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1.
Methods Mol Biol ; 2827: 351-362, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38985281

RESUMO

Since the term proteomics was coined by Marc Wilkins in 1994, there has been an explosion in the number of articles reporting the use of the proteomics technique. As the layers of biological organization and their regulation increase, the complexity of living beings increases. Thus, we go from the genome to tissues, cells, cellular compartments, and phenotypes and the complexity of the tools used to study this complexity also increases. Unlike the genome study, in the case of the proteome, we have a more complex panorama. We have a spatial and temporal proteome. Proteomics helps to answer complex biological questions since proteins' function depends on their molecular structure, subcellular localization, and posttranslational modifications. In this protocol, we describe a methodology to extract proteins using different methods, separating proteins by electrophoresis in double-dimensional gels and analyzing the gels using specialized software that allows obtaining information on the number and abundance of the proteins from the gels.


Assuntos
Coffea , Proteínas de Plantas , Proteômica , Proteômica/métodos , Proteínas de Plantas/metabolismo , Proteínas de Plantas/análise , Coffea/metabolismo , Coffea/química , Coffea/genética , Proteoma/análise , Eletroforese em Gel Bidimensional/métodos , Software
2.
Adv Rheumatol ; 64(1): 47, 2024 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-38872193

RESUMO

INTRODUCTION: Patients with psoriatic arthritis have some lipid metabolism changes and higher risk of metabolic syndrome (MetS) and cardiovascular diseases, regardless of traditional risk factors, suggesting that chronic inflammation itself plays a central role concerning the atherosclerosis. However, there is a lack of information regarding atherogenic pattern and lipoprotein subfractions burden in these individuals. AIM: To evaluate the HDL and LDL-cholesterol plasmatic levels and their subfractions after a nutritional intervention in patients with psoriatic arthritis (PsA). METHODS: This was a randomized, placebo-controlled clinical trial of a 12-week nutritional intervention. PsA patients were randomly assigned to 1-Placebo: 1 g of soybean oil daily, no dietetic intervention; 2-Diet + Supplementation: an individualized diet, supplemented with 604 mg of omega-3 fatty acids, three times a day; and 3-Diet + Placebo: individualized diet + 1 g of soybean oil. The LDL subfractions were classified as non-atherogenic (NAth), atherogenic (Ath) or highly atherogenic (HAth), whereas the HDL subfractions were classified as small, medium, or large particles, according to the current recommendation based on lipoproteins electrophoresis. RESULTS: A total of 91 patients were included in the study. About 62% of patients (n = 56) had an Ath or HAth profile and the main risk factors associated were male gender, longer skin disease duration and higher BMI. Thirty-two patients (35%) had a high-risk lipoprotein profile despite having LDL plasmatic levels below 100 mg/dL. The 12-week nutritional intervention did not alter the LDL subfractions. However, there were significant improvement of HDL subfractions. CONCLUSION: Recognizing the pro-atherogenic subfractions LDL pattern could be a relevant strategy for identifying PsA patients with higher cardiovascular risk, regardless total LDL plasmatic levels and disease activity. In addition, a short-term nutritional intervention based on supervised and individualized diet added to omega-3 fatty acids changed positively the HDLLARGE subfractions, while LDLLARGE subfraction was improved in hypercholesterolemic individuals. CLINICALTRIALS: gov identifier: NCT03142503 ( http://www. CLINICALTRIALS: gov/ ).


Assuntos
Artrite Psoriásica , HDL-Colesterol , LDL-Colesterol , Humanos , Artrite Psoriásica/dietoterapia , Artrite Psoriásica/sangue , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , LDL-Colesterol/sangue , HDL-Colesterol/sangue , Suplementos Nutricionais , Ácidos Graxos Ômega-3/administração & dosagem , Ácidos Graxos Ômega-3/sangue , Ácidos Graxos Ômega-3/uso terapêutico , Óleo de Soja/administração & dosagem , Aterosclerose/prevenção & controle , Aterosclerose/sangue
3.
J Chromatogr A ; 1729: 465035, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-38851029

RESUMO

This work presents the development, synthesis, and application of a layered double hydroxide (LDH) coupled to magnetic particles for the removal of antibiotics as tetracyclines (TC´s): tetracycline (TC), chlortetracycline (CT), oxytetracycline (OT), and doxycycline (DT) from milk samples. The LDH synthesis conditions, reaction time (30-90 min), molar ratios Mg2+/Al3+ (7:1-1:7), interlayer anion (NO3-, Cl-, CO32-, and dodecyl sulphate (DS-)) were evaluated. Under synthesis conditions (reaction time of 30 min, Mg2+/Al3+ molar ratio of 7:1, and DS- as interlayer anion), the LDH was coupled in a magnetic solid phase microextraction (MSPµE) methodology. At the optimal extraction conditions (pH 6, 5 min of contact time, 10 mg of adsorbent), a removal percentage of 99.0 % was obtained for each tetracycline. FTIR, TGA, SEM, and adsorption isotherms were employed to characterize the optimal adsorbent. Each experiment was corroborated by large-volume sample stacking capillary electrophoresis (LVSS-CE). The adsorbent was applied directly to positive milk samples (previously tested) for TC´s removal.


