Novel Electrospun Polymeric Nanofibers Loaded Different Medicaments as Drug Delivery Systems for Regenerative Endodontics.

BACKGROUND: A combination of antibiotics, including metronidazole (MET), ciprofloxacin (CIP), and minocycline (MINO), has been demonstrated to disinfect bacteria in necrotic teeth before regenerative processes. It has been presented clinically that antibiotic pastes may drive to possible stem cell death, creating difficulties in removing from the canal system, which can limit the regenerative procedure. This study was designed to (1) synthesize nanofibrous webs containing various concentrations of different medicaments (triple, double, and calcium hydroxide, Ca(OH)2), and (2) coat the electrospun fibrous gutta-percha (GP) cones. METHODS: Poly(vinylpyrrolidone) (PVP)-based electrospun fibrous webs were processed with low medicament concentrations. Scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) were carried out to investigate fiber morphology and antibiotic incorporation, and characterize GP-coated fibrous webs, respectively. The chemical and physical properties of dentine were determined via fourier transform infrared spectroscopy (FTIR) and Nano-SEM, respectively. The antimicrobial properties of the different fibrous webs were assessed against various bacteria by direct nanofiber/bacteria contact. Cytocompatibility was measured by applying the MTT method. RESULTS: The mean fiber diameter of the experimental groups of medicament-containing fibers ranged in the nm scale and was significantly smaller than PVP fibers. EDX analysis confirmed the presence of medicaments in the nanofibers. XPS analysis presented a complete coating of the fibers with GPs; FTIR and Nano-SEM showed no chemical and physical configuration of intracanal medicaments on the dentine surface. Meanwhile, nanofibrous webs led to a significant reduction in the percentage of viable bacteria compared to the negative control and PVP. CONCLUSION: Our findings suggest that TA-NFs, DA-NFs, and Ca(OH)2)-NFs coated GP cones have significant potential in eliminating intracanal bacteria, having cell-friendly behavior and clinical usage features.

Functional analysis of structural variants in single cells using Strand-seq.

Somatic structural variants (SVs) are widespread in cancer, but their impact on disease evolution is understudied due to a lack of methods to directly characterize their functional consequences. We present a computational method, scNOVA, which uses Strand-seq to perform haplotype-aware integration of SV discovery and molecular phenotyping in single cells by using nucleosome occupancy to infer gene expression as a readout. Application to leukemias and cell lines identifies local effects of copy-balanced rearrangements on gene deregulation, and consequences of SVs on aberrant signaling pathways in subclones. We discovered distinct SV subclones with dysregulated Wnt signaling in a chronic lymphocytic leukemia patient. We further uncovered the consequences of subclonal chromothripsis in T cell acute lymphoblastic leukemia, which revealed c-Myb activation, enrichment of a primitive cell state and informed successful targeting of the subclone in cell culture, using a Notch inhibitor. By directly linking SVs to their functional effects, scNOVA enables systematic single-cell multiomic studies of structural variation in heterogeneous cell populations.

COVID-19 pneumonia in patients with impaired fasting glucose, newly diagnosed diabetes and pre-existing diabetes: a tertiary center experience.

COVID-19 infection is known to increase mortality in patients with diabetes. We aim to demonstrate the differences in disease course and clinical outcomes of patients with COVID-19 regarding the presence of impaired fasting glucose, pre-existing diabetes mellitus (DM) or new-onset DM. 236 patients with positive reverse transcription-PCR tests for SARS-CoV-2 were included in this single-center, retrospective observational study between March 2020 and May 2021. Laboratory results, comorbidities, medications and imaging findings were noted. Logistic regression was used to estimate associated factors for admission to the intensive care unit (ICU). 43 patients with normal glucose, 53 with impaired fasting glucose, 60 with newly diagnosed DM, and 80 with pre-existing DM were classified. Patients with pre-existing DM had higher fasting glucose and glycated hemoglobin than the other groups (p<0.001 for all). Patients with newly diagnosed DM were more likely to need dexamethasone 6 mg (p=0.001). In both newly diagnosed diabetes and impaired fasting glucose groups, 250 mg methylprednisolone was needed at higher rates (p=0.002). Newly diagnosed DM had higher rates of intubation (21.6%) and more mortality (20.0%) (p=0.045 and p=0.028, respectively). Mortality and hospitalization in the ICU were lower in the group receiving antidiabetic treatment. The risk of ICU attendance was higher in patients with impaired fasting glucose (HR=1.71, 95% CI: 0.48 to 6.08) and newly diagnosed DM (HR=1.88, 95% CI: 0.57 to 6.17), compared with pre-existing DM and non-diabetics. Newly diagnosed DM and impaired fasting glucose are associated with increased mortality and intubation in inpatients with COVID-19.

Pediatric T-ALL type-1 and type-2 relapses develop along distinct pathways of clonal evolution.

