RESUMEN
Cytomegalovirus (CMV) mutations associated with antiviral resistance have become a major problem related to high mortality in kidney transplant patients. The aim of the study was to investigate mutations in the CMV genes UL97 and UL54 associated with antiviral resistance. A retrospective observational cohort study was carried out at Hospital Ophir Loyola (HOL), a reference in Kidney Transplantation. A total of 81 patients who underwent kidney transplantation were followed up between 2016 and 2018 were monitored for CMV viral load by performing qPCR. Sanger sequencing was performed on 66 patients. All CMV-positive kidney transplant recipients were included. Mutations were observed in 15 samples (22.72%) from patients. Most cases involved UL97 mutations. Mutation in UL54 without mutation in UL97 was detected in only 2 cases. Resistance mutations in UL97 were identified, such as M460V, L595S, H520Q, two co-mutations D465R + Del524 and A594P + D413A and a 3 codon deletion (del598-601). The search for mutations in the CMV genes identified mutations that confer resistance to conventional antivirals, such as ganciclovir and cidofovir, used in the treatment of these patients. Confirmation of the association with increased CMV viral load in transplanted patients, due to mutation in resistance genes, requires phenotypic analysis for confirmation purposes. These were the first findings in patients in northern Brazil that we know of.
Asunto(s)
Infecciones por Citomegalovirus , Trasplante de Riñón , Humanos , Antivirales/farmacología , Citomegalovirus/genética , Infecciones por Citomegalovirus/tratamiento farmacológico , Infecciones por Citomegalovirus/genética , Ganciclovir/farmacología , Mutación/genética , Estudios RetrospectivosRESUMEN
Background: Gastric cancer is one of the most incident types of cancer worldwide and presents high mortality rates and poor prognosis. MYC oncogene overexpression is a key event in gastric carcinogenesis and it is known that its protein positively regulates CDC25B expression which, in turn, plays an essential role in the cell division cycle progression. Menadione is a synthetic form of vitamin K that acts as a specific inhibitor of the CDC25 family of phosphatases. Methods: To better understand the menadione mechanism of action in gastric cancer, we evaluated its molecular and cellular effects in cell lines and in Sapajus apella, nonhuman primates from the new world which had gastric carcinogenesis induced by N-Methyl-N-nitrosourea. We tested CDC25B expression by western blot and RT-qPCR. In-vitro assays include proliferation, migration, invasion and flow cytometry to analyze cell cycle arrest. In in-vivo experiments, in addition to the expression analyses, we followed the preneoplastic lesions and the tumor progression by ultrasonography, endoscopy, biopsies, histopathology and immunohistochemistry. Results: Our tests demonstrated menadione reducing CDC25B expression in vivo and in vitro. It was able to reduce migration, invasion and proliferation rates, and induce cell cycle arrest in gastric cancer cell lines. Moreover, our in-vivo experiments demonstrated menadione inhibiting tumor development and progression. Conclusions: We suggest this compound may be an important ally of chemotherapeutics in the treatment of gastric cancer. In addition, CDC25B has proven to be an effective target for investigation and development of new therapeutic strategies for this malignancy.
Asunto(s)
Diterpenos/farmacología , Neoplasias Ováricas/genética , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Línea Celular Tumoral , Daño del ADN , Proteínas de Unión al ADN/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus/genética , Proteínas Represoras/genéticaRESUMEN
Artemisinin is a substance extracted from the Chinese plant Artemisia annua L. widely used in natural medicine for the treatment of various diseases. Artemether is a substance synthesized from artemisinin, and both drugs are commonly administered in the treatment of malaria. Although considered effective antimalarial drugs, very little is known about the genotoxic, cytotoxic and mutagenic effects of these drugs. Therefore, in the present study, we evaluated the genotoxic, mutagenic and cytotoxic effects of artemisinin (12.5, 25 and 50 µg/mL) and artemether (7.46; 14.92 and 29.84 µg/mL) in cultured human lymphocytes using the comet assay, the micronucleus test and the cytotoxicity assay for detection of necrosis and apoptosis by acridine orange/ethidium bromide staining. Our results showed a significant increase (p < 0.05) in the rate of DNA damage measured by comet assay and in the micronucleus frequency after treatment with both drugs. It was also observed that only artemisinin induced a statistically significant increase (p < 0.05) in the number of lymphocytes with death by necrosis 48 h after treatment. The results demonstrated that these two drugs induce mutagenic, genotoxic and cytotoxic effects in cultured human lymphocytes. Our data indicate the need for caution in the use of such drugs, since genotoxic/mutagenic effects may increase the risk of carcinogenesis.
