2019
Microbial associations are widespread across the insects. In the olive fruit fly Bactrocera oleae (Diptera: Tephritidae), vertically transmitted gut symbionts contribute to larval development inside the olive host, and to adult nutrition. Nevertheless, their effect on behavioural decisions of adults is unknown. In this study, we show that symbiotic bacteria affect oviposition behaviour in B. oleae. We studied the effect of different fruits as hosts and different gut-bacteria as gut-symbionts on oviposition attempts and fly development in B. oleae. Untreated flies that had native gut-symbionts attempted oviposition significantly more times than axenic flies as well as flies treated with medfly-associated Pantoea or Klebsiella bacteria. Axenic flies provided with a diet containing the homogenized gut of symbiotic flies recovered the same number of oviposition attempts as their symbiotic counterparts. As for as the different hosts, green olives (unripe) and grapes were preferred while black olives (ripe) elicited the least number of oviposition attempts, with an interactive effect of host and bacterial treatments. It appears that both the host attributes and the native gut-symbionts drive oviposition preference towards green olives in B. oleae. Moreover, both bacterial treatments and hosts significantly affected the development of B. oleae larvae. Though grapes elicited as many oviposition attempts as green olives, they yielded no pupae. Taken together, our results suggest that the intimate association between B. oleae and their gut-microbes, extends beyond nutritional support to behaviour. © 2019 Elsevier Ltd
Plants, especially perennials, growing in drylands and seasonally dry ecosystems are uniquely adapted to dry conditions. Legume shrubs and trees, capable of symbiotic dinitrogen (N 2 ) fixation, often dominate in drylands. However, the strategies that allow symbiotic fixation in these ecosystems, and their influence on the nitrogen cycle, are largely unresolved. We evaluated the climatic, biogeochemical and ontogenetic factors influencing nitrogen fixation in an abundant Mediterranean legume shrub, Calicotome villosa. We measured nodulation, fixation rate, nitrogen allocation and soil biogeochemistry in three field sites over a full year. A controlled experiment evaluated differences in plant regulation of fixation as a function of soil nutrient availability and seedling and adult developmental stages. We found a strong seasonal pattern, shifting between high fixation rates during the rainy season at flowering and seed-set times to almost none in the rainless season. Under controlled conditions, plants downregulated fixation in response to soil nitrogen availability, but this response was stronger in seedlings than in adult shrubs. Finally, we did not find elevated soil nitrogen under N 2 -fixing shrubs. We conclude that seasonal nitrogen fixation, regulation of fixation, and nitrogen conservation are key adaptations influencing the dominance of dryland legumes in the community, with broader consequences on the ecosystem nitrogen cycle. © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust
ABSTRACTMilk fat globule (MFG) size ranges over 3 orders of magnitude, from less than 200 nm to over 15 µm. The significance of MFG size derives from its tight association with its lipidome and proteome. More specifically, small MFG have relatively higher content of membrane compared with large globules, and this membrane exerts diverse positive health effects, as reported in human and animal studies. In addition, MFG size has industrial significance, as it affects the physicochemical and sensory characteristics of dairy products. Studies on the size regulation of MFG are scarce, mainly because various confounders indirectly affect MFG size. Because MFG size is determined before and during its secretion from mammary epithelial cells, studies on the size regulation of its precursors, the intracellular lipid droplets (LD), have been used as a proxy for understanding the mechanisms controlling MFG size. In this review, we provide evidence for 2 distinct mechanisms regulating LD size in mammary epithelial cells: co-regulation of fat content and triglyceride-synthesis capacity of the cells, and fusion between LD. The latter is controlled by the membrane's polar lipid composition and involves mitochondrial enzymes. Accordingly, this review also discusses MFG size regulation in the in vivo metabolic context, as MFG morphometric features are often modulated under conditions that involve animals' altered energy status.
