Bortezomib suppresses acute myelogenous leukaemia stem-like KG-1a cells via NF-κB inhibition and the induction of oxidative stress.

Acute myelogenous leukaemia (AML) originates and is maintained by leukaemic stem cells (LSCs) that are inherently resistant to antiproliferative therapies, indicating that a critical strategy for overcoming chemoresistance in AML therapy is to eradicate LSCs. In this work, we investigated the anti-AML activity of bortezomib (BTZ), emphasizing its anti-LSC potential, using KG-1a cells, an AML cell line with stem-like properties. BTZ presented potent cytotoxicity to both solid and haematological malignancy cells and reduced the stem-like features of KG-1a cells, as observed by the reduction in CD34- and CD123-positive cells. A reduction in NF-κB p65 nuclear staining was observed in BTZ-treated KG-1a cells, in addition to upregulation of the NF-κB inhibitor gene NFΚBIB. BTZ-induced DNA fragmentation, nuclear condensation, cell shrinkage and loss of transmembrane mitochondrial potential along with an increase in active caspase-3 and cleaved PARP-(Asp 214) level in KG-1a cells. Furthermore, BTZ-induced cell death was partially prevented by pretreatment with the pancaspase inhibitor Z-VAD-(OMe)-FMK, indicating that BTZ induces caspase-mediated apoptosis. BTZ also increased mitochondrial superoxide levels in KG-1a cells, and BTZ-induced apoptosis was partially prevented by pretreatment with the antioxidant N-acetylcysteine, indicating that BTZ induces oxidative stress-mediated apoptosis in KG-1a cells. At a dosage of 0.1 mg/kg every other day for 2 weeks, BTZ significantly reduced the percentage of hCD45-positive cells in the bone marrow and peripheral blood of NSG mice engrafted with KG-1a cells with tolerable toxicity. Taken together, these data indicate that the anti-LSC potential of BTZ appears to be an important strategy for AML treatment.

The evaluation of skin sensitization potential of the UVCB substance diisopentyl phthalate by in silico and in vitro methods.

Diisopentyl phthalate (DiPeP) is primarily used as a plasticizer or additive within the production of polyvinyl chloride (PVC), and has many additional industrial applications. Its metabolites were recently found in urinary samples of pregnant women; thus, this substance is of concern as relates to human exposure. Depending upon the nature of the alcohol used in its synthesis, DiPeP may exist either as a mixture consisting of several branched positional isomers, or as a single defined structure. This article investigates the skin sensitization potential and immunomodulatory effects of DiPeP CAS No. 84777-06-0, which is currently marketed and classified as a UVCB substance, by in silico and in vitro methods. Our findings showed an immunomodulatory effect for DiPeP in LPS-induced THP-1 activation assay (increased CD54 expression). In silico predictions using QSAR TOOLBOX 4.5, ToxTree, and VEGA did not identify DiPeP, in the form of a discrete compound, as a skin sensitizer. The keratinocyte activation (Key Event 2 (KE2) of the adverse outcome pathway (AOP) for skin sensitization) was evaluated by two different test methods (HaCaT assay and RHE assay), and results were discordant. While the HaCaT assay showed that DiPeP can activate keratinocytes (increased levels of IL-6, IL-8, IL-1α, and ILA gene expression), in the RHE assay, DiPeP slightly increased IL-6 release. Although inconclusive for KE2, the role of DiPeP in KE3 (dendritic cell activation) was demonstrated by the increased levels of CD54 and IL-8 and TNF-α in THP-1 cells (THP-1 activation assay). Altogether, findings were inconclusive regarding the skin sensitization potential of the UVCB DiPeP-disagreeing with the results of DiPeP in the form of discrete compound (skin sensitizer by the LLNA assay). Additional studies are needed to elucidate the differences between DiPeP isomer forms, and to better understand the applicability domains of non-animal methods in identifying skin sensitization hazards of UVCB substances.

Effects of dabigatran versus warfarin on 2-year cognitive outcomes in old patients with atrial fibrillation: results from the GIRAF randomized clinical trial.

