Faster growth with shorter antigens can explain a VSG hierarchy during African trypanosome infections:a feint attack by parasites

The parasitic African trypanosome, Trypanosoma brucei, evades the adaptive host immune response by a process of antigenic variation that involves the clonal switching of variant surface glycoproteins (VSGs). The VSGs that come to dominate in vivo during an infection are not entirely random, but display a hierarchical order. How this arises is not fully understood. Combining available genetic data with mathematical modelling, we report a VSG-length-dependent hierarchical timing of clonal VSG dominance in a mouse model, consistent with an inverse correlation between VSG length and trypanosome growth-rate. Our analyses indicate that, among parasites switching to new VSGs, those expressing shorter VSGs preferentially accumulate to a detectable level that is sufficient to trigger a targeted immune response. This may be due to the increased metabolic cost of producing longer VSGs. Subsequent elimination of faster-growing parasites then allows slower-growing parasites with longer VSGs to accumulate. This interaction between the host and parasite is able to explain the temporal distribution of VSGs observed in vivo. Thus, our findings reveal a length-dependent hierarchy that operates during T. brucei infection. This represents a ‘feint attack’ diversion tactic utilised by these persistent parasites to out-maneuver the host adaptive immune system

Subcellular localisation and identification of single atoms using quantitative scanning transmission electron microscopy

Determining the concentration of elements in subcellular structures poses a significant challenge. By locating an elemental species at high spatial resolution and with subcellular context, and subsequently quantifying it on an absolute scale, new information about cellular function can be revealed. Such measurements have not as yet been realised with existing techniques due to limitations on spatial resolution and inherent difficulties in detecting elements present in low concentrations. In this paper, we use scanning transmission electron microscopy (STEM) to establish a methodology for localising and quantifying high‐Z elements in a biological setting by measuring elastic electron scattering. We demonstrate platinum (Pt) deposition within neuronal cell bodies following in vivo administration of the Pt‐based chemotherapeutic oxaliplatin to validate this novel methodology. For the first time, individual Pt atoms and nanoscale Pt clusters are shown within subcellular structures. Quantitative measurements of elastic electron scattering are used to determine absolute numbers of Pt atoms in each cluster. Cluster density is calculated on an atoms‐per‐cubic‐nanometre scale, and used to show clusters form with densities below that of metallic Pt. By considering STEM partial scattering cross‐sections, we determine that this new approach to subcellular elemental detection may be applicable to elements as light as sodium. LAY DESCRIPTION: Heterogeneous elemental distributions drive fundamental biological processes within cells. While carbon, hydrogen, oxygen and nitrogen comprise by far the majority of living matter, concentrations and locations of more than a dozen other species must also be tightly controlled to ensure normal cell function. Oxaliplatin is a first‐line and adjuvant treatment for colorectal cancer. However, pain in the body's extremities (fingers and toes) significantly impairs clinical usage as this serious and persistent side effect impacts on both patient cancer care and quality of life. Annular dark‐field (ADF) imaging in the scanning transmission electron microscope (STEM) provides an image with strong atom‐number contrast and is sufficient to distinguish between different cell types and different organelles within the cells of the DRG. We also show that Pt may be imaged at the single atom level and be localised at very high resolution while still preserving a degree of ultrastructural context. The intrinsic image contrast generated is sufficient to identify these features without the need for heavy metal stains and other extensive processing steps which risk disturbing native platinum distributions within the tissue. We subsequently demonstrate that by considering the total elastic scattering intensity generated by nanometre‐sized Pt aggregations within the cell, the ADF STEM may be used to make a measurement of local concentration of Pt in units of atoms per cubic nanometre. We further estimate the minimum atomic number required to visualise single atoms in this setting, concluding that in similar samples it may be possible to detect species as light as sodium with atomic sensitivity

Observation of metal nanoparticles at atomic resolution in Pt-based cancer chemotherapeutics

The chemotherapeutics cisplatin and oxaliplatin are importanttools in the fight against cancer. Both compounds areplatinum complexes. Aberration-corrected scanning transmissionelectron microscopy using the annular dark-fieldimaging mode now routinely provides single-atom sensitivitywith atomic number contrast. Here, this imaging mode is usedto directly image the platinum within the two drugs in theirdried form on an amorphous carbon support film. The oxaliplatinis found to have wetted the supporting amorphous carbon,forming disordered clusters suggesting that the platinumhas remained within the complex. Conversely, the cisplatinsample reveals 1.8-nm-diameter metallic platinum clusters.The size and shape of the clusters do not appear to be dependenton drying rate nor formed by beam damage, which maysuggest that they were present in the original drug solution

