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1.
Sci Rep ; 10(1): 3171, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32081911

RESUMO

With the advent of personalized medicine, there is a movement to develop "smaller" and "smarter" microdevices that are able to distinguish similar cancer subtypes. Tumor cells display major differences when compared to their natural counterparts, due to alterations in fundamental cellular processes such as glycosylation. Glycans are involved in tumor cell biology and they have been considered to be suitable cancer biomarkers. Thus, more selective cancer screening assays can be developed through the detection of specific altered glycans on the surface of circulating cancer cells. Currently, this is only possible through time-consuming assays. In this work, we propose the "intelligent" Lab on Fiber (iLoF) device, that has a high-resolution, and which is a fast and portable method for tumor single-cell type identification and isolation. We apply an Artificial Intelligence approach to the back-scattered signal arising from a trapped cell by a micro-lensed optical fiber. As a proof of concept, we show that iLoF is able to discriminate two human cancer cell models sharing the same genetic background but displaying a different surface glycosylation profile with an accuracy above 90% and a speed rate of 2.3 seconds. We envision the incorporation of the iLoF in an easy-to-operate microchip for cancer identification, which would allow further biological characterization of the captured circulating live cells.

2.
Front Immunol ; 10: 2767, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31866996

RESUMO

Candida albicans causes systemic life-threatening infections, particularly in immunocompromised individuals, such as patients in intensive care units, patients undergoing chemotherapy, and post-surgical and neutropenic patients. The proliferation of invading Candida cells is mainly limited by the action of the human innate immune system, in which phagocytic cells play a fundamental role. This function is, however, limited in neutropenic patients, who rely mainly on the protective immunity mediated by macrophages. Macrophages have been shown to release extracellular DNA fibers, known as macrophage extracellular traps (METs), which can entrap and kill various microbes by a process called ETosis. In this study, we observed that, upon contact with C. albicans, macrophages became active in phagocyting and engulfing yeast cells. ETosis was induced in 6% of macrophages within the first 30 min of contact, and this percentage increased with the multiplicity of infection until a plateau was reached. After 2.5 h incubation, the presence of extracellular macrophage DNA was observed in approximately half of the cells. This study suggests that the formation of METs occurs before pyroptosis (first 6-8 h) and macrophage cell death (up to 24 h), and thus, METs could be included in models describing C. albicans-macrophage interactions. We also observed that macrophage ETosis and phagocytosis can occur simultaneously and that, in the first hours of infection, both processes are similarly important in controlling the proliferation of yeast cells, this being critical in neutropenic patients. Finally, it can also be concluded that, since C. albicans can degrade DNA, the structural component of METs, yeast extracellular DNase activity can be considered as an important virulence factor.

3.
Pathology ; 51(6): 593-599, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31466863

RESUMO

An association of well-differentiated gastroenteropancreatic neuroendocrine tumours (WD GEP NETs) with metabolic syndrome (MetS) was recently described. Yet no molecular mechanisms linking the two conditions are known. This study's aim was to identify putative molecular signatures linking WD GEP NETs and MetS to gain further insight into potential mechanisms for this association. Patients with WD GEP NETs (n=39), pancreatic (panNET) and gastro-intestinal (GI-NET), were clinically evaluated for presence of MetS. WD GEP NETs immunohistochemistry staining for Forkhead box protein M1 (FOXM1), insulin growth factor 1 receptor (IGF1R), Ki-67 and interleukin 6 (IL-6) was performed and quantified by computerised morphometric analysis. FOXM1, Ki-67, IGF1R or IL-6 expression in WD GEP NETs was not influenced by the presence of MetS. IL-6 peritumoural expression was higher in GI-NETs of patients with low HDL cholesterol (0.018±0.005% vs 0.030±0.005%, p=0.02). In GI-NETs, a higher IL-6 expression was also associated with disease progression (0.026±0.004% vs 0.016±0.002%, p=0.03). In WD GEP-NETs, MetS did not influence FOXM1, IGF1R and IL-6 expression. In GI-NETs, IL-6 expression was influenced by the MetS feature low HDL, and positively associated with disease progression. These data suggest that local and systemic inflammatory status can potentially modulate GI-NET behaviour.