BACKGROUND: Observational studies support a role for oral anticoagulation to reduce the risk of dementia in atrial fibrillation patients, but conclusive data are lacking. Since dabigatran offers a more stable anticoagulation, we hypothesized it would reduce cognitive decline when compared to warfarin in old patients with atrial fibrillation. METHODS: The GIRAF trial was a 24-month, randomized, parallel-group, controlled, open-label, hypothesis generating trial. The trial was done in six centers including a geriatric care unit, secondary and tertiary care cardiology hospitals in São Paulo, Brazil. We included patients aged ≥ 70 years and CHA2DS2-VASc score > 1. The primary endpoint was the absolute difference in cognitive performance at 2 years. Patients were assigned 1:1 to take dabigatran (110 or 150 mg twice daily) or warfarin, controlled by INR and followed for 24 months. Patients were evaluated at baseline and at 2 years with a comprehensive and thorough cognitive evaluation protocol of tests for different cognitive domains including the Montreal Cognitive Assessment (MoCA), Mini-Mental State Exam (MMSE), a composite neuropsychological test battery (NTB), and computer-generated tests (CGNT). RESULTS: Between 2014 and 2019, 5523 participants were screened and 200 were assigned to dabigatran (N = 99) or warfarin (N = 101) treatment. After adjustment for age, log of years of education, and raw baseline score, the difference between the mean change from baseline in the dabigatran group minus warfarin group was - 0.12 for MMSE (95% confidence interval [CI] - 0.88 to 0.63; P = 0.75), 0.05 (95% CI - 0.07 to 0.18; P = 0.40) for NTB, - 0.15 (95% CI - 0.30 to 0.01; P = 0.06) for CGNT, and - 0.96 (95% CI - 1.80 to 0.13; P = 0.02) for MoCA, with higher values suggesting less cognitive decline in the warfarin group. CONCLUSIONS: For elderly patients with atrial fibrillation, and without cognitive compromise at baseline that did not have stroke and were adequately treated with warfarin (TTR of 70%) or dabigatran for 2 years, there was no statistical difference at 5% significance level in any of the cognitive outcomes after adjusting for multiple comparisons. TRIAL REGISTRATION: Cognitive Impairment Related to Atrial Fibrillation Prevention Trial (GIRAF), NCT01994265 .

Regulation of Immune Cells by microRNAs and microRNA-Based Cancer Immunotherapy.

MicroRNAs (miRNAs) are small (~21 nucleotides) endogenous noncoding RNA molecules involved in the posttranscriptional regulation of gene expression. Modulation of gene expression by miRNAs occurs via base-pairing of the specific miRNA primary sequence to its corresponding target messenger RNA, which can be located either in the 3' untranslated region or within the coding sequence. This pairing can lead to either translational repression or cleavage of the mRNA, resulting in reduced levels of the target protein. MiRNAs are involved in mediating and controlling several interactions between immune and cancer cells and are also important regulators of immune responses. Increasing interest has focused on elucidating the role of miRNAs in the regulation of anticancer immune responses and how this could affect the efficacy of different cancer therapeutics. Indeed, immune responses have both pro- and anti-oncogenic effects, and functional interactions between immune and cancer cells in the tumor microenvironment are crucial in determining the course of cancer progression. Thus, understanding the role of miRNAs in controlling cancer immunity is important for revealing mechanisms that could be modulated to enhance the success of immunotherapy for patients with cancer. In this chapter, we discuss the involvement of miRNAs in the regulation of immune cells and potential therapeutic approaches in which miRNAs are used for cancer immunotherapy.

Masking ability of implant abutment substrates by using different ceramic restorative systems.

