RESUMO
Cancer is a disease with the highest mortality and morbidity rate worldwide. First-line drugs induce several side effects that drastically reduce the quality of life of people with this disease. Finding molecules to prevent it or generate less aggressiveness or no side effects is significant to counteract this problem. Therefore, this work searched for bioactive compounds of marine macroalgae as an alternative treatment. An 80% ethanol extract of dried Caulerpa sertularioides (CSE) was analyzed by HPLS-MS to identify the chemical components. CSE was utilized through a comparative 2D versus 3D culture model. Cisplatin (Cis) was used as a standard drug. The effects on cell viability, apoptosis, cell cycle, and tumor invasion were evaluated. The IC50 of CSE for the 2D model was 80.28 µg/mL versus 530 µg/mL for the 3D model after 24 h of treatment exposure. These results confirmed that the 3D model is more resistant to treatments and complex than the 2D model. CSE generated a loss of mitochondrial membrane potential, induced apoptosis by extrinsic and intrinsic pathways, upregulated caspases-3 and -7, and significantly decreased tumor invasion of a 3D SKLU-1 lung adenocarcinoma cell line. CSE generates biochemical and morphological changes in the plasma membrane and causes cell cycle arrest at the S and G2/M phases. These findings conclude that C. sertularioides is a potential candidate for alternative treatment against lung cancer. This work reinforced the use of complex models for drug screening and suggested using CSE's primary component, caulerpin, to determine its effect and mechanism of action on SKLU-1 in the future. A multi-approach with molecular and histological analysis and combination with first-line drugs must be included.
Assuntos
Caulerpa , Neoplasias Pulmonares , Humanos , Caulerpa/química , Qualidade de Vida , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Pontos de Checagem do Ciclo Celular , Neoplasias Pulmonares/metabolismo , Apoptose , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de CélulasRESUMO
The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-aptamers (C7/C9) binding to the SARS-CoV-2 Spike (S) protein were isolated but its performance in clinical samples remained to be tested. Here, 242 samples were collected through three different methods and subjected to florescence-linked aptamer assays (FLAA) based on C7/C9 aptamers through two readout protocols. Then, a step-by-step statistical approach which included agreement tests, proportion comparisons and binomial and multinomial logistic regressions was used to predict optimal conditions for the novel C7/C9 FLAA test. RTqPCR threshold cycles, symptoms onset and processing time were influential factors on FLAA test results. Naturally occurring mutations on S were also detected and analyzed. Aminoacidic substitutions D614G and T732A appeared relevant for aptamer recognition although further studies are necessary. The methodology presented here is the first step to determine the performance and diagnosis across a range of clinical contexts and it might serve as a base for a complete analysis applicable to other designs of new diagnostic tests.
RESUMO
Soil-transmitted helminths, such as Ascaris lumbricoides, are the most prevalent parasites globally. Optimal anthelmintic treatment for A. lumbricoides in endemically infected communities is challenged by several host-related and environmental factors influencing infection acquisition. We assessed the risk of A. lumbricoides (re)infection after treatment in a Venezuelan rural community. Individual merthiolate-iodine-formaldehyde-fixed faecal samples were collected from 224 persons before a single-dose pyrantel treatment and at 1, 3, 6, 9 and 15 months after treatment. Effects of age, sex and socioeconomic status (SES) on A. lumbricoides prevalence, eggs/gram faeces (EPG) and infection (re)acquisition were assessed using both generalised linear mixed-effects models and survival analysis. Pre-treatment A. lumbricoides prevalence was 39.7%. Higher prevalence was associated with younger age and lower SES. Higher EPG values were observed among young children. Median time to A. lumbricoides infection was six months after treatment: at 1, 3, 6, 9 and 15 months post-treatment, cumulative incidence was 6.7%, 18.9%, 34.6%, 42.2%, and 52.6%, respectively. Younger age, lower SES, and pre-treatment A. lumbricoides infection status showed significantly elevated hazard ratios. Mass drug administration protocols would benefit from considering these factors in selective treatment strategies and possibly more than just annual or biannual treatments in the target population.