4.
Int J Nanomedicine ; 14: 2349-2369, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31040661

RESUMO

Background: In view of the growing importance of nanotechnologies, the detection/identification of nanoparticles type has been considered of utmost importance. Although the characterization of synthetic/organic nanoparticles is currently considered a priority (eg, drug delivery devices, nanotextiles, theranostic nanoparticles), there are many examples of "naturally" generated nanostructures - for example, extracellular vesicles (EVs), lipoproteins, and virus - that provide useful information about human physiology or clinical conditions. For example, the detection of tumor-related exosomes, a specific type of EVs, in circulating fluids has been contributing to the diagnosis of cancer in an early stage. However, scientists have struggled to find a simple, fast, and low-cost method to accurately detect/identify these nanoparticles, since the majority of them have diameters between 100 and 150 nm, thus being far below the diffraction limit. Methods: This study investigated if, by projecting the information provided from short-term portions of the back-scattered laser light signal collected by a polymeric lensed optical fiber tip dipped into a solution of synthetic nanoparticles into a lower features dimensional space, a discriminant function is able to correctly detect the presence of 100 nm synthetic nanoparticles in distilled water, in different concentration values. Results and discussion: This technique ensured an optimal performance (100% accuracy) in detecting nanoparticles for a concentration above or equal to 3.89 µg/mL (8.74E+10 particles/mL), and a performance of 90% for concentrations below this value and higher than 1.22E-03 µg/mL (2.74E+07 particles/mL), values that are compatible with human plasmatic levels of tumor-derived and other types of EVs, as well as lipoproteins currently used as potential biomarkers of cardiovascular diseases. Conclusion: The proposed technique is able to detect synthetic nanoparticles whose dimensions are similar to EVs and other "clinically" relevant nanostructures, and in concentrations equivalent to the majority of cell-derived, platelet-derived EVs and lipoproteins physiological levels. This study can, therefore, provide valuable insights towards the future development of a device for EVs and other biological nanoparticles detection with innovative characteristics.


Assuntos
Tecnologia Biomédica/métodos , Técnicas Biossensoriais/métodos , Nanopartículas/química , Fibras Ópticas , Análise Discriminante , Exossomos/química , Vesículas Extracelulares , Análise de Fourier , Humanos , Polímeros/química , Soluções
5.
Opt Express ; 27(6): 8092-8111, 2019 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-30894786

RESUMO

Stimulated emission depletion (STED) fluorescence microscopy squeezes an excited spot well below the wavelength scale using a doughnut-shaped depletion beam. To generate a doughnut, a scale-free vortex phase modulation (2D-STED) is often used because it provides maximal transverse confinement and radial-aberration immunity (RAI) to the central dip. However, RAI also means blindness to a defocus term, making the axial origin of fluorescence photons uncertain within the wavelength scale provided by the confocal detection pinhole. Here, to reduce the uncertainty, we perturb the 2D-STED phase mask so as to change the sign of the axial concavity near focus, creating a dilated dip. By providing laser depletion power, the dip can be compressed back in three dimensions to retrieve lateral resolution, now at a significantly higher contrast. We test this coherent-hybrid STED (CH-STED) mode in x-y imaging of complex biological structures, such as the dividing cell. The proposed strategy creates an orthogonal direction in the STED parametric space that uniquely allows independent tuning of resolution and contrast using a single depletion beam in a conventional (circular polarization-based) STED setup.

