Ecologically sustainable chemical recommendations for agricultural pest control?

Effective pest control remains an essential part of food production, and it is provided both by chemicals and by natural enemies within agricultural ecosystems. These methods of control are often in conflict because of the negative impact of chemicals on natural enemies. There are already well-established approaches such as those provided by the International Organization for Biological and Integrated Control-Pesticides and Beneficial Organisms for testing, collecting, and publishing information on responses of natural enemies to chemicals based on laboratory responses of specific organisms; however, these tests do not assess the cumulative impact of chemical inputs across an entire season or consider impacts on the complex communities of natural enemies that can provide effective pest control on a farm. Here, we explore the potential of different approaches for assessing the impact of chemicals on agricultural ecosystems and we propose a simple metric for sustainable chemical use on farms that minimizes overall impact on beneficial groups. We suggest ways in which the effectiveness of metrics can be extended to include persistence and habitat features. Such metrics can assist farmers in developing targets for sustainable chemical use as demonstrated in the viticultural industry.     

Structural–functional insights of single and multi-domain Capsicum annuum protease inhibitors

Pin-II protease inhibitors (PIs) are the focus of research interest because of their large structural–functional diversity and relevance in plant defense. Two representative Capsicum annuum PI genes (CanPI-15 and -7) comprising one and four inhibitory repeat domains, respectively, were expressed and recombinant proteins were characterized. β-Sheet and unordered structure was found predominant in CanPI-15 while -7 also displayed the signatures of polyproline fold, as revealed by circular dichroism studies. Inhibition kinetics against bovine trypsin indicated three times higher potency of CanPI-7 (K_i ～ 57 μM) than -15 (～184 μM). Activity and structural stability of these CanPIs were revealed under various conditions of pH, temperature and denaturing agent. Structure prediction, docking studies with proteases and mass spectroscopy revealed the organization of multiple reactive site loops of multi domain PIs in space as well as the steric hindrances imposed while binding to proteases due to their close proximity.     

Macrophage fusion: are somatic and cancer cells possible partners?

Macrophages are present in all tissues and can fuse with themselves to differentiate into multinucleate osteoclasts or giant cells that play a central role in osteoporosis and chronic inflammatory diseases, respectively. Yet, the mechanism by which they fuse remains uncharacterized. The macrophage fusion receptor (MFR) and its ligand CD47 might mediate homotypic fusion of macrophages and allow for their recognition as 'self' before fusion. Although a novel process and controversial idea, macrophages might exploit a similar mechanism for fusion with somatic cells or tumor cells, with resultant organ repair or metastasis, respectively. Hence, macrophages might be the 'double-edged swords' of tissues.     

Macrophage fusion: are somatic and cancer cells possible partners?

The fusion of cells is a fundamental biological event that plays a central role in a variety of developmental and homeostatic processes. Macrophages are present in all tissues and can initiate interaction and fusion. The putative macrophage-fusion machinery is still poorly understood, but some of its components have been identified. Macrophages recognize each other as < self > in order to fuse but some essential questions remain: do macrophages fuse with somatic cells to repair tissues and organs? Do macrophages fuse with tumor cells to trigger metastasis? Agnès Vignery discusses these novel and challenging ideas in this review.     

Zn- and Cu-thioneins: a functional classification for metallothioneins?

This report intends to provide the reader with a deeper insight in the chemical, and extensively biological, characteristics of the metallothionein (MT) system. We have devoted nearly 20 years to the study of MTs and this has allowed us to form what we believe is a more complete picture of this peculiar family of metalloproteins. At the beginning of the 1990s, the landscape of this field was quite different from the overall picture we have now. Many researchers have contributed to the readjustment of this part of scientific knowledge. In our case, we implemented a unified method for obtaining MTs, for characterizing their metal-binding features, and for applying a unified research rationale. All this has helped to enlarge the initial picture that was mainly dominated by mammalian MT1/MT2 and yeast Cup1, by introducing approximately 20 new MTs. It has also allowed some characteristics to be clarified and examined in more detail, such as the cooperativity or the coexistence of multiple species in the metal-substitution reactions, the availability of Ag(I) or Cd(II) for use as respective probes for the Cu(I) and Zn(II) binding sites, the participation of chloride or sulfide ligands in the metal coordination spheres, and the feasibility of using in vitro data as representative of in vivo scenarios. Overall, the results yield enough data to consider new criteria for a proposal of classification of MTs based on MT metal-binding features, which complements the previous classifications, and that can shed light on the still controversial physiological functions of this peculiar superfamily of metalloproteins.     

Hybrid larch heterosis: for which traits and under which genetic control?

