RESUMEN
BACKGROUND: Breast cancer is the most commonly diagnosed cancer among women worldwide with limited treatment options. Artepillin C (3,5-diprenyl-4-hydroxycinnamic acid) is one of the main constituents of Brazilian propolis presenting different activities, including antitumoral effects against various types of cancer. OBJECTIVE: We evaluated the antitumoral potential and mechanisms of action of artepillin C against two distinct human breast cancer cell lines, MCF-7 and MDA-MB-231, to explore a new therapeutic candidate. METHODS: Cell viability was assessed by MTT assay and the long-term cytotoxicity was performed by clonogenic assay. The morphological changes were observed by light microscopy, analysis of cell death pathway by Annexin V FITC/propidium iodide (PI), lactate dehydrogenase (LDH) by colorimetry, DNA fragmentation by agarose gel and senescence by ß-galactosidase. Detection of total reactive oxygen species (ROS) by fluorescence microscopy and determination of mitochondrial transmembrane potential by flow cytometry were also performed. RESULTS: Artepillin C presented a strong and dose-time-dependent cytotoxic effect on MCF-7 and MDA-MB-231 cell lines, with cytotoxicity more evident in MCF-7. In both cancer cell lines, the clonogenic potential was significantly reduced and the morphology of the cells was changed. The treatment also induced death by necrosis and late apoptosis in MCF-7 and MDA-MB-231 and induced cell senescence in MCF-7. Also, artepillin C increased total ROS in both cancer cells and decreased mitochondrial membrane potential in MDA-MB-231 cells. CONCLUSION: Artepillin C presented antitumoral potential in two human breast cancer cell lines, MCF-7, and MDA-MB-231, suggesting a new promising option for the treatment and/or chemopreventive strategy for breast cancer.
Asunto(s)
Antineoplásicos , Neoplasias de la Mama , Fenilpropionatos , Própolis , Humanos , Femenino , Células MCF-7 , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Própolis/farmacología , Especies Reactivas de Oxígeno/metabolismo , Brasil , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Apoptosis , Línea Celular Tumoral , Proliferación CelularRESUMEN
BACKGROUND: Our previous proteomics data obtained from Candida albicans recovered after serial passage in a murine model of systemic infection revealed that Orf19.36.1 expression correlates with the virulence of the fungus. Therefore, the impact of ORF19.36.1 upon virulence was tested in this study. MATERIALS & METHODS: CRISPR-Cas9 technology was used to construct homozygous C. albicans orf19.36.1 null mutants and the phenotypes of these mutants examined in vitro (filamentation, invasion, adhesion, biofilm formation, hydrolase activities) and in vivo assays. RESULTS: The deletion of ORF19.36.1 did not significantly impact the phenotypes examined or the virulence of C. albicans in two infection models. CONCLUSION: These results suggest that, although Orf19.36.1 expression correlates with virulence, this protein is not essential for C. albicans pathobiology.
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Candida albicans , Candidiasis , Proteínas Fúngicas , Animales , Ratones , Candidiasis/microbiología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Virulencia/genéticaRESUMEN
Candida albicans is one of the major pathogens found in superficial and invasive infections. This fungus expresses several virulence factors and fitness attributes that are essential to the pathogenesis. In our previous study using a murine model of serial systemic candidiasis, virulence of the recovered C. albicans was enhanced and several virulence factors were also modified after five successive passages through mice (P1-P5). In this study, we aimed to correlate the different fungal morphologies, as well as the filamentation, invasion, and stress resistance abilities, of the cells recovered after passing through this model of infection with our previous findings regarding virulence. We obtained two colony morphology types from the recovered cells, differing in their peripheral filamentation. The morphotype 1, which presented zero to five filaments in the colony edge, was higher in P2, while morphotype 2, which presented more than five filaments in the colony edge, was predominant from P3 to P5. In general, morphotype 1 showed similar levels regarding filamentation in serum, invasion of agar and cells, and resistance to osmotic, oxidative, and thermal stress in all passages analyzed. The morphotype 2, however, exhibited an enhancement in these abilities over the passages. We observed an accordance with the increased virulence over the passages obtained in our previous study and the increased adaptability profile of morphotype 2. Therefore, we suggest that the behavior observed previously in the pathogenesis and virulence could be attributed, at least in part, to the greater presence and ability of morphotype 2.