6.
Front Microbiol ; 10: 284, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30858833

RESUMO

The pathogenic clade of the Sporothrix genus comprises the etiological agents of sporotrichosis, a worldwide emergent disease. Despite the growing understanding of their successful pathogen traits, there is little information on genome sizes and ploidy within the genus. Therefore, in this work, we evaluated the ploidy of four species of the Sporothrix genus, specifically Sporothrix brasiliensis, Sporothrix schenckii, Sporothrix globosa, and Sporothrix pallida. Through cell cycle analysis of the yeast-phase cells, we showed that the DNA content of G0/G1 cells was similar to the genome size determined by whole genome sequencing. Moreover, ploidy of S. schenckii, S. brasiliensis, and S. pallida that was determined by allele composition using next-generation sequencing (NGS) data is consistent with monomorphic positions at each allele. These data show that the analyzed strains of Sporothrix are haploid, or at least aneuploid, thereby laying the foundation for the development of a molecular toolbox for Sporothrix spp.

7.
Child Abuse Negl ; 83: 74-82, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30025306

RESUMO

Adolescence is a vulnerable period for mental health problems. Although child abuse and neglect (CAN) are known risk factors for some of them, it is not clear if the negative consequences on mental health also occur in families where CAN and a warm parent-child relationship coexist. The aim of this study is to explore this gap and investigate the effects of different types of CAN according to levels of warmth in the parent-child relationship on common mental disorders (CMD) in adolescence. This is a cross-sectional study encompassing 487 adolescents attending the ninth grade at 2 public and 4 private schools in Rio de Janeiro, Brazil. CAN was measured by the Child Trauma Questionnaire (CTQ), CMD by the General Health Questionnaire (GHQ-12) and the level of warmth in the parent-child relationship were recalled using short Egna Minnen Betraffände Uppfostran (s-EMBU-23). The adjusted separate effects of different types of CAN and a low level of warmth in the parent-child relationship, and both in tandem, were estimated using multivariate linear regression models. Results indicated that emotional abuse and neglect, physical abuse and neglect, and a low level of warmth in the parent-child relationship are important risk factors for CMD in adolescence. Nevertheless, in families where CAN coexist with a warm and affectionate parent-child relationship, the negative effects of CAN on mental health are attenuated. Evidence indicates that actions to prevent or interrupt CAN and improving parental practices could be effective strategies to reduce CMD in adolescence.


Assuntos
Maus-Tratos Infantis/psicologia , Emoções , Transtornos Mentais/etiologia , Relações Pais-Filho , Adolescente , Brasil , Criança , Estudos Transversais , Feminino , Humanos , Masculino , Saúde Mental , Pais/psicologia , Abuso Físico/psicologia , Fatores de Risco , Instituições Acadêmicas , Inquéritos e Questionários
8.
Front Microbiol ; 9: 1127, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29896184

RESUMO

Candida albicans is the main causative agent of candidiasis and one of the most frequent causes of nosocomial infections worldwide. In order to establish an infection, this pathogen supports effective stress responses to counter host defenses and adapts to changes in the availability of important nutrients, such as alternative carbon sources. These stress responses have clear implications on the composition and structure of Candida cell wall. Therefore, we studied the impact of lactate, a physiologically relevant carbon source, on the activity of C. albicans RLM1 transcriptional factor. RLM1 is involved in the cell wall integrity pathway and plays an important role in regulating the flow of carbohydrates into cell wall biosynthesis pathways. The role of C. albicans RLM1 in response to lactate adaptation was assessed in respect to several virulence factors, such as the ability to grow under cell wall damaging agents, filament, adhere or form biofilm, as well as to immune recognition. The data showed that growth of C. albicans cells in the presence of lactate induces the secretion of tartaric acid, which has the potential to modulate the TCA cycle on both the yeast and the host cells. In addition, we found that adaptation of C. albicans cells to lactate reduces their internalization by immune cells and consequent % of killing, which could be correlated with a lower exposure of the cell wall ß-glucans. In addition, absence of RLM1 has a minor impact on internalization, compared with the wild-type and complemented strains, but it reduces the higher efficiency of lactate grown cells at damaging phagocytic cells and induces a high amount of IL-10, rendering these cells more tolerable to the immune system. The data suggests that RLM1 mediates cell wall remodeling during carbon adaptation, impacting their interaction with immune cells.