RCD1 is a member of the plant-specific SRO protein family. Several SRO genes have been functionally identified in the regulation of abiotic stresses in Arabidopsis and other plant species. However, the function of SROs is largely unknown in apple (Malus×domestica). In this study, six MdSRO-encoding genes were isolated, categorized into two types and mapped to six chromosomes. The phylogenetic analysis demonstrated that the sequences of the AtSRO and MdSRO proteins are highly conserved. Subsequently, expression analysis showed that MdSRO genes had different expression profiles in different tissues and in response to various stresses. Finally, MdRCD1 was isolated for functional identification. The results showed that resistance to oxidation stress in apple calli was enhanced by MdRCD1 overexpression and weakened by MdRCD1 suppression. MdRCD1 also played a crucial role in the regulation of ROS homeostasis in transgenic apple calli and Arabidopsis. Ectopic expression of MdRCD1 significantly enhanced resistance to salt and oxidative stresses in transgenic lines. In addition, MdRCD1 also enhanced drought tolerance due to its influence on stomatal opening. Based on these results, we conclude that MdRCD1 is an important regulator in abiotic stress response.

     

Multiple receptor conformation docking and dock pose clustering as tool for CoMFA and CoMSIA analysis – a case study on HIV-1 protease inhibitors

Multiple receptors conformation docking (MRCD) and clustering of dock poses allows seamless incorporation of receptor binding conformation of the molecules on wide range of ligands with varied structural scaffold. The accuracy of the approach was tested on a set of 120 cyclic urea molecules having HIV-1 protease inhibitory activity using 12 high resolution X-ray crystal structures and one NMR resolved conformation of HIV-1 protease extracted from protein data bank. A cross validation was performed on 25 non-cyclic urea HIV-1 protease inhibitor having varied structures. The comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) models were generated using 60 molecules in the training set by applying leave one out cross validation method, r_loo² values of 0.598 and 0.674 for CoMFA and CoMSIA respectively and non-cross validated regression coefficient r² values of 0.983 and 0.985 were obtained for CoMFA and CoMSIA respectively. The predictive ability of these models was determined using a test set of 60 cyclic urea molecules that gave predictive correlation (r_pred²) of 0.684 and 0.64 respectively for CoMFA and CoMSIA indicating good internal predictive ability. Based on this information 25 non-cyclic urea molecules were taken as a test set to check the external predictive ability of these models. This gave remarkable out come with r_pred² of 0.61 and 0.53 for CoMFA and CoMSIA respectively. The results invariably show that this method is useful for performing 3D QSAR analysis on molecules having different structural motifs.     

Arbuscular mycorrhiza increased the activity of a biotrophic leaf pathogen – is a compensation possible?

Arbuscular mycorrhizal barley-plants were more susceptible to the obligate biotrophic shoot pathogen Erysiphe graminis f. sp. hordei. In experiments under greenhouse and open-air conditions on leaves of mycorrhizal plants, the sporulation rate of the mildew fungus was more than twice that on control plants. However, mycorrhizal plants suffered less than non-mycorrhizal plants in terms of grain number, ear yield and thousand-grain weight. Disease-yield-relationship analysis showed that the symbiosis neutralised the positive correlation between disease severity and yield loss (up to 25% infected leaf area tested). After mildew infection, nitrogen in ears of non-mycorrhizal barley was higher because of an impaired starch accumulation during grain filling. In mycorrhizal plants, leaf disease did not impair either the quantity or quality of grain yield. This improved compensation in mycorrhizal plants was related to maintained photosynthetic capacity and a delay in pathogen-induced senescence. Thus filling of long-term storage pools (fructans in internodes) and consequently reallocation of these reserves during grain filling was improved. The results suggest that higher availability of energy and material during grain formation, together with longer physiological activity, were the basis of yield maintenance and, therefore, expression of mycorrhiza-induced tolerance towards the pathogen.     

Can the sheep model fully represent the human model for the functional evaluation of cervical interbody fusion cages?

Sheep model is the most favourable choice for animal study for functional evaluation of the cervical fusion prostheses before clinical application; however, significantly large differences between sheep and human existed in terms of morphological characteristics and daily-activity motions. Questions should be raised as whether the differences between the two species have influence on the reliability of sheep model. Finite element models (FEM) of the cervical spinal system were built to characterize the differences between the two species with respect to the range of motion (ROM) and biomechanical behaviour, and experimental cadaver tests on both species were employed for validation purposes. Results indicate that sheep model represents the worst-case scenario of the human model with exaggerated stresses (up to 3 times more) and ROM (up to 10 times more). Moreover, sheep model is very sensitive to the variation of prostheses design, whilst human model does not, which denotes that the sheep model provides a rather amplified effect of a certain design for its biomechanical performance. Therefore, caution needs to be taken when sheep models were used as the animal model for functional evaluation over various design, and the FEM built in this study can be employed as an effective methodology for performance evaluation of cage prostheses of cervical spine.