Asunto(s)
Antimaláricos/toxicidad , Arteméter/toxicidad , Artemisininas/toxicidad , Linfocitos/efectos de los fármacos , Adulto , Antimaláricos/administración & dosificación , Apoptosis/efectos de los fármacos , Arteméter/administración & dosificación , Artemisininas/administración & dosificación , Ensayo Cometa , Daño del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Técnicas In Vitro , Linfocitos/patología , Masculino , Pruebas de Micronúcleos , Pruebas de Mutagenicidad , Mutágenos/administración & dosificación , Mutágenos/toxicidad , Necrosis/inducido químicamente , Adulto JovenRESUMEN
BACKGROUND/AIM: The evolution of gastric carcinogenesis remains largely unknown, as the regulatory mechanisms involved in the aggressiveness of gastric cancer are still poorly understood. Kinases are downstream modulators and effectors of various cell signaling cascades and play key roles in the development of neoplastic diseases. The objective of this study was to evaluate the expression of genes and proteins of the SRC family, including FYN, YES, BLK, FGR, LYN and SRC, in a model of intestinal gastric carcinogenesis generated by treating Cebus apella, a New World non-human primate, with N-methyl nitrosourea (MNU). MATERIALS AND METHODS: mRNA expression of genes was measured by real-time reverse transcription quantitative PCR (RT-qPCR) and protein expression was measured by western blotting in six Cebus apella treated with N-methyl-nitrosourea (MNU) for about 2.5 years. RESULTS: Elevated mRNA and protein expression mainly of the SRC and LYN kinases was observed. Their expression was gradually increasing as non-atrophic gastritis was evolving to gastric cancer. CONCLUSION: SRC family kinases play a key role in tumor progression and metastasis and may be a promising target for the treatment of gastric cancer.
Asunto(s)
Metilnitrosourea/efectos adversos , Neoplasias Gástricas/metabolismo , Familia-src Quinasas/genética , Familia-src Quinasas/metabolismo , Animales , Cebus , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica , Metástasis de la Neoplasia , Neoplasias Gástricas/inducido químicamente , Neoplasias Gástricas/genética , Regulación hacia ArribaRESUMEN
The potential mutagenic and histopathological effects of the hexavalent chromium were investigated in Lithobates catesbeianus tadpoles. These larvae (GS 25-31) were exposed to three nominal concentrations of potassium dichromate (4, 12, and 36â¯mgâ¯L-1) and 5â¯mgâ¯L-1 of Cyclophosphamide as a positive control (PC), for 24â¯h. A negative control (NC) was also added to the experiment. Our results showed that, in general, the micronuclei (MN) were less frequent than the erythrocyte nuclear abnormalities (ENA); there was a significant difference in the frequency of MN between the NC and all treated groups (pâ¯<â¯0.05) in a concentration-dependent curve, in addition the PC did not differ from the chromium treatments. Also, only PC and the group treated with potassium dichromate at 36â¯mgâ¯L-1 showed significantly higher frequencies of ENA than NC (pâ¯<â¯0.05). Chromium treatments promoted cell retention in the Sub-G1 phase and a decrease of cells in the S and G2/M phases indicating inhibition of the cell cycle. All treatments with chromium led to liver and kidney histopathological lesions, especially with 36â¯mgâ¯L-1 (greater number of lesions). In conclusion, hexavalent chromium was mutagenic to L. catesbeianus tadpoles and its toxic effects also resulted in anti-mitotic activity, besides inducing histopathological alterations in liver and kidney. Amphibians have been proven to be useful bioindicators, and we suggest that tadpoles of different species can be used to represent the environmental impacts in aquatic ecosystems.
Asunto(s)
Cromo/toxicidad , Dicromato de Potasio/toxicidad , Ranidae , Contaminantes Químicos del Agua/toxicidad , Animales , Anuros , Riñón/patología , Larva/efectos de los fármacos , Hígado/patología , Pruebas de Micronúcleos , Mitosis/efectos de los fármacos , Mutágenos , Rana catesbeianaRESUMEN
Five specimens of Plagioscion squamosissimus from Xingu River and ten specimens of Pellona castelnaeana from Mosqueiro Island, both in the State of Pará, Brazil, were examined to investigate the presence of anisakid nematodes, due to their zoonotic potential. Their parasitism indices and sites of infection were also determined. This is the first record of Anisakidae parasitizing Pellona castelnaeana. Four-hundred and eighty-four third-stage larvae (L3) of Anisakidae were found, of which 42 were found in P. squamosissimus and 442 in P. castelnaeana. The parasitism indices of the anisakid collected from the P. squamosissimus comprised prevalence (P) of 100%, mean infection intensity (MI) of 8.4, range of infection (RI) of 1- 13, mean abundance (MA) of 8.4, and infection site (IS) in the abdominal cavity. P. castelnaeana showed P = 100%, MI = 44.2, RI = 10-114, MA = 44.2, and IS = abdominal cavity, cecum and stomach. The life cycle of these nematodes can be completed in the Amazon basin, since the two hosts fish are part of the cetacean diet of the region, which participate as final hosts. These findings have an important consequence on epidemiology of anisakiasis, so attention should be extended to human protection against this public health risk.
