Batimastat Induces Cytotoxic and Cytostatic Effects in In Vitro Models of Hematological Tumors.

The role of metalloproteinases (MMPs) in hematological malignancies, like acute myeloid leukemia (AML), myelodysplastic neoplasms (MDS), and multiple myeloma (MM), is well-documented, and these pathologies remain with poor outcomes despite treatment advancements. In this study, we investigated the effects of batimastat (BB-94), an MMP inhibitor (MMPi), in single-administration and daily administration schemes in AML, MDS, and MM cell lines. We used four hematologic neoplasia cell lines: the HL-60 and NB-4 cells as AML models, the F36-P cells as an MDS model, and the H929 cells as a model of MM. We also tested batimastat toxicity in a normal human lymphocyte cell line (IMC cells). BB-94 decreases cell viability and density in a dose-, time-, administration-scheme-, and cell-line-dependent manner, with the AML cells displaying higher responses. The efficacy in inducing apoptosis and cell cycle arrests is dependent on the cell line (higher effects in AML cells), especially with lower daily doses, which may mitigate treatment toxicity. Furthermore, BB-94 activated apoptosis via caspases and ERK1/2 pathways. These findings highlight batimastat's therapeutic potential in hematological malignancies, with daily dosing emerging as a strategy to minimize adverse effects.

In Vitro Characterization of Reversine-Treated Gingival Fibroblasts and Their Safety Evaluation after In Vivo Transplantation.

Reversine is a purine derivative that has been investigated with regard to its biological effects, such as its anticancer properties and, mostly, its ability to induce the dedifferentiation of adult cells, increasing their plasticity. The obtained dedifferentiated cells have a high potential for use in regenerative procedures, such as regenerative dentistry (RD). Instead of replacing the lost or damaged oral tissues with synthetic materials, RD uses stem cells combined with matrices and an appropriate microenvironment to achieve tissue regeneration. However, the currently available stem cell sources present limitations, thus restricting the potential of RD. Based on this problem, new sources of stem cells are fundamental. This work aims to characterize mouse gingival fibroblasts (GFs) after dedifferentiation with reversine. Different administration protocols were tested, and the cells obtained were evaluated regarding their cell metabolism, protein and DNA contents, cell cycle changes, morphology, cell death, genotoxicity, and acquisition of stem cell characteristics. Additionally, their teratoma potential was evaluated after in vivo transplantation. Reversine caused toxicity at higher concentrations, with decreased cell metabolic activity and protein content. The cells obtained displayed polyploidy, a cycle arrest in the G2/M phase, and showed an enlarged size. Additionally, apoptosis and genotoxicity were found at higher reversine concentrations. A subpopulation of the GFs possessed stem properties, as supported by the increased expression of CD90, CD105, and TERT, the existence of a CD106+ population, and their trilineage differentiation capacity. The dedifferentiated cells did not induce teratoma formation. The extensive characterization performed shows that significant functional, morphological, and genetic changes occur during the dedifferentiation process. The dedifferentiated cells have some stem-like characteristics, which are of interest for RD.

Effect of oral hygiene in infants before dental eruption on Candida spp. colonization and the occurrence of oral candidiasis: A randomized clinical trial.

Background: To assess the influence of oral hygiene in infants before the primary tooth eruption on colonization by Candida spp. and the occurrence of oral candidiasis. Material and Methods: Fifty-six infants were randomly selected in their first 48 hours of life and allocated into 2 groups: Group I (Mothers were instructed to sanitize the oral cavity of the infant with gauze and filtered water once a day) and Group II (Mothers were instructed not to sanitize the oral cavity of the infant before the dental eruption). Data collection was performed one month after the birth of the infant, in their residence, including saliva collection for identification and quantification of Candida spp. Results: Colonization by Candida spp. species was found in 49.1% of the infants evaluated. There was no statistically significant difference between colonization by Candida spp. and intervention groups (p=0.947). 13.2% of the participants presented oral candidiasis during the first month of life, this prevalence was 15.4% in the control group and 11.1% in the intervention group, however, this difference was not significant (p=0.704). Conclusions: The Candida spp. colonization and the oral candidiasis occurrence, in the first month of the life of the infant, were not influenced by oral hygiene. Key words:Infants, oral hygiene, oral health, oral candidiasis.

AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells.

The non-homologous end joining pathway is vital for repairing DNA double-strand breaks (DSB), with DNA-dependent protein kinase (DNA-PK) playing a critical role. Altered DNA damage response (DDR) in chronic (CML) and acute myeloid leukemia (AML) offers potential therapeutic opportunities. We studied the therapeutic potential of AZD-7648 (DNA-PK inhibitor) in CML and AML cell lines. This study used two CML (K-562 and LAMA-84) and five AML (HEL, HL-60, KG-1, NB-4, and THP-1) cell lines. DDR gene mutations were obtained from the COSMIC database. The copy number and methylation profile were evaluated using MS-MLPA and DDR genes, and telomere length using qPCR. p53 protein expression was assessed using Western Blot, chromosomal damage through cytokinesis-block micronucleus assay, and γH2AX levels and DSB repair kinetics using flow cytometry. Cell density and viability were analyzed using trypan blue assay after treatment with AZD-7648 in concentrations ranging from 10 to 200 µM. Cell death, cell cycle distribution, and cell proliferation rate were assessed using flow cytometry. The cells displayed different DNA baseline damage, DDR gene expressions, mutations, genetic/epigenetic changes, and p53 expression. Only HEL cells displayed inefficient DSB repair. The LAMA-84, HEL, and KG-1 cells were the most sensitive to AZD-7648, whereas HL-60 and K-562 showed a lower effect on density and viability. Besides the reduction in cell proliferation, AZD-7648 induced apoptosis, cell cycle arrest, and DNA damage. In conclusion, these results suggest that AZD-7648 holds promise as a potential therapy for myeloid leukemias, however, with variations in drug sensitivity among tested cell lines, thus supporting further investigation to identify the specific factors influencing sensitivity to this DNA-PK inhibitor.

Autism Spectrum Disorder in Females and Borderline Personality Disorder: The Diagnostic Challenge.

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder characterized by pervasive deficits in communication and social interaction and patterns of repetitive, restrictive interests and/or stereotyped behaviors. Female sex/gender is not represented in the current conceptualization of ASD, and there is emerging evidence of a female phenotype. The etiology of ASD and borderline personality disorder (BPD) is not fully understood. Clinical observations suggest that ASD and BPD can overlap in clinical presentation and diagnostic characteristics, especially in female ASD cases. We report two clinical cases of two adolescent girls presenting overlap symptoms between ASD and BPD, raising questions about the female ASD phenotype and the potential misdiagnosis of ASD characteristics with BPD, as well as its impact on diagnosis and management. Diagnostic differentiation is crucial for targeted therapeutic interventions (psychopharmacological and psychosocial). Further studies are needed to enlighten the clinical similarities and diagnostic overlap between ASD females and BPD.

Monocytes in the Characterization of Pain in Palliative Patients with Severe Dementia-A Pilot Study.

In assessing and managing pain, when obtaining a self-report is impossible, therapeutic decision-making becomes more challenging. This study aimed to investigate whether monocytes and some membrane monocyte proteins, identified as a cluster of differentiation (CD), could be potential non-invasive peripheral biomarkers in identifying and characterizing pain in patients with severe dementia. We used 53 blood samples from non-oncological palliative patients, 44 patients with pain (38 of whom had dementia) and 0 without pain or dementia (controls). We evaluated the levels of monocytes and their subtypes, including classic, intermediate, and non-classic, and characterized the levels of specific phenotypic markers, namely CD11c, CD86, CD163, and CD206. We found that the relative concentrations of monocytes, particularly the percentage of classic monocytes, may be a helpful pain biomarker. Furthermore, the CD11c expression levels were significantly higher in patients with mixed pain, while CD163 and CD206 expression levels were significantly higher in patients with nociceptive pain. These findings suggest that the levels of monocytes, particularly the classic subtype, and their phenotype markers CD11c, CD163, and CD206 could serve as pain biomarkers in patients with severe dementia.