At some point early in the vertebrate lineage, two whole genome duplication events (1R, 2R) took place that allowed for the diversification and sub-/neo-functionalization of the glycoprotein hormones (GpHs). All jawed vertebrates possess the GpHs luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH), each of which are heterodimers with a common alpha subunit and unique beta subunits. In 2002, a novel glycoprotein hormone named thyrostimulin was described to have unique GpA2 and GpB5 subunits that were homologous to the vertebrate alpha and beta subunits. The presence of GpA2 and GpB5 in representative protostomes and deuterostomes indicates their ancestry in the GpH family. There are several reports of GpH subunit evolution, but none have included GpA2 and GpB5 for species in each major vertebrate class. Thus, we addressed the ancestry of two paralogous GpB5 subunits (GpB5a and GpB5b) that were previously only recognized in two teleost species. Our search for orthologous GpB5a and GpB5b sequences in representative vertebrates and phylogenetic analysis, in addition to the currently published evolutionary scenarios of the GpH family, supports that GpB5a and GpB5b are paralogs that arose from the first vertebrate whole genome duplication event (1R). Syntenic analysis supports lineage specific losses of GpB5a in chondrichthyes, basal actinopterygians, and tetrapods, and retention in coelacanth and teleosts. Additionally, we were unable to identify GpA2 transcripts from tilapia mRNA, suggesting that this species does not produce heterodimeric thyrostimulin. While the conserved or even species-specific functional role of thyrostimulin or its individual subunits are still unknown in vertebrates, the analyses presented here provide context for future studies on the functional divergence of the GpH family. © 2019 Hausken, Levavi-Sivan. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
At some point early in the vertebrate lineage, two whole genome duplication events (1R, 2R) took place that allowed for the diversification and sub-/neo-functionalization of the glycoprotein hormones (GpHs). All jawed vertebrates possess the GpHs luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH), each of which are heterodimers with a common alpha subunit and unique beta subunits. In 2002, a novel glycoprotein hormone named thyrostimulin was described to have unique GpA2 and GpB5 subunits that were homologous to the vertebrate alpha and beta subunits. The presence of GpA2 and GpB5 in representative protostomes and deuterostomes indicates their ancestry in the GpH family. There are several reports of GpH subunit evolution, but none have included GpA2 and GpB5 for species in each major vertebrate class. Thus, we addressed the ancestry of two paralogous GpB5 subunits (GpB5a and GpB5b) that were previously only recognized in two teleost species. Our search for orthologous GpB5a and GpB5b sequences in representative vertebrates and phylogenetic analysis, in addition to the currently published evolutionary scenarios of the GpH family, supports that GpB5a and GpB5b are paralogs that arose from the first vertebrate whole genome duplication event (1R). Syntenic analysis supports lineage specific losses of GpB5a in chondrichthyes, basal actinopterygians, and tetrapods, and retention in coelacanth and teleosts. Additionally, we were unable to identify GpA2 transcripts from tilapia mRNA, suggesting that this species does not produce heterodimeric thyrostimulin. While the conserved or even species-specific functional role of thyrostimulin or its individual subunits are still unknown in vertebrates, the analyses presented here provide context for future studies on the functional divergence of the GpH family.
At some point early in the vertebrate lineage, two whole genome duplication events (1R, 2R) took place that allowed for the diversification and sub-/neo-functionalization of the glycoprotein hormones (GpHs). All jawed vertebrates possess the GpHs luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH), each of which are heterodimers with a common alpha subunit and unique beta subunits. In 2002, a novel glycoprotein hormone named thyrostimulin was described to have unique GpA2 and GpB5 subunits that were homologous to the vertebrate alpha and beta subunits. The presence of GpA2 and GpB5 in representative protostomes and deuterostomes indicates their ancestry in the GpH family. There are several reports of GpH subunit evolution, but none have included GpA2 and GpB5 for species in each major vertebrate class. Thus, we addressed the ancestry of two paralogous GpB5 subunits (GpB5a and GpB5b) that were previously only recognized in two teleost species. Our search for orthologous GpB5a and GpB5b sequences in representative vertebrates and phylogenetic analysis, in addition to the currently published evolutionary scenarios of the GpH family, supports that GpB5a and GpB5b are paralogs that arose from the first vertebrate whole genome duplication event (1R). Syntenic analysis supports lineage specific losses of GpB5a in chondrichthyes, basal actinopterygians, and tetrapods, and retention in coelacanth and teleosts. Additionally, we were unable to identify GpA2 transcripts from tilapia mRNA, suggesting that this species does not produce heterodimeric thyrostimulin. While the conserved or even species-specific functional role of thyrostimulin or its individual subunits are still unknown in vertebrates, the analyses presented here provide context for future studies on the functional divergence of the GpH family.
Heroot L. Vahav, Pogoreltsev, Alla , Tulchinsky, Yuri , Fridman, Natalia , Börner, Armin , ו Gandelman, Mark . 6/13/2019.
“Synthesis And Characteristics Of Iridium Complexes Bearing N-Heterocyclic Nitrenium Cationic Ligands”. Organometallics, 38, 12, Pp. 2494-2501. .