Cinco espécimes de Plagioscion squamosissimus do rio Xingu e dez espécimes de Pellona castelnaeana da Ilha do Mosqueiro, ambos no Estado do Pará, Brasil, foram examinados para investigar a presença de nematodas Anisakidae, devido ao seu potencial zoonótico. Também foram determinados seus índices de parasitismo e sítios de infestação. Esse é o primeiro registro de Anisakidae parasitando Pellona castelnaeana. Foram encontradas 484 larvas de Anisakidae de terceiro estágio (L3), sendo 42 em P. squamosissimus e 442 em P. castelnaeana. Os índices parasitários de anisaquídeos coletados de P. squamosissimus compreenderam prevalência (P) de 100%, intensidade de infecção média (IM) de 8.4, amplitude de infecção (AI) de 1-13, abundância média (AM) de 8,4 e local de infecção (LI) na cavidade abdominal. P. castelnaeana apresentou P = 100%, IM = 44,2, AI = 10-114, AM = 44,2 e LI = cavidade abdominal, ceco e estômago. O ciclo de vida desses nematodas pode ser completo na bacia Amazônica, já que os dois hospedeiros fazem parte da dieta de cetáceos da região, que atuam como hospedeiros finais. Esses achados têm consequência importante na epidemiologia de anisaquíases, devendo haver mais atenção em relação à proteção humana contra esse problema de saúde pública.
Asunto(s)
Peces , Agua Dulce , ZoonosisRESUMEN
The goal of our study was to evaluate the effect of kaurenoic acid, obtained from copaiba oil resin, in gastric cancer (GC) and a normal mucosa of stomach (MNP01) cell lines. The compound was tested at concentrations of 2.5, 5, 10, 30 and 60µg/mL. Comet and micronucleus assays were used to access its potential genotoxicity in vitro. Moreover, we evaluated the effect of kaurenoic acid in cell cycle progression and in the transcription of genes involved in the control of the cell cycle: MYC, CCND1, BCL2, CASP3, ATM, CHK2 and TP53. Kaurenoic acid induced an increase on cell DNA damage or micronucleus frequencies on GC cell lines in a dose-dependent manner. The GC and MNP01 cell lines entering DNA synthesis and mitosis decreased significantly with kaurenoic acid treatment, and had an increased growth phase compared with non-treated cells. The treatment induced apoptosis (or necrosis) even at a concentration of 2.5µg/mL in relation to non-treated cells. GC cell lines presented reduced MYC, CCND1, BCL2 and CASP3 transcription while ATM, CHK2 and TP53 increased in transcription in relation to non-treated cells, especially at a concentration above 10µg/mL. The gene transcription in the MNP01 (non-treated non-cancer cell line) was designated as a calibrator for all the GC cell lines. In conclusion, our results showed that kaurenoic acid obtained from Copaifera induces DNA damage and increases the micronuclei frequency in a dose-dependent manner in GC cells, with a significant genotoxicity observed above the concentration of 5µg/mL. Moreover, this compound seems to be able to induce cell cycle arrest and apoptosis in GC cells.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Ciclo Celular/efectos de los fármacos , Diterpenos/farmacología , Mucosa Gástrica/citología , Neoplasias Gástricas , Antineoplásicos Fitogénicos/química , Línea Celular , Diterpenos/química , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Estructura Molecular , Pruebas de MutagenicidadRESUMEN
Maintaining water quality within tolerable limits is a basic need of the riverside communities in the Amazon. Using endemic aquatic organisms as biological models is useful for monitoring the environment. In this study, potential cytotoxic and genotoxic damages in Plagioscion squamosissimus (commonly known as silver croaker) from the Marajó Archipelago were evaluated using a flow cytometry assay and a survey of micronuclei (MN) frequency as well as other nuclear abnormalities (NA). P. squamosissimus specimens were collected at four locations in the Marajó Archipelago. Blood samples from these fish were used in the flow cytometry assay and piscine micronucleus test, and the resulting data were analyzed using analysis of variance (ANOVA). We did not observe a difference in the erythrocyte cell cycle distribution among the samples (P=0.9992), which suggests the absence of cytotoxic agent-induced apoptosis. The piscine micronucleus test exhibited differences in the samples from São Sebastião da Boa Vista (SSBV), and those from Anajás produced the highest mutagenicity indices. The MN frequencies were low for all groups, but the groups exhibited significantly different frequencies (P=0.0033). Reniform nuclei, nuclei with extensions, and lobed nuclei were combined and considered NA. The frequency differences for these NA were significant among sampling sites (P <0.0001). This report is the first to use flow cytometry in fish to evaluate cytotoxic agent-induced apoptosis. The micronucleus test results indicate the presence of pollutants that can change the genetic material of the fish studied. We also demonstrate that the Amazonian fish P. squamosissimus is important not only as a comestible species but also as an adequate model for biomonitoring in aquatic environments.
