Deciphering the regulatory landscapte of fetal and adult γδ T-cell development at single-cell resolution

γδ T cells with distinct properties develop in the embryonic and adult thymus and have been identified as critical players in a broad range of infections, antitumor surveillance, autoimmune diseases, and tissue homeostasis. Despite their potential value for immunotherapy, differentiation of γδ T cells in the thymus is incompletely understood. Here, we establish a high‐resolution map of γδ T‐cell differentiation from the fetal and adult thymus using single‐cell RNA sequencing. We reveal novel sub‐types of immature and mature γδ T cells and identify an unpolarized thymic population which is expanded in the blood and lymph nodes. Our detailed comparative analysis reveals remarkable similarities between the gene networks active during fetal and adult γδ T‐cell differentiation. By performing a combined single‐cell analysis of Sox13, Maf, and Rorc knockout mice, we demonstrate sequential activation of these factors during IL ‐17‐producing γδ T‐cell (γδT17) differentiation. These findings substantially expand our understanding of γδ T‐cell ontogeny in fetal and adult life. Our experimental and computational strategy provides a blueprint for comparing immune cell differentiation across developmental stages

Non-classical symmetries and the singular manifold method: A further two examples

This paper discusses two equations with the conditional Painleve property. The usefulness of the singular manifold method as a tool for determining the non-classical symmetries that reduce the equations to ordinary differential equations with the Painleve property is confirmed once moreComment: 9 pages (latex), to appear in Journal of Physics

PBP-A, a cyanobacterial dd-peptidase with high specificity for amidated muropeptides, exhibits pH-dependent promiscuous activity harmful to <em>Escherichia coli</em>

\ua9 The Author(s) 2024. Penicillin binding proteins (PBPs) are involved in biosynthesis, remodeling and recycling of peptidoglycan (PG) in bacteria. PBP-A from Thermosynechococcus elongatus belongs to a cyanobacterial family of enzymes sharing close structural and phylogenetic proximity to class A β-lactamases. With the long-term aim of converting PBP-A into a β-lactamase by directed evolution, we simulated what may happen when an organism like Escherichia coli acquires such a new PBP and observed growth defect associated with the enzyme activity. To further explore the molecular origins of this harmful effect, we decided to characterize deeper the activity of PBP-A both in vitro and in vivo. We found that PBP-A is an enzyme endowed with dd-carboxypeptidase and dd-endopeptidase activities, featuring high specificity towards muropeptides amidated on the d-iso-glutamyl residue. We also show that a low promiscuous activity on non-amidated peptidoglycan deteriorates E. coli’s envelope, which is much higher under acidic conditions where substrate discrimination is mitigated. Besides expanding our knowledge of the biochemical activity of PBP-A, this work also highlights that promiscuity may depend on environmental conditions and how it may hinder rather than promote enzyme evolution in nature or in the laboratory

KAT Ligation for Rapid and Facile Covalent Attachment of Biomolecules to Surfaces

The efficient and bioorthogonal chemical ligation reaction between potassium acyltrifluoroborates (KATs) and hydroxylamines (HAs) was used for the surface functionalization of a self-assembled monolayer (SAM) with biomolecules. An alkane thioether molecule with one terminal KAT group (S-KAT) was synthesized and adsorbed onto a gold surface, placing a KAT group on the top of the monolayer (KAT-SAM). As an initial test case, an aqueous solution of a hydroxylamine (HA) derivative of poly(ethylene glycol) (PEG) (HA-PEG) was added to this KAT-SAM at room temperature to perform the surface KAT ligation. Quartz crystal microbalance with dissipation (QCM-D) monitoring confirmed the rapid attachment of the PEG moiety onto the SAM. By surface characterization methods such as contact angle and ellipsometry, the attachment of PEG layer was confirmed, and covalent amide-bond formation was established by X-ray photoelectron spectroscopy (XPS). In a proof-of-concept study, the applicability of this surface KAT ligation for the attachment of biomolecules to surfaces was tested using a model protein, green fluorescent protein (GFP). A GFP was chemically modified with an HA linker to synthesize HA-GFP and added to the KAT-SAM under aqueous dilute conditions. A rapid attachment of the GFP on the surface was observed in real time by QCM-D. Despite the fact that such biomolecules have a variety of unprotected functional groups within their structures, the surface KAT ligation proceeded rapidly in a chemoselective manner. Our results demonstrate the versatility of the KAT ligation for the covalent attachment of a variety of water-soluble molecules onto SAM surfaces under dilute and biocompatible conditions to form stable, natural amide bonds

Animal models of chemotherapy-induced peripheral neuropathy: A machine-assisted systematic review and meta-analysis