Parthenolide Induces ROS-Mediated Apoptosis in Lymphoid Malignancies.

Lymphoid malignancies are a group of highly heterogeneous diseases frequently associated with constitutive activation of the nuclear factor kappa B (NF-κB) signaling pathway. Parthenolide is a natural compound used to treat migraines and arthritis and found to act as a potent NF-κB signaling inhibitor. This study evaluated in vitro parthenolide efficacy in lymphoid neoplasms. We assessed parthenolide metabolic activity in NCI-H929 (MM), Farage (GCB-DLBCL), Raji (BL), 697 and KOPN-8 (B-ALL), and CEM and MOLT-4 (T-ALL), by resazurin assay. Cell death, cell cycle, mitochondrial membrane potential (ΔΨmit), reactive oxygen species (ROS) and reduced glutathione (GSH) levels, activated caspase-3, FAS-ligand, and phosphorylated NF-κB p65 were evaluated using flow cytometry. CMYC, TP53, GPX1, and TXRND1 expression levels were assessed using qPCR. Our results showed that parthenolide promoted a metabolic activity decrease in all cell lines in a time-, dose-, and cell-line-dependent manner. The mechanism induced by parthenolide was demonstrated to be cell line dependent. Nonetheless, parthenolide promoted cell death by apoptosis with significant ROS increase (peroxides and superoxide anion) and GSH decrease combined with a ΔΨmit reduction across all studied cell lines. Despite the need to further understand parthenolide mechanisms, parthenolide should be considered as a possible new therapeutic approach for B- and T-lymphoid malignancies.

Screening for Autism Spectrum Disorders - Validation of the Portuguese Version of the Social Communication Questionnaire.

There are no assessment and screening tools for Autism Spectrum Disorders (ASD) validated for the Portuguese population. The Social Communication Questionnaire (SCQ) is an useful screening tool of ASD diagnosis. The main objectives of our study were to produce a Portuguese version of the SCQ (SCQ-PF), study its internal consistency, sensitivity and specificity in order to evaluate its validity as a screening instrument for ASD. We also wanted to study the impact of intellectual disability and verbal impairment and other mental disorders on SCQ-PF psychometric properties. The study included 211 children and adolescents, aged 4-17, divided in three groups: ASD Group (n = 96), Other Mental Disorders Group (OMD) (n = 63) and No Mental Disorders (NMD) Group (n = 52). Parents or other primary caregiver provided information on the SCQ items. The SCQ-PF score was significantly higher in the ASD group than in the other groups (p < 0.001). As to internal consistency, Cronbach's alpha was 87%. ASD subjects were distinguished from subjects without ASD (OMD and NMD Groups) and the area under the curve (AUC) was 0.897 (95% Confidence Interval: 0.852-0.943), for a cutoff of 14, which yielded the highest AUC, with values of sensitivity and specificity 0.76 and 0.93, respectively. These findings show that SCQ- PF with a cutoff of 14 is an acceptable and useful screening tool for ASD in the Portuguese population.

The Impact of Socio-Environmental, Individual, and Biological Factors on Oral Health-Related Quality of Life among Preschool Children: A Cohort Study with 3-Year Follow-Up.