STATEMENT OF PROBLEM: Information regarding the masking ability of ceramic crowns over different implant abutment materials is scarce. PURPOSE: The purpose of this in vitro study was to evaluate the masking ability of different monolithic or bilayer ceramic materials with different thicknesses over substrates indicated for implant restorations by using opaque and translucent evaluation pastes. MATERIAL AND METHODS: Disk-shaped specimens, shade A1 (VITA Classic; Ø10×1.5 to 2.5 mm), of different ceramics (a bilayer system [yttria-stabilized zirconia infrastructure+porcelain veneer: Zir+Pc] and monolithic systems [lithium disilicate under low, medium, or high translucency: LtLD, MtLD, or HtLD, respectively, and a high-translucent yttria-stabilized zirconia: HtZir]) were made (n=4). The color difference (ΔE00) was assessed by using the CIEDE2000 formula and considering the different ceramic systems over 5 implant abutment materials (A1 shade Zir [Zir A1]; white Zir [White Zir]; A1 low-translucency lithium disilicate [LD]; polyetheretherketone [PEEK]; and titanium [Ti]) when using 2 different evaluation pastes (translucent or opaque). The control comparison was the restorative material positioned over the Zir A1 substrate with a translucent evaluation paste. Statistical analysis was made by using a 2-way ANOVA and Tukey post hoc tests (α=.05) for ΔE00 data considering the restorative material and luting agent factors as their association. Additionally, ΔE00 data were qualitatively analyzed considering the acceptability and perceptibility thresholds. The translucency parameter (TP00) of each restorative material was evaluated, and data were submitted to 1-way ANOVA and Tukey post hoc tests (α=.05). RESULTS: The most predictable masking ability was seen with Zir+Pc regardless of the evaluation paste used. Nevertheless, under 1.5-mm thickness, Zir+Pc did not adequately mask Ti (ΔE00>1.77). Most monolithic ceramics did not mask discolored substrates (PEEK or Ti, ΔE00>1.77). The exception was HtZir, which presented acceptable masking ability over PEEK at 2.5-mm thickness with both evaluation pastes (ΔE00<1.77). Regardless of the restorative material thickness, Zir+Pc showed the lowest (P<.05) TP00 values (TP00=3.45 at 1.5-mm thickness; TP00=2.00 at 2.5-mm thickness), and HtLD presented the highest (P<.05, TP00=23.50 at 1.5-mm thickness; TP00=13.36 at 2.5-mm thickness). HtZir showed similar TP00 to MtLD at 1.5-mm thickness and similar TP00 to Zir+Pc when used at 2.5-mm thickness (P>.05). CONCLUSIONS: Monolithic ceramics should be used with caution over discolored implant abutments. Bilayer systems (Zir+Pc) were the most predictable approach to adequately masking discolored substrates such as PEEK or Ti. An increased restoration thickness provided higher masking ability for all restorative materials tested.

A novel lncRNA derived from an ultraconserved region: lnc-uc.147, a potential biomarker in luminal A breast cancer.

The human genome contains 481 ultraconserved regions (UCRs), which are genomic stretches of over 200 base pairs conserved among human, rat, and mouse. The majority of these regions are transcriptionally active (T-UCRs), and several have been found to be differentially expressed in tumours. Some T-UCRs have been functionally characterized, but of those few have been associated to breast cancer (BC). Using TCGA data, we found 302 T-UCRs related to clinical features in BC: 43% were associated with molecular subtypes, 36% with oestrogen-receptor positivity, 17% with HER2 expression, 12% with stage, and 10% with overall survival. The expression levels of 12 T-UCRs were further analysed in a cohort of 82 Brazilian BC patients using RT-qPCR. We found that uc.147 is high expressed in luminal A and B patients. For luminal A, a subtype usually associated with better prognosis, high uc.147 expression was associated with a poor prognosis and suggested as an independent prognostic factor. The lncRNA from uc.147 (lnc-uc.147) is located in the nucleus. Northern blotting results show that uc.147 is a 2,8 kb monoexonic trancript, and its sequence was confirmed by RACE. The silencing of uc.147 increases apoptosis, arrests cell cycle, and reduces cell viability and colony formation in BC cell lines. Additionally, we identifed 19 proteins that interact with lnc-uc.147 through mass spectrometry and demonstrated a high correlation of lnc-uc.147 with the neighbour gene expression and miR-18 and miR-190b. This is the first study to analyse the expression of all T-UCRs in BC and to functionally assess the lnc-uc.147.