The mechanisms underlying T-ALL relapse remain essentially unknown. Multilevel-omics in 38 matched pairs of initial and relapsed T-ALL revealed 18 (47%) type-1 (defined by being derived from the major ancestral clone) and 20 (53%) type-2 relapses (derived from a minor ancestral clone). In both types of relapse, we observed known and novel drivers of multidrug resistance including MDR1 and MVP, NT5C2 and JAK-STAT activators. Patients with type-1 relapses were specifically characterized by IL7R upregulation. In remarkable contrast, type-2 relapses demonstrated (1) enrichment of constitutional cancer predisposition gene mutations, (2) divergent genetic and epigenetic remodeling, and (3) enrichment of somatic hypermutator phenotypes, related to BLM, BUB1B/PMS2 and TP53 mutations. T-ALLs that later progressed to type-2 relapses exhibited a complex subclonal architecture, unexpectedly, already at the time of initial diagnosis. Deconvolution analysis of ATAC-Seq profiles showed that T-ALLs later developing into type-1 relapses resembled a predominant immature thymic T-cell population, whereas T-ALLs developing into type-2 relapses resembled a mixture of normal T-cell precursors. In sum, our analyses revealed fundamentally different mechanisms driving either type-1 or type-2 T-ALL relapse and indicate that differential capacities of disease evolution are already inherent to the molecular setup of the initial leukemia.

Comparing Antibiotic Pastes with Electrospun Nanofibers as Modern Drug Delivery Systems for Regenerative Endodontics.

Nanomaterials can be applied in different biomedical applications like diagnosis, treatment, and drug delivery due to their unique features. Using such materials in the endodontic treatment processes may prove challenging as these materials must exhibit antibacterial effects without posing any harm to the host cells. The approach involving nanofibers loaded with various antibacterial drugs offers a potential treatment method to enhance the elimination procedure of intracanal biofilms. Clinically, many models of bacterial biofilms were prepared under in vitro conditions for different aims. The process of drug delivery from polymeric nanofibers is based on the principle that the releasing ratio of drug molecules increases due to the increase in the surface area of the hosted structure. Our review discusses diverse approaches to loading/releasing drugs on/from nanofibers; we summarized many studies on electrospun nanofibers loaded with various drugs applied in the endodontic field. Moreover, we discussed both the advantages and the limitations of these modern endodontic treatment materials, comparing them with the traditional ones.

Aging of preleukemic thymocytes drives CpG island hypermethylation in T-cell acute lymphoblastic leukemia.

Cancer cells display DNA hypermethylation at specific CpG islands in comparison to their normal healthy counterparts, but the mechanism that drives this so-called CpG island methylator phenotype (CIMP) remains poorly understood. Here, we show that CpG island methylation in human T-cell acute lymphoblastic leukemia (T-ALL) mainly occurs at promoters of Polycomb Repressor Complex 2 (PRC2) target genes that are not expressed in normal or malignant T-cells and which display a reciprocal association with H3K27me3 binding. In addition, we revealed that this aberrant methylation profile reflects the epigenetic history of T-ALL and is established already in pre-leukemic, self-renewing thymocytes that precede T-ALL development. Finally, we unexpectedly uncover that this age-related CpG island hypermethylation signature in T-ALL is completely resistant to the FDA-approved hypomethylating agent Decitabine. Altogether, we here provide conceptual evidence for the involvement of a pre-leukemic phase characterized by self-renewing thymocytes in the pathogenesis of human T-ALL.

Chromatin accessibility landscape of pediatric T-lymphoblastic leukemia and human T-cell precursors.

We aimed at identifying the developmental stage at which leukemic cells of pediatric T-ALLs are arrested and at defining leukemogenic mechanisms based on ATAC-Seq. Chromatin accessibility maps of seven developmental stages of human healthy T cells revealed progressive chromatin condensation during T-cell maturation. Developmental stages were distinguished by 2,823 signature chromatin regions with 95% accuracy. Open chromatin surrounding SAE1 was identified to best distinguish thymic developmental stages suggesting a potential role of SUMOylation in T-cell development. Deconvolution using signature regions revealed that T-ALLs, including those with mature immunophenotypes, resemble the most immature populations, which was confirmed by TF-binding motif profiles. We integrated ATAC-Seq and RNA-Seq and found DAB1, a gene not related to leukemia previously, to be overexpressed, abnormally spliced and hyper-accessible in T-ALLs. DAB1-negative patients formed a distinct subgroup with particularly immature chromatin profiles and hyper-accessible binding sites for SPI1 (PU.1), a TF crucial for normal T-cell maturation. In conclusion, our analyses of chromatin accessibility and TF-binding motifs showed that pediatric T-ALL cells are most similar to immature thymic precursors, indicating an early developmental arrest.