Assuntos
Ascaríase , Helmintíase , Animais , Ascaríase/tratamento farmacológico , Ascaríase/epidemiologia , Ascaríase/parasitologia , Ascaris lumbricoides , Criança , Pré-Escolar , Fezes/parasitologia , Helmintíase/epidemiologia , Helmintíase/parasitologia , Humanos , Prevalência , População Rural , Solo/parasitologia , Venezuela/epidemiologiaRESUMO
The spontaneous interaction between human papillomavirus type 16 (HPV16) L1 virus-like particles (VLPs) and non-functionalized gold nanoparticles (nfGNPs) interferes with the nfGNPs' salt-induced aggregation, inhibiting the red-blue color shift in the presence of NaCl. Electron microscopy and competition studies showed that color-shift inhibition is a consequence of direct nfGNP-VLP interaction and, thus, may produce a negative impact on the virus entry cell process. Here, an in vitro infection system based on the HPV16 pseudovirus (PsV) was used to stimulate the natural infection process in vitro. PsVs carry a pseudogenome with a reporter gene, resulting in a fluorescent signal when PsVs infect a cell, allowing quantification of the viral infection process. Aggregation assays showed that nfGNP-treated PsVs also inhibit color shift in the presence of NaCl. High-resolution microscopy confirmed nfGNP-PsV complex formation. In addition, PsVs can interact with silver nanoparticles, suggesting a generalized interaction of metallic nanoparticles with HPV16 capsids. The treatment of PsVs with nfGNPs produced viral infection inhibition at a higher level than heparin, the canonical inhibitor of HPV infection. Thus, nfGNPs can efficiently interfere with the HPV16 cell entry process and may represent a potential active component in prophylactic formulations to reduce the risk of HPV infection.
Assuntos
Nanopartículas Metálicas , Proteínas Oncogênicas Virais , Infecções por Papillomavirus , Proteínas do Capsídeo/genética , Ouro/farmacologia , Ouro/uso terapêutico , Papillomavirus Humano 16/genética , Humanos , Nanopartículas Metálicas/uso terapêutico , Nanopartículas Metálicas/virologia , Infecções por Papillomavirus/prevenção & controle , Prata , Cloreto de Sódio/farmacologiaRESUMO
The rapid spread of SARS-CoV-2 infection throughout the world led to a global public health and economic crisis triggering an urgent need for the development of low-cost vaccines, therapies and high-throughput detection assays. In this work, we used a combination of Ideal-Filter Capillary Electrophoresis SELEX (IFCE-SELEX), Next Generation Sequencing (NGS) and binding assays to isolate and validate single-stranded DNA aptamers that can specifically recognize the SARS-CoV-2 Spike glycoprotein. Two selected non-competing DNA aptamers, C7 and C9 were successfully used as sensitive and specific biological recognition elements for the development of electrochemical and fluorescent aptasensors for the SARS-CoV-2 Spike glycoprotein with detection limits of 0.07 fM and 41.87 nM, respectively.
Assuntos
Aptâmeros de Nucleotídeos , COVID-19 , Aptâmeros de Nucleotídeos/genética , COVID-19/diagnóstico , Humanos , SARS-CoV-2/genética , Técnica de Seleção de Aptâmeros , Glicoproteína da Espícula de Coronavírus/análise , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/metabolismoRESUMO
ZO-2 is a peripheral tight junction (TJ) protein whose silencing in renal epithelia induces cell hypertrophy. Here, we found that in ZO-2 KD MDCK cells, in compensatory renal hypertrophy triggered in rats by a unilateral nephrectomy and in liver steatosis of obese Zucker (OZ) rats, ZO-2 silencing is accompanied by the diminished activity of LATS, a kinase of the Hippo pathway, and the nuclear concentration of YAP, the final effector of this signaling route. ZO-2 appears to function as a scaffold for the Hippo pathway as it associates to LATS1. ZO-2 silencing in hypertrophic tissue is due to a diminished abundance of ZO-2 mRNA, and the Sp1 transcription factor is critical for ZO-2 transcription in renal cells. Treatment of OZ rats with metformin, an activator of AMPK that blocks JNK activity, augments ZO-2 and claudin-1 expression in the liver, reduces the paracellular permeability of hepatocytes, and serum bile acid content. Our results suggest that ZO-2 silencing is a common feature of hypertrophy, and that ZO-2 is a positive regulator of the Hippo pathway that regulates cell size. Moreover, our observations highlight the importance of AMPK, JNK, and ZO-2 as therapeutic targets for blood-bile barrier dysfunction.
