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Abstract: Deleterious germ line RUNX1 variants cause the autosomal dominant familial platelet disorder with associated myeloid malignancy (FPDMM), characterized by thrombocytopenia, platelet dysfunction, and a predisposition to hematologic malignancies (HMs). We launched a FPDMM natural history study and, from January 2019 to December 2021, enrolled 214 participants, including 111 patients with 39 different RUNX1 variants from 45 unrelated families. Seventy of 77 patients had thrombocytopenia, 18 of 18 had abnormal platelet aggregometry, 16 of 35 had decreased platelet dense granules, and 28 of 55 had abnormal bleeding scores. Nonmalignant bone marrows showed increased numbers of megakaryocytes in 12 of 55 patients, dysmegakaryopoiesis in 42 of 55, and reduced cellularity for age in 30 of 55 adult and 17 of 21 pediatric cases. Of 111 patients, 19 were diagnosed with HMs, including myelodysplastic syndrome, acute myeloid leukemia, chronic myelomonocytic leukemia, acute lymphoblastic leukemia, and smoldering myeloma. Of those 19, 18 were relapsed or refractory to upfront therapy and referred for stem cell transplantation. In addition, 28 of 45 families had at least 1 member with HM. Moreover, 42 of 45 patients had allergic symptoms, and 24 of 30 had gastrointestinal (GI) symptoms. Our results highlight the importance of a multidisciplinary approach, early malignancy detection, and wider awareness of inherited disorders. This actively accruing, longitudinal study will genotype and phenotype more patients with FPDMM, which may lead to a better understanding of the disease pathogenesis and clinical course, which may then inform preventive and therapeutic interventions. This trial was registered at www.clinicaltrials.gov as #NCT03854318.

Germ line loss-of-function heterozygous mutations in the RUNX1 gene cause familial platelet disorder with associated myeloid malignancies (FPDMM) characterized by thrombocytopenia and a life-long risk of hematological malignancies. Although gene therapies are being considered as promising therapeutic options, current preclinical models do not recapitulate the human phenotype and are unable to elucidate the relative fitness of mutation-corrected and RUNX1-heterozygous mutant hematopoietic stem and progenitor cells (HSPCs) in vivo long term. We generated a rhesus macaque with an FPDMM competitive repopulation model using CRISPR/Cas9 nonhomologous end joining editing in the RUNX1 gene and the AAVS1 safe-harbor control locus. We transplanted mixed populations of edited autologous HSPCs and tracked mutated allele frequencies in blood cells. In both animals, RUNX1-edited cells expanded over time compared with AAVS1-edited cells. Platelet counts remained below the normal range in the long term. Bone marrows developed megakaryocytic dysplasia similar to human FPDMM, and CD34+ HSPCs showed impaired in vitro megakaryocytic differentiation, with a striking defect in polyploidization. In conclusion, the lack of a competitive advantage for wildtype or control-edited HSPCs over RUNX1 heterozygous-mutated HSPCs long term in our preclinical model suggests that gene correction approaches for FPDMM will be challenging, particularly to reverse myelodysplastic syndrome/ acute myeloid leukemia predisposition and thrombopoietic defects.