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Candida albicans , Candidiasis , Animales , Candidiasis/microbiología , Proteínas Fúngicas , Ratones , Virulencia , Factores de VirulenciaRESUMEN
BACKGROUND: Yeasts of the Candida parapsilosis complex have frequently been reported as agents of fungal infection in Brazil and worldwide, most of the cases are related to hospital-acquired infection. C. parapsilosis is the third most common cause of candidemia, and the hands of hospital workers as well as hospital surfaces have been suggested as possible sources. OBJECTIVES: In this study we verified the frequency of C. parapsilosis on the hands of workers and on surfaces in the adult intensive care unit (AICU) of a tertiary hospital in Paraná-Brazil. METHODS: Surface samples were collected with swabs moistened with saline, and a plastic bag with distilled water was used to collect samples from hands. The yeasts were identified by morphology, MALDI-TOF mass spectrometry and PCR-RFLP of the secondary alcohol dehydrogenase-encoding gene (SADH) after digestion with the restriction enzyme BanI. RESULTS AND CONCLUSIONS: A total of 223 yeast were found, of which 101 (45.29%) were identified as C. parapsilosis sensu stricto. Of these, 46.66% (n=35) were found on surfaces and 44.59% (n=66) on the hands of the employees. The analysis of C. parapsilosis strains by microsatellite loci (CP1, CP4, CP6 and B5) showed 80 different genotypes. Their antifungal susceptibility profile, evaluated by the microdilution broth method, revealed that C. parapsilosis was sensitive to amphotericin B, fluconazole and voriconazole, but not to micafungin. The results revealed the heterogeneity of the yeast population, suggesting that there is no common source of contamination in the AICU of this hospital.
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Candida parapsilosis/genética , Candida parapsilosis/aislamiento & purificación , Infección Hospitalaria/microbiología , Mano/microbiología , Personal de Salud/estadística & datos numéricos , Unidades de Cuidados Intensivos/estadística & datos numéricos , Adulto , Antifúngicos/farmacología , Candida parapsilosis/clasificación , Candida parapsilosis/efectos de los fármacos , Farmacorresistencia Fúngica , Variación Genética , Humanos , Espectrometría de Masas , Técnicas de Tipificación Micológica , Polimorfismo de Longitud del Fragmento de Restricción , Centros de Atención TerciariaRESUMEN
Vulvovaginal candidiasis (VVC) is a common vaginitis that affects women, especially in childbearing age, caused by Candida albicans in almost 80% of cases. Considering the limited drug arsenal available and the increasing fungal resistance profile, the search for new therapeutic sources with low toxicity and easy administration should be supported. Propolis has been used as a traditional medicine for multiple diseases, considering its particular composition and pharmaceutical properties that permits its wide applicability; it has also emerged as a potential antifungal agent. Thus, this study performed an in vitro and in vivo investigation into the efficacy of a new mucoadhesive thermoresponsive platform for propolis delivery (MTS-PRPe) in a preclinical murine model of VVC treatment caused by C. albicans. The methodologies involved chemical analysis, an assessment of the rheological and mucoadhesive properties of propolis formulations, in vitro and in vivo antifungal evaluations, histological evaluations and electron microscopy of the vaginal mucosa. The results demonstrated the antifungal activity of propolis extract and MTS-PRP against the standard strain and a fluconazole-resistant clinical isolate of C. albicans, in both in vitro and in vivo assays. These results were similar and even better, depending on the propolis concentration, when compared to nystatin. Thus, the formulation containing propolis exhibited good performance against C. albicans in a vulvovaginal candidiasis experimental model, representing a promising opportunity for the treatment of this infection.
