Belowground and aboveground herbivory differentially affect the transcriptome in roots and shoots of maize.

Plants recognize and respond to feeding by herbivorous insects by upregulating their local and systemic defenses. While defense induction by aboveground herbivores has been well studied, far less is known about local and systemic defense responses against attacks by belowground herbivores. Here, we investigated and compared the responses of the maize transcriptome to belowground and aboveground mechanical damage and infestation by two well-adapted herbivores: the soil-dwelling western corn rootworm Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) and the leaf-chewing fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). In responses to both herbivores, maize plants were found to alter local transcription of genes involved in phytohormone signaling, primary and secondary metabolism. Induction by real herbivore damage was considerably stronger and modified the expression of more genes than mechanical damage. Feeding by the corn rootworm had a strong impact on the shoot transcriptome, including the activation of genes involved in defense and development. By contrast, feeding by the fall armyworm induced only few transcriptional changes in the roots. In conclusion, feeding by a leaf chewer and a root feeder differentially affects the local and systemic defense of maize plants. Besides revealing clear differences in how maize plants respond to feeding by these specialized herbivores, this study reveals several novel genes that may play key roles in plant-insect interactions and thus sets the stage for in depth research into the mechanism that can be exploited for improved crop protection. Significance statement: Extensive transcriptomic analyses revealed a clear distinction between the gene expression profiles in maize plants upon shoot and root attack, locally as well as distantly from the attacked tissue. This provides detailed insights into the specificity of orchestrated plant defense responses, and the dataset offers a molecular resource for further genetic studies on maize resistance to herbivores and paves the way for novel strategies to enhance maize resistance to pests.

Comparative Screening of Mexican, Rwandan and Commercial Entomopathogenic Nematodes to Be Used against Invasive Fall Armyworm, Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is an important pest of maize originating from the Americas. It recently invaded Africa and Asia, where it causes severe yield losses to maize. To fight this pest, tremendous quantities of synthetic insecticides are being used. As a safe and sustainable alternative, we explore the possibility to control FAW with entomopathogenic nematodes (EPN). We tested in the laboratory whether local EPNs, isolated in the invasive range of FAW, are as effective as EPNs from FAW native range or as commercially available EPNs. This work compared the virulence, killing speed and propagation capability of low doses of forty EPN strains, representing twelve species, after placing them with second-, third- and sixth-instar caterpillars as well as pupae. EPN isolated in the invasive range of FAW (Rwanda) were found to be as effective as commercial and EPNs from the native range of FAW (Mexico) at killing FAW caterpillars. In particular, the Rwandan Steinernema carpocapsae strain RW14-G-R3a-2 caused rapid 100% mortality of second- and third-instar and close to 75% of sixth-instar FAW caterpillars. EPN strains and concentrations used in this study were not effective in killing FAW pupae. Virulence varied greatly among EPN strains, underlining the importance of thorough EPN screenings. These findings will facilitate the development of local EPN-based biological control products for sustainable and environmentally friendly control of FAW in East Africa and beyond.

The Role of Herbivore-induced Plant Volatiles in Trophic Interactions: The Swiss Connection.

It is increasingly evident that plants actively respond to the threats and challenges that they come to face while growing. This is particularly manifested in the dynamic responses to insect herbivory, especially in terms of the volatile compounds that the attacked plants emit. Indeed, many plants respond to insect-inflicted damage with the synthesis and release of volatile organic compounds. These emissions, commonly referred to as herbivore-induced plant volatiles (HIPVs), play important roles in the interactions between the emitting plants and their biotic environment. The odorous signal can be picked up and exploited by various organisms: neighbouring plants, herbivores and their natural enemies, such as predators and parasitoid wasps. Coincidence or not, scientists currently working in Switzerland have made numerous key contributions to the work in this field. By highlighting their work, we attempt to give a somewhat historic overview of this field of research.

Evolutionary changes in an invasive plant support the defensive role of plant volatiles.