This study assessed impact of socio-environmental, individual, and biological factors on the worsening and severe worsening of oral health-related quality of life (OHRQoL) among preschoolers and their families. A cohort study was conducted in Diamantina, Brazil, with 151 children between 1 and 3 years of age and their mothers, who were evaluated at baseline (2014) and re-evaluated after 3 years (2017). The children were clinically examined to assess the presence of dental caries, malocclusion, dental trauma, and enamel defects. The mothers answered the Early Childhood Oral Health Impact Scale (B-ECOHIS) and a questionnaire addressing individual characteristics of the child and socio-environmental factors. Extensive caries found in the follow-up (relative risk [RR] = 1.91; 95% confidence interval [CI] = 1.26-2.91) and failure to undergo the dental treatment recommended at baseline (RR = 2.49; 95% CI = 1.62-3.81) were associated with worsening of OHRQoL over 3 years. An increase in the number of children in the household (RR = 2.95; 95% CI = 1.06-8.25), occurrence of extensive caries in the follow-up (RR = 2.06; 95% CI = 1.05-4.07), and failure to undergo the dental treatment recommended at baseline (RR = 3.68; 95% CI = 1.96-6.89) were associated with a severe worsening of OHRQoL. In conclusion, the risk of worsening and severe worsening of OHRQoL was higher in preschoolers with extensive caries at follow-up and among those who did not undergo dental treatment. Furthermore, severe worsening of OHRQoL was also impacted by an increase in the number of children in the household.

Platelet Membrane Proteins as Pain Biomarkers in Patients with Severe Dementia.

Pain is one of the most frequent health problems, and its evaluation and therapeutic approach largely depend on patient self-report. When it is not possible to obtain a self-report, the therapeutic decision becomes more difficult and limited. This study aims to evaluate whether some membrane platelet proteins could be of value in pain characterization. To achieve this goal, we used 53 blood samples obtained from palliative patients, 44 with non-oncological pain and nine without pain. We observed in patients with pain a decrease in the percentage of platelets expressing CD36, CD49f, and CD61 and in the expression levels of CD49f and CD61 when compared with patients without pain. Besides that, an increase in the percentage of platelets expressing CD62p was observed in patients with pain. These results suggest that the levels of these platelet cluster differentiations (CDs) could have some value as pain biomarkers objectively since they are not dependent on the patient's participation. Likewise, CD40 seems to have some importance as a biomarker of moderate and/or severe pain. The identification of pain biomarkers such as CD40, CD49f, CD62p and CD61 can lead to an adjustment of the therapeutic strategy, contributing to a faster and more adequate control of pain and reduction in patient-associated suffering.

Alvespimycin Inhibits Heat Shock Protein 90 and Overcomes Imatinib Resistance in Chronic Myeloid Leukemia Cell Lines.

Heat shock protein 90 (HSP90) facilitates folding and stability and prevents the degradation of multiple client proteins. One of these HSP90 clients is BCR-ABL, the oncoprotein characteristic of chronic myeloid leukemia (CML) and the target of tyrosine kinase inhibitors, such as imatinib. Alvespimycin is an HSP90 inhibitor with better pharmacokinetic properties and fewer side effects than other similar drugs, but its role in overcoming imatinib resistance is not yet clarified. This work studied the therapeutic potential of alvespimycin in imatinib-sensitive (K562) and imatinib-resistant (K562-RC and K562-RD) CML cell lines. Metabolic activity was determined by the resazurin assay. Cell death, caspase activity, mitochondrial membrane potential, and cell cycle were evaluated by means of flow cytometry. Cell death was also analyzed by optical microscopy. HSPs expression levels were assessed by western blotting. Alvespimycin reduced metabolic activity in a time-, dose-, and cell line-dependent manner. Resistant cells were more sensitive to alvespimycin with an IC50 of 31 nM for K562-RC and 44 nM for K562-RD, compared to 50 nM for K562. This drug induced apoptosis via the mitochondrial pathway. In K562 cells, alvespimycin induced cell cycle arrest in G0/G1. As a marker of HSP90 inhibition, a significant increase in HSP70 expression was observed. Our results suggest that alvespimycin might be a new therapeutic approach to CML treatment, even in cases of resistance to imatinib.

Mothers' sense of coherence and oral health-related quality of life of preschoolers: a 3-year cohort study.

