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Journal of Immunology Research 2022The leading cause of mortality in patients with Marfan syndrome (MFS) is thoracic aortic aneurysm and dissection. Notch signaling is essential for vessel morphogenesis...
BACKGROUND
The leading cause of mortality in patients with Marfan syndrome (MFS) is thoracic aortic aneurysm and dissection. Notch signaling is essential for vessel morphogenesis and function. However, the role of Notch signaling in aortic pathology and aortic smooth muscle cell (SMC) differentiation in Marfan syndrome (MFS) is not completely understood.
METHODS
RNA-sequencing on ascending aortic tissue from a mouse model of MFS, , and wild-type controls was performed. Notch 3 expression and activation in aortic tissue were confirmed with real-time RT-PCR, immunohistochemistry, and Western blot. and wild-type mice were treated with a -secretase inhibitor, DAPT, to block Notch activation. Aortic aneurysms and rupture were evaluated with connective tissue staining, ultrasound, and life table analysis.
RESULTS
The murine RNA-sequencing data were validated with mouse and human MFS aortic tissue, demonstrating elevated Notch3 activation in MFS. Data further revealed that upregulation and activation of Notch3 were concomitant with increased expression of SMC contractile markers. Inhibiting Notch3 activation with DAPT attenuated aortic enlargement and improved survival of mice. DAPT treatment reduced elastin fiber fragmentation in the aorta and reversed the differentiation of SMCs.
CONCLUSIONS
Our data demonstrated that matrix abnormalities in the aorta of MFS are associated with increased Notch3 activation. Enhanced Notch3 activation in MFS contributed to aortic aneurysm formation in MFS. This might be mediated by inducing a contractile phenotypic change of SMC. Our results suggest that inhibiting Notch3 activation may provide a strategy to prevent and treat aortic aneurysms in MFS.
Topics: Animals; Aorta; Aortic Aneurysm; Diamines; Disease Models, Animal; Elastin; Fibrillin-1; Humans; Marfan Syndrome; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Molecular Targeted Therapy; Myocytes, Smooth Muscle; Receptor, Notch3; Thiazoles
PubMed: 35211631
DOI: 10.1155/2022/7538649 -
Advances in Clinical and Experimental... Feb 2022The sealer penetration into the dentinal tubules might be beneficial, especially in necrotic endodontic cases, as it provides the obstruction of the contaminated tubules.
BACKGROUND
The sealer penetration into the dentinal tubules might be beneficial, especially in necrotic endodontic cases, as it provides the obstruction of the contaminated tubules.
OBJECTIVES
To determine the effect of 3 final irrigants (sodium hypochlorite (NaOCl), alcohol and chlorhexidine (CHX)) on the penetration of an epoxy sealer into the dentinal tubules.
MATERIAL AND METHODS
The study was carried out on 60 single-canal human teeth with straight roots. The root canals were prepared to the ISO 40/04 size, using the Reciproc® instruments. The teeth were divided into 4 groups (n = 15). The canals in each group were irrigated according to the following scheme: group 1 (control) - 5.25% NaOCl; group 2 - smear layer removal (40% citric acid (CA) and 5.25% NaOCl) and 5.25% NaOCl; group 3 - smear layer removal (as in group 2), and 40% CA, water and 98% isopropyl alcohol; and group 4 - smear layer removal (as in group 2), and 40% CA, water and 2% CHX. The root canals were filled using the vertical condensation technique with gutta-percha and the porphyrin-labeled AH Plus™ sealer. After 3 days, 1-milimeter-thick cross-section slices were cut from the roots at a distance of 2 mm, 5 mm and 8 mm from the apex. The sections were imaged under a confocal microscope and the sealant penetration depth into the dentinal tubules was measured.
RESULTS
The longest resin tags in all parts of the roots were found in group 4 (CHX), and the shortest in group 1 (control). The mean depth of the sealer penetration (in micrometers) was as follows: 21, 22 and 23 (group 1); 201, 231 and 374 (group 2); 170, 232 and 280 (group 3); and 330, 408 and 638 (group 4) in the apical, middle and coronal parts, respectively.