It is increasingly evident that plants interact with their outside world through the production of volatile organic compounds,1-5 but whether the volatiles have evolved to serve in plant defense is still a topic of considerable debate.3,6-8 Unharmed leaves constitutively release small amounts of volatiles, but when the leaves are damaged by herbivorous arthropods, they emit substantially more volatiles. These herbivore-induced plant volatiles (HIPVs) attract parasitoids and predators that kill insect herbivores,9-12 and this can benefit the plants.13,14 As yet, however, there is no tangible evolutionary evidence that this tritrophic interplay contributes to the selection forces that have shaped the volatile emissions of plants.2,3,5-8,15 With this in mind, we investigated the evolutionary changes in volatile emissions in invasive common ragwort and the respective defensive roles of its constitutive and inducible volatiles. This Eurasian plant has invaded other continents, where it evolved for many generations in the absence of specialized herbivores and their natural enemies. We found that, compared to native ragworts, invasive plants release higher levels of constitutive volatiles but considerably lower levels of herbivore-induced volatiles. As a consequence, invasive ragwort is more attractive to a specialist moth but avoided by an unadapted generalist moth. Importantly, conforming to the indirect defense hypothesis, a specialist parasitoid was much more attracted to caterpillar-damaged native ragwort, which was reflected in higher parasitism rates in a field trial. The evolution of foliar volatile emissions appears to be indeed driven by their direct and indirect roles in defenses against insects.

Differential Levels of Fatty Acid-Amino Acid Conjugates in the Oral Secretions of Lepidopteran Larvae Account for the Different Profiles of Volatiles.

BACKGROUND: Plants have evolved sophisticated defense responses to insect herbivore attack, which often involve elicitors in the insects' oral secretions. The major eliciting compounds in insect oral secretions across different species and their potency in inducing volatile emissions have not yet been fully characterized and compared. RESULTS: Seven lepidopteran insects with variable duration of association with maize were selected, five species known as pests for a long time (Ostrinia furnacalis, Spodoptera exigua, Spodoptera litura, Mythimna separata, and Helicoverpa armigera) and two newly emerging pests (Athetis lepigone and Athetis dissimilis). Oral secretions of the newly emerging pests have the highest total contents of Fatty Acid-Amino Acid Conjugates (FACs), and their relative composition was well separated from that of the other five species in principal compound analysis. Redundancy analyses suggested that higher quantity of FACs was mainly responsible for the increases in maize volatiles, of which (E)-3,8-dimethyl-1,4,7-nonatriene (DMNT) and (E, E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (TMTT) were the most strongly inducible compounds. Adding FACs to the oral secretion of S. litura larvae significantly increased the emissions of TMTT and DMNT, confirming the key role of FACs in inducing volatile emissions in maize plants. Additional experiments with artificial diet spiked with linolenic acid suggested that variation in FACs is due to differences in internal FAC degradation and fatty acid excretion. CONCLUSION: Compared with two newly emerging pests A. lepigone and A. dissimilis, the long-term pests could diminish the volatile emission by maize through reducing the FAC content in their oral secretions, which may lower the risk of attracting natural enemies.

Insulin polymorphism induced by two polyphenols: new crystal forms and advances in macromolecular powder diffraction.

This study focuses on the polymorphism of human insulin (HI) upon the binding of the phenolic derivatives p-coumaric acid or trans-resveratrol over a wide pH range. The determination of the structural behaviour of HI via X-ray powder diffraction (XRPD) and single-crystal X-ray diffraction (SCXRD) is reported. Four distinct polymorphs were identified, two of which have not been reported previously. The intermediate phase transitions are discussed. One of the novel monoclinic polymorphs displays the highest molecular packing among insulin polymorphs of the same space group to date; its structure was elucidated by SCXRD. XRPD data collection was performed using a variety of instrumental setups and a systematic comparison of the acquired data is presented. A laboratory diffractometer was used for screening prior to high-resolution XRPD data collection on the ID22 beamline at the European Synchrotron Radiation Facility. Additional measurements for the most representative samples were performed on the X04SA beamline at the Swiss Light Source (SLS) using the MYTHEN II detector, which allowed the detection of minor previously untraceable impurities and dramatically improved the d-spacing resolution even for poorly diffracting samples.

Outcomes after spinal stenosis surgery by type of surgery in adults aged 60 years and older.