9.
Glia ; 66(9): 1826-1844, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29732611

RESUMO

During central nervous system development, oligodendrocytes form structurally and functionally distinct actin-rich protrusions that contact and wrap around axons to assemble myelin sheaths. Establishment of axonal contact is a limiting step in myelination that relies on the oligodendrocyte's ability to locally coordinate cytoskeletal rearrangements with myelin production, under the control of a transcriptional differentiation program. The molecules that provide fine-tuning of actin dynamics during oligodendrocyte differentiation and axon ensheathment remain largely unidentified. We performed transcriptomics analysis of soma and protrusion fractions from rat brain oligodendrocyte progenitors and found a subcellular enrichment of mRNAs in newly-formed protrusions. Approximately 30% of protrusion-enriched transcripts encode proteins related to cytoskeleton dynamics, including the junction mediating and regulatory protein Jmy, a multifunctional regulator of actin polymerization. Here, we show that expression of Jmy is upregulated during myelination and is required for the assembly of actin filaments and protrusion formation during oligodendrocyte differentiation. Quantitative morphodynamics analysis of live oligodendrocytes showed that differentiation is driven by a stereotypical actin network-dependent "cellular shaping" program. Disruption of actin dynamics via knockdown of Jmy leads to a program fail resulting in oligodendrocytes that do not acquire an arborized morphology and are less efficient in contacting neurites and forming myelin wraps in co-cultures with neurons. Our findings provide new mechanistic insight into the relationship between cell shape dynamics and differentiation in development.


Assuntos
Citoesqueleto de Actina/metabolismo , Diferenciação Celular/fisiologia , Proteínas Nucleares/metabolismo , Oligodendroglia/citologia , Oligodendroglia/metabolismo , Transativadores/metabolismo , Transcriptoma , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Técnicas de Cocultura , Regulação da Expressão Gênica , Neurônios/citologia , Neurônios/metabolismo , RNA Mensageiro/metabolismo , Ratos Wistar
10.
Med Mycol ; 56(5): 649-651, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29420774

RESUMO

Previous studies have correlated the severity of recurrent vulvovaginal Candida infections (VVC) and balanitis in patients from China with the presence of some dominant genotypes at the ORF RLM1. Here we tested VVC vs non-VVC isolates from Portugal, Brazil and Greece and, although the same genotypes were identified in VVC isolates, they were present in only five out of 150 strains. However, this analysis showed that VVC isolates presented a higher percentage of genotypes with similar high molecular weight alleles, in comparison with strains isolated from other biological sources.


Assuntos
Alelos , Candida albicans/genética , Candidíase Vulvovaginal/microbiologia , Variação Genética , Fatores de Transcrição/genética , Brasil , Candida albicans/classificação , Candida albicans/isolamento & purificação , Candidíase/microbiologia , DNA Fúngico/genética , Feminino , Proteínas Fúngicas/genética , Frequência do Gene , Estudos de Associação Genética , Genótipo , Grécia , Humanos , Repetições de Microssatélites , Peso Molecular , Portugal
11.
FEMS Yeast Res ; 17(6)2017 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-28910984

RESUMO

Succinic acid is a platform chemical that plays an important role as precursor for the synthesis of many valuable bio-based chemicals. Its microbial production from renewable resources has seen great developments, specially exploring the use of yeasts to overcome the limitations of using bacteria. The objective of the present work was to screen for succinate-producing isolates, using a yeast collection with different origins and characteristics. Four strains were chosen, two as promising succinic acid producers, in comparison with two low producers. Genome of these isolates was analysed, and differences were found mainly in genes SDH1, SDH3, MDH1 and the transcription factor HAP4, regarding the number of single nucleotide polymorphisms and the gene copy-number profile. Real-time PCR was used to study gene expression of 10 selected genes involved in the metabolic pathway of succinic acid production. Results show that for the non-producing strain, higher expression of genes SDH1, SDH2, ADH1, ADH3, IDH1 and HAP4 was detected, together with lower expression of ADR1 transcription factor, in comparison with the best producer strain. This is the first study showing the capacity of natural yeast isolates to produce high amounts of succinic acid, together with the understanding of the key factors associated, giving clues for strain improvement.


