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The photochemical degradation of metamitron (4-amino-4,5-dihydro-3-methyl-6-phenyl-1,2,4-triaz in-5-one) and imidacloprid (1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin- 2-ylideneamine) has been investigated by differential-pulse polarography (DPP) and micellar electrokinetic chromatography (MEKC); the degradation pathways of these pesticides were elucidated and their degradation products proposed. The electrochemical study of imidacloprid by DPP at different pH values demonstrated the occurrence of two different reduction processes; at pH6.8, two peaks at -0.90 V and -1.38 V, respectively, were obtained, which are related to the photochemical reduction processes. The photochemical degradation of imidacloprid caused by sunlight was polarographically monitored and its degradation products elucidated. The polarographic reduction of deaminometamitron (obtained by photochemical reduction of metamitron) yielded two peaks at -0.62 and -1.37 V, which are related to the reduction of the CN bonds. The effect of sunlight on the reduction of metamitron was monitored by DPP, and an increase of the concentration of the degradation products was observed with time. MEKC with UV-visible detection was used to separate the pesticides and the products of their photochemical degradation, which were identified in combination with DPP. [ABSTRACT FROM AUTHOR]

Background: Urine biomarkers are crucial for monitoring patient responses in treating urological pathologies, including non-muscle invasive bladder cancer (NMIBC). Yet, analysing urine biomarkers poses several challenges, including ensuring specimen stability during transportation and analytical processing. This prospective feasibility study aimed to investigate how urinary leukocytes and proteins are impacted by storing and refrigerating urine., Methods: Stability of leukocytes from four healthy donors (HD) spiked into urine supernatants was analyzed for up to 72 h at 4°C. Urine samples from five NMIBC patients undergoing BCG treatment were divided into two portions, followed by either immediate processing or overnight refrigeration. Urinary cell content and soluble factors were analyzed by multiparameter flow cytometry and Luminex®, respectively., Results: We confirmed the stability of healthy donor peripheral blood leukocytes spiked into cell-free urine supernatants from healthy donors or untreated bladder cancer patients for up to 72 h under refrigeration at + 5℃. Additionally, we conducted immune cell and proteomic analysis from urine samples obtained from five NMIBC patients receiving Bacillus Calmette-Guérin (BCG) therapy either processed immediately or after overnight refrigeration. We successfully demonstrated that leukocyte and protein composition remain stable in refrigerated urine from BCG-treated NMIBC for 24 h. This included granulocytes, monocytes, and T cells, as well as total protein, creatinine and 46 additional individual immune-related mediators., Conclusions: This work demonstrates the compatibility of refrigerated urine shipment from the collection sites to analytical laboratories with both immunophenotyping and proteomic analysis and establishes clear logistical benefits for numerous clinical settings focused on monitoring patient immune responses in the urine matrix., Competing Interests: Declarations. Ethics approval and consent to participate: The study was approved by the Health Research Authority, London—Stanmore Research Ethics Committee (19/LO/0179; 257743) and the Ethics Committee of the Canton de Vaud in Switzerland (#2019–00564). All patients provided written informed consent before taking part. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Living cures are cell-based, programmable therapies that integrate the latest learnings in immunology and synthetic biology. Although Adoptive Cell Therapies (ACTs) have transformed the treatment landscape of haematological malignancies by harnessing the powerful anti-tumour properties of immune cells, commercialisation and ensuring access is challenging. Their application in solid tumour treatment has been hindered by the immunosuppressive tumour microenvironment (TME) and its associated physical barriers. Conversely, bacterial immunotherapies offer cost-effective solutions by utilising tumour-colonising bacteria that trigger localised inflammatory responses within the TME. The authors briefly examine advancements in ACT and propose bacterial immunotherapies as an alternative or complementary treatment modality with potential use either as standalone therapies or in conjunction with other treatments., Competing Interests: At the time of submission, all authors were full time employees of Prokarium Ltd and received funding from Prokarium Ltd in the form of salary. This does not alter adherence to journal policies., (© 2024 Prokarium Ltd. Engineering Biology published by John Wiley & Sons Ltd on behalf of The Institution of Engineering and Technology.)