AIMS OF THE STUDY: Mobility disability due to spinal stenosis is common in the senior population and often surgery is warranted for patients with severe symptoms and neurological dysfunction. However, although current clinical guidelines recommend stabilisation surgery in addition to decompression in patients with spinal stenosis and instability due to degenerative spondylolisthesis, the relationship between outcomes and the specific type of surgery have not been well studied. We therefore assessed the postoperative recovery timeline for 12 months and compared patient-reported outcomes dependent on the extent of decompression and additional stabilisation among seniors undergoing spinal stenosis surgery. METHODS: We investigated 457 patients (mean age 76.0 ± 10.7 years, 58% women) from a consecutive cohort prior to spinal stenosis surgery. Follow-up was at 3 or 6months and at 12 months postoperatively. At each visit, pain, neurological dysfunction and disability were assessed using the North American Spine Society questionnaire. Repeated-measures analysis compared outcomes by type of surgery adjusting for baseline symptoms, gender, age, number of comorbidities, centre and year of surgery. RESULTS: Most improvement occurred within the first 3 to 6 months with little or no further improvement at 12 months. Over 12 months and in adjusted models, patients receiving one-segment versus multi-segment decompression experienced significantly greater reduction of pain (−49.2% vs −41.9%, p = 0.013) and neurological dysfunction (−37.1% vs −25.9%, p <0.0001), but only borderline greater reduction of disability (−32.7% vs −28.2%, p = 0.051). Moreover, reduction in pain and neurological function did not differ with or without additional stabilisation and extend of decompression. However, patients who received one-segment (−28.9%) or multi-segment (−28.3%) stabilisation experienced significantly less reduction in disability after surgery compared with those who were not stabilised (−34.1%, p <0.043). CONCLUSIONS: Among senior patients undergoing spinal stenosis surgery, recovery was largely complete by 3 to 6 months after surgery and differed little by type of surgery independently of symptoms prior to surgery and other covariates. However we could document a trend toward more improvement in particularly neurological dysfunction and disability with less invasive surgery.

Distinct Roles of Cuticular Aldehydes as Pheromonal Cues in Two Cotesia Parasitoids.

Cuticular compounds (CCs) that cover the surface of insects primarily serve as protection against entomopathogens, harmful substances, and desiccation. However, CCs may also have secondary signaling functions. By studying the role of CCs in intraspecific interactions, we may advance our understanding of the evolution of pheromonal communication in insects. We previously found that the gregarious parasitoid, Cotesia glomerata (L.), uses heptanal as a repellent pheromone to help avoid mate competition among sibling males, whereas another cuticular aldehyde, nonanal, is part of the female-produced attractive sex pheromone. Here, we show that the same aldehydes have different pheromonal functions in a related solitary parasitoid, Cotesia marginiventris (Cresson). Heptanal enhances the attractiveness of the female's sex pheromone, whereas nonanal does not affect a female's attractiveness. Hence, these common aldehydes are differentially used by the two Cotesia species to mediate, synergistically, the attractiveness of the main constituents of their respective sex pheromones. The specificity of the complete sex pheromone blend is apparently regulated by two specific, less volatile compounds, which evoke strong electroantennographic (EAG) responses. This is the first demonstration that volatile CCs have evolved distinct pheromonal functions to aid divergent mating strategies in closely related species. We discuss the possibility that additional compounds are involved in attraction and that, like the aldehydes, they are likely oxidative products of unsaturated cuticular hydrocarbons.

The Combined Use of an Attractive and a Repellent Sex Pheromonal Component by a Gregarious Parasitoid.

Gregarious parasitoids usually clump their cocoons together and the adults emerge in a synchronized fashion. This makes it easy for them to find mating partners and most copulations indeed take place at the natal patch. Yet, males should leave such sites when females are no longer receptive. As yet, this decision-making process and the possible involvement of pheromones were poorly understood. Here we report on a remarkable use of attractive and repellent pheromones of the well-studied gregarious parasitoid species Cotesia glomerata (L.) (Hymenoptera: Braconidae). Virgin C. glomerata females were found to release an attractive as well as a repellent compound, which in combination arrest males on the natal patch, but after mating the females stop the production of the attractant and the males are repelled. The repellent compound was identified as heptanal, which was also released by males, probably reducing male-male competition on the natal patch. We also confirmed that the sex ratio of the emerging wasps can vary considerably among patches, depending on the relative quality of hosts and the number of females that parasitize a host. The newly revealed use of attractive and repellent pheromone compounds by C. glomerata possibly helps maximize mating success under these variable conditions.

