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Abstract Bioactive cements based on tricalcium silicate have been introducedto the market for use in dentistry, with a variety of clinical applications. These cements are in contact with vital tissues such as dental pulp or periodontium in cases of unintentional extrusion; thus, it is important to know the genotoxicity and cytoxicity of these materials. The objective of this study was to evaluate the cytotoxicity and genotoxicity of bioactive sealers, Bio-C® Sealer and MTA Repair HP®, in human fibroblasts. Discs of bioactive sealers Bio-C® Sealer, and MTA Repair HP®, were prepared and set for 24h under sterile conditions. The discs were placed in culture medium at 2.5mg/mL inside a SRT6D roller mixer (Stuart, UK) at 60rpm for 24h. The eluates obtained were incubated for 24h with previously activated and cultured ATCC cell line fibroblasts at 80% confluence. The cytotoxicity was evaluated by Alamar Blue® and LIVE/ DEAD assays, as well as the analysis of the Tunel and Mitotracker assays to evaluate genotoxicity using the confocal laser-scanning microscope. In the Alamar Blue® assay, the Bio-C® Sealer presented a cell proliferation of 87%, while the MTA Repair HP® Sealer was 72%. A statistically significant difference was found between the MTA Repair HP® Sealant and the negative control (p=<0.001). Regarding the genotoxicity tests, in the Tunel assay, both materials stain the nucleus of the fibroblast cells exposed to the eluates, while in the Mitotracker assay, the MTA Repair HP® Sealer showed greater mitochondrial function than the Bio-C® Sealer. Calcium silicate-based sealers, Bio-C® Sealer and MTA Repair HP®, are not cytotoxic and have low genotoxicity.
Resumen Los cementos bioactivos a base de silicato tricálcico se introdujeron en el mercado para uso en odontología, con una variedad de aplicaciones clínicas. Estos cementos pueden estar en contacto con tejidos como la pulpa dental o el periodonto, en caso de extrusión no intencionada. Por lo tanto, es importante conocer la genotoxicidad y la citoxicidad de estos materiales. El objetivo de este estudio fue evaluar la citotoxicidad y genotoxicidad de los selladores bioactivos Bio-C® Sealer y MTA Repair HP® en fibroblastos humanos. Se prepararon discos de selladores bioactivos Bio-C® Sealer y MTA Repair HP® y se colocaron durante 24h en condiciones de esterilidad. Los discos se colocaron en medio de cultivo a 2,5mg/mL dentro de un mezclador de rodillos SRT6D (Stuart, Reino Unido) a 60rpm durante 24h. Los eluidos obtenidos se incubaron durante 24h con fibroblastos de la línea celular ATCC previamente activados y cultivados al 80% de confluencia. La citotoxicidad se evaluó mediante ensayos Alamar Blue® y LIVE/DEAD; así como el análisis de los ensayos Tunnel y Mitotracker para evaluar la genotoxicidad, utilizando el microscopio confocal láser de barrido. En el ensayo Alamar Blue®, el Sellador Bio-C® presentó una proliferación celular del 87%, mientras que el sellador MTA Repair HP® fue del 72%. Se encontró una diferencia estadísticamente significativa entre el sellador MTA Repair HP® con respecto al control negativo (p=<0.001). En cuanto a las pruebas de genotoxicidad, en el ensayo Tunel, ambos materiales tiñen el núcleo de las células fibroblásticas expuestas a los eluidos, mientras que el ensayo Mitotracker, el sellador MTA Repair HP®, mostró una mayor función mitocondrial que el Bio-C® Sealer. Los selladores a base de silicato de calcio, Bio-C® Sealer y MTA Repair HP® no son citotóxicos y tienen una baja genotoxicidad.
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1. The objective of this study was to determine the nutritional and energy values of four maize distiller's dried grains with solubles (DDGS) and one maize high protein distiller's dried grains (HP-DDG) from ethanol production plants in Brazil; to evaluate the digestibility, performance, nitrogen balance and energy values for broiler chickens fed diets containing these coproducts (Experiment I); and to evaluate the effects of xylanase inclusion in diets containing maize DDGS for broilers on energy availability, digestibility, nitrogen balance and gastrointestinal morphometry (Experiment II).2. For each experiment, 180 broiler chickens aged 17 and 30 days with initial weights of 450 ± 18 g and 1228 ± 33 g, respectively, were used; the chickens were distributed into 36 metabolism cages. The experimental design consisted of complete randomised blocks, with six replications per treatment and five birds per experimental unit. The treatments consisted of a basal diet (BD) and five test diets containing maize ethanol coproducts (Experiment I) one BD and five test diets containing DDGS with inclusions of 0, 8,000, 16,000, 24,000 and 32,000 BXU/kg xylanase (Experiment II). In Experiment I, HP-DDG and DDGS2 presented higher AME and AMEn values (14.1 and 13.9 MJ/kg and 13.4 and 13.3 MJ/kg, respectively), than did the other coproducts (p < 0.05). Compared with DDGS1 and DDGS3, DDGS4 and HP-DDG had higher digestible CP values (p < 0.05). In Experiment II, the inclusion of the enzyme quadratically affected the values of digestible CP and digestible EE (p < 0.05), with the maximum values occurring with the inclusion of 18 750 and 22,170 BXU/kg of xylanase, respectively.3. The digestible NDF and digestible MM values linearly increased with the inclusion of xylanase (p < 0.05). The addition of xylanase had no effect on gastrointestinal morphometry (p > 0.05). It was concluded that the inclusion of between 18,000 and 22,000 BXU/kg of xylanase resulted in better digestible CP and digestible EE values.
