Boo - Hoo - Hoo : You\u27re Gonna Cry When I\u27m Gone

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Enhanced transport of plant-produced rabies single chain antibody-RVG peptide fusion protein across an in cellulo blood-brain barrier device

The biomedical applications of antibody engineering are developing rapidly and have been expanded to plant expression platforms. In this study, we have generated a novel antibody molecule in planta for targeted delivery across the blood–brain barrier (BBB). Rabies virus (RABV) is a neurotropic virus for which there is no effective treatment after entry into the central nervous system. This study investigated the use of a RABV glycoprotein peptide sequence to assist delivery of a rabies neutralizing single-chain antibody (ScFv) across an in cellulo model of human BBB. The 29 amino acid rabies virus peptide (RVG) recognizes the nicotinic acetylcholine receptor (nAchR) at neuromuscular junctions and the BBB. ScFv and ScFv-RVG fusion proteins were produced in Nicotiana benthamiana by transient expression. Both molecules were successfully expressed and purified, but the ScFv expression level was significantly higher than that of ScFv-RVG fusion. Both ScFv and ScFv-RVG fusion molecules had potent neutralization activity against RABVin cellulo. The ScFv-RVG fusion demonstrated increased binding to nAchR and entry into neuronal cells, compared to ScFv alone. Additionally, a human brain endothelial cell line BBB model was used to demonstrate that plant-produced ScFv-RVGP fusion could translocate across the cells. This study indicates that the plant-produced ScFv-RVGP fusion protein was able to cross the in celluloBBB and neutralize RABV

Synthesis, XRD and HS-Analysis

An efficient microwave-assisted one-step synthetic route toward Mannich bases is developed from 4-hydroxyacetophenone and different secondary amines in quantitative yields, via a regioselective substitution reaction. The reaction takes a short time and is non-catalyzed and reproducible on a gram scale. The environmentally benign methodology provides a novel alternative, to the conventional methodologies, for the synthesis of mono- and disubstituted Mannich bases of 4-hydroxyacetophenone. All compounds were well-characterized by FT-IR, 1H NMR, 13C NMR, and mass spectrometry. The structures of 1-{4-hydroxy-3-[(morpholin-4-yl)methyl]phenyl}ethan-1-one (2a) and 1-{4-hydroxy-3-[(pyrrolidin-1-yl)methyl]phenyl}ethan-1-one (3a) were determined by single crystal X-ray crystallography. Compound 2a and 3a crystallize in monoclinic, P21/n, and orthorhombic, Pbca, respectively. The most characteristic features of the molecular structure of 2a is that the morpholine fragment adopts a chair conformation with strong intramolecular hydrogen bonding. Compound 3a exhibits intermolecular hydrogen bonding, too. Furthermore, the computed Hirshfeld surface analysis confirms H-bonds and π–π stack interactions obtained by XRD packing analyses

Measuring poly(acrylamide) flocculants in fresh water using inter-polymer complex formation

YesA novel detection method for poly(acrylamide) flocculants was developed using interpolymer complexation between flocculants and a probe (poly(acrylic acid-co-acenaphthylene)). This detection method was tested in fresh water situations and found to be robust against a range of contaminants; additionally it was capable of detecting levels of flocculant dosing below 1 mg l−1. This method is remarkably fast and requires little sample modification compared to existing methods of detection

Characterization and Generation of Male Courtship Song in Cotesia congregata (Hymenoptera: Braconidae)

Background Male parasitic wasps attract females with a courtship song produced by rapid wing fanning. Songs have been described for several parasitic wasp species; however, beyond association with wing fanning, the mechanism of sound generation has not been examined. We characterized the male courtship song of Cotesia congregata (Hymenoptera: Braconidae) and investigated the biomechanics of sound production. Methods and Principal Findings Courtship songs were recorded using high-speed videography (2,000 fps) and audio recordings. The song consists of a long duration amplitude-modulated “buzz” followed by a series of pulsatile higher amplitude “boings,” each decaying into a terminal buzz followed by a short inter-boing pause while wings are stationary. Boings have higher amplitude and lower frequency than buzz components. The lower frequency of the boing sound is due to greater wing displacement. The power spectrum is a harmonic series dominated by wing repetition rate ~220 Hz, but the sound waveform indicates a higher frequency resonance ~5 kHz. Sound is not generated by the wings contacting each other, the substrate, or the abdomen. The abdomen is elevated during the first several wing cycles of the boing, but its position is unrelated to sound amplitude. Unlike most sounds generated by volume velocity, the boing is generated at the termination of the wing down stroke when displacement is maximal and wing velocity is zero. Calculation indicates a low Reynolds number of ~1000. Conclusions and Significance Acoustic pressure is proportional to velocity for typical sound sources. Our finding that the boing sound was generated at maximal wing displacement coincident with cessation of wing motion indicates that it is caused by acceleration of the wing tips, consistent with a dipole source. The low Reynolds number requires a high wing flap rate for flight and predisposes wings of small insects for sound production

In‐situ microbially induced Ca2+‐alginate polymeric sealant for seepage control in porous materials

