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1.
Am J Primatol ; 86(7): e23635, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38738522

RESUMO

The golden lion tamarin (GLT) is an Endangered primate endemic to Brazil's lowland Atlantic Forest. After centuries of deforestation and capture for the pet trade, only a few hundred individuals survived, all in isolated forest fragments 85 km from Rio de Janeiro city. Intensive conservation actions, including reintroduction of zoo-born tamarins, increased numbers to about 3700 in 2014. The most severe yellow fever epidemic/epizootic in Brazil in 80 years reduced two of the largest GLT populations by over 90%. Herein we report the results of a 2023 survey of GLTs designed to examine the dynamics of population recovery following yellow fever. Results indicate that populations hard hit by yellow fever are recovering due in part to immigration from adjacent forest fragments. No local extirpations were observed. About 4800 GLTs live in the survey area. This represents a 31% increase since the baseline survey completed in 2014. Two factors explain most of the increase: four large areas that had no GLTs or very low-density populations in 2014 are now at moderate density (three areas) or low density (one area), explaining 71% of overall increase since 2014. Increase in forest area within our survey area may explain up to 16% of the increase in GLT numbers since 2014. Results of computer simulations suggest that strengthening forest connectivity will facilitate metapopulation resilience in the face of mortality factors such as yellow fever.


Assuntos
Leontopithecus , Dinâmica Populacional , Febre Amarela , Animais , Febre Amarela/epidemiologia , Brasil/epidemiologia , Doenças dos Macacos/epidemiologia , Espécies em Perigo de Extinção , Conservação dos Recursos Naturais , Feminino , Masculino
2.
Neurosurg Focus Video ; 9(2): V3, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37854644

RESUMO

A 58-year-old male was admitted to the authors' department due to cervicothoracic pain and disequilibrium. Physical examination evidenced sensory and motor deficits in the lower limbs. MRI evidenced an expansive intramedullary lesion compatible with ependymoma. The nuances of this surgical access and the management of intradural tumors are discussed.

3.
Bioprocess Biosyst Eng ; 46(5): 681-692, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36806976

RESUMO

Bioreactors can perform biochemical conversions mediated by biocatalysts, such as enzymes, animal cells, plants, and microorganisms. Among several existing models, airlift bioreactors are devices with the low shear environment and good mass transfer with low energy consumption, employed in several biochemical processes. The fluid flow is enabled through air injection by the sparger located at the bioreactor base. Despite its simple geometry compared with the conventional bioreactors, airlift performance can be optimized via geometrical modifications. Therefore, the objective of this work was to evaluate the effects of the addition of helical flow promoters, positioned in the riser and/or downcomer regions of an airlift of concentric tubes measuring the volumetric oxygen coefficient (kLa) and gas holdup. The results obtained by varying the gas flow rate from 1.0 to 4.0 vvm allowed the system evaluation of oxygen transfer and gas holdup. The inclusion of helical flow promoters increased the kLa, reaching up to 23% in oxygen transfer compared to tests without helicoids and up to 14% increase in the gas holdup. The inclusion of helical flow promotors was beneficial for all gas flow rates. Thus, including these flow promoters is an effective strategy to increase the oxygen transfer rate for bioprocess optimization.


Assuntos
Reatores Biológicos , Oxigênio , Oxigênio/química
4.
An Acad Bras Cienc ; 94(4): e20210262, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35946750

RESUMO

Cattle ranching is the primary land-use of deforested areas in the Brazilian Amazon. Deforestation precedes pasture establishment, implying tremendous amounts of greenhouse gas emissions caused by carbon stock losses. Despite several studies addressing carbon storage in forests, there is a lack of data regarding cultivated pastures. Hence, the estimation of greenhouse gas emissions associated with land-use change becomes uncertain. In this study, we assessed the carbon stock of cultivated pastures located in Rondônia, southwestern Brazilian Amazon. A total of 50 squared plots of 1 m² were randomly allocated in cattle ranching farms covered by Oxisols (Dystrophic Yellow and Dystrophic Red-Yellow Latosols). Carbon fraction ranged from 0.36 for belowground biomass to 0.45 gC.g-1 d.m. for aboveground biomass. The average total carbon stock was 5.17 MgC.ha-1, with non-significant differences when stratifying data by soil types. Considering data from the III Brazilian Inventory of Anthropogenic Emissions and Removals of Greenhouse Gases, our results suggested that land-use change from primary forests to cultivated pastures resulted in a loss of 192.54 MgC.ha-1, which corresponds to a net emission of 705.98 MgCO2eq.ha-1 to the atmosphere. This study provides valuable information to improve the Brazilian Inventory of Anthropogenic Emissions and Removals of Greenhouse Gases.