Essential Oil from Bark of Aniba parviflora (Meisn.) Mez (Lauraceae) Reduces HepG2 Cell Proliferation and Inhibits Tumor Development in a Xenograft Model.

Aniba parviflora (Meisn.) Mez (Lauraceae) is an aromatic plant of the Amazon rainforest, which has a tremendous commercial value in the perfumery industry; it is popularly used as flavoring sachets and aromatic baths. In Brazilian folk medicine, A. parviflora is used to treat victims of snakebites. Herein, we analyzed the chemical composition of A. parviflora bark essential oil (EO) and its effect on the growth of human hepatocellular carcinoma HepG2 cells in vitro and in vivo. EO was obtained by hydrodistillation and characterized by GC-MS and GC-FID. The main constituents of EO were linalool (16.3±3.15), α-humulene (14.5±2.41 %), δ-cadinene (10.2±1.09 %), α-copaene (9.51±1.12 %) and germacrene B (7.58±2.15 %). Initially, EO's cytotoxic effect was evaluated against five cancer cell lines (HepG2, MCF-7, HCT116, HL-60 and B16-F10) and one non-cancerous one (MRC-5), using the Alamar blue method after 72 h of treatment. The calculated IC50 values were 9.05, 22.04, >50, 15.36, 17.57, and 30.46 µg/mL, respectively. The best selectivity was for HepG2 cells with a selective index of 3.4. DNA Fragmentation and cell cycle distribution were quantified in HepG2 cells by flow cytometry after a treatment period of 24 and 48 h. The effect of EO on tumor development in vivo was evaluated in a xenograft model using C.B-17 SCID mice engrafted with HepG2 cells. In vivo tumor growth inhibition of HepG2 xenograft at the doses of 40 and 80 mg/kg were 12.1 and 62.4 %, respectively.

Fatigue performance of adhesively luted glass or polycrystalline CAD-CAM monolithic crowns.

STATEMENT OF PROBLEM: Data comparing the fatigue performance of adhesively luted glass or polycrystalline ceramic systems for computer-aided design and computer-aided manufacturing (CAD-CAM) are scarce. PURPOSE: The purpose of this in vitro study was to evaluate and compare the fatigue performance of monolithic crowns manufactured from glass or polycrystalline CAD-CAM ceramic systems adhesively luted to a dentin analog. MATERIALS AND METHODS: Fifty-four pairs of standardized preparations of dentin analog (NEMA Grade G10) and simplified ceramic crowns of 1.5-mm thickness were obtained with 3 ceramic materials: lithium disilicate (LD) glass-ceramic (IPS e.max CAD); zirconia-reinforced lithium silicate (ZLS) glass-ceramic (Vita Suprinity); and translucent yttrium fully stabilized polycrystalline zirconia (Trans YZ) (Prettau Anterior). The simplified crowns (n=15) were adhesively cemented onto the preparations and subjected to step-stress fatigue test (initial load of 400 N, 20 Hz, 10 000 cycles, followed by 100-N increment steps until failure). Collected data (fatigue failure load [FFL] and cycles for failure [CFF]) were submitted to survival analysis with the Kaplan-Meier and Mantel-Cox post hoc tests (α=.05) and to Weibull analysis (Weibull modulus and its respective 95% confidence interval). Failed crowns were submitted to fractography analysis. The surface characteristics of the internal surface (roughness, fractal dimension) of additional crowns were accessed, and the occlusal cement thickness obtained in each luted system was measured. RESULTS: Trans YZ crowns presented the highest values of FFL, CFF, and survival rates, followed by ZLS and LD (mean FFL: 1740 N>1187 N>987 N; mean CFF: 149 000>92 613>73 667). Weibull modulus and cement thickness were similar for all tested materials. LD presented the roughest internal surface, followed by ZLS (mean Ra: 226 nm>169 nm>93 nm). The LD and ZLS internal surfaces also showed higher fractal dimension, pointing to a more complex surface topography (mean fractal dimension: 2.242=2.238>2.147). CONCLUSIONS: CAD-CAM monolithic crowns of Trans YZ show the best fatigue performance. In addition, ZLS crowns also showed better performance than LD crowns.