Androgens in women, as well as in men, are intrinsic to maintenance of (i) reproductive competency, (ii) cardiac health, (iii) appropriate bone remodeling and mass retention, (iii) muscle tone and mass, and (iv) brain function, in part, through their mitigation of neurodegenerative disease effects. In recognition of the pluripotency of endogenous androgens, exogenous androgens, and selected congeners, have been prescribed off-label for several decades to treat low libido and sexual dysfunction in menopausal women, as well as, to improve physical performance. However, long-term safety and efficacy of androgen administration has yet to be fully elucidated. Side effects often observed include (i) hirsutism, (ii) acne, (iii) deepening of the voice, and (iv) weight gain but are associated most frequently with supra-physiological doses. By contrast, short-term clinical trials suggest that the use of low-dose testosterone therapy in women appears to be effective, safe and economical. There are, however, few clinical studies, which have focused on effects of androgen therapy on pre- and post-menopausal women; moreover, androgen mechanisms of action have not yet been thoroughly explained in these subjects. This review considers clinical effects of androgens on women's health in order to prevent chronic diseases and reduce cancer risk in gynecological tissues., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Objective: The aim of this paper is to present the results of a consensus meeting on the threshold property requirements for the clinical use of conventional glass-ionomer cements (GICs) for restorative indications., Methods: Twenty-one experts on GICs evaluated the results of tests on mechanical and optical properties of 18 different brands of restorative GICs: Bioglass R [B], Chemfil Rock [CR], Equia Forte [EF], Gold Label 2 [GL2], Gold Label 9 [GL9], Glass Ionomer Cement II [GI], Ionglass [IG], Ion Z [IZ], Ionomaster [IM], Ionofil Plus [IP], Ionostar Plus [IS], Ketac Molar Easymix [KM], Magic Glass [MG], Maxxion R [MA], Riva Self Cure [R], Vidrion R [V], Vitro Fil [VF] and Vitro Molar [VM]. All experiments were carried out by a team of researchers from Brazil and England following strict protocols, under the same laboratory conditions throughout, and maintaining data integrity., Results: There was consensus on: determining as primary properties of the material: compressive strength, microhardness, acid erosion and fluoride release, and as secondary properties: contrast ratio and translucency parameter, in order to rank the materials. Seven brands were below the thresholds for restorative indications: IZ, IM, IG, MA, VF, B and MG., Conclusions: Based on the primary properties adopted as being essential for restorative indications, the conventional restorative GICs that met the thresholds and could be considered suitable as long-term restorative materials were: EF, GI, GL9, KM, IP, GL2, IS, CR, V, VM and R. A decision-making process to select the best GIC must also include results from clinical trials., Clinical Significance: This study provides a ranking of GICs that could be considered suitable as long-term restorative materials based on their main properties., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Riverbank filtration is a commonly-used technology that improves water quality by passing river water through aquifers. In this study, a riverbank filtration site in Busan, South Korea, was investigated to understand the spatiotemporal evolution of high iron and sulfate concentrations observed in the riverbank-filtered water. Discrepancies between the nonreactive transport results and field measurements suggest that iron-sulfate-related geochemical reactions play a major role in the spatiotemporal evolution of the hydrochemical properties. Pyrite oxidation was hypothesized to be the main process driving the release of iron and sulfate. To test this hypothesis, a reactive transport model was developed, that implemented pyrite oxidation as a kinetic process and subsequent ferrous iron oxidation and ferric iron precipitation as equilibrium processes. The model accurately captured the temporal evolution of sulfate; however, iron concentrations were underestimated. Sensitivity tests revealed that adjusting reaction constants significantly improved the prediction of iron concentrations. The results of this study suggest that pyrite oxidation can affect the hydrochemistry of riverbank-filtered water and highlight the potential limitations of using theoretical reaction constants in field modeling applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The study aimed to evaluate the influence of cavosurface angle on stress concentration and gap formation in class V restorations. Cylindrical cavities 3 mm in diameter were prepared in forty-five