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Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Galinhas , Dieta , Digestão , Endo-1,4-beta-Xilanases , Zea mays , Animais , Galinhas/fisiologia , Galinhas/metabolismo , Zea mays/química , Ração Animal/análise , Digestão/efeitos dos fármacos , Fenômenos Fisiológicos da Nutrição Animal/efeitos dos fármacos , Dieta/veterinária , Endo-1,4-beta-Xilanases/metabolismo , Endo-1,4-beta-Xilanases/administração & dosagem , Masculino , Distribuição Aleatória , Etanol , Valor Nutritivo , Trato Gastrointestinal/metabolismo , Suplementos Nutricionais/análiseRESUMO
In this paper, we presented a mathematical model for tuberculosis with treatment for latent tuberculosis cases and incorporated social implementations based on the impact they will have on tuberculosis incidence, cure, and recovery. We incorporated two variables containing the accumulated deaths and active cases into the model in order to study the incidence and mortality rate per year with the data reported by the model. Our objective is to study the impact of social program implementations and therapies on latent tuberculosis in particular the use of once-weekly isoniazid-rifapentine for 12 weeks (3HP). The computational experimentation was performed with data from Brazil and for model calibration, we used the Markov Chain Monte Carlo method (MCMC) with a Bayesian approach. We studied the effect of increasing the coverage of social programs, the Bolsa Familia Programme (BFP) and the Family Health Strategy (FHS) and the implementation of the 3HP as a substitution therapy for two rates of diagnosis and treatment of latent at 1% and 5%. Based of the data obtained by the model in the period 2023-2035, the FHS reported better results than BFP in the case of social implementations and 3HP with a higher rate of diagnosis and treatment of latent in the reduction of incidence and mortality rate and in cases and deaths avoided. With the objective of linking the social and biomedical implementations, we constructed two different scenarios with the rate of diagnosis and treatment. We verified with results reported by the model that with the social implementations studied and the 3HP with the highest rate of diagnosis and treatment of latent, the best results were obtained in comparison with the other independent and joint implementations. A reduction of the incidence by 36.54% with respect to the model with the current strategies and coverage was achieved, and a greater number of cases and deaths from tuberculosis was avoided.
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Antituberculosos , Teorema de Bayes , Isoniazida , Tuberculose Latente , Cadeias de Markov , Conceitos Matemáticos , Método de Monte Carlo , Rifampina , Humanos , Brasil/epidemiologia , Incidência , Isoniazida/administração & dosagem , Antituberculosos/administração & dosagem , Rifampina/administração & dosagem , Rifampina/análogos & derivados , Rifampina/uso terapêutico , Tuberculose Latente/epidemiologia , Tuberculose Latente/tratamento farmacológico , Tuberculose Latente/mortalidade , Modelos Biológicos , Tuberculose/mortalidade , Tuberculose/epidemiologia , Tuberculose/tratamento farmacológico , Simulação por ComputadorRESUMO
1. The purpose of this study was to determine the metabolisable energy of high-protein distiller's dried grains with solubles (HP-DDGS) for meat quail (Coturnix coturnix coturnix; Experiment I) and evaluate the effects of dietary levels of HP-DDGS on animal performance, carcase characteristics, meat quality, and organ weights (Experiment II).2. In Experiment 1, 96 meat quail were distributed in a completely randomised design with two treatments (reference or test diet) and six replicates of eight birds. The experimental period consisted of 5 d adaptation, followed by 5 d total excreta collection. The experimental diets consisted of a reference (control) and a test diet formulated with 800 g/kg reference diet and 200 g/kg HP-DDGS.3. In Experiment 2, 612 meat quail were distributed in a completely randomised design fed one of six dietary levels of HP-DDGS (0, 85, 170, 255, 340, or 425 g/kg) as a replacement for soybean meal. At 42 d of age, birds were slaughtered and evaluated for carcase yield, organ weights, and meat quality.4. Apparent metabolisable energy values corrected for nitrogen retention of HP-DDGS were 12.5 and 12.3 MJ/kg for males and females, respectively.5. In the starter phase (1-21 d of age), increasing dietary HP-DDGS levels had a quadratic effect on body weight (BW) (P = 0.020) and body weight gain (BWG) (P = 0.019). The maximum BW and BWG values were estimated to be achieved with 296.0 and 296.2 g/kg dietary HP-DDGS, respectively. Overall (1-42 d of age), increasing dietary HP-DDGS levels in replacement of soybean meal did not affect animal performance, carcase yield, meat quality or organ weight in meat quail.6. It was concluded that dietary HP-DDGS can fully replace soybean meal in meat quail diets without affecting growth performance, carcase yield, meat quality or organ weight.