Infection with cerebral metacercariae of microphallid trematode parasites reduces reproductive output in the gammarid amphipod Gammarus insensibilis (Stock 1966) in UK saline lagoons

Saline lagoons are priority habitats in the United Kingdom supporting several protected specialist species. One specialist, the amphipod Gammarus insensibilis, is infected with behaviour-altering microphallid trematodes such as Microphallus papillorobustus. In saline lagoons around the coast of England (Gilkicker and Lymington–Keyhaven on the Hampshire coast and Moulton Marsh in Lincolnshire) there is variation in the prevalence of this parasite in the gammarid populations (0 at Salterns in the Lymington–Keyhaven lagoon system to 98% at Gilkicker). Infection intensity ranged from 0 to 20 metacercariae in individual amphipods. Higher infection intensity can alter the shape of the amphipod's head. Under experimental conditions respiration rate is significantly reduced in infected animals and reproductive output (expressed as early stage embryos mg g dry weight−1) is significantly lower in infected females. It is important to consider the role of host–parasite interactions in order to understand the ecology of specialist lagoon species such as G. insensibilis and their lagoon habitats

Infection with cerebral metacercariae of microphallid trematode parasites reduces reproductive output in the gammarid amphipod Gammarus insensibilis (Stock 1966) in UK saline lagoons

Saline lagoons are priority habitats in the United Kingdom supporting several protected specialist species. One specialist, the amphipod Gammarus insensibilis, is infected with behaviour-altering microphallid trematodes such as Microphallus papillorobustus. In saline lagoons around the coast of England (Gilkicker and Lymington–Keyhaven on the Hampshire coast and Moulton Marsh in Lincolnshire) there is variation in the prevalence of this parasite in the gammarid populations (0 at Salterns in the Lymington–Keyhaven lagoon system to 98% at Gilkicker). Infection intensity ranged from 0 to 20 metacercariae in individual amphipods. Higher infection intensity can alter the shape of the amphipod's head. Under experimental conditions respiration rate is significantly reduced in infected animals and reproductive output (expressed as early stage embryos mg g dry weight−1) is significantly lower in infected females. It is important to consider the role of host–parasite interactions in order to understand the ecology of specialist lagoon species such as G. insensibilis and their lagoon habitats

Nuclear DNA Replication in Trypanosomatids:There Are No Easy Methods for Solving Difficult Problems

In trypanosomatids, etiological agents of devastating diseases, replication is robust and finely controlled to maintain genome stability and function in stressful environments. However, these parasites encode several replication protein components and complexes that show potentially variant composition compared with model eukaryotes. This review focuses on the advances made in recent years regarding the differences and peculiarities of the replication machinery in trypanosomatids, including how such divergence might affect DNA replication dynamics and the replication stress response. Comparing the DNA replication machinery and processes of parasites and their hosts may provide a foundation for the identification of targets that can be used in the development of chemotherapies to assist in the eradication of diseases caused by these pathogens

A green solvent system for precursor phase-engineered sequential deposition of stable formamidinium lead triiodide for perovskite solar cells

Perovskite solar cells (PSCs) offer an efficient, inexpensive alternative to current photovoltaic technologies, with the potential for manufacture via high-throughput coating methods. However, challenges for commercial-scale solution-processing of metal-halide perovskites include the use of harmful solvents, the expense of maintaining controlled atmospheric conditions, and the inherent instabilities of PSCs under operation. Here, we address these challenges by introducing a high volatility, low toxicity, biorenewable solvent system to fabricate a range of 2D perovskites, which highly effective precursor phases for subsequent transformation to alpha-formamidinium lead triiodide (FAPbI3), fully processed under ambient conditions. PSCs utilising our FAPbI3 reproducibly show remarkable stability under illumination and elevated temperature (ISOS-L-2) and "damp heat" (ISOS-D-3) stressing, surpassing other state-of-the-art perovskite compositions. We determine that this enhancement is a consequence of the 2D precursor phase crystallisation route, which simultaneously avoids retention of residual low-volatility solvents (such as DMF and DMSO) and reduces the rate of degradation of FA+ in the material. Our findings highlight both the critical role of the initial crystallisation process in determining the operational stability of perovskite materials, and that neat FA+-based perovskites can be competitively stable despite the inherent metastability of the alpha-phase