Asunto(s)
Citotoxinas/toxicidad , Mutágenos/toxicidad , Perciformes , Contaminantes del Agua/toxicidad , Animales , Apoptosis/efectos de los fármacos , Brasil , Ciclo Celular/efectos de los fármacos , Daño del ADN , Monitoreo del Ambiente , Eritrocitos/efectos de los fármacos , Pruebas de Micronúcleos , Perciformes/genética , RíosRESUMEN
Mercury exhibits cytotoxic and mutagenic properties as a result of its effect on tubulin. This toxicity mechanism is related to the production of free radicals that can cause DNA damage. Methylmercury (MeHg) is one of the most toxic of the mercury compounds. It accumulates in the aquatic food chain, eventually reaching the human diet. Several studies have demonstrated that prolactin (PRL) may be differently affected by inorganic and organic mercury based on interference with various neurotransmitters involved in the regulation of PRL secretion. This study evaluated the cytoprotective effect of PRL on human lymphocytes exposed to MeHg in vitro, including observation of the kinetics of HL-60 cells (an acute myeloid leukemia lineage) treated with MeHg and PRL at different concentrations, with both treatments with the individual compounds and combined treatments. All treatments with MeHg produced a significant increase in the frequency of chromatid gaps, however, no significant difference was observed in the chromosomal breaks with any treatment. A dose-dependent increase in the mitotic index was observed for treatments with PRL, which also acts as a co-mitogenic factor, regulating proliferation by modulating the expression of genes that are essential for cell cycle progression and cytoskeleton organization. These properties contribute to the protective action of PRL against the cytotoxic and mutagenic effects of MeHg.
Asunto(s)
Contaminantes Ambientales/toxicidad , Compuestos de Metilmercurio/toxicidad , Prolactina/farmacología , Adulto , Células Cultivadas , Femenino , Células HL-60 , Humanos , Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
This study described the karyotype of Geophagus cf. proximus. Specimens were collected in Água Preta Lake, Parque Ambiental de Belém, Pará State, Brazil. The karyotype were 2n = 48 chromosomes (FN = 60: 12M/SM+36ST/A) and no sexual chromosome differentiation. C-banding showed centromeric staining in all chromosomes. The first chromosome pair, besides centromeric coloration, presented a totally heterochromatic long arm. The nucleolus organizer regions (NORs) were studied by means of AgNO3. NORs were located at the short arm of the second chromosome pair.
Este estudo descreve o cariótipo de Geophagus cf. proximus. Espécimes foram coletados no lago Água Preta, Parque Ambiental de Belém, Estado do Pará, Brasil. O cariótipo obtido apresentou 2n = 48 cromossomos (NF = 60: 12M/SM+36ST/A), sem diferenciação de cromossomos sexuais. O bandeamento C mostrou marcações centroméricas em todos os cromossomos. O primeiro par cromossômico, além da coloração centromérica, apresentou o braço longo totalmente heterocromático. As regiões organizadoras do nucléolo (RONs) foram estudadas por meio da coloração de AgNO3. As RONs foram encontradas no braço curto do segundo par de cromossomos.
Asunto(s)
Bandeo Cromosómico , CitogenéticaRESUMEN
The frequencies of micronuclei (MN) and morphological nuclear abnormalities (NA) in erythrocytes in the peripheral blood of tambaqui (Colossoma macropomum), treated with 2 mg.L(-1) methylmercury (MeHg), were analyzed. Two groups (nine specimens in each) were exposed to MeHg for different periods (group A - 24 h; group B - 120 h). A third group served as negative control (group C, untreated; n = 9). Although, when compared to the control group there were no significant differences in MN frequency in the treated groups, for NA, the differences between the frequencies of group B (treated for 120 h) and the control group were extremely significant (p < 0.02), thus demonstrating the potentially adverse effects of MeHg on C. macropomum erythrocytes after prolonged exposure.
RESUMEN
The frequencies of micronuclei (MN) and morphological nuclear abnormalities (NA) in erythrocytes in the peripheral blood of tambaqui (Colossoma macropomum), treated with 2 mg.L-1 methylmercury (MeHg), were analyzed. Two groups (nine specimens in each) were exposed to MeHg for different periods (group A - 24 h; group B - 120 h). A third group served as negative control (group C, untreated; n = 9). Although, when compared to the control group there were no significant differences in MN frequency in the treated groups, for NA, the differences between the frequencies of group B (treated for 120 h) and the control group were extremely significant (p < 0.02), thus demonstrating the potentially adverse effects of MeHg on C. macropomum erythrocytes after prolonged exposure.