Search

Presents a study which measured radiated emissions for simple digital circuits designed to operate with various logic families. Information on the emissions in the near and far fields; Difference in peak emissions between any two logic families; Use of test circuits to measure the propagation delay of each logic family; Results of this study.

A survey of blood lead concentrations is reported in a group of 199 young Walsall children (3-6 years of age). The geometric mean blood lead concentration was 0.47 mumol/l (range 0.2-1.6 mumol/l). There were no significant differences in blood lead concentrations between groups of children with different ages, sex, ethnic origin, or environment. [ABSTRACT FROM AUTHOR]

We recently demonstrated that vaccines comprising antigenic peptides conjugated to a glycolipid agonist, termed glycolipid-peptide (GLP) vaccines, efficiently generate substantial numbers of long-lived CD8 + liver-resident memory T (Trm) cells that are crucial for protection against malaria liver-stage infection. To understand the underlying mechanism, we examined the prerequisites for priming, differentiation, and secondary boosting of liver Trm cells using these GLP vaccines. Our study revealed that generation of long-lived liver Trm cells relies on CD8 + T cell priming by type 1 conventional dendritic (cDC1) cells, followed by post-priming exposure to a combination of vaccine-derived inflammatory and antigenic signals. Boosting of liver Trm cells is feasible using the same GLP vaccine, but a substantial delay is required for optimal responses due to natural killer T (NKT) cell anergy. Overall, our study unveils key requirements for the development of long-lived liver Trm cells, offering valuable insights for future vaccine design., Competing Interests: Declaration of interests G.F.P. and I.F.H. have been chief technical officer and chief scientific officer, respectively, of biotech start-up Avalia Immunotherapies Limited, and W.R.H. was a member of its Scientific Advisory Board. Avalia holds exclusive, worldwide license to patents related to aspects of the chemical design reported here. Avalia partially funded the study. W.R.H., I.F.H., L.E.H., B.J.C., and G.F.P. are inventors on patent application (WO2020231274) submitted by Victoria University of Wellington, Malcorp Biodiscoveries Limited, and Victoria Link Limited that covers the production of tissue-resident memory T cells with GLP vaccines. M.H.L., K.M.T., and I.C. are inventors on patents relating to Clec9A antibodies., (Copyright © 2025 The Author(s). Published by Elsevier Inc. All rights reserved.)

mRNA vaccines have recently generated significant interest due to their success during the COVID-19 pandemic. Their success is due to advances in mRNA design and encapsulation into ionizable lipid nanoparticles (iLNPs). This has highlighted the potential for the use of mRNA-iLNPs in other settings such as cancer, gene therapy, or vaccines for different infectious diseases. Here, we describe the production of mRNA-iLNPs using commercially available reagents that are suitable for use as vaccines and therapeutics. This article contains detailed protocols for the synthesis of mRNA by in vitro transcription with enzymatic capping and tailing and the encapsulation of the mRNA into iLNPs using the ionizable lipid DLin-MC3-DMA. DLin-MC3-DMA is often used as a benchmark for new formulations and provides an efficient delivery vehicle for screening mRNA design. The protocol also describes how the formulation can be adapted to other lipids. Finally, a stepwise methodology is presented for the characterization and quality control of mRNA-iLNPs, including measuring mRNA concentration and encapsulation efficiency, particle size, and zeta potential. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Synthesis of mRNA by in vitro transcription and enzymatic capping and tailing Basic Protocol 2: Encapsulation of mRNA into ionizable lipid nanoparticles Alternate Protocol: Small-scale encapsulation of mRNA using preformed vesicles Basic Protocol 3: Characterization and quality control of mRNA ionizable lipid nanoparticles., (© 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.)