Publisher's Version תקציר N-Heterocyclic nitrenium cations are isostructural and isoelectronic analogues of the ubiquitous N-heterocyclic carbene. We present the first examples of coordination of nitrenium ions to the Ir(I) and Ir(III) metal centers. This work includes rare complexes with cation–cation interactions between a positively charged ligand and an iridium ion. These species represent the first example of iridium complexes bearing any cationic ligand of group 15 elements analogous to the Arduengo carbene. These nitrenium-based monocationic and even dicationic Ir(I) complexes can smoothly oxidatively activate H2 at room temperature and ambient pressure. Utilizing our system, we were able to observe an Ir–dihydrogen σ-complex, which undergoes oxidative addition to yield a well-defined Ir(III) dihydride. Comparative studies of the analogous Rh(I)–nitrenium species, which exhibit reversible dihydrogen activation, are presented. New Ir complexes were fully characterized by multinuclear nuclear magnetic resonance (including 15N labeling) and X-ray crystallography.
Igor Schapiro, Gueye, Moussa , Paolino, Marco , Fusi, Stefania , Marchand, Gabriel , Haacke, Stefan , Martin, M. Elena , Huntress, Mark , Vysotskiy, Victor P. , Veryazov, Valera , Léonard, Jérémie , ו Olivucci, Massimo . 2019.
“Synthesis, Spectroscopy And Qm/Mm Simulations Of A Biomimetic Ultrafast Light-Driven Molecular Motor”. Photochem. Photobiol. Sci., 18, Pp. 2259-2269. doi:10.1039/C9PP00223E.
Publisher's Version תקציר A molecular motor potentially performing a continuous unidirectional rotation is studied by a multidisciplinary approach including organic synthesis, transient spectroscopy and excited state trajectory calculations. A stereogenic center was introduced in the N-alkylated indanylidene–pyrroline Schiff base framework of a previously investigated light-driven molecular switch in order to achieve the unidirectional CC rotary motion typical of Feringa’s motor. Here we report that the specific substitution pattern of the designed chiral molecule must critically determine the unidirectional efficiency of the light-induced rotary motion. More specifically, we find that a stereogenic center containing a methyl group and a hydrogen atom as substituents does not create a differential steric effect large enough to fully direct the motion in either the clockwise or counterclockwise direction especially along the E → Z coordinate. However, due to the documented ultrafast character and electronic circular dichroism activity of the investigated system, we find that it provides the basis for development of a novel generation of rotary motors with a biomimetic framework and operating on a picosecond time scale.
Synthetic biology lies on the interface between natural and artificial life. It consists of the assembly of natural biological components into artificially configured biological systems. A main focus of synthetic biology has been the engineering of new gene circuits that can produce artificial cellular functions. I propose to scale up this approach to include, beyond single cells and gene circuits, also entire multi-cellular organisms and the brain circuits that regulate their behavior. Such synthetic biology in the brain will offer new ways for understanding how brain connectivity relates to brain function, and could ultimately lead to futuristic technologies such as neuronally-programmed organic robots or biologically-based brain repair. As a first step towards this ambitious goal I have developed a technique for genetically inserting new synaptic connections into the nervous system of the nematode worm C. elegans, enabling the manipulation of information flow in the nervous system and the reprogramming of whole animal behavior in this organism. This approach may be expanded and adapted to other genetic models, and opens the way to possible new forms of artificial life. Such technology, if practiced responsibly, could offer considerable benefits to science, industry and medicine.
DOI
Synthetic biology lies on the interface between natural and artificial life. It consists of the assembly of natural biological components into artificially configured biological systems. A main focus of synthetic biology has been the engineering of new gene circuits that can produce artificial cellular functions. I propose to scale up this approach to include, beyond single cells and gene circuits, also entire multi-cellular organisms and the brain circuits that regulate their behavior. Such synthetic biology in the brain will offer new ways for understanding how brain connectivity relates to brain function, and could ultimately lead to futuristic technologies such as neuronally-programmed organic robots or biologically-based brain repair. As a first step towards this ambitious goal I have developed a technique for genetically inserting new synaptic connections into the nervous system of the nematode worm C. elegans, enabling the manipulation of information flow in the nervous system and the reprogramming of whole animal behavior in this organism. This approach may be expanded and adapted to other genetic models, and opens the way to possible new forms of artificial life. Such technology, if practiced responsibly, could offer considerable benefits to science, industry and medicine.