This paper presents a novel approach of using in-situ microbially induced Ca2+-alginate polymeric sealant for seepage control in porous materials. This process comprises two steps: (i) generation of insoluble calcium carbonate inside the pores of porous materials (such as sand) through a microbially induced carbonate precipitation (MICP) process in-situ and (ii) injection of sodium alginate for in-situ gelation via reaction between alginate and Ca2+ions. The experimental results showed that the hydraulic conductivity/permeability of sand decreased with the increase in alginate concentration. When 5% alginate was used with a CaCO3concentration of 0.18 g g-1sand, the permeability of the alginate-treated sand reduced from 5.0 × 10-4to 2.2 × 10-9 m s-1. The scanning electron microscopy images revealed that a film-type coating was formed around sand particles with spherical round crystals embedded. Furthermore, the in-situ formed Ca-alginate polymeric sealant can also be used for the removal of Cu2+ion and suspended particles from contaminated water by more than 90%. Built on the current research, the envisioned practical application of the proposed method may include clogging fractured rock, reducing seepage and prevent piping through dams, excavation dewatering, and forming barriers for remediating specific contaminants

Individual effect of recrystallisation nucleation sites on texture weakening in a magnesium alloy: Part 1- double twins

Recrystallised grain nucleation, grain growth and corresponding texture evolution in a cold-rolled rare earth containing WE43 Mg alloy during annealing at 490 �C was fully tracked using a quasi-in-situ electron backscatter diffraction method. The results show nucleation sites, such as double twins, can weaken the deformed texture and for the first time provide direct evidence that recrystallised grains originating from double twins can form the rare earth texture during annealing. Precipitation and recrystallisation occurred concurrently during most of the annealing period, with precipitates forming preferentially along prior grain and twin boundaries. These precipitates effectively retard the recrystallisation due to particle pinning leading to an excessively long time for the completion of recrystallisation. A large portion of recrystallised grains were observed to have 〈0001〉 poles tilted 20e45� away from the normal direction. The RE texture emerges during the nucleation of recrystallised grains and is maintained during subsequent uniform grain growth, which results in a stable RE texture being developed as recrystallisation progresses. The uniform grain growth could be attributed to solute drag suppressing the grain boundary mobility of those grains that had recrystallised with a basal texture and precipitate pinning restricting potential orientated grain growth

The Evolution of Compact Binary Star Systems

We review the formation and evolution of compact binary stars consisting of white dwarfs (WDs), neutron stars (NSs), and black holes (BHs). Binary NSs and BHs are thought to be the primary astrophysical sources of gravitational waves (GWs) within the frequency band of ground-based detectors, while compact binaries of WDs are important sources of GWs at lower frequencies to be covered by space interferometers (LISA). Major uncertainties in the current understanding of properties of NSs and BHs most relevant to the GW studies are discussed, including the treatment of the natal kicks which compact stellar remnants acquire during the core collapse of massive stars and the common envelope phase of binary evolution. We discuss the coalescence rates of binary NSs and BHs and prospects for their detections, the formation and evolution of binary WDs and their observational manifestations. Special attention is given to AM CVn-stars -- compact binaries in which the Roche lobe is filled by another WD or a low-mass partially degenerate helium-star, as these stars are thought to be the best LISA verification binary GW sources.Comment: 105 pages, 18 figure

Aqueous alteration of the Martian meteorite Northwest Africa 817: Probing fluid?rock interaction at the nakhlite launch site

The nakhlite meteorites characteristically contain iddingsite, a hydrous iron–magnesium silicate that formed by aqueous alteration on Mars. Iddingsite is most abundant in Northwest Africa (NWA) 817, and alteration products in this meteorite also have the lowest deuterium/hydrogen ratio of any nakhlite. Taken together, these distinctive properties could be interpreted to show that NWA 817 was altered under different physico‐chemical conditions than the other nakhlites and by liquid water from a separate reservoir. Here this interpretation is tested through a petrographic, mineralogical, chemical, and isotopic study of NWA 817. We find that its iddingsite occurs as olivine‐hosted veins of nanocrystalline smectite and Fe‐oxyhydroxide. Strong similarities in the mineralogy of iddingsite between NWA 817 and other nakhlites suggest that these meteorites were altered under comparable physico‐chemical conditions, with the Fe‐rich composition of NWA 817 olivine grains rendering them especially susceptible to aqueous alteration. Analyses of NWA 817 bulk samples by stepwise pyrolysis confirm that its iddingsite has unusually low deuterium/hydrogen ratios, but owing to terrestrial weathering of this meteorite, the hydrogen isotopic data cannot be used with confidence to infer the origin of Martian aqueous solutions. NWA 817 was most probably altered along with the other nakhlites over a short time period and in a common aqueous system. One interpretation of a correlation between the eruption ages of three of the nakhlites and the chemical composition of their iddingsite is that water originated from close to the surface of Mars and flowed through the nakhlite lava pile under the influence of gravity

Cryopreservation of human mucosal leukocytes

Background: Understanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible. Methods and Findings: To find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes. Cryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to the same cells tested fresh. Additionally, we tested whether it is better to cryopreserve endocervical cells on the cytobrush or in suspension. Conclusions: Leukocytes from cervicovaginal and colorectal tissues can be cryopreserved with good recovery of functional, viable cells using several different cryopreservation media. The number and volume of washes has an experimentally meaningful effect on the percentage of cells recovered. We provide a detailed, step-by-step protocol with best practices for cryopreservation of mucosal leukocytes