Assuntos
Perfilação da Expressão Gênica , Genômica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Ácido Succínico/metabolismo , Dosagem de Genes , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Proteínas de Saccharomyces cerevisiae/genética
12.
BMC Res Notes ; 10(1): 416, 2017 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-28821265

RESUMO

BACKGROUND: Poncet's disease is a rare syndrome characterized by articular impairment in a form of rare tuberculid. One of the theories of its cause involves an autoimmune response induced by the intravesical administration of the Calmette-Guerin Bacillus or the treatment of bladder carcinoma. Furthermore, there may be an appearance of oligoarticular or polyarticular arthritis, beginning 1-3 months after the start of therapy. Few physicians know the disease and the literature related to that syndrome is scarce and restricted to case reports, which contributes to its under diagnosis. CASE PRESENTATION: Female patient, 64 years old, Caucasian, in whom was noticed firstly dark urine, without haematuria or dysuria. Later felt also colic pain in the hypogastric region. Microscopically, the conclusive diagnosis was a high grade non-invasive papillary urothelial carcinoma. Thereupon, the treatment of the tumour began with transurethral resection technique and intravesical instillation of Calmette-Guérin Bacillus as adjuvant treatment. Eight months after the beginning of treatment, the lingering presence of the carcinoma was identified. Nevertheless, arthritis was identified through radiographs, after an increase in the clavicle capitation, right knee and left ankle in bone scintigraphy. Coinciding with the joint manifestations, the patient developed fever and purulent urethral discharge (culture was negative). Therefore, trying to investigate the cause of the arthritis, Purified Protein Derivate was taken, with reactive results. An increase of acute phase reactants was found, with other tests resulting normal: blood chemistry, Complete Blood Count, immunology and serology. Human Leukocyte Antigen typing by polymerase chain reaction revealed the presence of A24/AX, B44, B27, BW4/BW4, DQ7 and DQ5. Consequently, Poncet's disease was the diagnostic conclusion. The treatment with intravesical Calmette-Guérin Bacillus was immediately discontinued. The patient received corticosteroids associated with etoricoxib and isoniazid for 4 months, achieving disappearance of the inflammatory joint signs in 3 months. After 6 months, no joint pain recurrence or other manifestations suggesting active disease had been seen. CONCLUSIONS: Therefore, such diagnosis should be considered when confronted with an osteoarticular clinical picture in patients treated with intravesical Calmette-Guérin Bacillus, especially patients with HLA-B27 (+) and B7 (+), as Poncet's disease is a reactive arthritis.


Assuntos
Adjuvantes Imunológicos/efeitos adversos , Artrite Reativa/tratamento farmacológico , Carcinoma Papilar/diagnóstico , Tuberculose/tratamento farmacológico , Neoplasias da Bexiga Urinária/diagnóstico , Adjuvantes Imunológicos/administração & dosagem , Administração Intravesical , Corticosteroides/uso terapêutico , Artrite Reativa/induzido quimicamente , Artrite Reativa/diagnóstico , Carcinoma Papilar/patologia , Carcinoma Papilar/cirurgia , Etoricoxib , Feminino , Humanos , Isoniazida/uso terapêutico , Pessoa de Meia-Idade , Mycobacterium bovis/química , Mycobacterium bovis/imunologia , Piridinas/uso terapêutico , Sulfonas/uso terapêutico , Resultado do Tratamento , Tuberculose/induzido quimicamente , Tuberculose/diagnóstico , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/cirurgia
14.
J Mech Behav Biomed Mater ; 75: 41-49, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28692844