Assuntos
Proteínas Quinases Ativadas por AMP , Fígado Gorduroso , Proteína da Zônula de Oclusão-2/metabolismo , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Via de Sinalização Hippo , Hipertrofia , Ratos , Ratos Zucker , Proteínas de Junções ÍntimasRESUMO
BACKGROUND: Filaggrin (FLG) loss-of-function variants are major genetic risk factors for atopic dermatitis (AD), but these have not been studied in Latin American populations with and without AD. METHODS: FLG variants R501X and 2282del4 were genotyped in 275 Chilean adults with and without AD from the "Early origins of allergy and asthma" (ARIES) cohort and in 227 patients from an AD cohort based in Santiago, Chile. RESULTS: Among adults in the ARIES cohort, 3.3% were carriers of R501X and 2.9% of 2282del4 variants, all heterozygotes. In this cohort, 6.2% were FLG variant carriers: 11.1% of subjects reporting AD were carriers of FLG variants vs. 5.2% in those without AD (P = 0.13). In this first cohort, FLG variants were not significantly associated with asthma, allergic rhinitis, or food allergy. In the AD cohort, the prevalence of FLG variants was 7% for R501X, 2.2% for the 2282del4 variant, and 9.3% for the combined genotype. In this cohort, FLG variants were present in 15.5% of severe AD vs. 7.1% of mild-to-moderate AD subjects (P = 0.056). Evaluation of Chilean population from both cohorts combined (n = 502) revealed that FLG variants were not significantly associated with AD (OR = 1.92 [95% CI 0.95-3.9], P = 0.067) but were associated with asthma (OR = 2.16 [95% CI 1.02-4.56], P = 0.039). CONCLUSIONS: This is the first study to evaluate FLG loss-of-function variants R501X and 2282del4 in Latin American population, revealing a similar prevalence of these FLG variant carriers to that of European populations. Among Chileans, FLG variants were significantly associated with asthma but not AD.
Assuntos
Dermatite Atópica , Proteínas Filagrinas/genética , Adulto , Chile/epidemiologia , Dermatite Atópica/epidemiologia , Dermatite Atópica/genética , Predisposição Genética para Doença , Humanos , Mutação , PrevalênciaRESUMO
Cervical cancer is the leading cause of death by cancer in women from developing countries. Persistent infection with high-risk human papillomavirus (HPV) types 16 and 18 is a major risk factor for cervical carcinogenesis. Nevertheless, only a few women with morphologic expression of HPV infection progress into invasive disease suggesting the involvement of other factors in cervical carcinogenesis. MicroRNAs (miRNAs) are conserved small non-coding RNAs that negatively regulate gene expression including genes involved in fundamental biological processes and human cancer. Dysregulation of miRNAs has been widely reported in cervical cancer. This work focuses on reviewing the miRNAs affected during the HPV infection process, as well relevant miRNAs that contribute to the development and maintenance of malignant cervical tumor cells. Finally, we recapitulate on miRNAs that may be used to distinguish between healthy individuals from patients with precancerous lesions or cervical tumors.
RESUMO
Haloarchaea are extreme halophilic microorganisms belonging to the domain Archaea, phylum Euryarchaeota, and are producers of interesting antioxidant carotenoid compounds. In this study, four new strains of Haloarcula sp., isolated from saline lakes of the Atacama Desert, are reported and studied by high-resolution mass spectrometry (UHPLC-Q-Orbitrap-MS/MS) for the first time. In addition, determination of the carotenoid pigment profile from the new strains of Haloarcula sp., plus two strains of Halorubrum tebenquichense, and their antioxidant activity by means of several methods is reported. The effect of biomass on cellular viability in skin cell lines was also evaluated by MTT assay. The cholinesterase inhibition capacity of six haloarchaea (Haloarcula sp. ALT-23; Haloarcula sp. TeSe-41; Haloarcula sp. TeSe-51; Haloarcula sp. Te Se-89 and Halorubrum tebenquichense strains TeSe-85 and Te Se-86) is also reported for the first time. AChE inhibition IC50 was 2.96 ± 0.08 µg/mL and BuChE inhibition IC50 was 2.39 ± 0.09 µg/mL for the most active strain, Halorubrum tebenquichense Te Se-85, respectively, which is more active in BuCHe than that of the standard galantamine. Docking calculation showed that carotenoids can exert their inhibitory activity fitting into the enzyme pocket by their halves, in the presence of cholinesterase dimers.