Motor-cognitive dual-task performance of older women evaluated using Wii Balance Board.

BACKGROUND: Single- and dual-tasks are influenced by age-related impaired postural balance. Aim of this study was to analyze the Center of Pressure (CoP) oscillation during static balance in the presence or absence of cognitive task on older women. METHODS: Thirty-one healthy older women were assessed in a stand quiet position with open/closed eyes (single-task, OE and CE) and with cognitive task (dual-task, DT) through Wii Balance Board. Sway area, total displacement and CoP oscillation (CO) based on the number of times that CoP traveled through anteroposterior and mediolateral directions. Friedman test was used to compare OE, CE and DT. Dual-task interference percentage was used to quantify the cognitive load on balance whereas Spearman correlation coefficient was used to assess the association of cognitive domains and CO. RESULTS: The CO was significantly higher in DT than in single-tasks and participants were unable to maintain their limits of stability in mediolateral direction. The cost of DT interference was 30.5%, which is partially explained by the deviation of attention from postural control to spatial and temporal orientation. CONCLUSION: Our findings show that cognitive load during DT impairs balance in mediolateral direction, thus indicating the use of WBB to assess cognitive interference on postural control.

Potential of water lettuce (Pistia stratiotes L.) for phytoremediation: physiological responses and kinetics of zinc uptake.

Two greenhouse experiments were carried out to evaluate the phytoremediation potential, physiological responses and zinc (Zn) uptake kinetics of water lettuce (Pistia stratiotes L.). The phytoextraction experiment evaluated four doses of Zn (0.7 mg L-1 - represented the Zn in the nutrient solution, 1.8, 18 and 180 mg L-1 - corresponded to ten, hundred and a thousand times, respectively, the maximum permitted content for fresh water) at four different culture times (24, 48, 72 and 168 h). The Zn uptake kinetics of water lettuce were evaluated at two concentrations of Zn (1.8 and 18 mg L-1). The water lettuce attained the highest percentage removal at the lowest evaluated doses (0.7 and 1.8 mg L-1), reaching a maximum value of approximately 72% removal (when cultivated in 1.8 mg L-1 of Zn after 168 h of culture). The Zn uptake increased with culture time, increasing the synthesis of carotenoids at all doses evaluated. The highest doses of Zn resulted in a reduction in photosynthetic efficiency. The results showed a high potential of water lettuce to absorb and tolerate Zn, accumulating preferably in the roots, demonstrating that these plants are able to absorb large quantities of Zn in contaminated solution.

Comparison of cognitive functions among frail and prefrail older adults: a clinical perspective.

ABSTRACTObjective:To compare cognitive function among frail and prefrail older adults. DESIGN: Cross-sectional clinical study. PARTICIPANTS: Fifty-one non-institutionalized older individuals participated in this study. MEASUREMENTS: Cognitive functions were evaluated through Mini-Mental State Examination (Global Cognition), Digit Span Forward (short-term memory), Digit Span Backward (working memory), Verbal Fluency Test (semantic memory/executive function). Data were compared using parametric and non-parametric bivariate tests. Binary logistic regression was used to test a frailty prediction model. Statistical significance was defined as p ≤ 0.01 to compare groups. In the regression model, the p value was set to be ≤0.05. RESULTS: Statistically significant differences were observed in global cognition, and short-term memory between frail and prefrail individuals (p ≤ 0.01). Global cognition explained 14-19% of frailty's model. CONCLUSION: According to our findings, the evaluation of cognitive functions among older persons with frailty and prefrailty provides important complementary information to better manage frailty and its progression.

Anti-liver cancer activity in vitro and in vivo induced by 2-pyridyl 2,3-thiazole derivatives.