RESUMO

OBJECTIVES: The purpose of this in vitro study was to evaluate the bonding ability and monomer conversion of a universal adhesive system applied to dentin as functions of different curing times and storage. The results were compared among a variety of commercial adhesives. MATERIALS AND METHODS: Flat superficial dentin surfaces were exposed on human molars and assigned into one of the following adhesives (n = 15): total-etch Adper Single Bond 2 (SB) and Optibond Solo Plus (OS), self-etch Optibond All in One (OA) and Clearfil SE Bond (CSE), and Scotchbond Universal Adhesive in self-etch mode (SU). The adhesives were applied following the manufacturers' instructions and cured for 10, 20, or 40s. Specimens were processed for the microtensile bond strength (µTBS) test in accordance with the non-trimming technique and tested after 24h and 2 years. The fractured specimens were classified under scanning electron microscopy (SEM). Infrared (IR) spectra were obtained and monomer conversion (%) was calculated by comparing the aliphatic-to-aromatic IR absorption peak ratio before and after polymerization (n=5). Data were analyzed by 2-way ANOVA/Tukey's tests (α = 0.05). RESULTS: At 24-h evaluation, OA and CSE presented similar bond strength means irrespective of the curing time, whereas SB and SU exhibited significantly higher means when cured for 40s as did OS when cured for 20 or 40s (p < 0.05). At 2-year evaluation, only OA exhibited significantly higher bond strength when cured for 20 and 40s (p < 0.05). When the evaluation times were compared, OA also exhibited the same bonding ability when cured for longer periods of time (20 and 40s). All of the adhesives tested exhibited significantly lower monomer conversion when photoactivated according to the manufacturers' instructions (10s). CONCLUSIONS: Higher monomer conversions obtained with longer light exposure allow only higher immediate bond strength for most of the adhesives tested. After 2-year storage, only the self-etching adhesive Optibond All-In-One exhibited the same bonding ability when cured for longer periods of time.


Assuntos
Colagem Dentária , Adesivos Dentinários , Cura Luminosa de Adesivos Dentários , Teste de Materiais , Ataque Ácido Dentário , Dentina , Humanos , Resistência à Tração
15.
Nanomedicine (Lond) ; 11(24): 3205-3221, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27830593

RESUMO

AIM: Propose a nanoparticle for neuron-targeted retrograde gene delivery and describe a microfluidic-based culture system to provide insight into vector performance and safety. METHODS: Using compartmentalized neuron cultures we dissected nanoparticle bioactivity upon delivery taking advantage of (quantitative) bioimaging tools. RESULTS: Targeted and nontargeted nanoparticles were internalized at axon terminals and retrogradely transported to cell bodies at similar average velocities but the former have shown an axonal flux 2.7-times superior to nontargeted nanoparticles, suggesting an improved cargo-transportation efficiency. The peripheral administration of nanoparticles to axon terminals is nontoxic as compared with their direct administration to the cell body or whole neuron. CONCLUSION: A neuron-targeted nanoparticle system was put forward. Microfluidic-based neuron cultures are proposed as a powerful tool to investigate nanoparticle bio-performance.


Assuntos
Embrião de Mamíferos/metabolismo , Cadeias Pesadas de Imunoglobulinas/genética , Microfluídica/métodos , Nanopartículas/administração & dosagem , Neurônios/metabolismo , Plasmídeos/administração & dosagem , Animais , Axônios/metabolismo , Células Cultivadas , Quitosana/química , Embrião de Mamíferos/citologia , Embrião de Mamíferos/efeitos dos fármacos , Técnicas de Transferência de Genes , Terapia Genética , Nanopartículas/química , Neurônios/citologia , Neurônios/efeitos dos fármacos , Plasmídeos/genética , Ratos , Ratos Wistar
16.
Cad Saude Publica ; 32(8): e00179915, 2016 Aug 08.
Artigo em Português | MEDLINE | ID: mdl-27509553