Assuntos
Hidróxidos , Leite , Tetraciclinas , Leite/química , Animais , Tetraciclinas/isolamento & purificação , Tetraciclinas/análise , Tetraciclinas/química , Hidróxidos/química , Adsorção , Microextração em Fase Sólida/métodos , Antibacterianos/isolamento & purificação , Antibacterianos/química , Antibacterianos/análise , Dióxido de Silício/química
4.
Phytochem Anal ; 35(7): 1688-1694, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38925584

RESUMO

INTRODUCTION: 2,6-Disubstituted piperidin-3-ols are an important group of piperidine alkaloids found in species such as Senna spectabilis, whose main constituents include cassine and spectaline, compounds with relevant pharmacological activity. The analysis of these compounds is challenging due to the complexity of plant extracts and the absence of chromophores capable of absorbing ultraviolet (UV) radiation. OBJECTIVE: This paper presents a new analytical method to separate and quantify the non-UV-absorbing alkaloids present in ethanol extracts from S. spectabilis flowers using capillary zone electrophoresis (CZE) with indirect UV detection. METHODOLOGY: The optimized CZE method employs a background electrolyte containing 60 mM histidine (His), 15 mM α-cyclodextrin, 20% acetonitrile (ACN), and pH-adjusted to 4.7 with acetic acid (AcOH). RESULTS: The limit of detection (LOD) values was 10.2 and 13.9 mg L-1 for cassine and spectaline, respectively. For both analytes, the precision data were better than 2% of relative standard deviation (RSD) for migration times and peak areas. To evaluate the applicability of the developed method, ethanolic extracts from S. spectabilis flowers were prepared and analyzed. CONCLUSIONS: Thereby, the method proved to be efficient and complementary to conventional techniques, offering a cost-effective alternative in the quantification of the non-UV-absorbing piperidine alkaloids present in plant extracts.


Assuntos
Eletroforese Capilar , Etanol , Extratos Vegetais , Senna , Eletroforese Capilar/métodos , Extratos Vegetais/química , Extratos Vegetais/análise , Senna/química , Etanol/química , Alcaloides/análise , Limite de Detecção , Espectrofotometria Ultravioleta/métodos , Flores/química , Piperidinas/análise , Piperidinas/química
5.
J Chromatogr A ; 1720: 464809, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38490141

RESUMO

An ultrafast, efficient, and eco-friendly method combining magnetic solid phase extraction and capillary electrophoresis with diode array detection have been developed to determine ractopamine residues in food samples. A restricted access material based on magnetic and mesoporous molecularly imprinted polymer has been properly synthesized and characterized, demonstrating excellent selectivity and high adsorbent capacity. Short-end injection capillary electrophoresis method was optimized: 75 mM triethylamine pH 7 as BGE, -20 kV, 50 mbar by hydrodynamic injection during 8 s, and capillary temperature at 25 °C; reaching ultrafast ractopamine analysis (∼0.6 min) with good peak asymmetry, and free from interfering and/or baseline noise. After sample preparation optimization, the conditions were: 1000 µL of sample at pH 6, 20 mg of adsorbent, stirring time of 120 s, 250 µL of ultrapure water as washing solvent, 1000 µL of methanol: acetic acid (7: 3, v/v) as eluent, and the adsorbent can be reused four times. In these conditions, the analytical method showed recoveries around to 100 %, linearity ranged from 9.74 to 974.0 µg kg-1, correlation coefficient (r) ≥ 0,99 in addition to adequate precision, accuracy, and robustness. After proper validation, the method was successfully applied in the analysis ractopamine residues in bovine milk and bovine and porcine muscle.


Assuntos
Impressão Molecular , Polímeros Molecularmente Impressos , Fenetilaminas , Animais , Suínos , Extração em Fase Sólida/métodos , Eletroforese Capilar/métodos , Fenômenos Magnéticos , Impressão Molecular/métodos , Cromatografia Líquida de Alta Pressão/métodos
6.
J Environ Sci Health B ; 59(3): 88-97, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38189415

RESUMO

The relationship between bacterial diversity and the bioavailability of nutrients, toxic metals and the herbicide oxyfluorfen in a tropical vegetable growing area was evaluated. The study was conducted in a vegetable growing area located in the mountainous region of Rio de Janeiro (Brazil), and samples were collected in areas of vegetable cultivation and areas of environmental reserve. Fertility analyses and determination of the pseudototal levels of toxic metals in the soil samples were performed. The profile of the soil bacterial community was determined by amplification of the 16S rRNA gene and separation by DGGE. The results showed that the levels of toxic metals and elements associated with soil fertility were higher in vegetable production areas. These differences in the physical and chemical characteristics of the soil favored the presence of a greater number of OTUs in the cultivation areas (17.3-27 OTUs) than in the areas of environmental reserve (13-22 OTUs). Therefore, this study demonstrates that the presence of toxic metals and the herbicide oxyfluorfen and the increase in fertility in soils in areas with intensive vegetable cultivation resulting from the intensive management adopted in these areas promotes a differentiation of the bacterial profiles in soils in tropical vegetable growing areas.