A total of 24 hybrid compounds containing pyridyl and 1,3-thiazole moieties were screened against HL-60 (leukemia), MCF-7 (breast adenocarcinoma), HepG2 (hepatocellular carcinoma), NCI-H292 (lung carcinoma) human tumor cell lines and non-tumor cells (PBMC, human peripheral blood mononuclear cells). Most of them were highly potent in at least one cell line tested (IC50≤3µM), being HL-60 the most sensitive and HepG2 the most resistant cell line. Among them, TAP-07 and TP-07 presented cytotoxic activity in all tumor cell lines, including HepG2 (IC50 2.2 and 5.6µM, respectively) without antiproliferative effects to normal cells (PBMC) (IC50>30µM), making TAP-07 and TP-07, the compounds with the most favorable selectivity index. TAP-07 and TP-07 induced apoptosis in HepG2 cells and presented in vivo antitumor activity in hepatocellular xenograft cancer model in C.B-17 severe combined immunodeficient mice. Systemic toxicological verified by biochemical and histopathological techniques reveled no major signs of toxicity after treatment with TAP-07 and TP-07. Together the results indicated the anti-liver cancer activity of 2-pyridyl 2,3-thiazole derivatives.

Melipona mondury produces a geopropolis with antioxidant, antibacterial and antiproliferative activities.

Geopropolis is a special type of propolis produced by stingless bees. Several pharmacological properties have been described for different types of geopropolis, but there have been no previous studies of the geopropolis from Melipona mondury. In this study, we investigated the antioxidant, antibacterial, and antiproliferative activities of M. mondury geopropolis, and determined its chemical profile. The antioxidant activity was determined using in vitro ABTS·+, ·DPPH, and ß-carotene/linoleic acid co-oxidation methods. The antibacterial activity was determined using a microdilution method with Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant S. aureus. The antiproliferative effect was determined in tumor cell lines using the Alamar Blue assay. The chemical profile was obtained using UHPLC-MS and UHPLC-MS/MS. The butanolic fraction had the highest concentration of phenolic compounds and more potent antioxidant properties in all assays. This fraction also had bacteriostatic and bactericidal effects against all bacterial strains at low concentrations, especially S. aureus. The hexane fraction had the highest antiproliferative potential, with IC50 values ranging from 24.2 to 46.6 µg/mL in HL-60 (human promyelocytic leukemia cell) and K562 (human chronic myelocytic leukemia cell), respectively. Preliminary chemical analysis indicates the presence of terpenes and gallic acid in the geopropolis. Our results indicate the therapeutic potential of geopropolis from M. mondury against inflammatory, oxidative, infectious, and neoplastic diseases.

Cytotoxic potential of selected medicinal plants in northeast Brazil.

BACKGROUND: Great biodiversity is a highlight of Brazilian flora. In contrast, the therapeutic potentialities of most species used in folk medicine remain unknown. Several of these species are commonly used to treat cancer. In this study, we investigated the cytotoxic activity of 18 plants from 16 families that are found in the northeast region of Brazil. METHODS: The following species were studied: Byrsonima sericea DC. (Malpighiaceae), Cupania impressinervia Acev. Rodr. var. (revoluta) Radlk (Sapindaceae), Duranta repens Linn. (Verbenaceae), Helicostylis tomentosa (Poepp. & Endl) Rusby (Moraceae), Himatanthus bracteatus (A.DC.) Woodson (Apocynaceae), Ipomoea purga (Wender.) Hayne (Convolvulaceae), Ixora coccinea Linn. (Rubiaceae), Mabea piriri Aubl. (Euphorbiaceae), Miconia minutiflora (Melastomataceae), Momordica charantia L. (Cucurbitaceae), Ocotea glomerata (Nees) Mez (Lauraceae), Ocotea longifolia Kunth (Oreodaphne opifera Mart. Nees) (Lauraceae), Pavonia fruticosa (Mill.) Fawc. & Rendle (Malvaceae), Psychotria capitata Ruiz & Pav. (Rubiaceae), Schefflera morototoni (Aubl.) Maguire, Steyerm. & Frodin (Araliaceae), Solanum paludosum Moric. (Solanaceae), Xylopia frutescens Aubl. (Annonaceae) and Zanthoxylum rhoifolium Lam. (Rutaceae). Their dried leaves, stems, flowers or fruits were submitted to different solvent extractions, resulting in 55 extracts. After incubating for 72 h, the cytotoxicity of each extract was tested against tumor cell lines using the alamar blue assay. RESULTS: The B. sericea, D. repens, H. bracteatus, I. purga, I. coccinea, M. piriri, O. longifolia and P. capitata extracts demonstrated the most potent cytotoxic activity. The chloroform soluble fractions of D. repens flowers and the hexane extract of I. coccinea flowers led to the isolation of quercetin and a mixture of α- and ß-amyrin, respectively, and quercetin showed moderate cytotoxic activity. CONCLUSION: The B. sericea, D. repens, H. bracteatus, I. purga, I. coccinea, M. piriri, O. longifolia and P. capitata plants were identified as having potent cytotoxic effects. Further investigations are required to determine the mechanisms of cytotoxicity exhibited and their in vivo activities. This work reinforces the need to understand the therapeutics potentialities of Brazilian medicinal plants.