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Coturnix , Codorniz , Masculino , Feminino , Animais , Farinha , Galinhas , Dieta/veterinária , Proteínas Alimentares , Carne , Glycine max , Peso Corporal , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Grão Comestível , Zea maysRESUMO
O tecido ósseo é um tecido naturalmente capaz de se autorregenerar. Entretanto, defeitos ósseos de maiores dimensões têm tal processo dificultado e, em situações de difícil reparação espontânea necessitam de auxílio de enxertos ou materiais apropriados para dar suporte à neoformação óssea. Hidrogéis são excelentes materiais a serem utilizados para esse fim, uma vez que são capazes de mimetizar a maioria das propriedades dos tecidos conjuntivos. Recentemente, despertou-se um interesse significativo no desenvolvimento de hidrogéis híbridos que pudessem ser utilizados em várias aplicações biomédicas. Uma diversidade de partículas é combinada a esse hidrogéis resultando em melhores propriedades. Dessa forma, este estudo tem como objetivo avaliar a influência de um scaffold contendo ácido hialurônico associado a um biovidro experimental de origem vegetal - Equisetum hyemale (BG-Carb), na regeneração de um defeito ósseo em maxila de camundongos. Nesse estudo preliminar, camundongos C57BL/6 (n = 3 por grupo e tempo experimental) (CEUA #042/2022) tiveram seu primeiro molar superior esquerdo extraído. Em seguida, uma ponta diamantada KGS #2128 (KG Soresen®), acoplada a um motor em baixa rotação (5000 rpm) foi utilizada para a realização de um defeito com a remoção do septo alveolar. Parte dos animais tiveram os defeitos preenchidos exclusivamente por HyStem®-HP (hidrogel à base de ácido hialurônico) (2 µL), enquanto a outra parte recebeu BG-Carb (1 µg) disperso em HyStem®-HP (2 µL). Após a eutanásia, as amostram foram processadas para análise histológica de rotina para histomorfometria, graduação do processo inflamatório, vascularização e neoformação óssea após 7 e 21 dias pós-cirúrgicos. Os resultados foram analisados considerando o nível de significância de 5% (p < 0,05). Aos 7 dias, o grupo que recebeu a associação BG-Carb e HyStem®-HP apresentou maiores valores numéricos de neoformação óssea, todavia sem diferença significativa em relação ao uso exclusivo de HyStem®-HP (controle). Aos 21 dias, os grupos HyStem®-HP e BG- Carb associado a HyStem®-HP apresentaram resultados semelhantes tanto na quantificação de tecido ósseo como de conjuntivo. O escore total para graduação para cada material também foi semelhante aos 7 e 21 dias. Apesar disso, comparando-se cada material nos tempos 7 e 21 dias, o grupo HyStem®-HP apresentou aumento significativo de parâmetros qualitativos e quantitativos, demonstrando que a associação HyStem®-HP + BG-Carb apresentou neoformação óssea inicial mais robusta. A quantificação epitelial da região da cirurgia aos 21 dias em ambos os grupos revelou aumento significativo da espessura, continuidade e uniformidade do epitélio nos grupos tratados com o a associação dos biomateriais em relação ao controle. Nas condições experimentais deste estudo, tanto o uso do HyStem®-HP como do HyStem®-HP + BG-Carb podem promover neoformação óssea. No entanto, a associação do hidrogel à base de ácido hialurônico com o BG-Carb parece promover neoformação óssea inicial mais consistente e estimular a proliferação da mucosa adjacente à cirurgia com maior espessura do tecido epitelial.
Bone tissue is a naturally regenerative tissue. However, larger bone defects have a difficult time undergoing this process and require the assistance of grafts or appropriate materials to support new bone formation. Hydrogels are excellent materials to use for this purpose since they can mimic most of the properties of connective tissues. Recently, there has been significant interest in the development of hybrid hydrogels that can be used in various biomedical applications. A variety of particles can be combined with these hydrogels, resulting in better properties. Thus, this study aims to evaluate the influence of a scaffold containing hyaluronic acid associated with an experimental plant-derived bioactive glass - Equisetum hyemale (BG-Carb), on the regeneration of a mouse maxillary bone defect. In this preliminary study, C57BL/6 mice (n = 3 per group and experimental time) (CEUA # 042/2022) had their upper left first molar extracted. Then, a KGS # 2128 diamond bur (KG Soresen®), attached to a low- speed motor (5000 rpm), was used to create a defect with removal of the alveolar septum. Some of the animals had defects filled exclusively with HyStem®-HP (hyaluronic acid-based hydrogel) (2 µL), while the other part received BG-Carb (1 µg) dispersed in HyStem®-HP (2 µL). After euthanasia, the samples were processed for routine histological analysis for histomorphometry, grading of the inflammatory process, vascularization, and new bone formation after 7 and 21 post-surgical days. The results were analyzed considering a significance level of 5% (p < 0.05). At 7 days, the group that received the BG-Carb and HyStem®-HP association showed higher numerical values of new bone formation, but without a significant difference compared to the exclusive use of HyStem®-HP (control). At 21 days, the HyStem®-HP and BG- Carb groups associated with HyStem®-HP showed similar results in both bone and connective tissue quantification. The total score for grading for each material was also similar at 7 and 21 days. However, comparing each material at 7 and 21 days, the HyStem®-HP group showed a significant increase in qualitative and quantitative parameters, demonstrating that the HyStem®-HP + BG-Carb association presented a more robust initial bone formation. The epithelial quantification of the surgery region at 21 days in both groups revealed a significant increase in thickness, continuity, and uniformity of the epithelium in the groups treated with the association of biomaterials compared to the control. Under the experimental conditions of this study, both the use of HyStem®-HP and HyStem®-HP + BG-Carb can promote bone formation. However, the association of the hyaluronic acid-based hydrogel with BG-Carb appears to promote a more consistent initial bone formation and stimulate the proliferation of the mucosa adjacent to the surgery with greater thickness of the epithelial tissue.
