Signaling through TLR5 mitigates lethal radiation damage by neutrophil-dependent release of MMP-9

Acute radiation syndrome (ARS) is a major cause of lethality following radiation disasters. A TLR5 agonist, entolimod, is among the most powerful experimental radiation countermeasures and shows efficacy in rodents and non-human primates as a prophylactic (radioprotection) and treatment (radiomitigation) modality. While the prophylactic activity of entolimod has been connected to the suppression of radiation-induced apoptosis, the mechanism by which entolimod functions as a radiomitigator remains poorly understood. Uncovering this mechanism has significant and broad-reaching implications for the clinical development and improvement of TLR5 agonists for use as an effective radiation countermeasure in scenarios of mass casualty resulting from accidental exposure to ionizing radiation. Here, we demonstrate that in contrast to radioprotection, neutrophils are essential for the radiomitigative activity of entolimod in a mouse model of lethal ARS. Neutrophils express functional TLR5 and rapidly exit the bone marrow (BM), accumulate in solid tissues, and release MMP-9 following TLR5 stimulation which is accompanied by an increase in the number of active hematopoietic pluripotent precursors (HPPs) in the BM. Importantly, recombinant MMP-9 by itself has radiomitigative activity and, in the absence of neutrophils, accelerates the recovery of the hematopoietic system. Unveiling this novel TLR5-neutrophil-MMP-9 axis of radiomitigation opens new opportunities for the development of efficacious radiation countermeasures to treat ARS following accidental radiation disasters.

Identification of key proteins in the signaling crossroads between wound healing and cancer hallmark phenotypes

Wound healing (WH) and cancer seem to share common cellular and molecular processes that could work in a tight balance to maintain tissue homeostasis or, when unregulated, drive tumor progression. The “Cancer Hallmarks” comprise crucial biological properties that mediate the advancement of the disease and affect patient prognosis. These hallmarks have been proposed to overlap with essential features of the WH process. However, common hallmarks and proteins actively participating in both processes have yet to be described. In this work we identify 21 WH proteins strongly linked with solid tumors by integrated TCGA Pan-Cancer and multi-omics analyses. These proteins were associated with eight of the ten described cancer hallmarks, especially avoiding immune destruction. These results show that WH and cancer's common proteins are involved in the microenvironment modification of solid tissues and immune system regulation. This set of proteins, between WH and cancer, could represent key targets for developing therapies.

A case of fatal multidrug intoxication involving flualprazolam: distribution in body fluids and solid tissues

Purpose Designer benzodiazepines (DBZDs) increasingly emerged on the novel psychoactive substance (NPS) market in the last few years. They are usually sold as readily available alternatives to prescription benzodiazepines (BZDs) or added to counterfeit medicines. BZDs are generally considered relatively safe drugs due to the low risk of serious acute adverse effects in mono-intoxication, though e.g., alprazolam seems to display an elevated risk of respiratory depression. Here we report on a fatal intoxication involving the novel DBZD flualprazolam. Methods A complete postmortem examination was performed. General unknown screenings and analysis of drugs of abuse were performed on postmortem samples by immunoassay, gas chromatography–mass spectrometry and liquid chromatography–mass spectrometry. The standard addition method was employed to quantify flualprazolam in postmortem blood and tissues. Finally, a toxicological significance score (TSS) was assigned. Results Flualprazolam was detected in heart serum (25.4 ng/mL) and peripheral blood (21.9 ng/mL) as well as in urine, stomach contents, brain, liver and kidney (65.2–323 ng/g). The cause of death was deemed as central nervous system (CNS) and respiratory depression with agonal aspiration of stomach contents, in the setting of a multiple drug intake. Given the concentration levels of the co-consumed CNS depressants, the contribution of flualprazolam to the death was considered likely (TSS of 3). Conclusions Our results support that highly potent DBZDs like flualprazolam carry an elevated risk for unintended toxicity, especially in association with other CNS depressants. A multidisciplinary evaluation of fatalities remains mandatory, especially when pharmacological/toxicological data on intoxicating compounds are lacking. To our knowledge this is the first report of flualprazolam concentrations in solid tissues in human.