The aim of the present study was to assess whether mothers' sense of coherence (SOC) was a predictor of decline in oral health-related quality of life (OHRQoL) of preschoolers. A 3-year cohort study was conducted in Diamantina, Brazil. At baseline, 162 preschoolers aged one to three years were randomly selected from among children registered in local Primary Healthcare Units. In the first stage, mothers completed a sociodemographic questionnaire, the Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS), and the Brazilian short version of the Sense of Coherence Scale (SOC-13). The total score obtained from the SOC-13 was used to select exposed and unexposed children. Clinical examinations were performed to detect the presence of dental caries, traumatic dental injury, and malocclusion. At follow-up, mothers completed the sociodemographic questionnaire and the B-ECOHIS again. The incidence of severe dental caries and adherence to the proposed treatment at baseline were evaluated. A decline in OHRQoL was considered if there was an increase in the B-ECOHIS score of at least one unit. The chi-square test and Poisson regression were performed. A total of 151 preschoolers participated in the study, among whom 37.7% showed a decline in OHRQoL. Mothers' SOC was not associated with a negative impact on OHRQoL (RR = 1.24; 95%CI = 0.81-1.88), while the incidence of severe dental caries had a greater impact on the decline in OHRQoL (RR = 2.02; 95%CI = 1.29-3.16). Mothers' low SOC was not a predictor of decline in the OHRQoL of preschoolers after a 3-year follow-up period.

Family income modifies the association between frequent sugar intake and dental caries.

AIM: The aim of this cross-sectional study was to investigate whether family income modifies associations between dental caries and sex, age, mother's education, type of preschool, sugar intake, and toothbrushing. BACKGROUND: Dental caries is a multifactorial dyanamic disease primarily mediated by biofilm and sugar. DESIGN: A randomly selected sample of 308 Brazilian preschool children aged 1-3 years underwent a clinical oral examination for the assessment of moderate/extensive dental caries using codes 3-6 of the International Caries Detection and Assessment System. Mothers were asked to fill out a form addressing the child's demographic and socioeconomic characteristics as well as the frequency of sugar intake. Statistical analysis involved descriptive statistics, the chi-squared test, and Poisson regression models. RESULTS: The prevalence of moderate/extensive dental caries was 42.5%. The adjusted model revealed that within low-income families (<2 times the monthly minimum wage), the prevalence of dental caries was higher among children with a high frequency of sugar intake (≥ twice per day) than in those with a low frequency of sugar intake (< twice a day) (RR = 1.79; CI: 1.38-2.33). In families with higher income (≥2 times the monthly wage), no significant association between sugar intake and dental caries was, however, found. CONCLUSIONS: In conclusion, monthly family income can modify the association between the high frequency of sugar intake and dental caries.

Association between child's behaviour during first dental appointment and effectiveness of dental plaque removal by caregivers.

OBJECTIVE: To evaluate the association between the behaviour of children aged 1 to 4 years during their dental appointment and the effectiveness of dental plaque removal by caregivers. METHODS: This longitudinal study with intervention had the participation of 146 children (mean age = 34.89 months), 75 of whom (51.4%) showed positive behaviour (+ and ++) and 71 (48.6%), negative behaviour (- and - -). The children were evaluated at the first dental appointment, according to the Frankl scale. They were subjected to an assessment of oral conditions, and their plaque level was recorded (Quigley-Hein Index modified by Turesky) using the Evince® device. Caregivers received oral hygiene guidance. The dental plaque assessment was performed before giving the oral hygiene guidance and 14 days later. The statistical analysis included a descriptive assessment and the Wilcoxon test (p < 0.05). RESULTS: Mean dental plaque levels dropped significantly from the first to the second assessment (p < 0.001). The sample was divided according to the child's behaviour, observing that only the group of children with positive behaviour showed significantly less dental plaque in the second assessment (p < 0.001). CONCLUSION: The positive behaviour of children aged 1 to 4 years during the first dental appointments is associated with more effective dental plaque removal by caregivers.