     

Association of calpastatin with inactive calpain: a novel mechanism to control the activation of the protease?

It is generally accepted that the Ca(2+)-dependent interaction of calpain with calpastatin is the most relevant mechanism involved in the regulation of Ca(2+)-induced proteolysis. We now report that a calpain-calpastatin association can occur also in the absence of Ca(2+) or at very low Ca(2+) concentrations, reflecting the physiological conditions under which calpain retains its inactive conformational state. The calpastatin binding region is localized in the non-inhibitory L-domain containing the amino acid sequences encoded by exons 4-7. This calpastatin region recognizes a calpain sequence located near the end of the DII-domain. Interaction of calpain with calpastatins lacking these sequences becomes strictly Ca(2+)-dependent because, under these conditions, the transition to an active state of the protease is an obligatory requirement. The occurrence of the molecular association between Ca(2+)-free calpain and various recombinant calpastatin forms has been demonstrated by the following experimental results. Addition of calpastatin protected calpain from trypsin digestion. Calpain was coprecipitated when calpastatin was immunoprecipitated. The calpastatin molecular size increased following exposure to calpain. The two proteins comigrated in zymogram analysis. Furthermore, calpain-calpastatin interaction was perturbed by protein kinase C phosphorylation occurring at sites located at the exons involved in the association. At a functional level, calpain-calpastatin interaction at a physiological concentration of Ca(2+) represents a novel mechanism for the control of the amount of the active form of the protease potentially generated in response to an intracellular Ca(2+) influx.     

Sharing a predator: can an invasive alien pest affect the predation on a local pest?

Invasive species can strongly affect biotic interactions in ecosystems, interacting both directly and indirectly with local species. In European tomato greenhouses, the invasive alien pest Tuta absoluta may impact the population dynamics of other pests like whiteflies. Besides inducing damages to the host plant and competing for resources with local pests, this alien species may exert a predator-mediated interaction on local pests sharing common natural enemies. Biocontrol agents usually used against whiteflies may also prey upon T. absoluta and this could alter the dynamics of local pest populations. We evaluated possible resource competition and predator-mediated interactions in a system involving one mirid predator Macrolophus pygmaeus and two pests, T. absoluta and a local whitefly, Bemisia tabaci, on greenhouse tomatoes. Results showed that both resource competition and predator-mediated interactions occurred simultaneously. In the presence of the shared predator, there was a short-term positive effect of T. absoluta on B. tabaci [up to 5.9-fold increase of B. tabaci juveniles (egg + larvae) after four weeks]. However, in the long-term there was a negative predator-mediated interaction of T. absoluta on B. tabaci, i.e., after ten weeks the density of B. tabaci was 7.3-fold lower in the presence of the invasive pest. We emphasize the critical role of generalist predators in managing both local and invasive alien pest populations and that the strength and direction of predator-mediated indirect interactions can depend on the time scale considered.     

Microbial control in Asia: a bellwether for the future?

Advances and barriers faced by microbial control efforts in Asia offer instructive insights for microbial control in general. The papers in this series, which are based on plenary lectures given at the Society for Invertebrate Pathology 2006 meeting in Wuhan, China, explore the history and current status of microbial control in China, Japan, and Southeast Asia, and in doing so, bring to light the following key assumptions that deserve further examination; (1) the adoption rate of microbial control is well documented; (2) microbial control agents can compete directly with conventional insecticides; (3) microbial control agents are relatively easy and inexpensive to produce and develop; (4) patents will promote innovation and investor interest in microbial control. Alternative viewpoints are presented that can hopefully aid in future efforts to develop more safe and effective microbial control agents.     

Metallo-β-lactamases and a tug-of-war for the available zinc at the host–pathogen interface

Metallo-β-lactamases (MBLs) are zinc-dependent hydrolases that inactivate virtually all β-lactam antibiotics. The expression of MBLs by Gram-negative bacteria severely limits the therapeutic options to treat infections. MBLs bind the essential metal ions in the bacterial periplasm, and their activity is challenged upon the zinc starvation conditions elicited by the native immune response. Metal depletion compromises both the enzyme activity and stability in the periplasm, impacting on the resistance profile in vivo. Thus, novel inhibitory approaches involve the use of chelating agents or metal-based drugs that displace the native metal ion. However, newer MBL variants incorporate mutations that improve their metal binding abilities or stabilize the metal-depleted form, revealing that metal starvation is a driving force acting on MBL evolution. Future challenges require addressing the gap between in cell and in vitro studies, dissecting the mechanism for MBL metalation and determining the metal content in situ.     

