enzyme activity changes during leaf development and the inability to detect C₄ acid decarboxylases, and iii) by leaf CO₂ compensation values between 49 and 65 l of CO₂ 1^–1 and by other infrared gas exchange photosynthetic measurements. It is concluded that the leaf biochemistry of cassava follows the C₃ pathway of photosynthesis with no indication of a C₃-C₄ mechanism.However, cassava leaves exhibit several novel characteristics. Attached leaves have the ability to effectively partition carbon into sucrose with nearly 45% of the label in sucrose in about one min of ¹⁴CO₂ photosynthesis, contrasting with 34% in soybean (C₃) and 25% in pigweed (C₄). Cassava leaves displayed a strong preference for the synthesis of sucrose versus starch. Field grown cassava leaves exhibited high rates of photosynthesis and curvilinear responses to increasing sunlight irradiances with a tendency to saturate only at high irradiances, above 1500 mol m^–2 s^–1. Morphologically, the cassava leaf has papillose epidermal cells on its lower mesophyll surface that form fence-like arrangements encircling guard cells. It is proposed that the active synthesis of sugars has osmotic functions in the cassava plant and that the papillose epidermal cells function to maintain a healthy leaf water status in various environments.Abbreviations ADP adenosine diphosphate - Asp aspartate - BSA bovine serum albumin - CoA coenzyme A - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - FBP fructose-1,6-biphosphate - Gly glycine - HEPES N-2-hydroxyethylpiperazine-N-2-ethansulfonic acid - Mal malate - NAD nicotinamide adenine dinucleotide (oxidized form) - NADH nicotinamide adenine dinucleotide (reduced form) - NADP nicotinamide adenine dinucleotide phosphate (oxidized form) - PAR photosynthetic active radiation (400–700 nm) - PEP phosphenolpyruvate carboxylase - p-FBPase plastid fructose-1,6-biphosphatase - PGA 3-phosphoglyceric acid - PMSF phenylmethylsulfonyl fluoride - PVP polyvinylpyrrolidone - Rubisco ribulose-1,5-biphosphate carboxylase/oxygenase - RuBP ribulose-1,5-biphosphate - Ser serine - sugar-P sugar-phosphates     

Automatic Nuclei Segmentation in H&E Stained Breast Cancer Histopathology Images

The introduction of fast digital slide scanners that provide whole slide images has led to a revival of interest in image analysis applications in pathology. Segmentation of cells and nuclei is an important first step towards automatic analysis of digitized microscopy images. We therefore developed an automated nuclei segmentation method that works with hematoxylin and eosin (H&E) stained breast cancer histopathology images, which represent regions of whole digital slides. The procedure can be divided into four main steps: 1) pre-processing with color unmixing and morphological operators, 2) marker-controlled watershed segmentation at multiple scales and with different markers, 3) post-processing for rejection of false regions and 4) merging of the results from multiple scales. The procedure was developed on a set of 21 breast cancer cases (subset A) and tested on a separate validation set of 18 cases (subset B). The evaluation was done in terms of both detection accuracy (sensitivity and positive predictive value) and segmentation accuracy (Dice coefficient). The mean estimated sensitivity for subset A was 0.875 (±0.092) and for subset B 0.853 (±0.077). The mean estimated positive predictive value was 0.904 (±0.075) and 0.886 (±0.069) for subsets A and B, respectively. For both subsets, the distribution of the Dice coefficients had a high peak around 0.9, with the vast majority of segmentations having values larger than 0.8.     

The Role of γ-Secretase Activating Protein (GSAP) and Imatinib in the Regulation of γ-Secretase Activity and Amyloid-β Generation

γ-Secretase is a large enzyme complex comprising presenilin, nicastrin, presenilin enhancer 2, and anterior pharynx-defective 1 that mediates the intramembrane proteolysis of a large number of proteins including amyloid precursor protein and Notch. Recently, a novel γ-secretase activating protein (GSAP) was identified that interacts with γ-secretase and the C-terminal fragment of amyloid precursor protein to selectively increase amyloid-β production. In this study we have further characterized the role of endogenous and exogenous GSAP in the regulation of γ-secretase activity and amyloid-β production in vitro. Knockdown of GSAP expression in N2a cells decreased amyloid-β levels. In contrast, overexpression of GSAP in HEK cells expressing amyloid precursor protein or in N2a cells had no overt effect on amyloid-β generation. Likewise, purified recombinant GSAP had no effect on amyloid-β generation in two distinct in vitro γ-secretase assays. In subsequent cellular studies with imatinib, a kinase inhibitor that reportedly prevents the interaction of GSAP with the C-terminal fragment of amyloid precursor protein, a concentration-dependent decrease in amyloid-β levels was observed. However, no interaction between GSAP and the C-terminal fragment of amyloid precursor protein was evident in co-immunoprecipitation studies. In addition, subchronic administration of imatinib to rats had no effect on brain amyloid-β levels. In summary, these findings suggest the roles of GSAP and imatinib in the regulation of γ-secretase activity and amyloid-β generation are uncertain.