Revisiting the structure of a synthetic somatostatin analogue for peptide drug design.

Natural or artificially manufactured peptides attract scientific interest worldwide owing to their wide array of pharmaceutical and biological activities. X-ray structural studies are used to provide a precise extraction of information, which can be used to enable a better understanding of the function and physicochemical characteristics of peptides. Although it is vulnerable to disassociation, one of the most vital human peptide hormones, somatostatin, plays a regulatory role in the endocrine system as well as in the release of numerous secondary hormones. This study reports the successful crystallization and complete structural model of octreotide, a stable octapeptide analogue of somatostatin. Common obstacles in crystallographic studies arising from the intrinsic difficulties of obtaining a suitable single-crystal specimen were efficiently overcome as polycrystalline material was employed for synchrotron and laboratory X-ray powder diffraction (XPD) measurements. Data collection and preliminary analysis led to the identification of unit-cell symmetry [orthorhombic, P212121, a = 18.5453 (15), b = 30.1766 (25), c = 39.798 (4) Å], a process which was later followed by complete structure characterization and refinement, underlying the efficacy of the suggested (XPD) approach.

Combined use of herbivore-induced plant volatiles and sex pheromones for mate location in braconid parasitoids.

Herbivore-induced plant volatiles (HIPVs) are important cues for female parasitic wasps to find hosts. Here, we investigated the possibility that HIPVs may also serve parasitoids as cues to locate mates. To test this, the odour preferences of four braconid wasps - the gregarious parasitoid Cotesia glomerata (L.) and the solitary parasitoids Cotesia marginiventris (Cresson), Microplitis rufiventris Kokujev and Microplitis mediator (Haliday) - were studied in olfactometers. Each species showed attraction to pheromones but in somewhat different ways. Males of the two Cotesia species were attracted to virgin females, whereas females of M. rufiventris were attracted to virgin males. Male and female M. mediator exhibited attraction to both sexes. Importantly, female and male wasps of all four species were strongly attracted by HIPVs, independent of mating status. In most cases, male wasps were also attracted to intact plants. The wasps preferred the combination of HIPVs and pheromones over plant odours alone, except M. mediator, which appears to mainly use HIPVs for mate location. We discuss the ecological contexts in which the combined use of pheromones and HIPVs by parasitoids can be expected. To our knowledge, this is the first study to show that braconid parasitoids use HIPVs and pheromones in combination to locate mates.

Coxsackievirus B3 protease 3C: expression, purification, crystallization and preliminary structural insights.

Viral proteases are proteolytic enzymes that orchestrate the assembly of viral components during the viral life cycle and proliferation. Here, the expression, purification, crystallization and preliminary X-ray diffraction analysis are presented of protease 3C, the main protease of an emerging enterovirus, coxsackievirus B3, that is responsible for many cases of viral myocarditis. Polycrystalline protein precipitates suitable for X-ray powder diffraction (XRPD) measurements were produced in the presence of 22-28%(w/v) PEG 4000, 0.1 M Tris-HCl, 0.2 M MgCl2 in a pH range from 7.0 to 8.5. A polymorph of monoclinic symmetry (space group C2, unit-cell parameters a = 77.9, b = 65.7, c = 40.6 Å, ß = 115.9°) was identified via XRPD. These results are the first step towards the complete structural determination of the molecule via XRPD and a parallel demonstration of the accuracy of the method.

[The treatment of tuberculosis]. / Die Behandlung der Tuberkulose.