Cytotoxic Alkaloids from the Stem of Xylopia laevigata.

Xylopia laevigata (Annonaceae), known locally as "meiú" or "pindaíba", is widely used in folk medicine in Northeastern Brazil. In the present work, we performed phytochemical analyses of the stem of X. laevigata, which led to the isolation of 19 alkaloids: (-)-roemerine, (+)-anonaine, lanuginosine, (+)-glaucine, (+)-xylopine, oxoglaucine, (+)-norglaucine, asimilobine, (-)-xylopinine, (+)-norpurpureine, (+)-N-methyllaurotetanine, (+)-norpredicentrine, (+)-discretine, (+)-calycinine, (+)-laurotetanine, (+)-reticuline, (-)-corytenchine, (+)-discretamine and (+)-flavinantine. The in vitro cytotoxic activity toward the tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia) and HL-60 (human promyelocytic leukemia) and non-tumor peripheral blood mononuclear cells (PBMCs) was tested using the Alamar Blue assay. Lanuginosine, (+)-xylopine and (+)-norglaucine had the highest cytotoxic activity. Additionally, the pro-apoptotic effects of lanuginosine and (+)-xylopine were investigated in HepG2 cells using light and fluorescence microscopies and flow cytometry-based assays. Cell morphology consistent with apoptosis and a marked phosphatidylserine externalization were observed in lanuginosine- and (+)-xylopine-treated cells, suggesting induction of apoptotic cell death. In addition, (+)-xylopine treatment caused G2/M cell cycle arrest in HepG2 cells. These data suggest that X. laevigata is a potential source for cytotoxic alkaloids.

Antitumor Properties of the Essential Oil From the Leaves of Duguetia gardneriana.

Duguetia gardneriana, popularly known in the Brazilian northeast as "jaquinha", is a species belonging to the family Annonaceae. The aim of this work was to assess the chemical composition and antitumor properties of the essential oil from the leaves of D. gardneriana in experimental models. The chemical composition of the essential oil was analyzed via gas chromatography-flame ionization detector and gas chromatography-mass spectrometry. In vitro cytotoxic activity was determined in cultured tumor cells, and in vivo antitumor activity was assessed in B16-F10-bearing mice. The identified compounds were ß-bisabolene (80.99%), elemicin (8.04%), germacrene D (4.15%), and cyperene (2.82%). The essential oil exhibited a cytotoxic effect, with IC50 values of 16.89, 19.16, 13.08, and 19.33 µg/mL being obtained for B16-F10, HepG2, HL-60, and K562 cell lines, respectively. On the other hand, ß-bisabolene was inactive in all of the tested tumor cell lines (showing IC50 values greater than 25 µg/mL). The in vivo analysis revealed tumor growth inhibition rates of 5.37-37.52% at doses of 40 and 80 mg/kg/day, respectively. Herein, the essential oil from the leaves of D. gardneriana presented ß-bisabolene as the major constituent and showed cytotoxic and antitumor potential.

The dentist's role in identifying child abuse: an evaluation about experiences, attitudes, and knowledge.

