Genistein and Galantamine Combinations Decrease ß-Amyloid Peptide (1-42)-Induced Genotoxicity and Cell Death in SH-SY5Y Cell Line: an In Vitro and In Silico Approach for Mimic of Alzheimer's Disease.

Alzheimer's disease (AD) is the primary dementia-causing disease worldwide, involving a multifactorial combination of environmental, genetic, and epigenetic factors, with essential participation of age and sex. Biochemically, AD is characterized by the presence of abnormal deposition of beta amyloid peptide (Aß(1-42)), which in the brain is strongly correlated with oxidative stress, inflammation, DNA damage, and cholinergic impairment. The multiple mechanisms involved in its etiology create significant difficulty in producing an effective treatment. Neuroprotective properties of genistein and galantamine have been widely demonstrated through different mechanisms; however, it is unknown a possible synergistic neuroprotective effect against Aß(1-42). In order to understand how genistein and galantamine combinations regulate the mechanisms of neuroprotection, we conducted a set of bioassays in vitro to evaluate cell viability, clonogenic survival, cell death, and anti-genotoxicity. Through molecular docking and therapeutic viability assays, we analyzed the inhibitory activity exerted by genistein on three major protein targets (AChE, BChE, and NMDA) involved in AD. The results showed that genistein and galantamine afforded significant protection at higher concentrations; however, combinations of sub-effective concentrations of both compounds provided marked neuroprotection when they were combined. In silico approaches showed that genistein has higher scores than the positive controls and low toxicity levels; nevertheless, the therapeutic viability indicated that unlike galantamine, genistein cannot undergo the action by P glycoprotein (PGP) and probably may be unable to cross the blood-brain barrier. In conclusion, our results show that genistein and galantamine exert neuroprotective by decreasing genotoxicity and cell death. In silico analysis, suggest that genistein modulates positively the expression of AChE, BChE, and NMDA. In this context, a combination of two or more drugs could inspire an attractive therapeutic strategy.

Genotoxic evaluation of occupational exposure to antineoplastic drugs.

During the last years, several reports have provided evidence about adverse health effects on personal involved in Antineoplastic Drugs (ANPD) handling. ANPD has the ability to bind DNA, thus produce genotoxic damage. In this way, XRCC1 and XRCC3 proteins are necessary for efficient DNA repair and polymorphisms in this genes can be associated with an individual response to ANPD exposure. Therefore, the aim of this study was to evaluate genetic damage of occupational exposure to antineoplastic drugs and the possible effect of XRCC1 and XRCC3 polymorphisms in oncology employees from Bogotá, Colombia. Peripheral blood samples were obtained from 80 individuals, among exposed workers and healthy controls. The comet assay and Cytokinesis-block micronucleus cytome assay was performed to determinate genetic damage. From every sample DNA was isolated and genotyping for XRCC1 (Arg194Trp, Arg280His and Arg399Gln) and XRCC3 (Thr241Met) SNPs by PCR-RFLP. The exposed group showed a significant increase of comet assay results and micronucleus frequency, compared with unexposed group. It was observed a gender, exposure time and workplace effect on comet assay results. Our results showed no significant associations of comet assay results and micronucleus frequency with either genotype, allele, nor haplotype of XRCC1 and XRCC3 SNPs. The results suggest that occupational exposure to ANPD may lead to genotoxic damage and even be a risk to human health. To our knowledge, this is the first study to assess the genotoxic damage of occupational exposure to APND in South America.

Genotoxic effects in oral mucosal cells caused by the use of orthodontic fixed appliances in patients after short and long periods of treatment.

OBJECTIVE: This study aimed to evaluate the genotoxic effects in the oral epithelial cells of patients undergoing fixed orthodontic treatment and to compare these to a control group without treatment. The null hypothesis to be tested is that corrective orthodontic treatment at different periods does not cause genotoxic effects in patients. MATERIAL AND METHODS: An observational cross-sectional study including 74 patients enrolled in corrective orthodontic treatment and 21 control patients, between 11 and 35 years of age, of both genders, participated in the research. Patients undergoing treatment were divided into four treatment groups differentiated by treatment periods: G1, n = 21 (1 month to 12 months); G2, n = 21 (13 to 24 months); G3, n = 23 (25 to 48 months); and G4, n = 9 (over 48 months). Cells were collected by scraping the internal side of the cheek and subsequently placed in tubes containing 0.9% sodium chloride solution. The sample underwent evaluation for genotoxic effects by means of the micronucleus test (MNT). Bivariate analyses were performed using parametric tests (t test or ANOVA) and nonparametric tests (Chi-square test, Kruskal-Wallis test, Dunn post-test). The adopted level of significance was 5%. RESULTS: Statistically significant differences for any of the genotoxic abnormalities (binucleated, trinucleated, karyolysis, piknosis, nuclear buds) were not found except for karyolysis, which was higher in the control group than in G4 (p < 0.05). CONCLUSIONS: This study did not demonstrate evidence of genotoxic effects even after long periods of corrective orthodontic treatment. CLINICAL RELEVANCE: This study explores genotoxic effects in fixed orthodontic patients.