CONCLUSIONS
The final irrigation with CHX resulted in the deepest penetration of the epoxy sealer into the tubules. Isopropyl alcohol had the most negative impact on the sealer penetration into the tubules.
Topics: 2-Propanol; Chlorhexidine; Edetic Acid; Humans; Microscopy, Confocal; Root Canal Filling Materials; Root Canal Irrigants; Root Canal Preparation; Sodium Hypochlorite
PubMed: 35212490
DOI: 10.17219/acem/142991 -
Plant Physiology Mar 2022Growth promotion induced by the endosymbiont Piriformospora indica has been observed in various plants; however, except growth phytohormones, specific functional...
Growth promotion induced by the endosymbiont Piriformospora indica has been observed in various plants; however, except growth phytohormones, specific functional metabolites involved in P. indica-mediated growth promotion are unknown. Here, we used a gas chromatography-mass spectrometry-based untargeted metabolite analysis to identify tomato (Solanum lycopersicum) metabolites whose levels were altered during P. indica-mediated growth promotion. Metabolomic multivariate analysis revealed several primary metabolites with altered levels, with putrescine (Put) induced most significantly in roots during the interaction. Further, our results indicated that P. indica modulates the arginine decarboxylase (ADC)-mediated Put biosynthesis pathway via induction of SlADC1 in tomato. Piriformospora indica did not promote growth in Sladc1-(virus-induced gene silencing of SlADC1) lines of tomato and showed less colonization. Furthermore, using LC-MS/MS we showed that Put promoted growth by elevation of auxin (indole-3-acetic acid) and gibberellin (GA4 and GA7) levels in tomato. In Arabidopsis (Arabidopsis thaliana) adc knockout mutants, P. indica colonization also decreased and showed no plant growth promotion, and this response was rescued upon exogenous application of Put. Put is also important for hyphal growth of P. indica, indicating that it is co-adapted by both host and microbe. Taken together, we conclude that Put is an essential metabolite and its biosynthesis in plants is crucial for P. indica-mediated plant growth promotion and fungal growth.
Topics: Basidiomycota; Chromatography, Liquid; Gene Expression Regulation, Plant; Plant Roots; Putrescine; Tandem Mass Spectrometry
PubMed: 34791442
DOI: 10.1093/plphys/kiab536 -
International Endodontic Journal Aug 2020To evaluate the antimicrobial, toxicity and cleaning effectiveness of ethylenediaminetetraacetic acid (EDTA) and maleic acid (MA) alone and combined with cetrimide (CTR).
AIM
To evaluate the antimicrobial, toxicity and cleaning effectiveness of ethylenediaminetetraacetic acid (EDTA) and maleic acid (MA) alone and combined with cetrimide (CTR).
METHODOLOGY
Cytotoxic and genotoxic effects were assessed on Chinese hamster cells V79 using the MTT, clonogenic and micronucleus assays, respectively. The bacterial inhibitory and bactericidal concentrations (MIC and MBC, respectively) were determined on a strain of Enterococcus faecalis. Antimicrobial tests were performed on a biofilm model after treatment with the chelating agents by using a biofilm eradication concentration (MBEC) and confocal laser scanning microscope (CLSM) assays. Quantification of cell biomass and percentage of live and dead cells in the biomass were assessed for each group. The percentage reduction of accumulated hard-tissue debris (AHTD) after root canal preparation and final irrigation protocols was evaluated by micro-CT. Statistical tests of one-way analysis of variance (anova), Bonferroni test, Kruskal-Wallis test, Dunn's multiple comparison test and Wilcoxon matched-pairs signed-rank tests were used.