Single-molecule fluorescence detection (SMFD) experiments are useful in distinguishing sub-populations of molecular species when measuring heterogeneous samples. One experimental platform for SMFD is based on a confocal microscope, where molecules randomly traverse an effective detection volume. The non-uniformity of the excitation profile and the random nature of Brownian motion, produce fluctuating fluorescence signals. For these signals to be distinguished from the background, burst analysis is frequently used. Yet, the relation between the results of burst analyses and the underlying information of the diffusing molecules is still obscure and requires systematic assessment. In this work we performed three-dimensional Brownian motion simulations of SMFD, and tested the positions at which molecules emitted photons that passed the burst analysis criteria for different values of burst analysis parameters. The results of this work verify which of the burst analysis parameters and experimental conditions influence both the position of molecules in space when fluorescence is detected and taken into account, and whether these bursts of photons arise purely from single molecules, or not entirely. Finally, we show, as an example, the effect of bursts that are not purely from a single molecule on the accuracy in single-molecule Förster resonance energy transfer measurements.
Mamidi Samarasimhareddy, Mayer, Daniel , Metanis, Norman , Veprintsev, Dmitry , Hurevich, Mattan , ו Friedler, Assaf . 2019.
“A Targeted Approach For The Synthesis Of Multi-Phosphorylated Peptides: A Tool For Studying The Role Of Phosphorylation Patterns In Proteins”. Organic & Biomolecular Chemistry, 17, 42, Pp. 9284-9290. doi:10.1039/c9ob01874c.
תקציר
Protein phosphorylation barcodes, clusters of several phosphorylation sites within a short unfolded region, control many cellular processes. Existing biochemical methods used to study the roles of these barcodes suffer from low selectivity and provide only qualitative data. Chemically synthesized multiphosphopeptide libraries are selective and specific, but their synthesis is extremely difficult using the current peptide synthesis methods. Here we describe a new microwave assisted approach for synthesizing a library of multiphosphopeptides, using the C-terminus of rhodopsin as a proof of concept. Our approach utilizes multiple protocols for synthesizing libraries of multiphosphopeptides instead of the inefficient single protocol methods currently used. Using our approach we demonstrated the synthesis with up to seven phosphorylated amino acids, sometimes next to each other, an accomplishment that was impractical before. Synthesizing the Rhodopsin derived multiphosphopeptide library enabled dissecting the precise phosphorylation barcode required for the recruitment, activation and modulation of the conformation of Arrestin. Since phosphorylation barcodes modulate the activity of hundreds of GPCRs, synthesizing libraries of multiphosphopeptides is the method of choice for studying their molecular mechanisms of action. Our approach provides an invaluable tool for evaluating how protein phosphorylation barcodes regulate their activity.
Mamidi S., D., Mayer , N., Metanis , D., Veprintsev , M., Hurevich , ו A., Friedler . 2019.
“A Targeted Approach For The Synthesis Of Multi-Phosphorylated Peptides: A Tool For Studying The Role Of Phosphorylation Patterns In Proteins”. Org. Biomol. Chem. .
קישור תקציר Protein phosphorylation barcodes, clusters of several phosphorylation sites within a short unfolded region, control many cellular processes. Existing biochemical methods used to study the roles of these barcodes suffer from low selectivity and provide only qualitative data. Chemically synthesized multiphosphopeptides libraries are selective and specific, but their synthesis is extremely difficult using the current peptide synthesis methods. Here we decribe a new microwave assisted approach for synthesizing a library of multiphosphopeptides, using the C-terminus of rhodopsin as a proof of concept. Our approch utilizes multiple protocols for synthesizing libraries of multiphosphopeptides instead of the inefficent single protocol methods curretnly used. Using our approach we demonstrated the synthesis of with up to seven phosphorylated amino acids, sometimes next to each other, an accomplishment that was impractical before. Synthesizing the Rhodopsin derived multiphosphopeptide library enabled dissecting the precise phosphorylation barcode required for the recruitment, activation and modulation of the conformation of Arrestin. Since phosphorylation barcodes modulate the activity of hundreds of GPCRs, synthesizing libraries of multiphosphopeptides is the method of choice for studying their molecular mechanisms of action. Our approach provides an invaluable tool for evaluating how protein phosphorylation barcodes regulate their activity.
Samarasimhareddy Mamidi, Mayer, Daniel , Metanis, Norman , Veprintsev, Dmitry , Hurevich, Mattan , ו Friedler, Assaf . 2019.