RESUMO

The aim of this study was to assess the dimensional structure of the s-EMBU, used to measure parental educational practices in adolescents. The sample included 487 students from Greater Metropolitan Rio de Janeiro, Brazil, in 2013. Mean age was 14 years, and 47% were girls. The original dimensional structure was assessed by confirmatory factor analysis (CFA). The study also applied exploratory structural equation modeling (ESEM). CFA adjustment was unsatisfactory. In light of the ESEM, the most parsimonious solution concerned the three-factor model (RMSEA = 0.03, CFI = 0.97, and TLI = 0.96), but various items in the overprotection dimension showed higher factor loads in the rejection dimension. The items in the rejection and emotional warmth dimensions proved more appropriate. In Brazil, s-EMBU partially captures the proposed dimensions for measuring parental educational practices in adolescents. Emotional warmth had its dimensional structure confirmed, but rejection and especially overprotection require further refinement.


Assuntos
Relações Pais-Filho , Poder Familiar , Inquéritos e Questionários , Adolescente , Brasil , Comparação Transcultural , Análise Fatorial , Feminino , Humanos , Masculino , Pais , Fatores Socioeconômicos , Estudantes
17.
Food Chem ; 211: 509-20, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27283661

RESUMO

During must fermentation by Saccharomyces cerevisiae strains thousands of volatile aroma compounds are formed. The objective of the present work was to adapt computational approaches to analyze pheno-metabolomic diversity of a S. cerevisiae strain collection with different origins. Phenotypic and genetic characterization together with individual must fermentations were performed, and metabolites relevant to aromatic profiles were determined. Experimental results were projected onto a common coordinates system, revealing 17 statistical-relevant multi-dimensional modules, combining sets of most-correlated features of noteworthy biological importance. The present method allowed, as a breakthrough, to combine genetic, phenotypic and metabolomic data, which has not been possible so far due to difficulties in comparing different types of data. Therefore, the proposed computational approach revealed as successful to shed light into the holistic characterization of S. cerevisiae pheno-metabolome in must fermentative conditions. This will allow the identification of combined relevant features with application in selection of good winemaking strains.


Assuntos
Biologia Computacional , Fermentação , Variação Genética , Metaboloma , Saccharomyces cerevisiae/metabolismo , Vinho/microbiologia , Manipulação de Alimentos , Saccharomyces cerevisiae/genética , Vinho/análise
18.
Acta Biomater ; 39: 133-145, 2016 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-27150234

RESUMO

UNLABELLED: We evaluated the potential of a liposomal antigen delivery system (ADS) containing Candida albicans cell wall surface proteins (CWSP) in mediating protection against systemic candidiasis. Treatment of bone-marrow-derived dendritic cells with CWSP-loaded dioctadecyldimethylammonium bromide:monoolein (DODAB:MO) liposomes enhanced and prolonged their activation comparatively to free antigen, indicating that liposome-entrapped CWSP were released more sustainable. Therefore, we immunized mice with CWSP either in a free form or loaded into two different DODAB:MO liposome formulations, respectively designated as ADS1 and ADS2, prior to intravenous C. albicans infection. Immunization with ADS1, but not with ADS2, conferred significant protection to infected mice, comparatively to immunization with CWSP or empty liposomes as control. ADS1-immunized mice presented significantly higher serum levels of C. albicans-specific antibodies that enhanced phagocytosis of this fungus. In these mice, a mixed cytokine production profile was observed encompassing IFN-γ, IL-4, IL-17A and IL-10. Nevertheless, only production of IL-4, IL-17 and IL-10 was higher than in controls. In this study we demonstrated that DODAB:MO liposomes enhance the immunogenicity of C. albicans antigens and host protection in a murine model of systemic candidiasis. Therefore, this liposomal adjuvant could be a promising candidate to assess in vaccination against this pathogenic fungus. STATEMENT OF SIGNIFICANCE: This work describes the immunomodulation capacity of the previously validated antigen delivery system (ADS) composed by dioctadecyldimethylammonium bromide (DODAB) and monoolein (MO) lipids incorporating the cell wall surface proteins (CWSP) from C. albicans. Here, we not only present the ability of this system in facilitating antigen uptake by DCs in vitro, but also that this system induces higher levels of pro-inflammatory cytokines and opsonizing specific IgG antibodies in serum of mice immunized subcutaneously. We show that the ADS are efficient nanocarrier and modulate the immune response against intravenous C. albicans infection favoring mouse protection. In sum, we show that the incorporation of C. albicans antigens in DODAB:MO nanocarries are a promising vaccine strategy against C. albicans fungal infection.