We report a systematic review and meta-analysis of research using animal models of chemotherapy-induced peripheral neuropathy (CIPN). We systematically searched 5 online databases in September 2012 and updated the search in November 2015 using machine learning and text mining to reduce the screening for inclusion workload and improve accuracy. For each comparison, we calculated a standardised mean difference (SMD) effect size, and then combined effects in a random-effects meta-analysis. We assessed the impact of study design factors and reporting of measures to reduce risks of bias. We present power analyses for the most frequently reported behavioural tests; 337 publications were included. Most studies (84%) used male animals only. The most frequently reported outcome measure was evoked limb withdrawal in response to mechanical monofilaments. There was modest reporting of measures to reduce risks of bias. The number of animals required to obtain 80% power with a significance level of 0.05 varied substantially across behavioural tests. In this comprehensive summary of the use of animal models of CIPN, we have identified areas in which the value of preclinical CIPN studies might be increased. Using both sexes of animals in the modelling of CIPN, ensuring that outcome measures align with those most relevant in the clinic, and the animal's pain contextualised ethology will likely improve external validity. Measures to reduce risk of bias should be employed to increase the internal validity of studies. Different outcome measures have different statistical power, and this can refine our approaches in the modelling of CIPN

Silorane-based composite: Depth of cure, surface hardness, degree of conversion, and cervical microleakage in class II cavities

PubMedID: 21977956Objective: The purpose of this study was to determine the depth of cure, degree of conversion (DC), hardness, and cervical sealing ability of silorane-based composite (Filtek Silorane [FS; 3M, Seefeld, Germany]) and to compare with methacrylate-based composites (MBCs = Filtek Supreme XT [FSXT] and Filtek P60 [FP60]). Materials and Methods: The DC and hardness of every material were evaluated after 1, 7, and 30 days. The depth of cure was determined using the ISO 4049:2000 standard. Microleakage was evaluated by measuring dye penetration across the gingival wall in cross-sectioned specimens. Results: FS showed lower depth of cure than FSXT and FP60. The DC of FS was significantly lower when compared to FP60 and FSXT. FS exhibited lower hardness than both FSXT and FP60 after 1 day of storage. The hardness of FS remained unchanged during the storage period. FS showed reduced microleakage scores compared to FSXT and showed similar microleakage scores compared to FP60. Conclusions: In conclusion, the DC and cure depth of FS are lower than those of MBCs. However, FS revealed stable hardness in water that is comparable to MBCs. The sealing ability of FS is similar or even better than that of MBCs. © 2011 Wiley Periodicals, Inc

Silorane-based composite: Depth of cure, surface hardness, degree of conversion, and cervical microleakage in class II cavities

PubMed: 21977956Objective: The purpose of this study was to determine the depth of cure, degree of conversion (DC), hardness, and cervical sealing ability of silorane-based composite (Filtek Silorane [FS; 3M, Seefeld, Germany]) and to compare with methacrylate-based composites (MBCs = Filtek Supreme XT [FSXT] and Filtek P60 [FP60]). Materials and Methods: The DC and hardness of every material were evaluated after 1, 7, and 30 days. The depth of cure was determined using the ISO 4049:2000 standard. Microleakage was evaluated by measuring dye penetration across the gingival wall in cross-sectioned specimens. Results: FS showed lower depth of cure than FSXT and FP60. The DC of FS was significantly lower when compared to FP60 and FSXT. FS exhibited lower hardness than both FSXT and FP60 after 1 day of storage. The hardness of FS remained unchanged during the storage period. FS showed reduced microleakage scores compared to FSXT and showed similar microleakage scores compared to FP60. Conclusions: In conclusion, the DC and cure depth of FS are lower than those of MBCs. However, FS revealed stable hardness in water that is comparable to MBCs. The sealing ability of FS is similar or even better than that of MBCs. © 2011 Wiley Periodicals, Inc

Validation of the Medical Research Council and a newly developed prognostic index in patients with malignant glioma: How useful are prognostic indices in routine clinical practice?

Although different prognostic indices for malignant gliomas have been developed, their validity outside of clinical trials has not been widely tested. The aim of this study was to determine whether the Medical Research Council (MRC) brain tumour prognostic index was able to stratify patients for survival managed in routine practice, and secondly to compare the results with our newly developed prognostic score which included tumour grade and only 3 prognostic groups. The MRC and the new prognostic index were calculated for a group of 119 adult patients with malignant glioma managed by surgical resection/biopsy and post-operative radiotherapy. For the MRC and new score, 6 and 3 prognostic groups were defined, respectively. For all patients median survival was 11 (2-66) months. The overall survival rate at 12 and 24 months were 43% and 18%, respectively. The MRC median and two-year survival rates were 14 months and 26% for a score of 1-10, 14 months and 27% for a score of 11-15, 13 months and 22% for a score of 16-20, 8 months and 10% for a score of 21-25, 8 months and 0% for those scoring 26-33. There was only one patient in the 34-38 group. For the new prognostic index, median and two-year survival rates were respectively 16 and 26%; 12 and 23%; 8 and 7% for the good, intermediate and poor prognostic groups. Both indices were significant factors for survival in univariate analysis (MRC index, p = 0.0089, new index p = 0.0002), but not in multivariate analysis. Both the MRC and our newly devised prognostic score were able to separate patients into good and poor prognostic groups, which may aid in treatment decisions, although there was less differentiation between the MRC groups especially over the first year. Both scores use routinely available factors. However, inclusion of tumour grade in the new score may be an advantage over the MRC index