“A Targeted Approach For The Synthesis Of Multi-Phosphorylated Peptides: A Tool For Studying The Role Of Phosphorylation Patterns In Proteins”. Organic & Biomolecular Chemistry, 17, 42, Pp. 9284–9290.
M. Ben-David-Naim, Dagan, A. , Grad, E. , Aizik, G. , Nordling-David, M.M. , Clyne, A.M. , Granot, Z. , ו Golomb, G.. 2019.
“Targeted Sirna Nanoparticles For Mammary Carcinoma Therapy”. Cancers, 11, 4. doi:10.3390/cancers11040442.
Publisher's Version Non-viral, polymeric-based, siRNA nanoparticles (NPs) have been proposed as promising gene delivery systems. Encapsulating siRNA in targeted NPs could confer improved biological stability, extended half-life, enhanced permeability, effective tumor accumulation, and therapy. In this work, a peptide derived from apolipoprotein B100 (ApoB-P), the protein moiety of low-density lipoprotein, was used to target siRNA-loaded PEGylated NPs to the extracellular matrix/proteoglycans (ECM/PGs) of a mammary carcinoma tumor. siRNA against osteopontin (siOPN), a protein involved in breast cancer development and progression, was encapsulated into PEGylated poly(d,l-lactic-co-glycolic acid) (PLGA) NPs using the double emulsion solvent diffusion technique. The NPs obtained possessed desired physicochemical properties including 200 nm size, a neutral surface charge, and high siOPN loading of 5 µg/mg. ApoB-P-targeted NPs exhibited both enhanced binding to isolated ECM and internalization by MDA-MB-231 human mammary carcinoma cells, in comparison to non-targeted NPs. Increased accumulation of the targeted NPs was achieved in the primary mammary tumor of mice xenografted with MDA-MB-231 mammary carcinoma cells as well as in the lungs, one of the main sites affected by metastases. siOPN NPs treatment resulted in significant inhibition of tumor growth (similar bioactivity of both formulations), accompanied with significant reduction of OPN mRNA levels ( 40% knockdown of mRNA levels). We demonstrated that targeted NPs possessed enhanced tumor accumulation with increased therapeutic potential in mice models of mammary carcinoma © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
A. Pauletti, Terrone, G. , Shekh-Ahmad, T. , Salamone, A. , Ravizza, T. , Rizzi, M. , Pastore, A. , Pascente, R. , Liang, L.-P. , Villa, B.R. , Balosso, S. , Abramov, A.Y. , van Vliet, E.A. , Giudice, E.D. , Aronica, E. , Patel, M. , Walker, M.C. , ו Vezzani, A.. 2019.
“Targeting Oxidative Stress Improves Disease Outcomes In A Rat Model Of Acquired Epilepsy”. Brain, 142, 7. doi:10.1093/brain/awz130.
Publisher's Version B. de Roos, Aura, A.-M. , Bronze, M. , Cassidy, A. , Conesa, M.-T.G. , Gibney, E.R. , Greyling, A. , Kaput, J. , Kerem, Zohar , Knežević, N. , Kroon, P. , Landberg, R. , Manach, C. , Milenkovic, D. , Rodriguez-Mateos, A. , Tomás-Barberán, F.A. , van de Wiele, T. , ו Morand, C. . 2019.
“Targeting The Delivery Of Dietary Plant Bioactives To Those Who Would Benefit Most: From Science To Practical Applications”. European Journal Of Nutrition, 58, Pp. 65-73. doi:10.1007/s00394-019-02075-5.
Publisher's Version תקציר Background: A healthy diet and optimal lifestyle choices are amongst the most important actions for the prevention of cardiometabolic diseases. Despite this, it appears difficult to convince consumers to select more nutritious foods. Furthermore, the development and production of healthier foods do not always lead to economic profits for the agro-food sector. Most dietary recommendations for the general population represent a “one-size-fits-all approach” which does not necessarily ensure that everyone has adequate exposure to health-promoting constituents of foods. Indeed, we now know that individuals show a high variability in responses when exposed to specific nutrients, foods, or diets. Purpose: This review aims to highlight our current understanding of inter-individual variability in response to dietary bioactives, based on the integration of findings of the COST Action POSITIVe. We also evaluate opportunities for translation of scientific knowledge on inter-individual variability in response to dietary bioactives, once it becomes available, into practical applications for stakeholders, such as the agro-food industry. The potential impact from such applications will form an important impetus for the food industry to develop and market new high quality and healthy foods for specific groups of consumers in the future. This may contribute to a decrease in the burden of diet-related chronic diseases. © 2019, The Author(s).