Caliphruria subedentata (Amaryllidaceae) decreases genotoxicity and cell death induced by ß-amyloid peptide in SH-SY5Y cell line.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by neuritic plaques (NPs), and neurofibrillary tangles (NFTs). ß-Amyloid peptide 1-4 2 (Aß(1-42)) is the principal component of NPs and is associated with oxidative stress, as well as dysfunction of cholinergic neurotransmission system and cell death. Nevertheless, one of the most promising therapeutic approaches for patients with AD is based on the pharmacological intervention to increases acetylcholine levels and reduces oxidative stress in AD brain. Previous studies have indicated that alkaloids from Amaryllidaceae family exhibit a wide range of biological activities. The purpose of this study was to investigate whether C. subedentata extract may modulate Aß(1-42)- induced genotoxicity in SH-SY5Y cell line. Here, we conducted a set of bioassays to measure: viability, clonogenic survival, cell death, chromosome damage and DNA strand breaks. The results showed that Aß(1-42) significantly inhibited cell viability through necrosis rather than apoptosis, increased the percentage of DNA damage and caused mitochondrial morphological alterations. Treatment with the C. subedentata extract led to a significant recovery of cell survival, decreased necrotic cell death and exerted an induction of antigenotoxic effects; additionally, the extract caigused inhibition of acetylcholinesterase (AChE). The present study confirms neuroprotective activities of C. subedentata belonging Amaryllidaceae family and provide a novel information to clarify the mechanisms by which the extracts decrease DNA damage levels induced by Aß(1-42).

Genotoxic and cytotoxic effects of Haas appliance in exfoliated buccal mucosa cells during orthodontic treatment.

OBJECTIVES: To evaluate the genotoxic and cytotoxic effects of Haas appliances through micronuclei test and cytogenetic damage analysis in buccal mucosa epithelial cells of patients undergoing orthodontic treatment. MATERIALS AND METHODS: Twenty-eight patients, 6-12 years of age and of both genders, who required a Haas appliance for the correction of a posterior crossbite were included. Epithelial cells from the mucosa were collected by gently scraping the inside of both the right and left cheeks. The cells were collected before the insertion of the appliance (T0), 1 month after the device was installed (T1), and again 3 months after the appliance was immobilized (T2). The cells were processed to obtain slides. Feulgen/Fast Green was used as the staining method, and the number of normal, karyolytic, pyknotic, nuclear buds, bi/trinucleated, and micronucleus cells were counted under light microscopy. Cellular abnormalities were evaluated with parametric and nonparametric tests for comparison of the means by analysis of variance testing, Tukey posttest, or the Kruskal-Wallis test and then by Dunn's posttest. The significance level was 5%. RESULTS: There were no statistically significant changes in the micronuclei in the evaluated periods ( P > .05). Nuclear buds increased at T1 ( P < .05), returning to baseline levels at T2. Other abnormalities (cariolytic, pyknotic, and bi/trinucleated cells) showed a significant increase at T1 and T2 ( P < .0001). CONCLUSIONS: The Haas appliance did not cause an increase in micronuclei in cells of the buccal mucosa. However, statistically significant increases in cariolytic, pyknotic, and bi/trinucleated cells were observed during treatment, suggesting possible DNA damage.

Role of GSK3ß in breast cancer susceptibility.

BACKGROUND: Breast cancer is one of the principal causes of death among Brazilian women, so it is a challenge to find new and specific early diagnostic markers, using simple and fast procedures. GSK3ß gene is an important Wnt signaling regulator involved in ß-Catenin degradation. Wnt signaling is associated with initiation and progression process in many tumor types, and alterations in ß-Catenin explain only a small proportion of aberrant signaling found in breast cancer, indicating that other Wnt signaling components and/or regulators as GSK3ß may be involved. OBJECTIVE: The aim of this study was to evaluate the genetic, epigenetic and transcriptional alterations of GSK3ß in breast cancer. METHODS: Peripheral blood samples from 204 breast cancer and healthy women were collected. Assessment of rs334558 polymorphism was performed by PCR-RFLP, promoter methylation profiles analysis by MS-PCR and qPCR was used to determine GSK3ß expression levels. RESULTS: The rs334558 polymorphism showed a strong association with aggressive cancer. A significant increase was observed in GSK3ß expression level respect to hormone receptors status and tumor size. CONCLUSION: The results indicated an inverse relationship between GSK3ß performance and tumor progression. This is the first study to relate GSK3ß gene with breast cancer in Brazilian population.