Assuntos
Carbono , Gases de Efeito Estufa , Animais , Brasil , Carbono/análise , Bovinos , Conservação dos Recursos Naturais , Florestas
5.
Braz J Microbiol ; 53(3): 1263-1269, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35426068

RESUMO

Immunological assays to detect SARS-CoV-2 Spike Receptor Binding Domain (RBD) antigen seroconversion in humans are important tools to monitor the levels of protecting antibodies in the population in response to infection and/or immunization. Here we describe a simple, low cost, and high throughput Ni2+ magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in Escherichia coli. A 6xHis-tagged Spike S1 RBD was expressed in E. coli and purified by affinity chromatography. The protein was mobilized on the surface of Ni2+ magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population.


Assuntos
COVID-19 , SARS-CoV-2 , Anticorpos Antivirais , Escherichia coli/genética , Humanos , Imunoensaio/métodos , Imunoglobulina G , Fenômenos Magnéticos , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/genética
6.
Anal Biochem ; 631: 114360, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34481802

RESUMO

To monitor the levels of protecting antibodies raised in the population in response to infection and/or to immunization with SARS-CoV-2, we need a technique that allows high throughput and low-cost quantitative analysis of human IgG antibodies reactive against viral antigens. Here we describe an ultra-fast, high throughput and inexpensive assay to detect SARS-CoV-2 seroconversion in humans. The assay is based on Ni2+ magnetic particles coated with His tagged SARS-CoV-2 antigens. A simple and inexpensive 96 well plate magnetic extraction/homogenization process is described which allows the simultaneous analysis of 96 samples and delivers results in 7 min with high accuracy.


Assuntos
Anticorpos Antivirais/sangue , Teste Sorológico para COVID-19/métodos , COVID-19/diagnóstico , Imunoglobulina G/sangue , SARS-CoV-2/isolamento & purificação , Anticorpos Antivirais/imunologia , Antígenos Virais/sangue , Antígenos Virais/imunologia , COVID-19/sangue , COVID-19/imunologia , Teste Sorológico para COVID-19/economia , Ensaio de Imunoadsorção Enzimática/economia , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Imunoglobulina G/imunologia , Imãs/química , Níquel/química , SARS-CoV-2/imunologia , Sensibilidade e Especificidade , Soroconversão , Fatores de Tempo
7.
Braz J Microbiol ; 52(4): 2069-2073, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34342836

RESUMO

Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here, the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 RT-qPCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen-coated plates were stable for up to 3 months at 4 °C. The ELISA method described is ready for mass production and will be an additional tool to track COVID-19 cases.


Assuntos
COVID-19 , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Ensaio de Imunoadsorção Enzimática , Soroconversão , Anticorpos Antivirais/sangue , COVID-19/diagnóstico , COVID-19/imunologia , Humanos , Imunidade Humoral , Proteínas do Nucleocapsídeo/genética , Fosfoproteínas/imunologia , Sensibilidade e Especificidade
8.
Mol Cell Endocrinol ; 535: 111393, 2021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34245846

RESUMO

Reproductive cancers in both genders represent serious health problems, whose incidence has significantly risen over the past decades. Although considerable differences among reproductive cancers exist, we aimed to identify similar signaling pathways and key molecular oncomarkers shared among six human reproductive cancers that can advance the current knowledge of cancer biology to propose new strategies for more effective therapies. Using a computational analysis approach, here we uncover aberrant miRNAs-mRNAs networks shared in six reproductive tumor types, and identify common molecular mechanisms strictly associated with cancer promotion and aggressiveness. Based on the fact that estrogenic and androgenic signaling pathways were most active in prostate and breast cancers, we further demonstrated that both androgen and estrogen deprivation therapy are capable of regulating the expression of the same key molecular sensors associated with endoplasmic reticulum dysfunction and cell cycle in these cancers. Overall, our data reveal a potential mechanistic framework of cellular processes that are shared among reproductive cancers, and particularly, highlight the importance of hormonal deprivation in breast and prostate cancers and potentially new biomarkers of response to these therapeutic approaches.