Malaria is caused by Plasmodium species transmitted by Anopheles mosquitoes. Following a mosquito bite, Plasmodium sporozoites migrate from skin to liver, where extensive replication occurs, emerging later as merozoites that can infect red blood cells and cause symptoms of disease. As liver tissue-resident memory T cells (Trm cells) have recently been shown to control liver-stage infections, we embarked on a messenger RNA (mRNA)-based vaccine strategy to induce liver Trm cells to prevent malaria. Although a standard mRNA vaccine was unable to generate liver Trm or protect against challenge with Plasmodium berghei sporozoites in mice, addition of an agonist that recruits T cell help from type I natural killer T cells under mRNA-vaccination conditions resulted in significant generation of liver Trm cells and effective protection. Moreover, whereas previous exposure of mice to blood-stage infection impaired traditional vaccines based on attenuated sporozoites, mRNA vaccination was unaffected, underlining the potential for such a rational mRNA-based strategy in malaria-endemic regions., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Full-spectrum flow cytometry is now routinely used in many laboratories internationally, and the demand for this technology is rapidly increasing. With capacity to use larger and more complex staining panels, standardized protocols are required for optimal panel design and analysis. Importantly, for ex vivo analysis, tissue preparation methods also need to be optimized to ensure samples are truly representative of tissues in situ. This is particularly relevant given the recent interest in adaptive immune cells that form residency in specific organs. Here we provide optimized protocols for tissue processing and phenotyping of memory T cells and natural killer T (NKT) cell subsets from liver, lung, spleen, and lymph node using full-spectrum flow cytometry. We provide a 21-color antibody panel for identification of different memory subsets, including tissue-resident memory T (T RM ) cells, which are increasingly regarded as important effectors in adaptive immunity. We show that processing procedures can affect outcomes, with liver T RM cells particularly sensitive to heat, such that accurate evaluation requires fast processing at defined temperatures. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Processing mouse liver for flow cytometric analysis of memory T and NKT cell subsets Basic Protocol 2: Processing mouse spleen for flow cytometric analysis of memory T and NKT cell subsets Basic Protocol 3: Processing mouse lungs for flow cytometric analysis of memory T and NKT cell subsets Basic Protocol 4: Processing mouse lymph nodes for flow cytometric analysis of memory T and NKT cell subsets Basic Protocol 5: Staining and flow cytometric analysis of samples for memory T and NKT cell subsets Support Protocol: Obtaining cell counts from flow cytometry data., (© 2022 Wiley Periodicals LLC.)

Objective: Evidence supports a bidirectional relationship between stress and asthma exacerbations in children, suggesting that interventions to reduce stress may improve both psychosocial quality of life and disease course. Here, we examine the feasibility of a stress management intervention for 7- to 12-year-olds with asthma., Methods: Two trials were conducted. Cohort 1 (n = 11) was recruited from the community and attended intervention sessions at an urban university. Cohort 2 (n = 7) was school based and recruited from an African American charter school. Six individual intervention sessions focused on psychoeducation about asthma, stress, and emotions; problem-solving and coping skills training; and relaxation training paired with physiological feedback. Pre- and post-intervention stress, mood, and lung function data were collected. Satisfaction surveys were administered after intervention completion., Results: The intervention was rated as highly acceptable by participating families. Feasibility was much stronger for the school-based than the university-based recruitment mechanism. Initial efficacy data suggest that both cohorts showed pre- to post-intervention improvements in lung function, perceived stress, and depressed mood., Conclusion: Findings provide evidence for the feasibility of offering asthma-related stress management training in a school setting. Initial findings offer support for future, large-scale efficacy studies.

Purpose: To investigate the effect of the combined treatment of photodynamic therapy and specific VEGF165 inhibition with pegaptanib sodium (Macugen; Eyetech Pharmaceuticals, Lexington, MA) on ocular neovascularization., Methods: Photodynamic therapy's (PDT's) effects on the integrity of pegaptanib sodium were analyzed by HPLC, a VEGF165-binding assay, and a VEGF165-induced tissue factor gene expression assay. The effects of mono- or combined treatment on vessel growth and regression were determined in a murine corneal neovascularization model. The effects of combined treatment on vessel growth were also determined in a murine choroidal neovascularization model., Results: PDT did not affect the chemical composition of pegaptanib sodium nor the efficacy of pegaptanib sodium in the inhibition of VEGF165 binding to Flt-1 and VEGF165-induced gene expression. In an animal model of effects on existing ocular neovascular lesions (corneal neovascularization), PDT monotherapy yielded an initial regression of these vessels, but there followed a rapid regrowth. In contrast, pegaptanib sodium monotherapy yielded little regression but potently abrogated further vessel growth. The combination of pegaptanib sodium and PDT resulted in the regression of the neovascular lesions, as observed with PDT alone, but also prevented significant vessel regrowth, leading to a significantly greater reduction in lesion size than did each monotherapy. In addition, there was a significantly greater effect of the combination of pegaptanib sodium and PDT on lesion size in choroidal neovascularization than with each monotherapy. Pretreatment with pegaptanib sodium appeared to decrease the efficacy of PDT-induced vessel regression in corneal neovascularization, and as such the enhanced efficacy over monotherapy when the agents were delivered simultaneously was not observed., Conclusions: Although the combined simultaneous treatment of ocular neovascularization with PDT and pegaptanib sodium may provide a more effective approach for the regression and overall treatment of CNV associated with AMD, the order of addition of these treatments may play a role in achieving optimal efficacy.

This paper considers the spread of the 1918 influenza virus as it occurred among Inupiat communities on Alaska's Seward Peninsula. Documented historical sources concerning the epidemic among the Native communities are scarce and contain limited information. In contrast, oral histories of the event provide not only information about community mortality rates, but they also corroborate the few written documents that discuss the epidemic. Additionally, it has been found that the dispersal of the virus on the peninsula can only be accurately reconstructed with the aid of oral testimony. Data useful to both anthropologists and epidemiologists can be derived from this approach to historical analysis.