RESUMO
The Let-7:LIN28 regulatory loop is a paradigm in miRNA regulation. LIN28 harbors two RNA binding domains, which interact with well-conserved sequences in pre-let-7 RNAs, the GNGAY and the GGAG motifs. Here, the differential binding between LIN28B and pre-let-7 members was associated with the structural characteristics of the pre-let-7 family mapped by SHAPE, uncovering diverse structural patterns within pre-let-7 members. Pre-let-7 mutants supported a relevant role of the GGAG motif location and the preE-stem stability for the interaction with LIN28B. Based on these results, we propose a core RNA structure for LIN28B interaction.
Assuntos
MicroRNAs/química , MicroRNAs/metabolismo , Precursores de RNA/química , Precursores de RNA/metabolismo , Proteínas de Ligação a RNA/metabolismo , Sequência de Bases , Humanos , MicroRNAs/genética , Modelos Moleculares , Conformação de Ácido Nucleico , Ligação Proteica , Precursores de RNA/genéticaRESUMO
OBJECTIVES: To analyze the fundamentals of the global health agenda from 1944 to 2018, especially regarding Universal Health Coverage, in order to unveil its relations with capital accumulation in health services and to contribute to world social mobilization to change this tendency. METHODS: A historical study was carried out based on a purposeful selection of primary sources on the global health agenda from multilateral organizations and secondary sources about the changes of capitalism from the study period. RESULTS: The global health agenda changed from the state responsibility for health to an insurance healthcare system based on markets. The medical-industrial complex pressured national economies, broke postwar pacts, and urged economic globalization. The neoliberal, neoclassical, and neo-institutional discourse that promoted a new state-market relationship eased the new capital accumulation in healthcare into financial and cognitive capitalism. CONCLUSIONS: Understanding these relationships allows us to provide elements for social mobilization geared to transform the healthcare sector toward a new vision of health with a nature-society relationship that contributes to socially constructing human and environmental health, rather than gaining profits based on illness and chronic suffering.
Assuntos
Atenção à Saúde/economia , Saúde Global/economia , Saúde Global/história , Serviços de Saúde/economia , Política , Cobertura Universal do Seguro de Saúde/economia , Cobertura Universal do Seguro de Saúde/história , Cobertura Universal do Seguro de Saúde/legislação & jurisprudência , Atenção à Saúde/história , Atenção à Saúde/legislação & jurisprudência , Atenção à Saúde/estatística & dados numéricos , Saúde Global/legislação & jurisprudência , Saúde Global/estatística & dados numéricos , Serviços de Saúde/história , Serviços de Saúde/legislação & jurisprudência , Serviços de Saúde/estatística & dados numéricos , História do Século XX , História do Século XXI , Humanos , Cobertura Universal do Seguro de Saúde/estatística & dados numéricosAssuntos
Antipsicóticos/efeitos adversos , Haloperidol/efeitos adversos , Hipnóticos e Sedativos/administração & dosagem , Lorazepam/administração & dosagem , Transtornos Psicóticos , Adulto , Antidepressivos/uso terapêutico , Antipsicóticos/administração & dosagem , Feminino , Haloperidol/administração & dosagem , Humanos , Transtornos Psicóticos/diagnóstico , Transtornos Psicóticos/tratamento farmacológico , Recidiva , Risperidona/uso terapêutico , Sertralina/uso terapêutico , Distúrbios do Início e da Manutenção do Sono/tratamento farmacológico , Distúrbios do Início e da Manutenção do Sono/etiologiaRESUMO
This dataset includes data obtained at the Atmospheric Microphysics and Radiation Laboratory (LAMAR) of the Huancayo Observatory (12.04° S, 75.32° W, 3313 m ASL). Two Parsivel2 and two tipping bucket rain gauges are used in this dataset which are operating together since 2018. Data is given in NetCDF format, including two types of files, one NetCDF for precipitation totals and another which contains Parsivel2 data. This data set was collected in the complex topography conditions of the tropical Andes, and its potential use is to study the microphysics of orographic rainfall, atmospheric models and rainfall estimation algorithms.