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Apiterapia/métodos , Candidiasis Vulvovaginal/terapia , Sistemas de Liberación de Medicamentos/métodos , Própolis/uso terapéutico , Adhesivos , Animales , Antifúngicos/uso terapéutico , Candida albicans/efectos de los fármacos , Femenino , Ratones , Ratones Endogámicos BALB C , Pruebas de Sensibilidad Microbiana , Própolis/administración & dosificación , ReologíaRESUMEN
At present, cervical cancer is the fourth leading cause of cancer among women worldwide with no effective treatment options. In this study we aimed to evaluate the efficacy of hypericin (HYP) encapsulated on Pluronic® P123 (HYP/P123) photodynamic therapy (PDT) in a comprehensive panel of human cervical cancer-derived cell lines, including HeLa (HPV 18-positive), SiHa (HPV 16-positive), CaSki (HPV 16 and 18-positive), and C33A (HPV-negative), compared to a nontumorigenic human epithelial cell line (HaCaT). Were investigated: (i) cell cytotoxicity and phototoxicity, cellular uptake and subcellular distribution; (ii) cell death pathway and cellular oxidative stress; (iii) migration and invasion. Our results showed that HYP/P123 micelles had effective and selective time- and dose-dependent phototoxic effects on cervical cancer cells but not in HaCaT. Moreover, HYP/P123 micelles accumulated in endoplasmic reticulum, mitochondria and lysosomes, resulting in photodynamic cell death mainly by necrosis. HYP/P123 induced cellular oxidative stress mainly via type II mechanism of PDT and inhibited cancer cell migration and invasion mainly via MMP-2 inhibition. Taken together, our results indicate a potentially useful role of HYP/P123 micelles as a platform for HYP delivery to more specifically and effectively treat cervical cancers through PDT, suggesting they are worthy for in vivo preclinical evaluations.
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Antineoplásicos/administración & dosificación , Nanopartículas , Perileno/análogos & derivados , Fotoquimioterapia/métodos , Neoplasias del Cuello Uterino/tratamiento farmacológico , Antracenos , Antineoplásicos/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Sistemas de Liberación de Medicamentos , Femenino , Células HeLa , Humanos , Micelas , Invasividad Neoplásica , Estrés Oxidativo/efectos de los fármacos , Perileno/administración & dosificación , Perileno/farmacología , Poloxaleno/química , Factores de Tiempo , Neoplasias del Cuello Uterino/patologíaRESUMEN
Paracoccidioidomycosis (PCM) is a neglected disease present in Latin America with difficulty in treatment and occurrence of serious sequelae. Thus, the development of alternative therapies is imperative. In the current work, two oxadiazole compounds (LMM5 and LMM11) presented fungicidal activity against Paracoccidioides spp. The minimum inhibitory and fungicidal concentration values ranged from 1 to 32 µg/mL, and a synergic effect was observed for both compounds when combined with Amphotericin B. LMM5 and LMM11 were able to reduce CFU counts (≥2 log10) on the 5th and 7th days of time-kill curve, respectively. The fungicide effect was confirmed by fluorescence microscopy (FUN-1/FUN-2). The hippocratic screening and biochemical analysis were performed in Balb/c male mice that received a high dose of each compound, and the compounds showed no in vivo toxicity. The treatment of experimental PCM with the new oxadiazoles led to significant reduction in CFU (≥1 log10). Histopathological analysis of the groups treated exhibited control of inflammation, as well as preserved lung areas. These findings suggest that LMM5 and LMM11 are promising hits structures, opening the door for implementing new PCM therapies.
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Antifúngicos/farmacología , Oxadiazoles/farmacología , Paracoccidioides/efectos de los fármacos , Anfotericina B/farmacología , Animales , Antifúngicos/administración & dosificación , Recuento de Colonia Microbiana , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Histocitoquímica , Pulmón/microbiología , Pulmón/patología , Masculino , Ratones Endogámicos BALB C , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Oxadiazoles/administración & dosificación , Paracoccidioidomicosis/tratamiento farmacológico , Paracoccidioidomicosis/microbiología , Paracoccidioidomicosis/patología , Resultado del TratamientoRESUMEN
In recent decades, the prognosis for burn patients has improved considerably with the development of specialized care. The acellular dermal matrix (ADM) is a totally artificial acellular device that functions to control water loss, prevent penetration by bacteria and allow migration of endothelial cells and fibroblasts from patient tissues. However, little is known about its effectiveness against yeasts. The present study evaluated the capacity of colonization and migration of some human commensal yeasts. Three clinical isolates from skin scales, identified as Candida parapsilosis, Candida glabrata and Rhodotorula mucilaginosa, were used. Their ability to cross the ADM was evaluated. After three days, all isolates had crossed the ADM. C. parapsilosis showed the lowest growth, while R. mucilaginosa showed intermediate and C. glabrata the highest growth. In the plates incubated for seven days, the growth of C. parapsilosis and C. glabrata increased by 1 log over the third day. All isolates have the capacity to colonize and migrate through the matrix, increasing the potential risk to burn patients, who can develop severe and even fatal infections by invasive fungi.