Microleakage of high viscosity glass-ionomer and glass-carbomer with and without coating before and after hydrothermal aging / Microinfiltracao de ionomero de vidro de alta viscosidade e carbômero de vidro com e sem revestimento antes e depois de envelhecimento hidroterma

Objective: The objective of this study was to evaluate the microleakage patterns of GIC and GGC with and without their protective surface coatings on enamel and dentin margins before and after aging. Material and Methods: Two rectangular cavities (height: 2 mm; width: 3 mm; depth: 1.5 mm) were prepared on each tooth at the cemento-enamel junction were prepared on human permanent molars (N=56) and the teeth were randomly assigned to be restored with one of the following: a) high viscosity glass-ionomer cement (GIC) (EQUIA Fil, C Corp., Tokyo, Japan) (n=28), b) glasscarbomer cement (GCC) (Glass Carbomer Products, Leiden, The Netherlands) (n=28). Half of the teeth were further divided into two groups where one group received protective surface coating (SC) (G-Coat Plus, GC Corp) (n=14) and the other group did not (n=14). Half of the teeth were stored for 24 hours (n=7), and the other half was thermocycled (5000 cycles, 5-55°C) (n=7). For microleakage analysis, the teeth were immersed in 5% methylene blue dye for 24 hours, sectioned into two equal halves. Microleakage patterns were evaluated using stereomicroscope and scored on a scale of 0-3 (0: No dye penetration, 1: Dye penetration less than half of the axial wall, 2: Dye penetration more than half the axial wall, 3: Dye penetration spreading along the axial wall). Data were analyzed using Kruskal-Wallis tests at the significance level of 0.05. Results: Compared to 24 h storage, after thermocycling, surface coating on GIC decreased microleakage significantly compared to GCC (p=0.046) but not for GCC. In the thermocycled groups, coated GIC showed significantly less leakage at the enamel margin but no significant difference was found with both GIC and GCC in the dentin margins. Conclusion: The application of surface coating significantly reduced the microleakage scores of GIC but not GCC, within the enamel margins only. (AU)

Objetivo: O objetivo deste estudo foi avaliar os padrões de microinfiltração de GIC e GGC com e sem seus revestimentos protetores superficiais nas margens de esmalte e dentina antes e após o envelhecimento. Material e Métodos: duas cavidades retangulares (altura: 2 mm; largura: 3 mm; profundidade: 1,5 mm) foram preparadas em cada dente na junção cemento-esmalte de molares permanentes humanos (N = 56), sendo aleatoriamente designados para serem restaurados com um dos seguintes: a) cimento de ionômero de vidro (GIC) de alta viscosidade (EQUIA Fil, C Corp., Tóquio, Japão) (n = 28), b) cimento de vidrocarbômero (GCC) Carbomer Products, Leiden, Holanda) (n = 28). Metade dos dentes foram divididos em dois grupos, onde um grupo recebeu revestimento protetor de superfície (SC) (G-Coat Plus, GC Corp) (n = 14) e o outro grupo não (n = 14). Metade dos dentes foram armazenados por 24 horas (n = 7), e a outra metade foi termociclada (5000 ciclos, 5-55 ° C) (n = 7). Para análise de microinfiltração, os dentes foram imersos em corante azul de metileno a 5% por 24 horas, seccionados em duas metades iguais. Os padrões de microinfiltração foram avaliados usando estereomicroscópio e pontuados numa escala de 0-3 (0: Sem penetração de corante; 1: penetração de corante inferior à metade da parede axial; 2: penetração de corante mais do que metade da parede axial; 3: penetração de corante ao longo da parede axial). Os dados foram analisados pelo teste de Kruskal-Wallis ao nível de significância de 0,05. Resultados: em comparação com o armazenamento de 24 h, após a termociclagem, o revestimento de superfície no GIC diminuiu significativamente a microinfiltração em comparação com o GCC (p = 0,046), mas não para o GCC. Nos grupos termociclados, o GIC revestido apresentou significativamente menos infiltração na margem do esmalte, mas não houve diferença significativa para o GIC e o GCC nas margens dentinárias. Conclusão: A aplicação do revestimento de superfície reduziu significativamente os escores de microinfiltração do GIC, mas não do GCC, apenas nas margens do esmalte. (AU)

PDX models recapitulate the genetic and epigenetic landscape of pediatric T-cell leukemia.

We compared 24 primary pediatric T-cell acute lymphoblastic leukemias (T-ALL) collected at the time of initial diagnosis and relapse from 12 patients and 24 matched patient-derived xenografts (PDXs). DNA methylation profile was preserved in PDX mice in 97.5% of the promoters (ρ = 0.99). Similarly, the genome-wide chromatin accessibility (ATAC-Seq) was preserved remarkably well (ρ = 0.96). Interestingly, both the ATAC regions, which showed a significant decrease in accessibility in PDXs and the regions hypermethylated in PDXs, were associated with immune response, which might reflect the immune deficiency of the mice and potentially the incomplete interaction between murine cytokines and human receptors. The longitudinal approach of this study allowed an observation that samples collected from patients who developed a type 1 relapse (clonal mutations maintained at relapse) preserved their genomic composition; whereas in patients who developed a type 2 relapse (subset of clonal mutations lost at relapse), the preservation of the leukemia's composition was more variable. In sum, this study underlines the remarkable genomic stability, and for the first time documents the preservation of the epigenomic landscape in T-ALL-derived PDX models.