Mothers' sense of coherence and oral health-related quality of life of preschoolers: a 3-year cohort study

Abstract The aim of the present study was to assess whether mothers' sense of coherence (SOC) was a predictor of decline in oral health-related quality of life (OHRQoL) of preschoolers. A 3-year cohort study was conducted in Diamantina, Brazil. At baseline, 162 preschoolers aged one to three years were randomly selected from among children registered in local Primary Healthcare Units. In the first stage, mothers completed a sociodemographic questionnaire, the Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS), and the Brazilian short version of the Sense of Coherence Scale (SOC-13). The total score obtained from the SOC-13 was used to select exposed and unexposed children. Clinical examinations were performed to detect the presence of dental caries, traumatic dental injury, and malocclusion. At follow-up, mothers completed the sociodemographic questionnaire and the B-ECOHIS again. The incidence of severe dental caries and adherence to the proposed treatment at baseline were evaluated. A decline in OHRQoL was considered if there was an increase in the B-ECOHIS score of at least one unit. The chi-square test and Poisson regression were performed. A total of 151 preschoolers participated in the study, among whom 37.7% showed a decline in OHRQoL. Mothers' SOC was not associated with a negative impact on OHRQoL (RR = 1.24; 95%CI = 0.81-1.88), while the incidence of severe dental caries had a greater impact on the decline in OHRQoL (RR = 2.02; 95%CI = 1.29-3.16). Mothers' low SOC was not a predictor of decline in the OHRQoL of preschoolers after a 3-year follow-up period.

Risk indicators of untreated dental caries incidence among preschoolers: a prospective longitudinal study.

The aim of the present study was to evaluate the influence of socioeconomic factors, oral conditions and the impact of OHRQoL as possible risk indicators related to the incidence of untreated dental caries in preschool children two years after an initial examination. A prospective longitudinal study was performed with a sample of 288 preschool children allocated to two groups at baseline (T0): caries free (n = 144) and with untreated dental caries (n = 144). Untreated dental caries was determined through clinical examinations performed by a calibrated dentist at T0 (Kappa > 0,89) and T1 (two years after the baseline) (Kappa > 0,91) using the dmft criteria. Parents/caregivers answered a socioeconomic questionnaire and the Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS) at T0 and T1. Mann-Whitney test and hierarchically adjusted Poisson regression models were used (95%CI, p < 0,05). The incidence of untreated dental caries was 41.3%. Low (RR = 1.63; 95%CI:1.18-2.26; p < 0.001) and high severity of untreated dental caries (RR = 1.92; 95%CI:1.36-2.72; p < 0.001), monthly household income less than two times the Brazilian minimum salary (RR = 1.79; 95%CI:1.04-3.25; p = 0.042) and overall B-ECOHIS score (RR = 1.03; 95%CI:1.02-1.05; p < 0.001) at T0 were risk indicators for the incidence of untreated dental caries among the preschool children. In conclusion, the incidence of untreated dental caries was high and the higher severity of untreated dental caries, the lower monthly income and the higher the B-ECOHIS score (indicating a negative impact on quality of life) were risk indicators to the developing of new lesions of untreated dental caries after 2 years.

Parvifloron D-based potential therapy for glioblastoma: Inducing apoptosis via the mitochondria dependent pathway.

Glioblastoma (GB) is the most malignant and frequent primary tumor of the central nervous system. The lack of diagnostic tools and the poor prognosis associated with this tumor type leads to restricted and limited options of treatment, namely surgical resection and radio-chemotherapy. However, despite these treatments, in almost all cases, patients experience relapse, leading to survival rates shorter than 5 years (∼15-18 months after diagnosis). Novel therapeutic approaches are urgently required (either by discovering new medicines or by repurposing drugs) to surpass the limitations of conventional treatments and improve patients' survival rate and quality of life. In the present work, we investigated the antitumor potential of parvifloron D (ParvD), a drug lead of natural origin, in a GB cell line panel. This natural drug lead induced G2/M cell cycle arrest and apoptosis via activation of the intrinsic mitochondria-dependent pathway. Moreover, the necessary doses of ParvD to induce pronounced inhibitory effects were substantially lower than that of temozolomide (TMZ, first-line treatment) required to promote comparable effects. Therefore, ParvD may have the potential to overcome the resistance related to TMZ and contribute to the pursuit of hopeful treatments based on ParvD as a drug lead for future chemotherapeutics.

