Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and offers potential for smartphone-based detection of larval insecticide resistance

Pyrethroid-impregnated nets have contributed significantly to halving the burden of malaria but resistance threatens their future efficacy and the pipeline of new insecticides is short. Here we report that an invertebrate automated phenotyping platform (INVAPP), combined with the algorithm Paragon, provides a robust system for measuring larval motility in Anopheles gambiae (and An. coluzzi) as well as Aedes aegypti with the capacity for high-throughput screening for new larvicides. By this means, we reliably quantified both time- and concentration-dependent actions of chemical insecticides faster than using the WHO standard larval assay. We illustrate the effectiveness of the system using an established larvicide (temephos) and demonstrate its capacity for library-scale chemical screening using the Medicines for Malaria Venture (MMV) Pathogen Box library. As a proof-of-principle, this library screen identified a compound, subsequently confirmed to be tolfenpyrad, as an effective larvicide. We have also used the INVAPP / Paragon system to compare responses in larvae derived from WHO classified deltamethrin resistant and sensitive mosquitoes. We show how this approach to monitoring larval response to insecticides can be adapted for use with a smartphone camera application and therefore has potential for further development as a simple portable field-assay with associated real-time, geo-located information to identify hotspots

Imperfect and askew: A review of asymmetric genitalia in araneomorph spiders (Araneae: Araneomorphae)

Bilateral asymmetry in the genitalia is a rare but widely dispersed phenomenon in the animal tree of life. In arthropods, occurrences vary greatly from one group to another and there seems to be no common explanation for all the independent origins. In spiders, genital asymmetry appears to be especially rare. Most known species show almost perfectly symmetrical genitals with the right and left sides being mirror images of each other. However, some examples of asymmetric genitalia have been studied and many other reports are scattered in the taxonomic literature. Based on a broad literature survey, we found several species in thirteen families with evidence of genital asymmetry, mostly expressed only in females. Our review suggests that spider genital asymmetries, although rare, are more common than previously thought and taxonomic descriptions and illustrations are a useful but not entirely reliable tool for studying them. Here we also report on directional asymmetry in the liocranid spider Teutamus politus, the first known case of morphologically asymmetric male genitals in Entelegynae spiders. Generalities, evolution and categorization of asymmetry in spiders are further discussed.Animal science

5@5 - a 5 GeV energy threshold array of imaging atmospheric Cherenkov telescopes at 5 km altitude

We discuss the concept and the performance of a powerful future ground-based astronomical instrument - a stereoscopic array of several large imaging atmospheric Cherenkov telescopes installed at a very high mountain elevation of about 5 km a.s.l. or more - for the study of the gamma-ray sky at energies from several GeV to 100 GeV.Comment: 33 pages, 25 figures, the revised version accepted for publication in Astroparticle Physic

Development of COVID-19 monoclonal antibodies and recombinant proteins as reagents for biomedical research and diagnostic test

Since SARS-COV-2 virus spread worldwide and COVID-19 turned rapidly into a pandemic illness, the necessity for vaccines and diagnostic tests became crucial. The viral surface is decorated with Spike, the major antigenic determinant and main target for vaccine development. Within Spike, the receptor binding domain (RBD), constitutes the main target of highly neutralizing antibodies found in COVID-19 convalescent plasma. Besides vaccination, another important aspect of Spike (and RBD) is their use as immunogen for the development of poli- and monoclonal antibodies (mAbs) for therapeutic and diagnostic purposes. Here we report the development and preliminary biochemical characterization of a set of monoclonal antibodies against the Spike RBD domain along with the recombinant expression of two mayor COVID-19 protein reagents: the viral Spike RBD domain and the extracellular domain of the human receptor ACE2. RBD and the extracellular domain of ACE2 (aa 1-740) were obtained through transient gene transfection (TGE) in two different mammalian cell culture systems: HEK293T adherent monolayers and Expi293 suspension cultures. Due to its low cost and ease scale-up, all transfections were carried with polyethyleneimine (PEI). Expressed proteins were purified from culture supernatants by immobilized metal affinity chromatography. Anti-RBD mAbs were developed from two different immunization schemes: one aimed to elicit antibodies with viral neutralizing potential, and the other with the ability to recognize denatured RBD for routinary lab immunoassays. To achieve this, the first group of mice was immunized with RBD in aluminium salts (RBD/Al) and the other with RBD emulsified in Freunds adyuvant (RBD/FA). Polyclonal and monoclonal antibody reactivities against native or denatured RBD forms were then assessed by ELISA. Complete RBD denaturation was followed by intrinsic fluorescence spectral changes upon different physicochemical stress treatments. As expected, RBD/Al immunized mice developed an antibody response shifted to native RBD while those immunized with RBD/FA showed a high response against both forms of the protein. In accordance with the observed polyclonal response, RBD/FA derived mAbs recognize both, native and denatured RBD. On the contrary, hybridomas generated from the RBD/Al protocol mostly recognize RBD in its native state. Further ELISA binding assays revealed that all RBD/FA derived mAbs can form a trimeric complex with ACE2 and RBD, denoting they would not have viral neutralizing activity. ELISA competition assays with the RBD/ACE2 complex aimed to determine the neutralization potential of the RBD/Al derived mAbs are under way. Overall, the anti-Spike RBD mAbs and the recombinant RBD and ACE2 proteins presented here constitute valuable tools for diverse COVID-19 academic research projects and local immunity surveillance testing.Fil: Acuña Intrieri, M. E. Universidad Nacional de San Martin. Centro de Rediseño E Ingenieria de Proteinas.; ArgentinaFil: Deriane, M.A. Universidad Nacional de San Martin. Centro de Rediseño E Ingenieria de Proteinas.; ArgentinaFil: Miller, C.. Universidad Nacional de San Martin. Centro de Rediseño E Ingenieria de Proteinas.; ArgentinaFil: Czibener, Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Correa, E.. No especifíca;Fil: Cragnaz, L.. No especifíca;Fil: Guerra, L.. No especifíca;Fil: Rodriguez, S.. No especifíca;Fil: Goldbaum, F.A.. Universidad Nacional de San Martin. Centro de Rediseño E Ingenieria de Proteinas.; ArgentinaFil: Seigelchifer, M.. No especifíca;Fil: Comerci, Diego José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Montagna, Georgina Nuri. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Cerutti, Maria Laura. Universidad Nacional de San Martin. Centro de Rediseño E Ingenieria de Proteinas.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaLVII Annual Meeting of the Argentine Society for Biochemistry and Molecular Biology Research y XVI Annual Meeting of the Argentinean Society for General MicrobiologyVirtualArgentinaSociedad Argentina de Investigación Bioquímica y Biología MolecularAsociación Civil de Microbiología Genera