Assuntos
Neoplasias da Mama/genética , Biologia Computacional/métodos , Neoplasias do Endométrio/genética , Redes Reguladoras de Genes , MicroRNAs/genética , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Ovarianas/genética , Neoplasias da Próstata/genética , Neoplasias Testiculares/genética , Neoplasias Uterinas/genética , Antagonistas de Androgênios/farmacologia , Antagonistas de Androgênios/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Bases de Dados Genéticas , Neoplasias do Endométrio/tratamento farmacológico , Antagonistas de Estrogênios/farmacologia , Antagonistas de Estrogênios/uso terapêutico , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Humanos , Masculino , Neoplasias Embrionárias de Células Germinativas/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias da Próstata/tratamento farmacológico , Análise de Sobrevida , Neoplasias Testiculares/tratamento farmacológico , Neoplasias Uterinas/tratamento farmacológico
9.
Bioprocess Biosyst Eng ; 44(8): 1721-1732, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33821325

RESUMO

The production of biocompounds through the cultivation of filamentous microorganisms is mainly affected by Oxygen Transfer Rate (OTR) and shear rate ([Formula: see text]) conditions. Despite efforts have been made to evaluate the effect of operating variables (impeller speed, N; and airflow rate, ϕair) on clavulanic acid production, no analysis regarding the effect of OTR and [Formula: see text] was made. Then, the aim of this study was to evaluate the dissociated effect of physical phenomena such as oxygen transfer and shear rate in the production of clavulanic acid from Streptomyces clavuligerus using a stirred tank bioreactor. Streptomyces clavuligerus cultivations were performed at five different OTR and [Formula: see text] conditions by manipulating the operating conditions (N, ϕair, and gas inlet composition). Cultivations performed at equal impeller speed (600 rpm, similar [Formula: see text]) using oxygen enrichment, showed that CA productivity (ProdCA) was positively affected by OTR increase. Subsequently, the different shear conditions (achieved by varying the impeller speed) lead to an increase in CA production levels. Despite both OTR and shear rate positively enhanced CA productivity, [Formula: see text] exhibited the highest impact: an increase of 145% in OTRinitial enhanced the clavulanic acid productivity of about 29%, while an increment in the shear rate of 134% raised the ProdCA in 53%.


Assuntos
Ácido Clavulânico/química , Microbiologia Industrial/métodos , Oxigênio/química , Streptomyces/metabolismo , Reatores Biológicos , Biotecnologia/métodos , Meios de Cultura , Desenho de Equipamento , Resistência ao Cisalhamento , Fatores de Tempo
10.
Preprint em Inglês | medRxiv | ID: ppmedrxiv-21255396

RESUMO

A technique allowing high throughput, fast and low-cost quantitative analysis of human IgG antibodies reacting to SARS-CoV-2 antigens will be required to understand the levels of protecting antibodies in the population raised in response to infections and/or to immunization. We described previously a fast, simple, and inexpensive Ni2+ magnetic bead immunoassay which allowed detection of human antibodies reacting against the SARS-CoV-2 nucleocapsid protein using a minimal amount of serum or blood. A major drawback of the previously described system was that it only processed 12 samples simultaneously. Here we describe a manually operating inexpensive 96 well plate magnetic extraction / homogenization process which allows high throughput analysis delivering results of 96 samples in chromogenic format in 12 minutes or in fluorescent ultrafast format which takes only 7 minutes. We also show that His tag antigen purification can be performed on the fly while loading antigens to the Ni2+ magnetic beads in a process which takes only 12 min reducing the pre analytical time and cost. Finally, we show that the magnetic bead immunoassay is antigen flexible and can be performed using either Nucleocapsid, Spike or Spike RBD. The method performed with low inter and intra assay variability using different antigens and detection modes and was able to deliver >99.5% specificity and >95% sensitivity for a cohort of 203 pre pandemic and 63 COVID-19 positive samples.

11.
Preprint em Inglês | medRxiv | ID: ppmedrxiv-21252100

RESUMO

Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 PCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen coated plates were stable for up to 3 months at 4{degrees}C). The ELISA method described is ready to mass production and will be an additional toll to track COVID-19 cases.