Genetic Variants of ABC and SLC Transporter Genes and Chronic Myeloid Leukaemia: Impact on Susceptibility and Prognosis.

Solute carrier (SLC) and ATP-binding cassette (ABC) transporters comprise a variety of proteins expressed on cell membranes responsible for intrusion or extrusion of substrates, respectively, including nutrients, xenobiotics, and chemotherapeutic agents. These transporters mediate the cellular disposition of tyrosine kinase inhibitors (TKIs), and their genetic variants could affect its function, potentially predisposing patients to chronic myeloid leukaemia (CML) and modulating treatment response. We explored the impact of genetic variability (single nucleotide variants-SNVs) of drug transporter genes (ABCB1, ABCG2, SLC22A1, and SLC22A5) on CML susceptibility, drug response, and BCR-ABL1 mutation status. We genotyped 10 SNVs by tetra-primers-AMRS-PCR in 198 CML patients and 404 controls, and assessed their role in CML susceptibility and prognosis. We identified five SNVs associated with CML predisposition, with some variants increasing disease risk, including TT genotype ABCB1 (rs1045642), and others showing a protective effect (GG genotype SLC22A5 rs274558). We also observed different haplotypes and genotypic profiles associated with CML predisposition. Relating to drug response impact, we found that CML patients with the CC genotype (rs2231142 ABCG2) had an increased risk of TKI resistance (six-fold). Additionally, CML patients carrying the CG genotype (rs683369 SLC22A1) presented a 4.54-fold higher risk of BCR-ABL1 mutations. Our results suggest that drug transporters' SNVs might be involved in CML susceptibility and TKI response, and predict the risk of BCR-ABL1 mutations, highlighting the impact that SNVs could have in therapeutic selection.

Antitumor Effect of Brusatol in Acute Lymphoblastic Leukemia Models Is Triggered by Reactive Oxygen Species Accumulation.

Acute lymphoblastic leukemia (ALL) is one of the most common hematological malignancies at pediatric ages and is characterized by different chromosomal rearrangements and genetic abnormalities involved in the differentiation and proliferation of lymphoid precursor cells. Brusatol is a quassinoid plant extract extensively studied due to its antineoplastic effect through global protein synthesis and nuclear factor erythroid 2-related factor-2 (NRF2) signaling inhibition. NRF2 is the main regulator of cellular antioxidant response and reactive oxygen species (ROS), which plays an important role in oxidative stress regulation. This study aimed to evaluate the effect of brusatol in in vitro models of ALL. KOPN-8 (B-ALL), CEM (T-ALL), and MOLT-4 (T-ALL) cell lines were incubated with increasing concentrations of brusatol, and the metabolic activity was evaluated using the resazurin assay. Flow cytometry was used to evaluate cell death, cell cycle, mitochondrial membrane potential (Δψmit), and to measure ROS and reduced glutathione (GSH) levels. Our results show that brusatol promoted a decrease in metabolic activity in ALL cell lines in a time-, dose-, and cell-line-dependent manner. Brusatol induced a cytostatic effect by cell cycle arrest in G0/G1 in all cell lines; however, cell death mediated by apoptosis was only observed in T-ALL cells. Brusatol leads to an oxidative stress imbalance by the increase in ROS levels, namely, superoxide anion. Redox imbalance and cellular apoptosis induced by brusatol are highly modulated by mitochondria disruption as a decrease in mitochondrial membrane potential is detected. These data suggest that brusatol might represent a new therapeutic approach for acute lymphoblastic leukemia, particularly for ALL T-cell lineage.