RESULTS
Cetrimide alone as well as in combination with EDTA and MA at dilutions of 1/10 and 1/100 was significantly more toxic as compared to untreated controls (P < 0.001). All tested mixtures were nontoxic at a dilution of 1/1000. EDTA retained a weak inhibitory and bactericidal effect against planktonic cells, whilst MA inhibited cells growth and killed 99.9% of the cells when diluted. CTR revealed the most prominent effect, being inhibitory and bactericidal, also when diluted. Cetrimide alone or combined with EDTA was able to remove, respectively, 40% (P < 0.01) and 60% (P < 0.001) of the entire biomass after 1 min. Conversely, MA alone and in combination with CTR did not have a significant effect on biomass reduction. After final irrigation, the AHTD volume was significantly decreased in all groups (P < 0.05). EDTA + CTR and MA + CTR were associated with a significant reduction in the percentage of AHTD on the entire root canal compared to the same solutions without surfactant.
CONCLUSIONS
7% MA was less cytotoxic in comparison with 17% EDTA. The addition of cetrimide to EDTA and MA removed accumulated hard-tissue debris effectively from the canal walls and increased their antimicrobial activity when compared to the same solutions without detergents.
Topics: Animals; Anti-Infective Agents; Chelating Agents; Cricetinae; Dental Pulp Cavity; Edetic Acid; Root Canal Irrigants; Root Canal Preparation; Sodium Hypochlorite
PubMed: 32344451
DOI: 10.1111/iej.13314 -
Scientific Reports Jan 2021Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 pandemic outbreak is a betacoronavirus named SARS-CoV-2. For public health interventions, a...
Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 pandemic outbreak is a betacoronavirus named SARS-CoV-2. For public health interventions, a diagnostic test with high sensitivity and specificity is required. The gold standard protocol for diagnosis by the Word Health Organization (WHO) is RT-PCR. To detect low viral loads and perform large-scale screening, a low-cost diagnostic test is necessary. Here, we developed a cost-effective test capable of detecting SARS-CoV-2. We validated an auxiliary protocol for molecular diagnosis with the SYBR Green RT-PCR methodology to successfully screen negative cases of SARS-CoV-2. Our results revealed a set of primers with high specificity and no homology with other viruses from the Coronovideae family or human respiratory tract pathogenic viruses, presenting with complementarity only for rhinoviruses/enteroviruses and Legionella spp. Optimization of the annealing temperature and polymerization time led to a high specificity in the PCR products. We have developed a more affordable and swift methodology for negative SARS-CoV-2 screening. This methodology can be applied on a large scale to soften panic and economic burden through guidance for isolation strategies.
Topics: Benzothiazoles; COVID-19; COVID-19 Nucleic Acid Testing; DNA, Single-Stranded; Diamines; Enterovirus; Genome, Viral; Humans; Organic Chemicals; Phylogeny; Polymerase Chain Reaction; Quinolines; Real-Time Polymerase Chain Reaction; Rhinovirus; SARS-CoV-2; Sensitivity and Specificity; Temperature; Viral Load
PubMed: 33500453
DOI: 10.1038/s41598-021-81245-0 -
Microbial Cell Factories Nov 2022Polyamide (nylon) is an important material, which has aroused plenty of attention from all aspects. PA 5.4 is one kind of nylon with excellent property, which consists...
BACKGROUND
Polyamide (nylon) is an important material, which has aroused plenty of attention from all aspects. PA 5.4 is one kind of nylon with excellent property, which consists of cadaverine and succinic acid. Due to the environmental pollution, bio-production of cadaverine and succinic acid has been more attractive due to the less pollution and environmental friendliness. Microbes, like Escherichia coli, has been employed as cell factory to produce cadaverine and succinic acid. However, the accumulation of cadaverine will cause severe damage on cells resulting in inhibition on cell growth and cadaverine production. Herein, a novel two stage co-production of succinic acid and cadaverine was designed based on an efficient thermos-regulated switch to avoid the inhibitory brought by cadaverine.