12.
Animals (Basel) ; 11(2)2021 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-33494489

RESUMO

The whole corn germ (WCG), due to its desirable nutritional characteristics, has been studied as feed for ruminants. This study aimed to evaluate the effects of WCG inclusion as a linoleic acid source in diets for feedlot lambs on carcass characteristics, physicochemical composition, sensory attributes, and fatty acid profile of the meat. Forty non-castrated, crossbreed Dorper x Santa Inês lambs were distributed in a completely randomized design to evaluate the inclusion levels (0, 30, 60, 90, and 120 g/kg dry matter (DM)) of whole corn germ (WCG) in the diet. The dietary inclusion of WCG did not influence (p > 0.05) the weight gain and carcass characteristics, with the exception of the subcutaneous fat thickness (p < 0.01), which was higher in animals fed diets with higher levels of WCG. Lightness (L *; p = 0.04), yellowness (b *; p < 0.01), shear force (p = 0.04), linoleic fatty acid concentrations (p = 0.03), and total polyunsaturated fatty acids (p = 0.04) had a quadratic increase due to WCG inclusion in the diets. The use of up to 120 g/kg DM of WCG in lamb diets does not affect the carcass characteristics, physicochemical composition, and sensory attributes of the meat. Despite this, the best polyunsaturated fatty acid profile in lambs' meat is obtained using 76.7 g/kg DM of WCG.

13.
ACS Sens ; 6(3): 703-708, 2021 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-33496577

RESUMO

Immunological methods to detect SARS-CoV-2 seroconversion in humans are important to track COVID-19 cases and the humoral response to SARS-CoV-2 infections and immunization to future vaccines. The aim of this work was to develop a simple chromogenic magnetic bead-based immunoassay which allows rapid, inexpensive, and quantitative detection of human antibodies against SARS-CoV-2 in serum, plasma, or blood. Recombinant 6xHis-tagged SARS-CoV-2 Nucleocapsid protein was mobilized on the surface of Ni2+ magnetic beads and challenged with serum or blood samples obtained from controls or COVID-19 cases. The beads were washed, incubated with anti-human IgG-HPR conjugate, and immersed into a solution containing a chromogenic HPR substrate. Bead transfer and homogenization between solutions was aided by a simple low-cost device. The method was validated by two independent laboratories, and the performance to detect SARS-CoV-2 seroconversion in humans was in the same range as obtained using the gold standard immunoassays ELISA and Luminex, though requiring only a fraction of consumables, instrumentation, time to deliver results, and volume of sample. Furthermore, the results obtained with the method described can be visually interpreted without compromising accuracy as demonstrated by validation at a point-of-care unit. The magnetic bead immunoassay throughput can be customized on demand and is readily adapted to be used with any other 6xHis tagged protein or peptide as antigen to track other diseases.


Assuntos
Anticorpos Antivirais/sangue , Teste Sorológico para COVID-19 , COVID-19/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , SARS-CoV-2/imunologia , COVID-19/sangue , COVID-19/imunologia , Humanos , Imunoglobulina G/imunologia , Fenômenos Magnéticos
14.
Food Chem ; 344: 128603, 2021 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-33234437

RESUMO

Enzymatic lipophilization is an important process to extend the use of anthocyanins in lipidic media. In this work delphinidin 3-O-sambubioside (Dp3sam) isolated from Hibiscus sabdariffa L. flower was esterified with octanoic acid using Candida antarctica lipase B. The physical-chemical properties of the new lipophilic pigment were studied by UV-vis spectroscopy. Dp3sam with chloride, acetate and formate as counter ions were employed to study the lipophilization reaction. The hydrolysis of the reagent was avoided with a formate counter ion and the expected product was achieved with a noteworthy change of solubility. 1D and 2D NMR characterization of Dp3sam-C8 confirmed that the lipophilization took place at the primary alcohol of the glucoside moiety. Overall, the Dp3sam-C8 ester presents a stabilization of the quinoidal base (blue color) at neutral or moderate alkaline pH, which foresees a potential use of this pigment as a broad kind of industries on lipo-soluble formulations.


Assuntos
Antocianinas/análise , Ácidos Graxos/metabolismo , Proteínas Fúngicas/metabolismo , Hibiscus/química , Lipase/metabolismo , Acilação , Antocianinas/isolamento & purificação , Antocianinas/metabolismo , Biocatálise , Cromatografia Líquida de Alta Pressão , Cor , Dissacarídeos/química , Hibiscus/metabolismo , Concentração de Íons de Hidrogênio , Espectrometria de Massas , Extratos Vegetais/química , Extração em Fase Sólida
15.
Enzyme Microb Technol ; 139: 109587, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32732036