The prokaryotic cytoskeleton: a putative target for inhibitors and antibiotics?

In the recent decade, our view on the organization of the bacterial cell has been revolutionized by the identification of cytoskeletal elements. Most bacterial species have structural homologs of actin and tubulin that assemble into dynamic, filamentous structures at precisely defined sub-cellular locations. The essential cell division protein FtsZ forms a dynamic ring at mid-cell and is similar in its structure to tubulin. Proteins of the MreB family, which are structural homologs of actin, assemble into helical or straight filaments in the bacterial cytoplasm. As in eukaryotic cells, the bacterial cytoskeleton drives essential cellular processes such as cell division, cell wall growth, DNA movement, protein targeting, and alignment of organelles. Different high-throughput assays have been developed to search for inhibitors of components of the bacterial cytoskeleton. Cell-based assays for the detection of cell division inhibitors as well as FtsZ GTPase assays led to the identification of several compounds that inhibit the polymerization of FtsZ, by this blocking bacterial cell division. Such inhibitors might not only be valuable tools for basic research, but might also lead to novel therapeutic agents against pathogenic bacteria. For example, the polyphenol dichamanetin, the 2-alkoxycarbonylaminopyridine SRI-3072, and the benzophenanthridine alkaloid sanguinarine inhibit the GTPase activity of FtsZ and exhibit antimicrobial activity.     

Death waits for no man – Does it wait for a virus? How enteroviruses induce and control cell death

Enteroviruses (EVs) are the most common human viral pathogens. They cause a variety of pathologies, including myocarditis and meningoencephalopathies, and have been linked to the onset of type I diabetes. These pathologies result from the death of cells in the myocardium, central nervous system, and pancreas, respectively. Understanding the role of EVs in inducing cell death is crucial to understanding the etiologies of these diverse pathologies. EVs both induce and delay host cell death, and their exquisite control of this balance is crucial for their success as human viral pathogens. Thus, EVs are tightly involved with cell death signaling pathways and interact with host cell signaling at multiple points. Here, we review the literature detailing the mechanisms of EV-induced cell death. We discuss the mechanisms by which EVs induce cell death, the signaling pathways involved in these pathways, and the strategies by which EVs antagonize cell death pathways. We also discuss the role of cell death in both the resulting pathology in the host and in the facilitation of viral spread.     

Which predatory mite can control both a dominant mite pest,Tetranychus urticae,and a latent mite pest,Eotetranychus asiaticus,on strawberry?

After biological control of Tetranychus urticae using Phytoseiulus persimilis, a latent mite pest, Eotetranychus asiaticus, was found on strawberries growing in a plastic greenhouse in western Japan. To determine whether the release ofP. persimilis, an exotic natural enemy of T.urticae, enhanced the occurrence of the latent pest, the efficiency of P. persimilis in controlling E.asiaticus was compared with the efficiency of two indigenous phytoseiid mites, Amblyseius californicus and A. womersleyi, under laboratory conditions. Phytoseiuluspersimilis and A. californicus attacked not onlyT. urticae but also E. asiaticus. However, the predators produced very few eggs and their offspring rarely reached adulthood when fed on E. asiaticus, so their potential as control agents for the latent mite appears to be low. In contrast, A.womersleyi feeding on E. asiaticus reproduced as well as when fed on T. urticae, and exterminated both species of spider mite. This suggests that A. womersleyihas greater potential as a biological control agent in crops where both species may occur. This revised version was published online in August 2006 with corrections to the Cover Date.     

SARS-CoV-2 main protease inhibitors: What is moving in the field of peptides and peptidomimetics?

The COVID-19 pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still affecting people worldwide. Despite the good degree of immunological protection achieved through vaccination, there are still severe cases that require effective antivirals. In this sense, two specific pharmaceutical preparations have been marketed already, the RdRp polymerase inhibitor molnupiravir and the main viral protease inhibitor nirmatrelvir (commercialized as Paxlovid, a combination with ritonavir). Nirmatrelvir is a peptidomimetic acting as orally available, covalent, and reversible inhibitor of SARS-CoV-2 main viral protease. The success of this compound has revitalized the search for new peptide and peptidomimetic protease inhibitors. This highlight collects some selected examples among those recently published in the field of SARS-CoV-2.