This study sought to investigate dentists' experiences, attitudes, and knowledge in recognizing and reporting suspected cases of child abuse. It was designed as a cross-sectional study across dental practices. Data were collected through a self-administered questionnaire, which was distributed randomly to 500 dentists. The questionnaire investigated characteristics of the population, education concerning child abuse, experience and attitudes in reporting suspected cases, and the ability to recognize signs of abuse. Approximately 60% of the dentists responded to the survey. Among the dentists who participated in the survey, 87% believed that recognizing child abuse is important, however, 63.2% reported that they did not know how to act in such situations, and 44.2% were unaware of the proper child protection authorities to contact. Among the dentists surveyed, 94.7% reported they did not receive enough education concerning child abuse in their undergraduate studies. While 31.3% of dentists suspected child abuse among their patients, 84% reported their suspicions to the proper authorities. The reason cited most often (33.3%) for not reporting suspected abuse was the fear of litigation and its potential impact on their practice. Only 34.2% of dentists demonstrated knowledge about the potential signs of child abuse.

Effect of erosive conditions on different sealant materials used in paediatric dentistry.

To evaluate the effect of acidic challenge on erosion depth and topographic characteristics of different materials used as occlusal sealants. Two hundred specimens of five sealant materials (Fuji IX, Ketac Molar, Fuji II, Equia and Clinpro) and forty bovine teeth enamel samples (control) were prepared and exposed to acidic challenge. The specimens were immersed in four different solutions (orange juice, coke drink, citric acid or distilled water) under mildly shaken conditions for 3 days. The erosion depth profiles were measured using a profilometer and Scanning Electron Microscope (SEM). Two-way ANOVA with Tukey post-hoc test was performed to evaluate the interactions. Sealant material and acidic challenge had significant effects on erosion depth. Among the materials, Fuji II presented the highest mean of erosion depth after immersion in orange juice, coke drink, and citric acid. All materials groups presented higher erosion depth values after immersion in the citric acid solution, except Clinpro. Bovine enamel presented higher erosion depth values compared to all materials when submitted to erosive challenge. Sealant materials submitted to the acidic challenge presented different degrees of erosion and topographic modification; however, they are less susceptible to erosion than bovine teeth enamel.

Emetine induces oxidative stress, cell differentiation and NF-κB inhibition, suppressing AML stem/progenitor cells.

Acute myeloid leukemia (AML) is a fatal malignancy of the blood and bone marrow. Leukemic stem cells (LSCs) are a rare subset of leukemic cells that promote the development and progression of AML, and eradication of LSCs is critical for effective control of this disease. Emetine is an FDA-approved antiparasitic drug with antitumor properties; however, little is known about its potential against LSCs. Herein, we explored the antileukemic potential of emetine, focusing on its effects on AML stem/progenitor cells. Emetine exhibited potent cytotoxic activity both in hematologic and solid cancer cells and induced AML cell differentiation. Emetine also inhibited AML stem/progenitor cells, as evidenced by decreased expression of CD34, CD97, CD99, and CD123 in KG-1a cells, indicating anti-AML stem/progenitor cell activities. The administration of emetine at a dosage of 10 mg/kg for two weeks showed no significant toxicity and significantly reduced xenograft leukemic growth in vivo. NF-κB activation was reduced in emetine-treated KG-1a cells, as shown by reduced phospho-NF-κB p65 (S529) and nuclear NF-κB p65. DNA fragmentation, YO-PRO-1 staining, mitochondrial depolarization and increased levels of active caspase-3 and cleaved PARP (Asp214) were detected in emetine-treated KG-1a cells. Moreover, treatment with the pancaspase inhibitor Z-VAD(OMe)-FMK partially prevented the apoptotic cell death induced by emetine. Emetine treatment also increased cellular and mitochondrial reactive oxygen species, and emetine-induced apoptosis in KG-1a cells was partially prevented by the antioxidant N-acetylcysteine, indicating that emetine induces apoptosis, at least in part, by inducing oxidative stress. Overall, these studies indicate that emetine is a novel potential anti-AML agent with promising activity against stem/progenitor cells, encouraging the development of further studies aimed at its clinical application.