The incidence of tuberculosis decreases. However, clinical cases frequently raise questions, mainly in the context of contact tracing or when antimicrobial resistance is suspected. Empiric standard treatment consists of rifampin, isoniazid, ethambutol and pyrazinamide. This initial regimen aims to reduce the number of pathogenic germs while the consolidation therapy should eradicate the remaining pathogens. Treatment duration and adherence are crucial for cure. For the treatment of latent tuberculosis the traditional 6 to 9 months isoniazid regimen is still the treatment of choice. In complex cases such as tuberculosis in immunocompromised patients or if resistant tuberculosis is suspected, patients should be referred to a specialized center.

Exceptional Use of Sex Pheromones by Parasitoids of the Genus Cotesia: Males Are Strongly Attracted to Virgin Females, but Are No Longer Attracted to or Even Repelled by Mated Females.

Sex pheromones have rarely been studied in parasitoids, and it remains largely unknown how male and female parasitoids locate each other. We investigated possible attraction (and repellency) between the sexes of two braconid wasps belonging to the same genus, the gregarious parasitoid, Cotesia glomerata (L.), and the solitary parasitoid, Cotesia marginiventris (Cresson). Males of both species were strongly attracted to conspecific virgin females. Interestingly, in C. glomerata, the males were repelled by mated females, as well as by males of their own species. This repellency of mated females was only evident hours after mating, implying a change in pheromone composition. Males of C. marginiventris were also no longer attracted, but not repelled, by mated females. Females of both species showed no attraction to the odors of conspecific individuals, male or female, and C. glomerata females even appeared to be repelled by mated males. Moreover, the pheromones were found to be highly specific, as males were not attracted by females of the other species. Males of Cotesia glomerata even avoided the pheromones of female Cotesia marginiventris, indicating the recognition of non-conspecific pheromones. We discuss these unique responses in the context of optimal mate finding strategies in parasitoids.

Differential performance and parasitism of caterpillars on maize inbred lines with distinctly different herbivore-induced volatile emissions.

Plant volatiles induced by insect feeding are known to attract natural enemies of the herbivores. Six maize inbred lines that showed distinctly different patterns of volatile emission in laboratory assays were planted in randomized plots in the Central Mexican Highlands to test their ability to recruit parasitic wasps under field conditions. The plants were artificially infested with neonate larvae of the fall armyworm Spodoptera frugiperda, and two of its main endoparasitoids, Campoletis sonorensis and Cotesia marginiventris, were released in the plots. Volatiles were collected from equally treated reference plants in the neighbourhood of the experimental field. The cumulative amount of 36 quantified volatile compounds determined for each line was in good accordance with findings from the laboratory; there was an almost 15-fold difference in total emission between the two extreme lines. We found significant differences among the lines with respect to the numbers of armyworms recovered from the plants, their average weight gain and parasitism rates. Average weight of the caterpillars was negatively correlated with the average total amount of volatiles released by the six inbred lines. However, neither total volatile emission nor any specific single compound within the blend could explain the differential parasitism rates among the lines, with the possible exception of (E)-2-hexenal for Campoletis sonorensis and methyl salicylate for Cotesia marginiventris. Herbivore-induced plant volatiles and/or correlates thereof contribute to reducing insect damage of maize plants through direct plant defence and enhanced attraction of parasitoids, alleged indirect defence. The potential to exploit these volatiles for pest control deserves to be further evaluated.

Continuous monitoring of electrode-skin impedance mismatch during bioelectric recordings.

In bioelectric recordings, an electrode-skin impedance mismatch leads to a reduced common-mode rejection ratio (CMRR) of the amplifier. For this reason, the impedance of each individual electrode-skin contact is usually measured prior to a recording. The measurement circuit itself degrades the CMRR and is switched off during the bioelectric recording. In this paper, we present a new method that allows to monitor the electrode-skin impedance in a continuous way without reducing the CMRR of the amplifier. The new method is based on an additional common-mode signal that is superimposed on the bioelectric signal.

An improved method to continuously monitor the electrode-skin impedance during bioelectric measurements.

In bioelectric recordings the electrode-skin impedance mismatch reduces the common mode rejection ratio (CMRR) of the amplifier. For this reason, the impedance of each individual electrode-skin contact is usually measured prior to a recording. The measurement circuit itself degrades the CMRR and is switched off during the bioelectric recording. In this work we present a new method that allows to monitor the electrode-skin impedance in a continuous way without reducing the CMRR of the amplifier. The new method is based on an additional common mode signal which is superimposed on the bioelectric signal.