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Materiais Biocompatíveis , Regeneração Óssea , Hidrogéis , Ácido Hialurônico , MaxilaRESUMO
BACKGROUND: The high prevalence and persistence of Helicobacter pylori (H. pylori) infection, as well as the diversity of pathologies related to it, suggest that the virulence factors used by this microorganism are varied. Moreover, as its proteome contains 340 hypothetical proteins, it is important to investigate them to completely understand the mechanisms of its virulence and survival. We have previously reported that the hypothetical protein HP0953 is overexpressed during the first hours of adhesion to inert surfaces, under stress conditions, suggesting its role in the environmental survival of this bacterium and perhaps as a virulence factor. AIM: To investigate the expression and localization of HP0953 during adhesion to an inert surface and against gastric (AGS) cells. METHODS: Expression analysis was performed for HP0953 during H. pylori adhesion. HP0953 expression at 0, 3, 12, 24, and 48 h was evaluated and compared using the Kruskal-Wallis equality-of-populations rank test. Recombinant protein was produced and used to obtain polyclonal antibodies for immunolocalization. Immunogold technique was performed on bacterial sections during adherence to inert surfaces and AGS cells, which was analyzed by transmission electron microscopy. HP0953 protein sequence was analyzed to predict the presence of a signal peptide and transmembrane helices, both provided by the ExPASy platform, and using the GLYCOPP platform for glycosylation sites. Different programs, via, I-TASSER, RaptorX, and HHalign-Kbest, were used to perform three-dimensional modeling. RESULTS: HP0953 exhibited its maximum expression at 12 h of infection in gastric epithelium cells. Immunogold technique revealed HP0953 localization in the cytoplasm and accumulation in some peripheral areas of the bacterial body, with greater expression when it is close to AGS cells. Bioinformatics analysis revealed the presence of a signal peptide that interacts with the transmembrane region and then allows the release of the protein to the external environment. The programs also showed a similarity with the Tip-alpha protein of H. pylori. Tip-alpha is an exotoxin that penetrates cells and induces tumor necrosis factor alpha production, and HP0953 could have a similar function as posttranslational modification sites were found; modifications in turn require enzymes located in eukaryotic cells. Thus, to be functional, HP0953 may necessarily need to be translocated inside the cell where it can trigger different mechanisms producing cellular damage. CONCLUSION: The location of HP0953 around infected cells, the probable posttranslational modifications, and its similarity to an exotoxin suggest that this protein is a virulence factor.
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Infecções por Helicobacter , Helicobacter pylori , Proteínas de Bactérias/metabolismo , Células Epiteliais/metabolismo , Epitélio/metabolismo , Exotoxinas/metabolismo , Mucosa Gástrica/patologia , Infecções por Helicobacter/microbiologia , Humanos , Sinais Direcionadores de Proteínas , Proteoma/metabolismo , Proteínas Recombinantes/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fatores de Virulência/metabolismoRESUMO
Psoralen (PSO) and 5-methoxypsoralen (5-MOP) are widely used drugs in oral photochemotherapy against vitiligo and major bioactive components of root bark extract of Brosimum gaudichaudii Trécul (EBGT), previously standardized by LC-MS. However, the exceptionally low water solubility of these psoralens can cause incomplete and variable bioavailability limiting their applications and patient adherence to treatment. Therefore, the purpose of this work was to investigate the effects of 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) inclusion complex on the solubility and jejunal permeability of PSO and 5-MOP from EBGT. Characterization of inclusion complexes were evaluated by current methods in nuclear magnetic resonance studies on aqueous solution, Fourier transform infrared spectroscopy, thermal analysis, and scanning electron microscopy in solid state. Ex vivo rat jejunal permeability was also investigated and compared for both pure psoralens and plant extract formulation over a wide HP-ß-CD concentration range (2.5 to 70 mM). Phase solubility studies of the PSO- and 5-MOP-HP-ß-CD inclusion complex showed 1:1 inclusion complex formation with small stability constants (Kc < 500 M−1). PSO and 5-MOP permeability rate decreased after adding HP-ß-CD by 6- and 4-fold for pure standards and EBGT markers, respectively. Nevertheless, the complexation with HP-ß-CD significantly improved solubility of PSO (until 10-fold) and 5-MOP (until 31-fold). As a result, the permeability drop could be overcome by solubility augmentation, implying that the HP-ß-CD inclusion complexes with PSO, 5-MOP, or EBGT can be a valuable tool for designing and developing novel oral drug product formulation containing these psoralens for the treatment of vitiligo.