A green solvent enables precursor phase engineering of stable formamidinium lead triiodide perovskite solar cells

Perovskite solar cells (PSCs) offer an efficient, inexpensive alternative to current photovoltaic technologies, with the potential for manufacture via high-throughput coating methods. However, challenges for commercial-scale solution-processing of metal-halide perovskites include the use of harmful solvents, the expense of maintaining controlled atmospheric conditions, and the inherent instabilities of PSCs under operation. Here, we address these challenges by introducing a high volatility, low toxicity, biorenewable solvent system to fabricate a range of 2D perovskites, which we use as highly effective precursor phases for subsequent transformation to α-formamidinium lead triiodide (α-FAPbI3), fully processed under ambient conditions. PSCs utilising our α-FAPbI3 reproducibly show remarkable stability under illumination and elevated temperature (ISOS-L-2) and “damp heat” (ISOS-D-3) stressing, surpassing other state-of-the-art perovskite compositions. We determine that this enhancement is a consequence of the 2D precursor phase crystallisation route, which simultaneously avoids retention of residual low-volatility solvents (such as DMF and DMSO) and reduces the rate of degradation of FA+ in the material. Our findings highlight both the critical role of the initial crystallisation process in determining the operational stability of perovskite materials, and that neat FA+-based perovskites can be competitively stable despite the inherent metastability of the α-phase

Major Surface Glycoproteins of Insect Forms of Trypanosoma brucei Are Not Essential for Cyclical Transmission by Tsetse

Procyclic forms of Trypanosoma brucei reside in the midgut of tsetse flies where they are covered by several million copies of glycosylphosphatidylinositol-anchored proteins known as procyclins. It has been proposed that procyclins protect parasites against proteases and/or participate in tropism, directing them from the midgut to the salivary glands. There are four different procyclin genes, each subject to elaborate levels of regulation. To determine if procyclins are essential for survival and transmission of T. brucei, all four genes were deleted and parasite fitness was compared in vitro and in vivo. When co-cultured in vitro, the null mutant and wild type trypanosomes (tagged with cyan fluorescent protein) maintained a near-constant equilibrium. In contrast, when flies were infected with the same mixture, the null mutant was rapidly overgrown in the midgut, reflecting a reduction in fitness in vivo. Although the null mutant is patently defective in competition with procyclin-positive parasites, on its own it can complete the life cycle and generate infectious metacyclic forms. The procyclic form of T. brucei thus differs strikingly from the bloodstream form, which does not tolerate any perturbation of its variant surface glycoprotein coat, and from other parasites such as Plasmodium berghei, which requires the circumsporozoite protein for successful transmission to a new host

A green solvent enables precursor phase engineering of stable formamidinium lead triiodide perovskite solar cells

Perovskite solar cells (PSCs) offer an efficient, inexpensive alternative to current photovoltaic technologies, with the potential for manufacture via high-throughput coating methods. However, challenges for commercial-scale solution-processing of metal-halide perovskites include the use of harmful solvents, the expense of maintaining controlled atmospheric conditions, and the inherent instabilities of PSCs under operation. Here, we address these challenges by introducing a high volatility, low toxicity, biorenewable solvent system to fabricate a range of 2D perovskites, which we use as highly effective precursor phases for subsequent transformation to α-formamidinium lead triiodide (α-FAPbI3), fully processed under ambient conditions. PSCs utilising our α-FAPbI3 reproducibly show remarkable stability under illumination and elevated temperature (ISOS-L-2) and “damp heat” (ISOS-D-3) stressing, surpassing other state-of-the-art perovskite compositions. We determine that this enhancement is a consequence of the 2D precursor phase crystallisation route, which simultaneously avoids retention of residual low-volatility solvents (such as DMF and DMSO) and reduces the rate of degradation of FA+ in the material. Our findings highlight both the critical role of the initial crystallisation process in determining the operational stability of perovskite materials, and that neat FA+-based perovskites can be competitively stable despite the inherent metastability of the α-phase