RESULTS
The fermentation process was divided into two phase, one for cell growth and lysine production and the other for cadaverine and succinic acid synthesis. The genes of ldhA and ackA were deleted to construct succinic acid pathway in cadaverine producer strain. Then, a thermal switch system based on pR/pL promoter and CI857 was established and optimized. The fermentation conditions were investigated that the optimal temperature for the first stage was determined as 33 ℃ and the optimal temperature for the second stage was 39 ℃. Additionally, the time to shifting temperature was identified as the fermentation anaphase. For further enhance cadaverine and succinic acid production, a scale-up fermentation in 5 L bioreactor was operated. As a result, the titer, yield and productivity of cadaverine was 55.58 g/L, 0.38 g/g glucose and 1.74 g/(L·h), respectively. 28.39 g/L of succinic acid was also obtained with yield of 0.19 g/g glucose.
CONCLUSION
The succinic acid metabolic pathway was constructed into cadaverine producer strain to realize the co-production of succinic acid and cadaverine. This study provided a novel craft for industrial co-production of cadaverine and succinic acid.
Topics: Cadaverine; Succinic Acid; Escherichia coli; Nylons; Glucose
PubMed: 36419122
DOI: 10.1186/s12934-022-01965-4 -
Analytical Chemistry Dec 2023Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a...
Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a quantitative method of hydroxyl radical generation via Fe(II)-ethylenediaminetetraacetic acid (EDTA)-catalyzed Fenton chemistry that provides ready access to protein oxidative footprinting using equipment commonly found in research and process control laboratories. Robust and reproducible dose-dependent oxidation of protein samples is observed and quantitated by mass spectrometry with as fine a single residue resolution. An oxidation analysis of lysozyme provides a readily accessible benchmark for our method. The efficacy of our oxidation method is demonstrated by mapping the interface of a RAS-monobody complex, the surface of the NIST mAb, and the interface between PRC2 complex components. These studies are executed using standard laboratory tools and a few pennies of reagents; the mass spectrometry analysis can be streamlined to map the protein structure with single amino acid residue resolution.
Topics: Edetic Acid; Hydroxyl Radical; Proteins; Protein Footprinting; Oxidative Stress; Oxidation-Reduction
PubMed: 38049117
DOI: 10.1021/acs.analchem.3c02319 -
BMC Oral Health Jun 2024Surface tension and contact angle properties, which play a crucial role in determining the effectiveness of irrigation solutions in penetrating dentin surfaces and... (Comparative Study)
Comparative Study
BACKGROUND
Surface tension and contact angle properties, which play a crucial role in determining the effectiveness of irrigation solutions in penetrating dentin surfaces and dentin tubules, are highly important for the development of new irrigation solutions and their preferences. The aim of the current study was to compare the surface tension and contact angle properties of different irrigation solutions used in endodontics, both on the dentin surface and within dentin tubules.
METHODS
In this study, the contact angles and surface tensions of 5.25% sodium hypochlorite (NaOCl), 17% ethylenediaminetetraacetic acid (EDTA), 2% chlorhexidine (CHX), 5% boric acid (BA), 0.02% hypochlorous acid (HOCl), 0.2% chlorine dioxide (ClO), Biopure MTAD, QMix solutions, and distilled water (control group) were measured. Measurements were conducted using a goniometer device (Attension Theta Lite Tensiometer, Biolin Scientific, USA), employing the sessile drop method for contact angle measurements on pre-prepared dentin surfaces, and the pendant drop method for surface tension.
RESULTS
Contact angle measurements revealed no statistically significant differences between the contact angle values of MTAD, ClO, and CHX or between NaOCl, QMix, BA, and HOCl (p > 0.05). However, EDTA exhibited a significantly greater contact angle than did MTAD, ClO, CHX, NaOCl, QMix, BA, and HOCl (p < 0.05). Furthermore, the contact angle of dentin with distilled water was greater than that with all other solutions tested (p < 0.05). Surface tension measurements revealed that the surface tension values of QMix and MTAD were statistically similar (p > 0.05). CHX exhibited lower surface tension than distilled water and HOCl (p < 0.05), and it also had lower surface tension than ClO, NaOCl, and BA (p < 0.05). Additionally, the surface tension of the samples treated with EDTA was greater than that of all other solutions tested (p < 0.05).