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Furocumarinas , Moraceae , Vitiligo , beta-Ciclodextrinas , 2-Hidroxipropil-beta-Ciclodextrina/química , Animais , Varredura Diferencial de Calorimetria , Permeabilidade , Extratos Vegetais/farmacologia , Ratos , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X , beta-Ciclodextrinas/químicaRESUMO
Herein were tested 7 hydrophobic-polar sequences in two types of 2D-square space lattices, homogeneous and correlated, the latter simulating molecular crowding included as a geometric boundary restriction. Optimization of 2D structures was carried out using a variant of Dill's model, inspired by convex function, taking into account both hydrophobic (Dill's model) and polar interactions, including more structural information to reach better folding solutions. While using correlated networks, degrees of freedom in the folding of sequences were limited; as a result in all cases, more successful structural trials were found in comparison to a homogeneous lattice. The majority of employed sequences were designed by our workgroup, two of them were folded with other approaches, and another is a modified version of a previous sequence, initial forms of the other two have been employed but without taking into account polar-polar contributions. Three of them are newly proposed, intended to test the conjoint hydrophobic-hydrophobic and polar-polar contributions in crowded spaces. One sequence turned out to be the most difficult of the seven folded, this perhaps due to intrinsic (i) degrees of freedom and (ii) motifs of the expected 2D HP structure. Meanwhile two-sequence, although optimal folding was not achieved for neither of the two approaches, folding with correlated network approach not only produced better results than homogeneous space, but for them the best values found with crowding were very close to the expected optimal fitness. In general, five sequences were better folded with medium lattice units for correlated media; instead, another two sequences were better folded with a bit larger degree of lattice unit, revealing that depending on the degrees of freedom and particular folding, motifs in each sequence would require tuned crowding to achieve better folding. Therefore, the main goal herein was to obtain a modified 2D HP lattice model to mimic folding of proteins or secondary structures, like ß-sheets, taking into account both hydrophobic-hydrophobic and polar-polar interactions, and fold them in a crowded environment. This simple but enough construction would be conducted to determine the needed information to fold sequences in a sort of a minimal but complete heuristic model. Finally, we claim that all folded sequences into crowded spaces achieve better results than homogeneous ones.
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Dobramento de Proteína , Proteínas , Simulação por Computador , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Conformação Proteica , Proteínas/químicaRESUMO
BACKGROUND: Chronic lymphocytic leukaemia (CLL) is a neoplasm of B-cells characterized by variable prognosis. Exploring the proteome of CLL cells may provide insights into the disease. Therefore, eleven proteomics experiments were conducted on eleven primary CLL samples. RESULTS: We reported a CLL proteome consisting of 919 proteins (false discovery rate (FDR) 1%) whose identification was based on the sequencing of two or more distinct peptides (FDR of peptide sequencing 1%). Mass spectrometry-based protein identification was validated for four different proteins using Western blotting and specific antibodies in different CLL samples. Small sizes of nucleolin (~57 kDa and ~68 kDa) showed a potential association with good prognosis CLL cells (n = 8, p < 0.01). Compared with normal B-cells, CLL cells over-expressed thyroid hormone receptor-associated protein 3 (THRAP3; n = 9; p = 0.00007), which is implicated in cell proliferation; and heterochromatin protein 1-binding protein 3 (HP1BP3; n = 10; p = 0.0002), which promotes cell survival and tumourogenesis. A smaller form of HP1BP3, which may correspond to HP1BP3 isoform-2, was specifically identified in normal B-cells (n = 10; p = 0.0001). HP1BP3 and THRAP3 predicted poor prognosis of CLL (p 0.05). Consistently, THRAP3 and HP1BP3 were found to be associated with cancer-related pathways (p 0.05). CONCLUSIONS: Our findings add to the known proteome of CLL and confirm the prognostic importance of two novel cancer-associated proteins in this disease.
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Leucemia Linfocítica Crônica de Células B , Biomarcadores Tumorais/análise , Espectrometria de Massas , Fatores de Transcrição/análise , Proteínas Nucleares/análise , Western Blotting , Cromatografia Líquida , Proteômica , Proteínas de Ligação a DNA/análiseRESUMO
OBJECTIVE: To improve the diagnostic efficiency of current tests for auditory processing disorders (APDs) by creating new test signals using digital filtering methods. METHODS: We conducted a prospective study from August 1, 2014, to August 31, 2019, using 3 low speech redundancy tests with novel test signals that we created with specially designed digital filters: the binaural resynthesis test and the low pass and high pass filtered speech tests. We validated and optimized these new tests, then applied them to healthy individuals across different age groups to examine how age affected performance and to children with APD before and after acoustically controlled auditory training (ACAT) to assess clinical improvement after treatment. RESULTS: We found a progressive increase in performance accuracy with less restrictive filters (P<.001) and with increasing age for all tests (P<.001). Our results suggest that binaural resynthesis and auditory closure mature at similar rates. We also demonstrate that the new tests can be used for the diagnosis of APD and for the monitoring of ACAT effects. Interestingly, we found that patients having the most severe deficits also benefited the most from ACAT (P<.001). CONCLUSION: We introduce a method that substantially improves current diagnostic tools for APD. In addition, we provide information on auditory processing maturation in normal development and validate that our method can detect APD-related deficits and ACAT-induced improvements in auditory processing.