bovine incisors, changing only the angle of the bur in relation to the flat surface of the tooth. The cavities maintained the same volume (17.67 mm³). The samples were divided according to the cavosurface angle, into three groups (n = 15): 90°, 120°, 135°. After adhesive application (Futurabond U, VOCO), the cavity was filled with bulk placement of a resin composite (GrandioSO, VOCO). The teeth were analyzed with stereomicroscopy. Data of marginal gap formation were statistically analyzed with a one-way analysis of variance (ANOVA) followed by Tukey tests (significance level: α = 0.05). Finite element analysis (FEA) was used to study residual stress in these geometries and to correlate those stresses with experimentally measured gap formation. The elastic modulus and polymerization shrinkage were determined for FEA. Residual shrinkage stresses were expressed in maximum principal stress (MPS). There was a significant difference in the gap formation among the groups (p = 0.001). A significantly lower marginal gap formation was found for 120° and 135° angles, with no significant difference between them. The cavosurface angle at 90° caused substantially higher stresses, in the restoration interface, with greater marginal gap. For the 120° and 135° angles, the stress concentrations were smaller and were located in the dental structure. The cavosurface angle influenced the marginal gap formation and stress concentration., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate influence of protease inhibitors on degradation of sound, sclerotic and caries-affected dentin. Thirty-nine molars were used, thirteen for each dentin condition. Three slices were obtained from each tooth, each one immersed in the following different solutions for 1 h: 1) artificial saliva; 2) 2% chlorhexidine; 3) 0.5% epigallocatechin-gallate (EGCG). After immersion, samples were subjected to enzymatic degradation challenge (collagenase from Clostridium histolyticum). Nanohardness (HIT) and elastic modulus (Er) were measured before and after enzymatic challenge, as well as the ultimate tensile strength (UTS). Results of UTS, HIT and Er tests were submitted to repeated measurements ANOVA and Tukey post-test (α = 0.05). Higher values of UTS were found for sound dentin in control (40.30 ± 21.38 MPa) and 0.5% EGCG (30.05 ± 19.67 MPa) groups. Before degradation, higher values of HIT (0.237 ± 0.062 GPa) and Er (5.58 ± 1.75 GPa) were found for 0.5% EGCG group in caries-affected dentin. After degradation, 2% chlorhexidine group had higher values of HIT and Er for sound (0.134 ± 0.020 GPa and 3.57 ± 0.40 GPa) and sclerotic (0.201 ± 0.048 GPa and 4.30 ± 0.56 GPa) dentin. The 2% chlorhexidine showed best effect increasing HIT and Er, mainly for sclerotic dentin, before/after enzymatic degradation. The 0.5% EGCG showed better effect on HIT and Er in caries-affected dentin, before enzymatic degradation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Introduction: This study evaluated the efficacy of photodynamic inactivation (PDI) with hematoporphyrin IX (H) and modified hematoporphyrin IX (MH) at 10 μmol/L, using a blue light-emitting diode (LED), fluence of 75 J/cm, 2 over planktonic cultures and biofilm of Streptococcus mutans (UA 159)., Methods: Suspensions containing 107 cells/mL were tested under different experimental conditions: a) H and LED (H+L+), b) MH and LED (MH+L+), c) only LED (P-L+), d) only H (H+L-), e) only MH (MH+L-), and f) control group, no LED or photosensitizer treatment (P-L-). The study also evaluated the effect of PDI on S mutans biofilm on metallic or ceramic brackets bonded on specimens of human teeth. The strains were seeded onto Mitis salivarius-bacitracin-sacarose agar to determine the number of colony-forming units., Results: H and MH under LED irradiation were effective on planktonic cultures (P <0.0001). H and MH (H+L+ and MH+L+) caused a reduction of 3.80 and 6.78 log 10  CFU/mL. PDI with the use of H or MH and LED exerted a strong antimicrobial effect over S mutans showing 54% and 100% reduction, respectively. PDI on S mutans biofilm on metallic and ceramic brackets with the use of H was not effective (P = 0.0162, P = 0.1669), however, MH caused a significant reduction of 44% and 53% of the cell count on metallic and ceramic brackets, respectively (P = 0.0020, P = 0.004)., Conclusions: In vitro planktonic cultures with the use of H or MH and LED exerted significant antimicrobial activity. No effect was observed on S mutans biofilm on either bracket type with the use of H, MH showed better results, suggesting a promising use against dental caries and white spot lesions., (Copyright © 2018 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Antibiotic-containing polymer-based nanofibers (hereafter referred to as scaffolds) have demonstrated great potential for