RESUMO

The hydrodynamic environment in bioreactors affects the oxygen transfer rate and the shear conditions during microbial cultivations. Therefore, assessment of the effect of the hydrodynamic environment on cellular morphology can contribute to favoring the production of metabolites of interest. The aim of this work was to use image analysis in order to quantify the fragmentation of Aspergillus niger pellets in a conventional bioreactor operated using different impeller speeds, air flow rates, and impeller configurations including Rushton turbines and Elephant Ear impellers, with evaluation of the influence of the hydrodynamic environment on the production of cellulolytic enzymes. An empirical kinetic model was proposed to describe the dynamics of pellet fragmentation and quantify the shear conditions. The results showed that the agitation speed affected the dynamics of pellet fragmentation in two ways, by accelerating the damage process and by increasing the magnitude of the fragmentation. Both endoglucanase and ß-glucosidase production exhibited a linear relationship with the pellet fragmentation percentage, which was directly related to the shear conditions. Interestingly, ß-glucosidase production was favored under high shear conditions, while the highest endoglucanase production occurred under low shear conditions. These findings may be useful for defining suitable systems and operating conditions for the production of metabolites including enzymes in bioreactors, as well as defining conditions that favour a specific pre-determined enzyme cocktail.


Assuntos
Aspergillus niger/enzimologia , Reatores Biológicos , Celulase/biossíntese , beta-Glucosidase/biossíntese , Fermentação , Hidrodinâmica , Cinética
16.
Bioprocess Biosyst Eng ; 42(6): 995-1008, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30848359

RESUMO

The shear rate is an important bioreactor parameter that needs to be evaluated due to its impact on microorganism morphology and viability, and consequently on bioproduct formation. Airlift bioreactors, classified as low-shear devices, are used as an alternative to conventional stirred-tank reactors. Considerable efforts have been made to characterize the shear environments in airlift bioreactors, using the average shear rate ([Formula: see text]) as a key parameter. However, there is no agreement among the values obtained in different studies, which can differ even in orders of magnitude. The methodologies used to obtain [Formula: see text] in the different studies could be the reason for the lack of agreement among them. In this work, [Formula: see text] in a concentric tube airlift bioreactor was evaluated using computational fluid dynamics (CFD), as well as based on universal velocity profiles for liquid flows in smooth pipes and annuli. Good agreement was obtained between the CFD-based average shear rates and the values obtained from universal velocity profiles, indicating that CFD simulation is a valuable tool for [Formula: see text] prediction.


Assuntos
Reatores Biológicos , Hidrodinâmica , Modelos Químicos , Resistência ao Cisalhamento
17.
Anticancer Agents Med Chem ; 18(15): 2070-2077, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30147015

RESUMO

INTRODUCTION: The understanding of cancer has evolved into a complex disease, which has heterogeneous characteristics between different patients, and more than that, a broad range of genetically distinct cells in the same tumor. Chronic and non-transmissible diseases such as cancer have become even more important worldwide and advances in their characterization and properties are running out. In a suitable setting, early diagnosis of cancer and proper treatment are essential to overcome the barriers of tumor healing and quality assurance of the patient's life. The studies included in the analysis were independently retrieved by the authors. The text words included "Non-Small Cell Lung Cancer," "NSCLC," "biomarkers in NSCLC" and "gene in NSCLC"." We were used to perform a systematic literature search in the PubMed and Web of Science databases. There was limit on the start date for published articles (2001), and the search ended in December 2017. For a more comprehensive analysis, articles were only in English. Non-small Cell Lung Cancer (NSCLC) Microenvironment: This review focuses on lung cancer, especially NSCLC, one of the most worldwide common cancer types, showing the importance of tumor microenvironment in this disease. NSCLC Biomarkers: This review shows a molecular aspect of NSCLC biomarkers used in clinical approaches, such as EGFR, KRAS, MET, indicating mutations that are crucial for cancer progression and related to treatment properties. We also describe 11 drugs commonly used in cancer treatment that are correlated with the patient genetic profile. CONCLUSION: This review highlights the importance of molecular diagnosis of mutations in lung cancer, driving to individualized therapy with the intent of getting a better response to treatment and improving patients' quality of life.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/terapia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/terapia , Medicina de Precisão , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Humanos , Neoplasias Pulmonares/diagnóstico , Microambiente Tumoral
18.
Food Funct ; 9(3): 1726-1735, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29493669