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Dermis Acelular/microbiología , Quemaduras/complicaciones , Quemaduras/microbiología , Levaduras/crecimiento & desarrollo , Quemaduras/patología , Candida glabrata/crecimiento & desarrollo , Candida glabrata/patogenicidad , Candida parapsilosis/crecimiento & desarrollo , Candida parapsilosis/patogenicidad , Interacciones Huésped-Patógeno , Humanos , Rhodotorula/crecimiento & desarrollo , Rhodotorula/patogenicidad , Factores de Riesgo , Piel/lesiones , Piel/microbiología , Piel/patología , Levaduras/aislamiento & purificación , Levaduras/patogenicidadRESUMEN
PURPOSE: Cervical cancer is characterized as an important public health problem. According to latest estimates, cancer of the cervix is the fourth most common cancer among women. Due to its high prevalence, the search for new and efficient drugs to treat this infection is continuous. The progression of HPV-associated cervical cancer involves the expression of two viral proteins, E6 and E7, which are rapidly degraded by the ubiquitin-proteasome system through the increase in reactive oxygen species generation. Vitamins are essential to human substances, participate in the regulation of metabolism, and facilitate the process of energy transfer. METHODS: Some early studies have indicated that vitamin K3 exerts antitumor activity by inducing cell death by apoptosis through an increase in the generation of reactive oxygen species. Thus, we evaluated the antiproliferative effect and a likely mechanism of action of vitamin K3 against cervical epithelial cells transformed by HPV 16 (SiHa cells) assessing the production of total ROS, the mitochondrial membrane potential, the cell morphology, the cell volume, and the cell membrane integrity. RESULTS: Our results show that vitamin K3 induces an increase in ROS production in SiHa cells, triggering biochemical and morphological events, such as depolarization of mitochondrial membrane potential and decreasing cell volume. CONCLUSION: Our data showed that vitamin K3 generates an oxidative imbalance in SiHa cells, leading to mechanisms that induce cell death by apoptosis.
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Células Epiteliales/metabolismo , Papillomavirus Humano 16/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Vitamina K 3/metabolismo , Línea Celular Tumoral , Femenino , Humanos , Neoplasias del Cuello Uterino/patologíaRESUMEN
INTRODUCTION AND HYPOTHESIS: Acute uncomplicated lower urinary tract infections (UTI) and vulvovaginal candidiasis (VVC) both occur frequently in women. Although VVC is believed to commonly occur after antibiotic therapy, few studies have demonstrated this association. Thus, the aim of the study was to estimate the prevalence of colonization by Candida spp. and VVC after norfloxacin (NOR) use for UTI and the effects on the vaginal microbiota and inflammatory process. METHODS: This was a prospective cohort study of women with culture-proven UTI who were treated with NOR (antibiotic group). The control group consisted of women with noninfectious diseases or in preventive care. Candida vaginal infections were monitored both clinically and mycologically at baseline and at the follow-up evaluation. RESULTS: All women showed UTI remission after NOR treatment, and no woman in either group, antibiotic and control, showed symptoms of VVC. Both groups showed similar ratios of a positive Candida culture at baseline (6.7 % and 12.8 %, respectively) and at follow-up (3.3 % and 8.5 %, respectively) (p = 0.2768 and p = 0.5035, respectively). The antibiotic group showed no increased risk of Candida colonization or VVC after NOR treatment compared with the control group [odds ratio (OR) 0.556, 95 % confidence interval (CI) 0.2407-10.05]. CONCLUSIONS: NOR was effective for UTI treatment, did not increase the risk of vaginal colonization by Candida or VVC, and did not lead to major disturbances of the vaginal microbiota.