their use in regenerative endodontics from both an antimicrobial and cytocompatibility perspective. This study sought to evaluate in vitro the effects of ciprofloxacin (CIP)-containing polymer scaffolds against Enterococcus faecalis biofilms., Methods: Human mandibular incisors were longitudinally sectioned to prepare radicular dentin specimens. Sterile dentin specimens were distributed in 24-well plates and inoculated with E. faecalis for biofilm formation. Infected dentin specimens were exposed to 3 groups of scaffolds, namely polydioxanone (PDS) (control), PDS + 5 wt% CIP, and PDS + 25 wt% CIP for 2 days. Colony-forming units (CFU/mL) (n = 10) and scanning electron microscopy (SEM) (n = 2) were performed to quantitatively and qualitatively assess the antimicrobial effectiveness, respectively., Results: PDS scaffold containing CIP at 25 wt% showed maximum bacteria elimination with no microbial growth, differing statistically (P < .05) from the control (PDS) and from PDS scaffold containing CIP at 5 wt%. Statistical differences (P < .05) were also seen for the CFU/mL data between pure PDS (5.92-6.02 log CFU/mL) and the PDS scaffold containing CIP at 5 wt% (5.39-5.87 log CFU/mL). SEM images revealed a greater concentration of bacteria on the middle third of the dentin specimen after 5 days of biofilm formation. On scaffold exposures, SEM images showed similar results when compared with the CFU/mL data. Dentin specimens exposed to PDS + 25 wt% CIP scaffolds displayed a practically bacteria-free surface., Conclusions: On the basis of the data presented, newly developed antibiotic-containing electrospun scaffolds hold promise as an intracanal medicament to eliminate biofilm/infection before regenerative procedures., (Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

CBFβ and RUNX1 form a DNA-binding heterodimer and are both required for hematopoietic stem cell (HSC) generation in mice. However, the exact role of CBFβ in the production of HSCs remains unclear. Here, we generated and characterized 2 zebrafish cbfb null mutants. The cbfb(-/-) embryos underwent primitive hematopoiesis and developed transient erythromyeloid progenitors, but they lacked definitive hematopoiesis. Unlike runx1 mutants, in which HSCs are not formed, nascent, runx1(+)/c-myb(+) HSCs were formed in cbfb(-/-) embryos. However, the nascent HSCs were not released from the aorta-gonad-mesonephros (AGM) region, as evidenced by the accumulation of runx1(+) cells in the AGM that could not enter circulation. Moreover, wild-type embryos treated with an inhibitor of RUNX1-CBFβ interaction, Ro5-3335, phenocopied the hematopoietic defects in cbfb(-/-) mutants, rather than those in runx1(-/-) mutants. Finally, we found that cbfb was downstream of the Notch pathway during HSC development. Our data suggest that runx1 and cbfb are required at 2 different steps during early HSC development. CBFβ is not required for nascent HSC emergence but is required for the release of HSCs from AGM into circulation. Our results also indicate that RUNX1 can drive the emergence of nascent HSCs in the AGM without its heterodimeric partner CBFβ.

The mechanisms of cardiovascular protective effects of ghrelin and its synthetic analogs are still largely unknown. Our first aim was to ascertain whether or not natural and synthetic ligands of GHS-R1a are capable of interfering with the activity of the renin-angiotensin system. Second, since polymorphisms in the ACE gene have been associated with Alzheimer's dementia (AD) and ACE is potentially involved in brain β-amyloid degradation, we also investigated the state of ghrelin axis and inflammatory markers in patients with AD and vascular dementia (VaD). Desacyl ghrelin, hexarelin, EP80317, and GHRP-6 all significantly inhibited ACE activity in vitro; by comparison, the efficacies of ghrelin and MK-0677 were significantly lower, suggesting that ACE-inhibiting activity is unrelated to ligand affinity to GHS-R1a. ACE was capable of cleaving Aβin vitro, reducing its ability to aggregate in fibrillar Aβ. Interestingly, this protective effect of ACE was blunted by enalapril but not hexarelin or EP80317. Desacyl ghrelin levels were lower in VaD subjects compared with AD and control subjects, whereas ghrelin and TNF-α levels were similar in all groups. VaD subjects demonstrated greater levels of mRNA for GHS-R1a, PPAR-γ and CD36 in peripheral blood lymphocytes compared with other groups. In conclusion, some GHSs are effective ACE-inhibitors, and this activity may contribute to their cardiovascular effects. Hexarelin or EP80317 do not inhibit the N-domain of ACE, which is also involved in the metabolism of β-amyloid, suggesting the possibility of developing new antihypertensive drugs with improved therapeutic potential., (Copyright © 2012 Elsevier Ltd. All rights reserved.)