RESUMO

The lack of digestibility of certain gluten proteins is essential in the development of celiac disease (CD). Gluten proteins are remarkably resistant to luminal and brush-border proteolysis owing to their high proline and glutamine content. Consequently, large fragments remain intact after digestion exerting toxic effects. Intestinal brush-border membrane vesicles (BBMV) have been described as having strong proteolytic activity mainly through prolyl endopeptidase enzymes. The purpose of this work was to monitor the gastrointestinal digestion of specific CD epitopes by means of an in vitro gastrointestinal digestion model that included incubation with brush-border membrane enzymes. Gluten hydrolysates were characterized by mass spectrometry and the immunologic peptides were tracked by searching the main T-cell stimulating epitopes which have been widely described. The immunologic potential of gluten hydrolysates was further analysed by enzyme-linked immunosorbent assay (ELISA). The results showed that the composition of gluten hydrolysates depended on the digestion time and protein structural characteristics. On the other hand, the main T-cell stimulating epitopes formed during hydrolysis depend on the precursor protein. Glutenin oligopeptides were degraded faster whereas gliadin, mainly α-gliadin oligopeptides, remained intact for a long time. MS-based analysis showed that the formation of the epitopes from γ-gliadin and ω-gliadin or glutenin was favoured but they were generally degraded during the gastrointestinal treatment. However, the peptides containing the epitope PFPQPQLPY (α-gliadin) remained intact even after 180 min of digestion time. Overall, from all the epitopes tracked, PFPQPQLPY was the most resistant to in vitro BBMV digestion.


Assuntos
Glutens/química , Peptídeos/química , Triticum/química , Doença Celíaca/metabolismo , Digestão , Epitopos/química , Epitopos/metabolismo , Glutens/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Espectrometria de Massas , Peptídeos/metabolismo , Proteólise
19.
Food Funct ; 8(8): 2712-2721, 2017 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-28703824

RESUMO

Immunogenic gluten peptides trigger Celiac Disease (CD), an adaptive immune response in genetically predisposed individuals. Given the structural similarity between all gluten proteins their individual CD influence is not clear. Hence, the extraction, separation and characterization of wheat gluten proteins have become relevant to measure their individual potential immunoreactivity. Wheat proteins were extracted from commercial wheat flour and further isolated by preparative HPLC. The resulting richest gliadin sub-fractions were characterized by nano-LC-MS/MS following a shotgun proteomic approach in order to identify the prolamins in the original commercial wheat flour. It was found that the gliadin extract was additionally composed of glutenins and avenin-like proteins. Accurate prolamin identification has emerged as a need to delve deep into the influence of each fraction on the onset of celiac disease. After protein characterization, the immunoreactivity towards the main epitope related to CD was verified by ELISA and western blotting for several different gluten fractions.


Assuntos
Doença Celíaca/etiologia , Farinha/análise , Prolaminas/química , Triticum/química , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Farinha/efeitos adversos , Humanos , Prolaminas/efeitos adversos , Prolaminas/imunologia , Espectrometria de Massas em Tandem , Triticum/efeitos adversos , Triticum/imunologia
20.
Food Funct ; 8(5): 2030-2037, 2017 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-28492692

RESUMO

This study was designed to evaluate the pharmacokinetics of Port and table red wine anthocyanins in healthy men. Volunteers were recruited to drink 250 mL of a table red wine (221 mg of anthocyanins) and 150 mL of young Port red wine (49 mg of anthocyanins). Venous blood was collected from participants at 0, 15, 30, 60 and 120 min after wine ingestion. Urine samples were collected at baseline and at 120 min. Anthocyanins and anthocyanin metabolites in plasma and urine samples were quantified by HPLC-DAD and tentatively identified by LC-MS. Red wine anthocyanins were detected in their intact forms in both plasma and urine samples, but the glucuronylated metabolites of peonidin and malvidin (PnGlucr and MvGlucr) were the two main derivatives detected after both red wine consumptions. For the first time, and supported by the synthesis of Mv3Glucr, the main pathway followed by Mv3glc after absorption was described and involves anthocyanidin conjugation with glucuronic acid after glucose removal. Despite the lower total content of anthocyanins ingested when volunteers drank Port wine, no differences were observed in the plasma Cmax of MvGlucr and PnGlucr after table and Port red wine consumption. The relative bioavailability of anthocyanins in Port wine was 96.58 ± 5.74%, compared to the anthocyanins present in red wine. In conclusion, both Port and table red wines are good sources of bioavailable anthocyanins.


Assuntos
Antocianinas/sangue , Antocianinas/química , Vinho/análise , Adulto , Cromatografia Líquida de Alta Pressão , Estudos Cross-Over , Humanos , Masculino , Adulto Jovem
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