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Antibacterianos/farmacología , Candida/aislamiento & purificación , Candidiasis Vulvovaginal/epidemiología , Microbiota/efectos de los fármacos , Norfloxacino/farmacología , Vagina/microbiología , Enfermedad Aguda , Adolescente , Adulto , Antibacterianos/uso terapéutico , Brasil/epidemiología , Estudios de Casos y Controles , Recuento de Colonia Microbiana , Femenino , Humanos , Persona de Mediana Edad , Norfloxacino/uso terapéutico , Prevalencia , Estudios Prospectivos , Infecciones Urinarias/tratamiento farmacológico , Adulto JovenRESUMEN
Vulvovaginal candidiasis (VVC) is characterized by an infection of the vulva and vagina, mainly caused by Candida albicans, a commensal microorganism that inhabits the vaginal, digestive, and respiratory mucosae. Vulvovaginal candidiasis affects approximately 75% of women, and 5% develop the recurrent form (RVVC). The aim of the present study was to evaluate whether neutrophils microbicidal response is triggered when activated with RVVC isolates caused by C. albicans. Our results showed that RVVC isolates induced neutrophil migration but significantly decrease the microbicidal activity of neutrophils, compared with VVC and ASS isolates. The microbicidal activity of neutrophils is highly dependent on the production of reactive oxygen species/reactive nitrogen species (ROS/RNS). However, this isolate induced detoxification of ROS/RNS produced by neutrophils, reflected by the high level of thiol groups and by the oxygen consumption. Therefore, RVVC isolates induced biochemical changes in the inflammatory response triggered by neutrophils, and these effects were mainly related to the detoxification of ROS/RNS through the thioredoxin reductase (TR), a key antioxidant enzyme in fungi. This might be one of the resistance mechanisms triggered by RVVC caused by C. albicans.
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Candida albicans/inmunología , Proteínas Fúngicas/inmunología , Neutrófilos/inmunología , Reductasa de Tiorredoxina-Disulfuro/inmunología , Vagina/inmunología , Candida albicans/patogenicidad , Candidiasis Vulvovaginal/microbiología , Movimiento Celular , Citotoxicidad Inmunológica , Femenino , Proteínas Fúngicas/metabolismo , Humanos , Ácido Hipocloroso/metabolismo , Neutrófilos/microbiología , Cultivo Primario de Células , Recurrencia , Compuestos de Sulfhidrilo/inmunología , Compuestos de Sulfhidrilo/metabolismo , Reductasa de Tiorredoxina-Disulfuro/metabolismo , Vagina/microbiologíaRESUMEN
OBJECTIVES: To determine the frequency of yeast in the oral cavity of patients with chronic renal failure, undergoing hemodialysis (PCRFH); identification and antifungal susceptibility profile of yeast and demographic profile of patients. METHODS: We performed mouthwash in 146 PCRFH; the rinse fluid was collected and cultured, yeasts grown were identified by phenotypic and molecular methods. The antifungal susceptibility profile was determined against nystatin, amphotericin B, fluconazole, voriconazole, and caspofungin based in Clinical and Laboratory Standards Institute (document M27-A3). RESULTS: Positive culture was observed in 39% of patients, of whom 53% were women; the median of dialysis time was 2.9 years. The age of the colonized patients varied between 26 and 84 years, with a median of 52.5 years. PCRFH over 45 years were significantly more colonized (P = 0.0108) as well as denture wearers (84.0%). We isolated 81 yeasts, predominantly Candida albicans (63%) followed by Candida glabrata. In general, yeasts were sensitive to the evaluated antifungal agents, but there was significant variation in the minimum inhibitory concentration, especially among non-C. albicans Candida (NCAC) compared to fluconazole, caspofungin, and amphotericin B. NCAC required significantly higher concentrations of fluconazole (P < 0.01). CONCLUSION: The rate of colonization by yeasts in PCRFH was high, and there was variability in species distribution and antifungal susceptibility profile. These results are little known in this group of patients and are important for controlling the risk of developing invasive fungal infections.