RESUMO
Prior to and during the process of human labor, maternal circulating leukocytes infiltrate the maternal-fetal interface (choriodecidua) and become activated resembling choriodecidual leukocytes. Since, there is no evidence comparing maternal circulating and choriodecidual leukocytes, herein, we characterized their transcriptome and explored the biological processes enriched in choriodecidual leukocytes. From women undergoing spontaneous term labor we isolated circulating and choriodecidual leukocytes, performed microarray analysis (n = 5) and qRT-PCR validation (n = 9) and interaction network analysis with up-regulated genes. We found 270 genes up-regulated and only 17 genes down-regulated in choriodecidual leukocytes compared to maternal circulating leukocytes. The most up-regulated genes were CCL18, GPNMB, SEPP1, FN1, RNASE1, SPP1, C1QC, and PLTP. The biological processes enriched in choriodecidual leukocytes were cell migration and regulation of immune response, chemotaxis, and humoral immune responses. Our results show striking differences between the transcriptome of choriodecidual and maternal circulating leukocytes. Choriodecidual leukocytes are enriched in immune mediators implicated in the spontaneous process of labor at term.
Assuntos
Decídua/metabolismo , Trabalho de Parto/genética , Leucócitos/metabolismo , Transcriptoma , Adulto , Decídua/citologia , Feminino , Humanos , Trabalho de Parto/sangue , Trabalho de Parto/metabolismo , GravidezRESUMO
The miR-143/145 cluster is down-regulated in cervical tumor cells suggesting a role in tumorigenesis including cytoskeleton remodeling, a key event for tumor progression. The aim of the present work was to determine the role of miR-143/145 in the modulation of the myosin regulator phospho-myosin light chain (pMLC). HeLa monolayer and tridimensional cultures were transfected with miR-143 or miR-145 mimics inhibiting cell viability, proliferation, migration and invasion, mainly through miR-145. MiR-145 transfection increased pMLC levels by targeting the MYPT1 subunit of the regulatory myosin phosphatase. MYPT1 knockdown by siRNAs reproduced miR-145 effects suggesting miR-145 as a tumor suppressor through MYPT1 targeting, leading to a subsequent increase of pMLC levels with implications for cervical cell viability, migration and invasion.
Assuntos
Técnicas de Cultura de Células , Movimento Celular , MicroRNAs/metabolismo , Fosfatase de Miosina-de-Cadeia-Leve/metabolismo , Sequência de Bases , Movimento Celular/genética , Sobrevivência Celular/genética , Técnicas de Silenciamento de Genes , Células HeLa , Humanos , Queratinócitos/metabolismo , MicroRNAs/genética , Cadeias Leves de Miosina/metabolismo , Invasividade Neoplásica , Fosforilação , RNA Interferente Pequeno/metabolismo , Esferoides Celulares/metabolismo , Ensaio Tumoral de Célula-TroncoRESUMO
Hypothyroxinemia (Hpx) is a thyroid hormone deficiency (THD) condition highly frequent during pregnancy, which although asymptomatic for the mother, it can impair the cognitive function of the offspring. Previous studies have shown that maternal hypothyroidism increases the severity of experimental autoimmune encephalomyelitis (EAE), an autoimmune disease model for multiple sclerosis (MS). Here, we analyzed the immune response after EAE induction in the adult offspring gestated in Hpx. Mice gestated in Hpx showed an early appearance of EAE symptoms and the increase of all parameters of the disease such as: the pathological score, spinal cord demyelination, and immune cell infiltration in comparison to the adult offspring gestated in euthyroidism. Isolated CD4+CD25+ T cells from spleen of the offspring gestated in Hpx that suffer EAE showed reduced capacity to suppress proliferation of effector T cells (TEff) after being stimulated with anti-CD3 and anti-CD28 antibodies. Moreover, adoptive transfer experiments of CD4+CD25+ T cells from the offspring gestated in Hpx suffering EAE to mice that were induced with EAE showed that the receptor mice suffer more intense EAE pathological score. Even though, no significant differences were detected in the frequency of Treg cells and IL-10 content in the blood, spleen, and brain between mice gestated in Hpx or euthyroidism, T cells CD4+CD25+ from spleen have reduced capacity to differentiate in vitro to Treg and to produce IL-10. Thus, our data support the notion that maternal Hpx can imprint the immune response of the offspring suffering EAE probably due to a reduced capacity to trigger suppression. Such "imprints" on the immune system could contribute to explaining as to why adult offspring gestated in Hpx suffer earlier and more intense EAE.