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Epilepsy is a neurological disorder of genetic or environmental origin characterized by recurrent spontaneous seizures. A rodent model of temporal lobe epilepsy is induced by a single administration of pilocarpine, a non-selective cholinergic muscarinic receptor agonist. The molecular changes associated with pilocarpine-induced seizures are still poorly described. Epigenetic multiprotein complexes that regulate gene expression by changing the structure of chromatin impose transcriptional memories. Among the epigenetic enzymes relevant to the epileptogenic process is lysine-specific demethylase 1 (LSD1, KDM1A), which regulates the expression of genes that control neuronal excitability. LSD1 forms complexes with the CoREST family of transcriptional corepressors, which are molecular bridges that bring HDAC1/2 and LSD1 enzymes to deacetylate and demethylate the tail of nucleosomal histone H3. To test the hypothesis that LSD1-complexes are involved in initial modifications associated with pilocarpine-induced epilepsy, we studied the expression of main components of LSD1-complexes and the associated epigenetic marks on isolated neurons and the hippocampus of pilocarpine-treated mice. Using a single injection of 300 mg/kg of pilocarpine and after 24 h, we found that protein levels of LSD1, CoREST2, and HDAC1/2 increased, while CoREST1 decreased in the hippocampus. In addition, we observed increased histone H3 lysine 9 di- and trimethylation (H3K9me2/3) and decreased histone H3 lysine 4 di and trimethylation (H3K4me2/3). Similar findings were observed in cultured hippocampal neurons and HT-22 hippocampal cell line treated with pilocarpine. In conclusion, our data show that muscarinic receptor activation by pilocarpine induces a global repressive state of chromatin and prevalence of LSD1-CoREST2 epigenetic complexes, modifications that could underlie the pathophysiological processes leading to epilepsy.
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A comparison exercise of Latin American and Caribbean Secondary Standards Dosimetry Laboratories (SSDLs) was jointly organized by the International Atomic Energy Agency (IAEA) and the Ionizing Radiation Metrology Laboratory at the Federal University of Pernambuco (LMRI-DEN/UFPE). This exercise was organized during an IAEA regional meeting on the review and update of calibration capabilities in Latin America, held in Recife, during the period from 23 to 27 April 2018 under the technical cooperation project ME-RLA 9085-170572. Fifteen participating SSDLs were required to irradiate optically stimulated personal dosimeters in terms of the personal dose equivalent Hp(10) in137Cs radiation quality. In addition, the IAEA Dosimetry Laboratory in Seibersdorf, Austria, and the National Physical Laboratory in Teddington, Middlesex, UK participated in this exercise as reference institutes. Each participant received 10 dosimeters that were hand-carried directly to the SSDL. Two nominal dose values of 2 mSv and 4 mSv were selected for this exercise. The participants irradiated the dosimeters using the setup and the procedures which are normally used in their standard laboratory for Hp(10) dosimeter irradiations. The dosimeters were evaluated as they were received by the coordinating laboratory, using a single BeOSL Reader. The results show that, except for one laboratory, the differences between the dosimeter reading and the assigned values were within 10%; this is consistent with the expanded uncertainty. The results indicate that most of the participant laboratories have a good capability to irradiate personal dosimeters in the quantity Hp(10).
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Monitoramento de Radiação , Proteção Radiológica , Calibragem , Humanos , Laboratórios , América Latina , Dosímetros de RadiaçãoRESUMO
Neurocysticercosis (NC) presents two broad clinical entities: extraparenchymal (EP-NC) and parenchymal (P-NC). Using ELISA methodology, we demonstrate autoantibodies to tubulin and the Major oligodendrocyte glycoprotein (MOG) in the CSF of most, but not all, EP-NC samples. Levels of these autoantibodies were considerably reduced or absent in the P-NC samples. There was a striking correlation between levels of anti-tubulin and anti-MOG, and the significant correlation between the levels of autoantibodies and cellularity in the CSF, suggests that stimulation of the autoantibody response may be a function of cerebral inflammation. A hypothetical model to describe the pathogenesis of EP-NC is presented.