Enteric fever is a major global healthcare issue caused largely by Salmonella enterica serovars Typhi and Paratyphi A. The objective of this study was to develop a novel, bivalent oral vaccine capable of protecting against both serovars. Our approach centred on genetically engineering the attenuated S. Typhi ZH9 strain, which has an excellent safety record in clinical trials, to introduce two S. Paratyphi A immunogenic elements: flagellin H:a and lipopolysaccharide (LPS) O:2. We first replaced the native S. Typhi fliC gene encoding flagellin with the highly homologous fliC gene from S. Paratyphi A using Xer-cise technology. Next, we replaced the S. Typhi rfbE gene encoding tyvelose epimerase with a spacer sequence to enable the sustained expression of O:2 LPS and prevent its conversion to O:9 through tyvelose epimerase activity. The resulting new strain, ZH9PA, incorporated these two genetic changes and exhibited comparable growth kinetics to the parental ZH9 strain. A formulation containing both ZH9 and ZH9PA strains together constitutes a new bivalent vaccine candidate that targets both S. Typhi and S. Paratyphi A antigens to address a major global healthcare gap for enteric fever prophylaxis. This vaccine is now being tested in a Phase I clinical trial (NCT04349553).

Pentraxin 3 (PTX3) is an inflammatory mediator acting as a fluid-phase pattern recognition molecule and playing an essential role in innate immunity and matrix remodeling. Inflammatory mediators also contribute to skeletal homeostasis, operating at multiple levels in physiological and pathological conditions. This study was designed to investigate the role of PTX3 in physiological skeletal remodeling and bone healing. Micro-computed tomography (μCT) and bone histomorphometry of distal femur showed that PTX3 gene-targeted female and male mice ( ptx3 -/- ) had lower trabecular bone volume than their wild-type ( ptx3 +/+ ) littermates (BV/TV by μCT: 3.50 ± 1.31 vs 6.09 ± 1.17 for females, p  < 0.0001; BV/TV 9.06 ± 1.89 vs 10.47 ± 1.97 for males, p  = 0.0435). In addition, μCT revealed lower trabecular bone volume in second lumbar vertebra of ptx3 -/- mice. PTX3 was increasingly expressed during osteoblast maturation in vitro and was able to reverse the negative effect of fibroblast growth factor 2 (FGF2) on osteoblast differentiation. This effect was specific for the N -terminal domain of PTX3 that contains the FGF2-binding site. By using the closed transversal tibial fracture model, we found that ptx3 -/- female mice formed significantly less mineralized callus during the anabolic phase following fracture injury compared to ptx3 +/+ mice (BV/TV 17.05 ± 4.59 vs 20.47 ± 3.32, p  = 0.0195). Non-hematopoietic periosteal cells highly upregulated PTX3 expression during the initial phase of fracture healing, particularly CD51 + and αSma + osteoprogenitor subsets, and callus tissue exhibited concomitant expression of PTX3 and FGF2 around the fracture site. Thus, PTX3 supports maintenance of the bone mass possibly by inhibiting FGF2 and its negative impact on bone formation. Moreover, PTX3 enables timely occurring sequence of callus mineralization after bone fracture injury. These results indicate that PTX3 plays an important role in bone homeostasis and in proper matrix mineralization during fracture repair, a reflection of the function of this molecule in tissue homeostasis and repair.

Many body surfaces harbor organ-specific γδ T cell compartments that contribute to tissue integrity. Thus, murine dendritic epidermal T cells (DETCs) uniquely expressing T cell receptor (TCR)-Vγ5 chains protect from cutaneous carcinogens. The DETC repertoire is shaped by Skint1, a butyrophilin-like (Btnl) gene expressed specifically by thymic epithelial cells and suprabasal keratinocytes. However, the generality of this mechanism has remained opaque, since neither Skint1 nor DETCs are evolutionarily conserved. Here, Btnl1 expressed by murine enterocytes is shown to shape the local TCR-Vγ7(+) γδ compartment. Uninfluenced by microbial or food antigens, this activity evokes the developmental selection of TCRαβ(+) repertoires. Indeed, Btnl1 and Btnl6 jointly induce TCR-dependent responses specifically in intestinal Vγ7(+) cells. Likewise, human gut epithelial cells express BTNL3 and BTNL8 that jointly induce selective TCR-dependent responses of human colonic Vγ4(+) cells. Hence, a conserved mechanism emerges whereby epithelia use organ-specific BTNL/Btnl genes to shape local T cell compartments., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Long pentraxin 3 (PTX3) is a non-redundant component of the humoral arm of innate immunity. The present study was designed to investigate the interaction of PTX3 with Neisseria meningitidis. PTX3 bound acapsular meningococcus, Neisseria-derived outer membrane vesicles (OMV) and 3 selected meningococcal antigens (GNA0667, GNA1030 and GNA2091). PTX3-recognized microbial moieties are conserved structures which fulfil essential microbial functions. Ptx3-deficient mice had a lower antibody response in vaccination protocols with OMV and co-administration of PTX3 increased the antibody response, particularly in Ptx3-deficient mice. Administration of PTX3 reduced the bacterial load in infant rats challenged with Neisseria meningitidis. These results suggest that PTX3 recognizes a set of conserved structures from Neisseria meningitidis and acts as an amplifier/endogenous adjuvant of responses to this bacterium.