Production and characterization of anti-human interferon γ receptor antibody fragments that inhibit cytokine binding to the receptor

Three single-chain antibody fragments that recognize the extracellular human interferon γ receptor α-chain (IFNγR), and inhibit the binding of human IFNγ, have been produced in Escherichia coli. These fragments are derived from murine anti-receptor monoclonal antibodies, and comprise the variable heavy (VH) domain linked to the variable light (VL) chain through a 15 amino acid linker [(GGGGS)3]. Using surface plasmon resonance technology (BIAcore), the soluble proteins were shown to retain a high affinity for recombinant IFNγR, and by radioimmunoassay to possess high inhibitory activity towards IFNγ-binding to human Raji cells. The antibody fragments most likely recognize epitopes that overlap the cytokine binding site on the receptor surface. Attempts to dissect further the antibodies to isolated VH- and VL-chains and to synthetic linear and cyclic peptides derived from the individual complementarity determining regions failed to afford fragments with significant IFNγR binding affinity. Nevertheless, these native-like variable region fragments and petidomimetics derived from them are of interest in the design of novel IFNγR antagonist

Sudlow site II of human serum albumin remains functional after gold nanocluster encapsulation : a fluorescence-based drug binding study of L-Dopa

Fluorescent protein-encapsulated gold nanoclusters (AuNCs) offer a non-toxic means of sensing and imaging biological phenomena on the nanoscale. However, the biofunctionality of proteins encapsulating AuNCs has not been fully elucidated to date. Here we studied the biofunctionality of the second major drug binding site (Sudlow II) of Human Serum Albumin (HSA) encapsulated AuNCs after AuNC synthesis. L-Dopa, a fluorescent drug molecule associated with the clinical treatment of Parkinson’s disease, which commonly binds to the Sudlow II site, was used to study the availability of the site before and after AuNC synthesis through changes to its fluorescence characteristics. L-Dopa was observed using its intrinsic fluorescence to readily bind to HSA-AuNCs complexes. Interestingly, the fluorescence emission intensity of AuNCs linearly increased with L-Dopa concentration while exciting the AuNC directly at 470 nm, Using a 400 nMHSA-AuNC solution, L-Dopa was rapidly detected at a limit of 300 pM, indicating that HSA-AuNCs fluorescence is extremely sensitive to molecular binding at the Sudlow II binding site. Future research may be able to utilize this sensitivity to improve the fluorescence characteristics of AuNCs within HSA-AuNCs for imaging and sensing including drug binding studies

Optimal omegas – barriers and novel methods to narrow omega-3 gaps. A narrative review

Copyright © 2024 Derbyshire, Birch, Bonwick, English, Metcalfe and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Dietary intakes of omega-3 long chain polyunsaturated fatty acids (O3LC-PUFAs) such as eicosapentaenoic and docosahexaenoic acid are central to development and health across the life course. O3LC-PUFAs have been linked to neurological development, maternal and child health and the etiology of certain non-communicable diseases including age-related cognitive decline, cardiovascular disease, and diabetes. However, dietary inadequacies exist in the United Kingdom and on a wider global scale. One predominant dietary source of O3LC-PUFAs is fish and fish oils. However, growing concerns about overfishing, oceanic contaminants such as dioxins and microplastics and the trend towards plant-based diets appear to be acting as cumulative barriers to O3LC-PUFAs from these food sources. Microalgae are an alternative provider of O3LC-PUFA-rich oils. The delivery of these into food systems is gaining interest. The present narrative review aims to discuss the present barriers to obtaining suitable levels of O3LC-PUFAs for health and wellbeing. It then discusses potential ways forward focusing on innovative delivery methods to utilize O3LC-PUFA-rich oils including the use of fortification strategies, bioengineered plants, microencapsulation, and microalgae

Evidence for mechanical and chemical alteration of iron‐nickel meteorites on Mars: Process insights for Meridiani Planum

The weathering of meteorites found on Mars involves chemical and physical processes that can provide clues to climate conditions at the location of their discovery. Beginning on sol 1961, the Opportunity rover encountered three large iron meteorites within a few hundred meters of each other. In order of discovery, these rocks have been assigned the unofficial names Block Island, Shelter Island, and Mackinac Island. Each rock presents a unique but complimentary set of features that increase our understanding of weathering processes at Meridiani Planum. Significant morphologic characteristics interpretable as weathering features include (1) a large pit in Block Island, lined with delicate iron protrusions suggestive of inclusion removal by corrosive interaction; (2) differentially eroded kamacite and taenite lamellae in Block Island and Shelter Island, providing relative timing through crosscutting relationships with deposition of (3) an iron oxide–rich dark coating; (4) regmaglypted surfaces testifying to regions of minimal surface modification, with other regions in the same meteorites exhibiting (5) large‐scale, cavernous weathering (in Shelter Island and Mackinac Island). We conclude that the current size of the rocks is approximate to their original postfall contours. Their morphology thus likely results from a combination of atmospheric interaction and postfall weathering effects. Among our specific findings is evidence supporting (1) at least one possible episode of aqueous acidic exposure for Block Island; (2) ripple migration over portions of the meteorites; (3) a minimum of two separate episodes of wind abrasion; alternating with (4) at least one episode of coating‐forming chemical alteration, most likely at subzero temperatures