Assuntos
Autoanticorpos/líquido cefalorraquidiano , Encéfalo/diagnóstico por imagem , Glicoproteína Mielina-Oligodendrócito/líquido cefalorraquidiano , Neurocisticercose/líquido cefalorraquidiano , Neurocisticercose/diagnóstico por imagem , Tubulina (Proteína)/líquido cefalorraquidiano , Animais , Biomarcadores/líquido cefalorraquidiano , Equador/epidemiologia , Humanos , México/epidemiologia , Neurocisticercose/epidemiologia , Tecido Parenquimatoso/metabolismo , Fragmentos de Peptídeos/líquido cefalorraquidiano , SuínosRESUMO
Understanding the packaging of DNA into chromatin has become a crucial aspect in the study of gene regulatory mechanisms. Heterochromatin establishment and maintenance dynamics have emerged as some of the main features involved in genome stability, cellular development, and diseases. The most extensively studied heterochromatin protein is HP1a. This protein has two main domains, namely the chromoshadow and the chromodomain, separated by a hinge region. Over the years, several works have taken on the task of identifying HP1a partners using different strategies. In this review, we focus on describing these interactions and the possible complexes and subcomplexes associated with this critical protein. Characterization of these complexes will help us to clearly understand the implications of the interactions of HP1a in heterochromatin maintenance, heterochromatin dynamics, and heterochromatin's direct relationship to gene regulation and chromatin organization.
Assuntos
Proteínas Cromossômicas não Histona/metabolismo , Eucromatina/metabolismo , Heterocromatina/metabolismo , Animais , Homólogo 5 da Proteína Cromobox , Proteínas Cromossômicas não Histona/química , Proteínas Cromossômicas não Histona/genética , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Regulação da Expressão Gênica , Instabilidade Genômica , Humanos , Elementos Isolantes , Filogenia , Ligação Proteica , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre ProteínasRESUMO
Taenia solium is the most common parasite infection of the brain, causing neurocysticercosis and typically found in rural communities with free-ranging pigs. Identification of transmission in rural areas is essential for its control. Risk factors and transmission of the parasite were evaluated in three rural Venezuelan communities (Valle del Rio and Potrero Largo, Cojedes state; and Palmarito, Portuguesa state) by a questionnaire (112 households) and coprological (492 samples) and serological (433 human and 230 porcine sera) analysis, respectively. Typical risk factors were found in all three communities: free-foraging pig husbandry, deficient sanitary conditions, high open defecation and ignorance of the parasite life cycle. Coprological examinations revealed a high level of soil-transmitted parasites. Importantly, two T. solium adult worm carriers were identified in each of the three communities. Anti-metacestode antibodies and the HP10 secreted metacestode glycoprotein were detected at significant levels in human and porcine sera in Valle del Rio, Potrero Largo and Palmarito. In conclusion, these communities may be considered to be endemic for taeniasis/cysticercosis, and the instigation of an appropriate control programme is recommended.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Cisticercose/epidemiologia , População Rural , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/parasitologia , Teníase/epidemiologia , Adulto , Animais , Antígenos de Helmintos/análise , Cisticercose/imunologia , Características da Família , Fezes/parasitologia , Humanos , Fatores de Risco , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/imunologia , Taenia solium/imunologia , Teníase/imunologia , VenezuelaRESUMO
CONTEXT: Little is known about the association between haptoglobin level and cardiometabolic traits. A previous genome-wide association study identified rs2000999 in the HP gene as the stronger genetic contributor to serum haptoglobin level in European populations. OBJECTIVE AND DESIGN: We investigated the association of HP rs2000999 with serum haptoglobin and childhood and adult obesity in up to 540/697 and 592/691 Mexican cases and controls, respectively. Anthropometric and biochemical data were collected. Serum haptoglobin was measured by an immunoturbidimetry assay. HP rs2000999 was genotyped using the TaqMan technology. Mendelian randomization analysis was performed using the Wald and inverse variance weighting methods. RESULTS: Haptoglobin level was positively associated with childhood and adult obesity. HP rs2000999 G allele was positively associated with haptoglobin level in children and adults. HP rs2000999 G allele was positively associated with childhood but not adult obesity. The association between HP rs2000999 and childhood obesity was removed after adjusting for haptoglobin level. In a Mendelian randomization analysis, haptoglobin level genetically predicted by HP rs2000999 showed a significant causal effect on childhood obesity by the Wald and inverse variance weighting methods. CONCLUSION: Our data provide evidence for the first time for a causal positive association between serum haptoglobin level and childhood obesity in the Mexican population. Our study contributes to the genetic elucidation of childhood obesity and proposes haptoglobin as an important biomarker and treatment target for obesity.