Assuntos
Encefalomielite Autoimune Experimental/etiologia , Encefalomielite Autoimune Experimental/metabolismo , Hipotireoidismo/complicações , Exposição Materna/efeitos adversos , Complicações na Gravidez , Efeitos Tardios da Exposição Pré-Natal , Transferência Adotiva , Animais , Biomarcadores , Diferenciação Celular , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/patologia , Encefalomielite Autoimune Experimental/terapia , Feminino , Hipotireoidismo/sangue , Hipotireoidismo/etiologia , Imunofenotipagem , Ativação Linfocitária , Contagem de Linfócitos , Metimazol/administração & dosagem , Metimazol/efeitos adversos , Camundongos , Proteína Básica da Mielina/metabolismo , Fenótipo , Gravidez , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Tireotropina/sangue , Tiroxina/sangueRESUMO
BACKGROUND: The let-7 microRNAs (miRNAs) are frequently dysregulated in carcinogenic processes, including cervical cancer. LIN28 proteins regulate let-7 biogenesis by binding to conserved sequences within the pre-miRNA structure. Nevertheless, recent research has shown that some let-7 miRNAs may escape LIN28 regulation. OBJECTIVE: Correlate pre-let-7 miRNAs and LIN28B levels in cervical cell lines with different malignancy and HPV content. METHODS: Pre-let-7 levels were determined by RTqPCR. LIN28B and other let-7 targets were analyzed by immunoblot. In silico tools were used to correlate let-7 and LIN28B expression and to analyze prelet- 7 sequences and structures. RESULTS: Lin28B protein was detected in all tested cell lines although it was more expressed in tumor cell lines. High levels of pre-let-7c/f-1 and pre-miR-98 were present in almost all cell lines regardless malignancy and LIN28B expression. Pre-let-7g/i were mainly expressed in tumor cell lines, pre-let-7e and pre-let-7-a3 were absent in all cell lines and pre-let-7a-2 showed indistinct expression. LIN28B showed positive correlation with pre-let-7i/g/f-1 and pre-miR-98 in tumor cell lines, suggesting escape from regulation. Sequence alignment and analysis of pre-let-7 miRNAs showed distinctive structural features within the preE region that may influence the ideal pre-let-7 structuring for LIN28B interaction. Short preE-stems were present in pre-let-7 that may escape LIN28B regulation, but long preEstems were mostly associated with high-level pre-let-7 miRNAs. CONCLUSION: The observed differences of pre-let-7 levels in cervical cell lines may be the result of alternative preE structuring affecting interaction with LIN28B thus resulting in differential let-7 regulation.
Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Proteínas de Ligação a RNA/genética , Neoplasias do Colo do Útero/genética , Pareamento de Bases , Sequência de Bases , Sequência Conservada , Feminino , Humanos , MicroRNAs/metabolismo , Proteínas de Ligação a RNA/metabolismo , Alinhamento de Sequência , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: Aberrant miRNA expression is associated with the development of several diseases including cervical cancer. Dysregulation of miR-125a-5p is present in a plethora of tumors, but its role in cervical cancer is not well understood. OBJECTIVE: The aim was to analyze the expression profile of miR-125a-5p in tumor and immortal cell lines with further target prediction, validation and function analysis. METHODS: MiR-125a-5p expression was determined by real-time RT-PCR from nine cervical cell lines. In silico tools were used to find target transcripts with an miR-125-5p complementary site within the 3'UTR region. Further target selection was based on gene ontology annotation and ΔG analysis. Target validation was performed by transfection of synthetic miR-125a-5p mimics and luciferase assays. Functional evaluation of miR-125a-5p on migration was performed by transwell migration assays. RESULTS: Differential miR-125a-5p expression was observed between immortal and tumor cells regardless of the human papillomavirus (HPV) content. Thermodynamic and ontological analyses showed Microtubule-Affinity-Regulating Kinase1 (MARK1) as a putative target for miR-125a-5p. An inverse correlation was observed among miR-125a-5p expression and MARK1 protein levels in tumor but not in immortal cells. Luciferase assays showed direct miR-125a-5p regulation over MARK1 through recognition of a predicted target site within the 3'-UTR. HeLa and C-33A cervical tumor cells enhanced migration after transfection with miR-125a-5p mimics and stimulation of cell migration was reproduced by siRNA-mediated inhibition of MARK1. CONCLUSION: The results showed MARK1 as a novel functional target for miR-125a-5p with implications on cell migration of tumor cervical cancer cells.
Assuntos
Movimento Celular , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Proteínas Serina-Treonina Quinases/metabolismo , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Regiões 3' não Traduzidas , Proliferação de Células , Feminino , Humanos , Proteínas Serina-Treonina Quinases/genética , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/metabolismoRESUMO
Resumen: Introducción: El síndrome de la cimitarra consiste en una rara malformación, caracterizada por una conexión anómala parcial de una o ambas venas pulmonares derechas a la vena cava inferior, hipoplasia de pulmón derecho y circulación sistémica desde la aorta descendente. El síndrome de la cimitarra en ocasiones se asocia con otras malformaciones congénitas, entre las que se incluye la persistencia del conducto arterioso (PCA). Caso clínico: Paciente de sexo femenino de cuatro años de edad con síndrome de la cimitarra, variedad ''adulto'', asociado con PCA sintomático. Se realizó exitosamente oclusión del conducto mediante un asa arteriovenosa femoral con un dispositivo Amplatzer® PDA, sin complicaciones. Conclusiones: El manejo del síndrome de la cimitarra es complejo y amerita de un estudio hemodinámico completo para determinar el tratamiento adecuado. La hipertensión arterial pulmonar es un factor de mal pronóstico.
Abstract: Background: Scimitar syndrome consists in a rare malformation characterized by a partial abnormal connection in one or both right pulmonary veins to the inferior vena cava, right lung hypoplasia and systemic circulation from the descending aorta. Scimitar syndrome is occasionally associated to other congenital malformations, such as patent ductus arteriosus (PDA). Case report: We report a 4-year-old patient with ''adult'' variety of scimitar syndrome associated to symptomatic PDA, which was successfully occluded using retrograde guidewire-established femoral arteriovenous loop with an Amplatzer® PDA occluder, without complications. Conclusions: Scimitar syndrome is complex and requires a complete hemodynamic study for the determination of the appropriate treatment. Pulmonary arterial hypertension is a factor associated with poor prognosis.
Assuntos
Pré-Escolar , Feminino , Humanos , Síndrome de Cimitarra/cirurgia , Derivação Arteriovenosa Cirúrgica/métodos , Permeabilidade do Canal Arterial/cirurgia , Síndrome de Cimitarra/fisiopatologia , Derivação Arteriovenosa Cirúrgica/instrumentação , Permeabilidade do Canal Arterial/patologia , Dispositivo para Oclusão SeptalRESUMO
BACKGROUND: Scimitar syndrome consists in a rare malformation characterized by a partial abnormal connection in one or both right pulmonary veins to the inferior vena cava, right lung hypoplasia and systemic circulation from the descending aorta. Scimitar syndrome is occasionally associated to other congenital malformations, such as patent ductus arteriosus (PDA). CASE REPORT: We report a 4-year-old patient with "adult" variety of scimitar syndrome associated to symptomatic PDA, which was successfully occluded using retrograde guidewire-established femoral arteriovenous loop with an Amplatzer® PDA occluder, without complications. CONCLUSIONS: Scimitar syndrome is complex and requires a complete hemodynamic study for the determination of the appropriate treatment. Pulmonary arterial hypertension is a factor associated with poor prognosis.