CONCLUSION
The direct linear relationship between the surface tension of liquids and contact angles on different surfaces may not always hold true, and these values should be considered independently for each solution on various surfaces. Considering the contact angles and surface tension properties of irrigation solutions with root canal dentin, it can be suggested for clinical use that ClO could be recommended over NaOCl, and similarly, BA could be recommended over EDTA.
Topics: Root Canal Irrigants; Dentin; Surface Tension; Humans; Sodium Hypochlorite; Edetic Acid; Hypochlorous Acid; Surface Properties
PubMed: 38867263
DOI: 10.1186/s12903-024-04453-w -
Journal of Feline Medicine and Surgery Feb 2021The aim of this study was to assess the reliability of an EDTA-based method for measuring cat blood bioamines.
OBJECTIVES
The aim of this study was to assess the reliability of an EDTA-based method for measuring cat blood bioamines.
METHODS
Test 1 involved collecting blood samples from seven university laboratory cats. The samples were transferred to EDTA, heparin and plain tubes to determine concentrations of four bioamines (serotonin, dopamine, epinephrine [adrenaline] and norepinephrine [noradrenaline]). Correlation of measurements performed on EDTA plasma, with those performed on heparinized plasma or serum were assessed by intraclass correlation coefficients (ICCs). In test 2, blood samples from five owned cats were collected and stored in EDTA tubes and divided between duplicate Eppendorf tubes labeled as different cats for blinding purposes and analyzed independently for the same four bioamines as in test 1. Reliability of concentration determination for these duplicates was assessed by ICCs and coefficients of variation (CVs).
RESULTS
In test 1, there was no significant correlation between the EDTA plasma serotonin and serum serotonin concentrations. There was also no significant correlation between EDTA plasma and heparin plasma concentrations for either epinephrine or norepinephrine. There was a statistically significant but weak correlation between EDTA plasma and heparin plasma dopamine concentrations. In test 2, there was no correlation for repeat-analyzed serotonin and epinephrine concentrations. Although there were statistically significant correlations for dopamine and norepinephrine, CVs for each analyte were in excess of 30%.
CONCLUSIONS AND RELEVANCE
Before any further attempt is made to measure and report on neurotransmitter concentrations in domestic cats, it is essential that the robustness of the methodology is carefully validated and the data presented.
Topics: Animals; Cats; Edetic Acid; Neurotransmitter Agents; Reproducibility of Results
PubMed: 32478638
DOI: 10.1177/1098612X20924916 -
International Journal of Pharmaceutics Apr 2023Therapeutic liquid mixtures, as deep eutectic systems, are considered a sustainable strategy that can be useful for the modification and enhancement of the...
Therapeutic liquid mixtures, as deep eutectic systems, are considered a sustainable strategy that can be useful for the modification and enhancement of the pharmacokinetics and pharmacodynamics of different active ingredients. In this study, we assessed the stability and antibacterial activity of therapeutic liquid formulations prepared with anti-tuberculosis drugs. Tuberculosis therapy presents various pitfalls related, for example, to the administration of prolonged regimens of multiple drugs, different severe adverse effects, low compliance of the patient to treatment and the development of drug resistance. During this study, it was possible to assess the physicochemical stability of the formulations for 6 months, by polarized optical microscopy, H NMR and FTIR-ATR. Furthermore, the mixtures present an antibacterial effect against a drug-susceptible Mycobacterium tuberculosis strain (H37Rv). This was particularly evident for the mixtures with ethambutol incorporated, making them interesting to pursue with further studies and evaluation of clinical applicability. Upon infection, it was also observed that a single and higher dose appears to be more effective than lower separate doses, which could allow the production of patient-friendly formulations.
Topics: Humans; Tuberculosis; Antitubercular Agents; Mycobacterium tuberculosis; Ethambutol
PubMed: 36965645
DOI: 10.1016/j.ijpharm.2023.122862