Towards a Realistic Neutron Star Binary Inspiral: Initial Data and Multiple Orbit Evolution in Full General Relativity

This paper reports on our effort in modeling realistic astrophysical neutron star binaries in general relativity. We analyze under what conditions the conformally flat quasiequilibrium (CFQE) approach can generate ``astrophysically relevant'' initial data, by developing an analysis that determines the violation of the CFQE approximation in the evolution of the binary described by the full Einstein theory. We show that the CFQE assumptions significantly violate the Einstein field equations for corotating neutron stars at orbital separations nearly double that of the innermost stable circular orbit (ISCO) separation, thus calling into question the astrophysical relevance of the ISCO determined in the CFQE approach. With the need to start numerical simulations at large orbital separation in mind, we push for stable and long term integrations of the full Einstein equations for the binary neutron star system. We demonstrate the stability of our numerical treatment and analyze the stringent requirements on resolution and size of the computational domain for an accurate simulation of the system.Comment: 22 pages, 18 figures, accepted to Phys. Rev.

Mobile DNA in Old World monkeys: A glimpse through the rhesus macaque genome

The completion of the draft sequence of the rhesus macaque genome allowed us to study the genomic composition and evolution of transposable elements in this representative of the Old World monkey lineage, a group of diverse primates closely related to humans. The L1 family of long interspersed elements appears to have evolved as a single lineage, and Alu elements have evolved into four currently active lineages. We also found evidence of elevated horizontal transmissions of retroviruses and the absence of DNA transposon activity in the Old World monkey lineage. In addition, ∼100 precursors of composite SVA (short interspersed element, variable number of tandem repeat, and Alu) elements were identified, with the majority being shared by the common ancestor of humans and rhesus macaques. Mobile elements compose roughly 50% of primate genomes, and our findings illustrate their diversity and strong influence on genome evolution between closely related species

Metastability, negative specific heat and weak mixing in classical long-range many-rotator system

We perform a molecular dynamical study of the isolated $d=1$ classical Hamiltonian ${\cal H} = {1/2} \sum_{i=1}^N L_i^2 + \sum_{i \ne j} \frac{1-cos(\theta_i-\theta_j)}{r_{ij}^\alpha} ;(\alpha \ge 0)$, known to exhibit a second order phase transition, being disordered for $u \equiv U/N{\tilde N} \ge u_c(\alpha,d)$ and ordered otherwise ($U\equiv$ total energy and ${\tilde N} \equiv \frac{N^{1-\alpha/d}-\alpha/d}{1-\alpha/d}$). We focus on the nonextensive case $\alpha/d \le 1$ and observe that, for $u<u_c$, a basin of attraction exists for the initial conditions for which the system quickly relaxes onto a longstanding metastable state (whose duration presumably diverges with $N$ like ${\tilde N}$) which eventually crosses over to the microcanonical Boltzmann-Gibbs stable state. The temperature associated with the (scaled) average kinetic energy per particle is lower in the metastable state than in the stable one. It is exhibited for the first time that the appropriately scaled maximal Lyapunov exponent $\lambda_{u<u_c}^{max}(metastable) \propto N^{-\kappa_{metastable}} ;(N \to \infty)$, where, for all values of $\alpha/d$, $\kappa_{metastable}$ numerically coincides with {\it one third} of its value for $u>u_c$, hence decreases from 1/9 to zero when $\alpha/d$ increases from zero to unity, remaining zero thereafter. This new and simple {\it connection between anomalies above and below the critical point} reinforces the nonextensive universality scenario.Comment: 9 pages and 4 PS figure