Assuntos
Predisposição Genética para Doença , Haptoglobinas/genética , Obesidade Infantil/genética , Polimorfismo de Nucleotídeo Único , Alelos , Estudos de Casos e Controles , Criança , Feminino , Genótipo , Haptoglobinas/análise , Humanos , Masculino , Análise da Randomização Mendeliana , México , Obesidade Infantil/sangueRESUMO
Immunodiagnosis has a supportive role in the diagnosis of neurocysticercosis (NCC). The aim of this study was to compare the validity of seven immunodiagnostic tests among serum samples from 58 patients with NCC, 26 patients with neurological diseases other than NCC, and 15 healthy controls. One test for viable parasite detection (HP10 antigen assay) and six for antibody detection were evaluated. For the entire sample, sensitivities ranged from 55.2% (NOVALISA) to 81.0% (enzyme-linked immunosorbent assay [ELISA] Taenia solium antibody), with the sensitivity of the latter test significantly higher than that of the in-house ELISA Taenia crassiceps, NOVALISA, enzyme-linked immunoelectrotransfer blot (EITB) CDC, and HP10. Overall, specificities were high, ranging from 85.4% (ELISA Ts) to 97.1% (NOVALISA), with no statistically significant differences. Detection of HP10 antigen was significantly associated with the presence of vesicular parasites. The simple and low-cost ELISA Taenia solium antibody Ab instead of EITB is recommended to support NCC diagnosis in both rural and hospital settings in Mexico.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Testes Diagnósticos de Rotina/métodos , Neurocisticercose/diagnóstico , Taenia solium/imunologia , Adulto , Animais , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Testes Imunológicos/métodos , Masculino , México , Neurocisticercose/imunologia , População Rural , Sensibilidade e EspecificidadeRESUMO
RESUMEN Los materiales a base de silicato de calcio han demostrado ser bioactivos debido a su capacidad para producir apatita carbonatada biológicamente compatible. El objetivo de este estudio fue analizar la bioactividad de Biodentine™ y MTA Repair HP® en contacto con discos de dentina humana, que se obturaron y dividieron aleatoriamente para formar cuatro grupos: grupo 1 Biodentine™, grupo 2 MTA Repair HP®, grupo control positivo MTA Angelus® y grupo control negativo IRM®, los cuales se incubaron en solución PBS durante 10 días, para posterior análisis por medio de MEB-EDS y Espectroscopía Raman. Los tres materiales a base de silicato de calcio analizados en este estudio demostraron ser bioactivos pues al entrar en contacto con una solución a base de fosfato desencadenaron la precipitación inicial de fosfato de calcio amorfo, que actúa como precursor durante la formación de apatita carbonatada.
ABSTRACT Calcium silicate-based materials have been shown to be bioactive due to their ability to produce biologically compatible carbonated apatite. The objective of this study was to analyze the bioactivity of Biodentine ™ and MTA Repair HP® in contact with human dentine discs, which were sealed and divided randomly to form four groups: group 1 Biodentine™, group 2 MTA Repair HP®, positive control group MTA Angelus® and negative control group IRM®, which were incubated in PBS solution for 10 days, for a subsequent analysis by means of MEB-EDS and Raman spectroscopy. The three calcium-based materials analyzed in this study proved to be bioactive because upon contact with a phosphate-based solution they were triggered at the onset of amorphous calcium phosphate, as the precursor during the formation of carbonated apatite.
Assuntos
Apatitas/análise , Análise Espectral Raman , Calcarea Silicata/análise , Materiais Dentários/análise , Sinergismo FarmacológicoRESUMO
Roads and highways can affect the spread of insect-borne diseases by limiting or amplifying the spatiotemporal distribution of vectors, pathogens, and hosts, which can, in turn, lead to the creation of a nidus of infection. The aim of this study was to compare the diversity (richness and abundance) of phlebotomine sand flies in household and forest edge environments found along two different segments of an Amazonian highway. Sampling was conducted along the northern and southern portions of highway BR-319, in Amazonas State, Brazil. At each sampling point, Hoover Pugedo traps were set in indoor and outdoor habitats, and at forests edges, and captures were made between 06:00 pm and 06:00 am. A total of 1,189 sand flies were captured and 48 species were identified. As expected, a greater number of species and individuals were captured in forest edge environments. Permutational Multivariate Analyses of Variance (PERMANOVA) and Permutational Analyses of Multivariate Dispersions (PERMDISP) analyses showed that sand fly fauna differed significantly among habitats, but no variance in species composition was observed between the two road segments. Some of the captured species were species that have been implicated as vectors of Leishmania spp. Ross, 1903 (Kinetoplastida: Trypanosomatidae).
Assuntos
Biodiversidade , Psychodidae , Animais , Brasil , Florestas , Habitação , Densidade DemográficaRESUMO
The benzopyran HP1, a compound isolated from Hypericum polyanthemum, has demonstrated significant opioid-mediated antinociceptive activity after its oral administration. Despite the pharmacological potential, the poor aqueous solubility limits the oral absorption of this compound. For this reason, HP1 has been alternatively incorporated in lipid-based drug delivery systems. Given that nanoemulsions showed higher antinociceptive action than the free compound in a previous report, in this study, the main objective was to investigate the intestinal transport mechanisms of this system. The Ussing chamber model and rat jejunum were selected for this purpose. The apparent permeability coefficient of HP1 increased approximately 5.3 times after its incorporation in nanoemulsions. Considering that the absorptive transport of HP1 was significantly higher than the secretory transport, the participation of active transporters was suggested. The amount of HP1 in the acceptor chamber was reduced during permeability assays performed at 4⯰C, supporting the hypothesis that active transporters are involved in the intestinal transport of this compound. The amount of free fatty acids released from nanoemulsion was approximately 60% after 90â¯min, demonstrating that part of this system is disassembled before absorption. Nanoemulsion constituents would be able to form new structures with biological constituents, leading to a rapid solubilization of HP1. A mucoadhesion rate of 50% was achieved by nanoemulsion after 30â¯min, which would also contribute to explain the higher absorption of this system. The particle size of the nanoemulsion is also compatible with endocytosis-mediated transport. Taken together, these results suggest that nanoemulsions containing HP1 could be efficiently delivered to humans considering that different absorption mechanisms are exploited.