Increased expression of Ectodysplasin A2 Receptor EDA2R is the most remarkable and ubiquitous aging-related transcriptional hallmark

We report that increased mRNA-expression of Ectodysplasin-A2-Receptor (EDA2R) is the most remarkable aging-associated transcriptional perturbation in humans, and that its induction is confirmed in murine-models of Hutchinson-Gilford progeria syndrome and in other species. Up-regulation of EDA2R occurs in all solid-tissues and is complemented by the progressive transcriptional increase of its ligand in bloodstream. Strengthening of EDA2R/EDA-A2 signaling may exacerbate inflammation in ageing individuals which might be prevented by EDA2R/EDA-A2 blockade.

PROSPECTS FOR MANUFACTURING FRAMES OF METAL CERAMIC STRUCTURES OF DENTAL PROSTHESES BY THE METHOD OF SELECTIVE LASER SINTERING

The article presents the results of evaluating the prototyping of metal-ceramic structures made by casting and selective laser sintering. To achieve this goal, 27 patients with fabricated metal-ceramic crowns and bridgeworks were examined. Two study groups were created. The first group included 14 patients for whom dental prosthesis frameworks were made by casting. The second group consisted of 13 people, for whom the frames of dentures were made by the method of selective laser sintering. The analysis of clinical effectiveness was carried out according to the following criteria: 1- precision of the felling of prosthesis frame to solid tissues of abutment teeth; 2- the condition of the marginal periodontium; 3- the integrity of the ceramic cladding. The results of the study showed that the precision to solid tooth tissues of metal-ceramic dental prostheses made by laser selective sintering is higher than of frames made by casting. In accordance to the second criteria the best results were also shown by the participants of the second group. No chipping of the ceramic veneer from the metal-ceramic denture frameworks made by laser selective sintering was found. Thus, dentures which frameworks are made by the method of selective laser sintering are characterized by a higher objective assessment of their precision to the solid tissues of the abutment teeth. Lesions of the marginal periodontium both inflammatory and dystrophic were less in the participants of the second group. The frequency of defects in the coating of metal-ceramic dentures was significantly lower in the case of fabrication frameworks by laser selective sintering.

The standard addition method and its validation in forensic toxicology

Purpose In the quantitative forensic toxicological analyses using instruments, major methods to be employed are conventional matrix-matched calibration method (MMCM). However, nowadays, the needs for using the standard addition methods (SAM) are increasing. In spite of this situation, there are no reports of the guidelines for the validations of SAM. In this review, the principle, how to perform it, advantages, disadvantages, reported application data, and the details of validation procedures for the SAM are described. Methods Various databases such as SciFinder, Google and Google Scholar were utilized to collect relevant reports referring to the SAM. The long experiences of our research group on the SAM were also included in this review. Results Although the experimental procedures for the SAM are much more laborious than those of the MMCM, the SAM is essential to quantify target xenobiotic(s) in special matrices such as human solid tissues or biles, which remarkably interfere with the usual quantitative analyses. The validation methods for the SAM have been also proposed for the cases in the absence of the blank matrices. Conclusions To our knowledge, this is the first presentation of detailed SAM procedure and its validation, which will facilitate the use of the SAM in forensic toxicology. Especially for its validation, new simple methods have been proposed.

Vesicle-bound regulatory RNAs are associated with tissue aging

Previous work on murine models and human demonstrated global as well as tissue-specific molecular aging trajectories in solid tissues and body fluids(1-8). Extracellular vesicles like exosomes play a crucial role in communication and information exchange in between such systemic factors and solid tissues(9,10). We sequenced freely circulating and vesicle-bound small regulatory RNAs in mice at five time points across the average life span from 2 to 18 months. Intriguingly, each small RNA class exhibits unique aging patterns, which showed differential signatures between vesicle-bound and freely circulating molecules. In particular, tRNA fragments showed overall highest correlation with aging which also matched well between sample types, facilitating age prediction with non-negative matrix factorization (86% accuracy). Interestingly, rRNAs exhibited inverse correlation trajectories between vesicles and plasma while vesicle-bound microRNAs (miRNAs) were exceptionally strong associated with aging. Affected miRNAs regulate the inflammatory response and transcriptional processes, and adipose tissues show considerable effects in associated gene regulatory modules. Finally, nanoparticle tracking and electron microscopy suggest a shift from overall many small to fewer but larger vesicles in aged plasma, potentially contributing to systemic aging trajectories and affecting the molecular aging of organs.