Low-noise two-wired buffer electrodes for bioelectric amplifiers.

Active buffer electrodes are known to improve the immunity of bioelectric recordings against power line interferences. A survey of published work reveals that buffer electrodes are almost exclusively designed using operational amplifiers (opamps). In this paper, we discuss the advantage of utilizing a single transistor instead. This allows for a simple electrode, which is small and requires only two wires. In addition, a single transistor adds considerably less noise when compared to an opamp with the same power consumption. We then discuss output resistance and gain as well as their respective effect on the common mode rejection ratio (CMRR). Finally, we demonstrate the use of two-wired buffer electrodes for a bioelectric amplifier.

A pseudodifferential amplifier for bioelectric events with DC-offset compensation using two-wired amplifying electrodes.

Most wired active electrodes reported so far have a gain of one and require at least three wires. This leads to stiff cables, large connectors and additional noise for the amplifier. The theoretical advantages of amplifying the signal on the electrodes right from the source has often been described, however, rarely implemented. This is because a difference in the gain of the electrodes due to component tolerances strongly limits the achievable common mode rejection ratio (CMRR). In this paper, we introduce an amplifier for bioelectric events where the major part of the amplification (40 dB) is achieved on the electrodes to minimize pick-up noise. The electrodes require only two wires of which one can be used for shielding, thus enabling smaller connecters and smoother cables. Saturation of the electrodes is prevented by a dc-offset cancelation scheme with an active range of +/- 250 mV. This error feedback simultaneously allows to measure the low frequency components down to dc. This enables the measurement of slow varying signals, e.g., the change of alertness or the depolarization before an epileptic seizure normally not visible in a standard electroencephalogram (EEG). The amplifier stage provides the necessary supply current for the electrodes and generates the error signal for the feedback loop. The amplifier generates a pseudodifferential signal where the amplified bioelectric event is present on one lead, but the common mode signal is present on both leads. Based on the pseudodifferential signal we were able to develop a new method to compensate for a difference in the gain of the active electrodes which is purely software based. The amplifier system is then characterized and the input referred noise as well as the CMRR are measured. For the prototype circuit the CMRR evaluated to 78 dB (without the driven-right-leg circuit). The applicability of the system is further demonstrated by the recording of an ECG.

Evaluating the induced-odour emission of a Bt maize and its attractiveness to parasitic wasps.

The current discussion on the safety of transgenic crops includes their effects on beneficial insects, such as parasitoids and predators of pest insects. One important plant trait to consider in this context is the emission of volatiles in response to herbivory. Natural enemies use the odours that result from these emissions as cues to locate their herbivorous prey and any significant change in these plant-provided signals may disrupt their search efficiency. There is a need for practical and reliable methods to evaluate transgenic crops for this and other important plant traits. Moreover, it is imperative that such evaluations are done in the context of variability for these traits among conventional genotypes of a crop. For maize and the induction of volatile emissions by caterpillar feeding this variability is known and realistic comparisons can therefore be made. Here we used a six-arm olfactometer that permits the simultaneous collection of volatiles emitted by multiple plants and testing of their attractiveness to insects. With this apparatus we measured the induced odour emissions of Bt maize (Bt11, N4640Bt) and its near-isogenic line (N4640) and the attractiveness of these odours to Cotesia marginiventris and Microplitis rufiventris, two important larval parasitoids of common lepidopteran pests. Both parasitoid species were strongly attracted to induced maize odour and neither wasp distinguished between the odours of the transgenic and the isogenic line. Also wasps that had previously experienced one of the odours during a successful oviposition divided their choices equally between the two odours. However, chemical analyses of collected odours revealed significant quantitative differences. The same 11 compounds dominated the blends of both genotypes, but the isogenic line released a larger amount of most of these. These differences may be due to altered resource allocation in the transgenic line, but it had no measurable effect on the wasps' behaviour. All compounds identified here had been previously reported for maize and the differential quantities in which they were released fall well within the range of variability observed for other maize genotypes.