• We report the development of a custom designed MEMS micro-hotplate capable of operating at high temperatures (up to 700 ̊C). • The MEMS micro-hotplate can be directly mounted in a slide holder of a fluorescent confocal microscope. • Temperature-sensitive nanosensors (550 nm diameter) were synthesised by covalently linking rhodamine B to a silica sol–gel matrix. • Nanosensors were thermo-optically characterised using the MEMS device and confocal microscopy. • The temperature dependent fluorescence response of the nanosensors was found to operate over wide range, up to 145 ̊C.

Biologic therapeutics are the medicines of the future and are destined to transform the approaches by which the causes and symptoms of diseases are cured and alleviated. These approaches will be accelerated through the development of novel strategies that target multiple pharmacologically active sites using a combination of different biologics, or mixtures of biologics and small molecule therapeutics. However, for this potential to be realised, advancements in co-formulation strategies for biologic therapeutics must be established. This review describes the current and emerging developments within this field and highlights the challenges and potential solutions, that will pave-the-way towards their clinical translation.

The nematode Pristionchus pacificus has been established as a model for comparative studies using the well known Caenorhabditis elegans as a reference. Despite their relatedness, previous studies have revealed highly divergent development and a number of morphological differences including the lack of a pharyngal structure, the grinder, used to physically lyse the ingested bacteria in C. elegans. To complement current knowledge about developmental and ecological differences with a better understanding of their feeding modes, we have sequenced the intestinal transcriptomes of both nematodes. In total, we found 464 intestine-enriched genes in P. pacificus and 724 in C. elegans, of which the majority (66 %) has been identified by previous studies. Interestingly, only 15 genes could be identified with shared intestinal enrichment in both species, of which three genes are Hedgehog signaling molecules supporting a highly conserved role of this pathway for intestinal development across all metazoa. At the level of gene families, we find similar divergent trends with only five families displaying significant intestinal enrichment in both species. We compared our data with transcriptomic responses to various pathogens. Strikingly, C. elegans intestine-enriched genes showed highly significant overlaps with pathogen response genes whereas this was not the case for P. pacificus, indicating shifts in pathogen susceptibility that might be explained by altered feeding modes. Our study reveals first insights into the evolution of feeding systems and the associated changes in intestinal gene expression that might have facilitated nematodes of the P. pacificus lineage to colonize new environments. These findings deepen our understanding about how morphological and genomic diversity is created during the course of evolution.

Silica nanoparticles (SNPs) have been used as favoured platforms for sensor, drug delivery and biological imaging applications, due to their ease of synthesis, size-control and bespoke physico-chemical properties. In this study, we have developed a protocol for the synthesis of size-tuneable SNPs, with diameters ranging from 20 nm to 500 nm, through the optimisation of experimental components required for nanoparticle synthesis. This protocol was also used to prepare fluorescent SNPs, via covalent linkages of fluorophores, to the nanoparticle matrix using 3-aminopropyltriethoxysilane (APTES). This enabled the fabrication of ratiometric, fluorescent, pH-sensitive nanosensors (75 nm diameter) composed SNPs covalently linked to two pH-sensitive fluorescent dyes Oregon Green (OG) and 5(6)-carboxyfluorescein (FAM) and a reference fluorescent dye 5-(6)-carboxytetramethylrhodamine (TAMRA), extending the dynamic range of measurement from pH 3.5 to 7.5. In addition, size-tuneable, core–shell SNPs, covalently linked to a fluorescent TAMRA core were synthesised to investigate distance-dependant fluorescence quenching between TAMRA and black hole quencher 2 (BHQ2®) using nanometre-sized silica shells as physical spacers. The results showed a significant fluorescence quenching could be observed over greater distances than that reported for the classical distance-dependent molecular fluorescence quenching techniques, e.g. the Förster (fluorescence) resonance energy transfer (FRET). The methods and protocols we have detailed in this manuscript will provide the basis for the reproducible production of size tunable SNPs, which will find broad utility in the development of sensors for biological applications.

Necator americanus, a haematophagous hookworm parasite, infects ~10% of the world’s population and is considered to be a significant public health risk. Its lifecycle has distinct stages, permitting its successful transit from the skin via the lungs (L₃) to the intestinal tract (L₄ maturing to adult). It has been hypothesised that the L₃ larval sheath, which is shed during percutaneous infection (exsheathment), diverts the immune system to allow successful infection and reinfection in endemic areas. However, the physicochemical properties of the L₃ larval cuticle and sheath, which are in direct contact with the skin and its immune defences, are unknown. In the present study, we controlled exsheathment, to characterise the sheath and underlying cuticle surfaces in situ, using atomic force microscopy (AFM) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). AFM revealed previously unseen surface area enhancing nano-annuli exclusive to the sheath surface and confirmed greater adhesion forces exist between cationic surfaces and the sheath, when compared to the emergent L₃ cuticle. Furthermore, ToF-SIMS elucidated different chemistries between the surfaces of the cuticle and sheath which could be of biological significance. For example, the phosphatidylglycerol rich cuticle surface may support the onward migration of a lubricated infective stage, while the anionic and potentially immunologically active heparan sulphate rich deposited sheath could result in the diversion of immune defences to an inanimate antigenic nidus. We propose that our initial studies into the surface analysis of this hookworm provides a timely insight into the physicochemical properties of a globally important human pathogen at its infective stage and anticipate that the development and application of this analytical methodology will support translation of these findings into a biological context. Necator americanus is an intestinal hookworm parasite of humans that is commonly found in tropical and sub-tropical climates. N. americanus infections can be treated effectively with anthelmintic drug therapy; however, in endemic areas re-infection quickly returns. Chronic hookworm infection can lead to intestinal blood loss, iron deficiency anaemia, malnutrition and physical and intellectual impairment. N. americanus surfaces may possess key physicochemical properties that permit successful host infection. Therefore, we harnessed controlled exsheathment of infective axenic L₃ larva to investigate the physicochemical properties of the emergent cuticle and deposited sheath, using atomic force microscopy (AFM) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Our results provide an early insight into the differential physicochemical properties of these bio-surfaces, allowing the development of a hypothesis as to how these chemistries may be involved in infection and immune evasion. This new analytical platform will allow us to test this hypothesis and translate our findings into an immuno-biological context.