Assuntos
Antígenos de Fungos , Candida albicans/imunologia , Candidíase Invasiva/prevenção & controle , Proteínas Fúngicas , Vacinas Fúngicas , Glicerídeos , Imunização/métodos , Compostos de Amônio Quaternário , Animais , Antígenos de Fungos/química , Antígenos de Fungos/imunologia , Antígenos de Fungos/farmacologia , Candidíase Invasiva/imunologia , Feminino , Proteínas Fúngicas/química , Proteínas Fúngicas/imunologia , Proteínas Fúngicas/farmacologia , Vacinas Fúngicas/química , Vacinas Fúngicas/imunologia , Vacinas Fúngicas/farmacologia , Glicerídeos/química , Glicerídeos/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/farmacologia
19.
Clin Exp Optom ; 99(4): 336-41, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27146845

RESUMO

BACKGROUND: The purpose of this study was to investigate the effect of three months of soft contact lens wear on conjunctival goblet cell density and epithelial cell morphology. METHODS: This was a longitudinal clinical trial. Conjunctival impression cytology was performed on the superior palpebral conjunctiva in fifty-four eyes of twenty-seven neophyte contact lens wearers before and after three months of contact lens wear. Goblet cell density was determined by optical microscopy and epithelial cell morphology was classified according to the Tseng classification. Changes in goblet cell density as well as epithelial cell grading were determined. The effects of lens material and wearing modality on cytological changes were also investigated. RESULTS: Goblet cell density reduced significantly by 85 ± 151 cells/mm(2) (p < 0.001) after three months of contact lens wear. Reduction in goblet cell density was associated with lens materials; it was higher in conventional hydrogel lenses in comparison to silicone-hydrogel lenses (p = 0.008). The highest reduction in goblet cell density was found with Nelfilcon A lens wear (p = 0.002) and the lowest with Comfilcon A lens wear (p = 0.414). There was no statistically significant difference in grading of epithelial metaplasia before and after three months of contact lens wear (p = 0.075). Age was not correlated with the reduction in goblet cell density (r = -0.196, p = 0.160) but it was associated with the change in epithelial cell morphology (p = 0.036). CONCLUSION: Three months of soft contact lens wear statistically significantly reduced goblet cell density; however, no significant changes were found in the grading of epithelial metaplasia. Contact lenses with lower oxygen permeability, higher Young modulus and higher thickness highly affected the conjunctival cytology.


Assuntos
Túnica Conjuntiva/patologia , Lentes de Contato Hidrofílicas/efeitos adversos , Células Caliciformes/patologia , Adulto , Fatores Etários , Contagem de Células , Células Epiteliais/patologia , Feminino , Humanos , Estudos Longitudinais , Masculino , Projetos Piloto , Estudantes
20.
Cell Rep ; 15(3): 490-498, 2016 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-27068466

RESUMO

The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings.


Assuntos
Axônios/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Proteínas dos Microfilamentos/metabolismo , Animais , Transporte Axonal , Citoesqueleto/metabolismo , Cones de Crescimento/metabolismo , Hipocampo/patologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Degeneração Neural/metabolismo , Degeneração Neural/patologia
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