The Role and Mechanism of Erythrocyte Invasion by Francisella tularensis

Francisella tularensis is an extremely virulent bacterium that can be transmitted naturally by blood sucking arthropods. During mammalian infection, F. tularensis infects numerous types of host cells, including erythrocytes. As erythrocytes do not undergo phagocytosis or endocytosis, it remains unknown how F. tularensisinvades these cells. Furthermore, the consequence of inhabiting the intracellular space of red blood cells (RBCs) has not been determined. Here, we provide evidence indicating that residing within an erythrocyte enhances the ability of F. tularensis to colonize ticks following a blood meal. Erythrocyte residence protected F. tularensis from a low pH environment similar to that of gut cells of a feeding tick. Mechanistic studies revealed that the F. tularensis type VI secretion system (T6SS) was required for erythrocyte invasion as mutation of mglA (a transcriptional regulator of T6SS genes), dotU, or iglC (two genes encoding T6SS machinery) severely diminished bacterial entry into RBCs. Invasion was also inhibited upon treatment of erythrocytes with venom from the Blue-bellied black snake (Pseudechis guttatus), which aggregates spectrin in the cytoskeleton, but not inhibitors of actin polymerization and depolymerization. These data suggest that erythrocyte invasion by F. tularensis is dependent on spectrin utilization which is likely mediated by effectors delivered through the T6SS. Our results begin to elucidate the mechanism of a unique biological process facilitated by F. tularensis to invade erythrocytes, allowing for enhanced colonization of ticks

Monitoring Influenza Activity in the United States: A Comparison of Traditional Surveillance Systems with Google Flu Trends

Google Flu Trends was developed to estimate US influenza-like illness (ILI) rates from internet searches; however ILI does not necessarily correlate with actual influenza virus infections.Influenza activity data from 2003-04 through 2007-08 were obtained from three US surveillance systems: Google Flu Trends, CDC Outpatient ILI Surveillance Network (CDC ILI Surveillance), and US Influenza Virologic Surveillance System (CDC Virus Surveillance). Pearson's correlation coefficients with 95% confidence intervals (95% CI) were calculated to compare surveillance data. An analysis was performed to investigate outlier observations and determine the extent to which they affected the correlations between surveillance data. Pearson's correlation coefficient describing Google Flu Trends and CDC Virus Surveillance over the study period was 0.72 (95% CI: 0.64, 0.79). The correlation between CDC ILI Surveillance and CDC Virus Surveillance over the same period was 0.85 (95% CI: 0.81, 0.89). Most of the outlier observations in both comparisons were from the 2003-04 influenza season. Exclusion of the outlier observations did not substantially improve the correlation between Google Flu Trends and CDC Virus Surveillance (0.82; 95% CI: 0.76, 0.87) or CDC ILI Surveillance and CDC Virus Surveillance (0.86; 95%CI: 0.82, 0.90).This analysis demonstrates that while Google Flu Trends is highly correlated with rates of ILI, it has a lower correlation with surveillance for laboratory-confirmed influenza. Most of the outlier observations occurred during the 2003-04 influenza season that was characterized by early and intense influenza activity, which potentially altered health care seeking behavior, physician testing practices, and internet search behavior

Immigration and the Common Profit: Native Cloth Workers, Flemish Exiles, and Royal Policy in Fourteenth-Century London

Drawing on a wide variety of published and unpublished sources, this article reconstructs a crucial episode in the relationship between the English Crown, its native subjects and the kingdom’s immigrant population during the later Middle Ages. Determined that their presence would boost the development of the local textile industries, Edward III encouraged high numbers of skilled Flemish cloth workers who had been exiled from their home county at the start of the 1350s to settle in the realm. Most of them took up residence in London, where they produced higher-quality cloth for the domestic market and, probably, for export. Soon, however, the immigrants’ activities conflicted with the privileges that had structured the capital’s economic life for centuries. Their work was contested by London’s native weavers who, since the middle of the twelfth century, had enjoyed the sole right to produce cloth in the city. Hoping that the control over the immigrants’ activities would help them to overcome the crisis in the market for lower-quality textiles they were struggling with, the natives petitioned the king to obtain the incorporation of the Flemish weavers into their guild for over twenty-five years. Yet, arguing that the Flemings’ contribution benefited the common profit of the whole kingdom in a way that transcended the interests of any particular group, the Crown rejected all their requests and avoided every attempt at discussion. Each time political communication broke down, the native weavers took out their frustrations by physically attacking their Flemish counterparts. These incidents became increasingly violent during the years leading up to the Peasants’ Revolt in 1381 and came to a dramatic conclusion during the rebellion itself

In vitro and in vivo evaluation of a single chain antibody fragment generated in planta with potent rabies neutralisation activity.

Rabies causes more than 60,000 human deaths annually in areas where the virus is endemic. Importantly, rabies is one of the few pathogens for which there is no treatment following the onset of clinical disease with the outcome of infection being death in almost 100% of cases. Whilst vaccination, and the combination of vaccine and rabies immunoglobulin treatment for post-exposure administration are available, no tools have been identified that can reduce or prevent rabies virus replication once clinical disease has initiated. The search for effective antiviral molecules to treat those that have already developed clinical disease associated with rabies virus infection is considered one of the most important goals in rabies research. The current study assesses a single chain antibody molecule (ScFv) based on a monoclonal antibody that potently neutralises rabies in vitro as a potential therapeutic candidate. The recombinant ScFv was generated in Nicotiana benthamiana by transient expression, and was chemically conjugated (ScFv/RVG) to a 29 amino acid peptide, specific for nicotinic acetylcholine receptor (nAchR) binding in the CNS. This conjugated molecule was able to bind nAchR in vitro and enter neuronal cells more efficiently than ScFv. The ability of the ScFv/RVG to neutralise virus in vivo was assessed using a staggered administration where the molecule was inoculated either four hours before, two days after or four days after infection. The ScFv/RVG conjugate was evaluated in direct comparison with HRIG and a potential antiviral molecule, Favipiravir (also known as T-705) to indicate whether there was greater bioavailability of the ScFv in the brains of treated mice. The study indicated that the approach taken with the ScFv/RVG conjugate may have utility in the design and implementation of novel tools targetting rabies virus infection in the brain