Assuntos
Éteres Difenil Halogenados , Poluentes do Solo , Solo , Solo/química , Verduras , RNA Ribossômico 16S/genética , Brasil , Nutrientes/análise , Microbiologia do Solo , Poluentes do Solo/toxicidade , Poluentes do Solo/análise
7.
Electrophoresis ; 45(3-4): 300-309, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37946567

RESUMO

This paper analyzes the role of the diffusion coefficient in the movement of analytes that can reversibly react with a selector given a product in the presence of drift. The problem mimics the movement of enantiomers in a capillary electrophoresis experiment. As is well known, the signal in the capillary must be sharp enough to make a good determination of the effective mobility of the analytes being analyzed. The essence of the technique is based on fast interconversion rates. Therefore, the effective diffusion coefficient must be negligible during the experiment. In the present work, an exact expression for both the apparent mobility and the diffusion coefficient is obtained. This is done by writing the rate equations governing the process and solving them using the generating function technique. The effective mobility coincides with the Wren and Rowe equation, whereas the diffusion coefficient allows us to determine the values of the parameters to be taken into account so that this quantity is minimal or close to zero. On the other hand, the numerical solution of the kinetic equations and Monte Carlo simulations allow us to follow the signal in the capillary and to determine its space-time evolution.


Assuntos
Eletroforese Capilar , Eletroforese Capilar/métodos , Estereoisomerismo , Cinética , Método de Monte Carlo , Difusão
8.
Electrophoresis ; 45(1-2): 35-54, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37946578

RESUMO

This review covers the know-how of the Grupo de Química Analítica e Quimiometria regarding the analysis of fatty acids by capillary electrophoresis acquired over its 20 years of existence. Therefore, the fundamentals, advantages, and applications of this technique for analyzing different fatty acids in samples such as food, oils, cosmetics, and biological matrices are presented and discussed. Capillary electrophoresis is, thus, shown as an interesting and valuable separation technique for the target analysis of these analytes as an alternative to the gas chromatography coupled to flame ionization detection, as it offers advantages over the latter such as low analysis times, low sample and reagent consumption, the use of a nondedicated column, and simpler sample preparation. In addition, the methods shown in this literature review can be useful for quality control, adulteration, and health-related research by regulatory agencies.


Assuntos
Eletroforese Capilar , Ácidos Graxos , Ácidos Graxos/análise , Eletroforese Capilar/métodos , Cromatografia Gasosa/métodos , Óleos , Contaminação de Medicamentos
9.
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1558588

RESUMO

Introducción: la anemia de células falciformes es una enfermedad genética autosómica recesiva considerada la enfermedad hereditaria más frecuente en Cuba. El Programa Cubano de prevención de anemia por hematíes falciformes, se basa en el pesquisaje mediante estudio de electroforesis de hemoglobina a todas las gestantes, el cual es aplicado desde el año 1983. Objetivo: describir el impacto del Programa Cubano de prevención de anemia por hematíes falciformes en Granma, en el período de 1987-2021. Métodos: se realizó un estudio observacional, ambipestivo, de serie de casos en las gestantes pertenecientes a la provincia Granma en el período 1987-2021. Se revisaron las estadísticas de los departamentos municipales y provincial. Se analizaron las variables como frecuencia de portadoras, variantes de hemoglobina, cobertura de esposos estudiados, diagnóstico prenatal molecular y casos positivos. Para el análisis estadístico se emplearon medidas de resumen como frecuencias absolutas y relativas. Resultados: se comprobó que la frecuencia de portadoras de la provincia es de 3,9 %. La variante más frecuente es la Hb AS. Se estudiaron 62 % de los esposos. Se realizó el 70,3 % de los diagnósticos moleculares, de los cuales 21,3 % fue positivo optando por la terminación voluntaria de la gestación en el 69 % de ellos. Conclusiones: se logró impacto del Programa Cubano de prevención de anemia por hematíes falciformes en el período de 1987-2021 en cuanto a la detección de portadoras de la enfermedad, frecuencia de diagnósticos prenatales moleculares, enfermos e interrupciones por esta causa.


Introduction: Sickle cell anemia is an autosomal recessive genetic disease. It is considered the most common hereditary disease in Cuba. The Cuban sickle cell anemia prevention Program is based on the screening of all pregnant women by hemoglobin electrophoresis. This program has been applied since 1983. Objective: To describe the impact of the Cuban sickle cell anemia prevention Program during the period 1987-2021. Methods: An observational study was conducted on a series of cases of pregnant women belonging to the province of Granma in the period from 1987 to 2021. Municipal and provincial statistics were reviewed. Variables such as carrier frequency, hemoglobin variants, coverage of studied spouses, molecular prenatal diagnosis, and positive cases were analyzed. Summary measures such as absolute and relative frequencies were used for statistical analysis. Results: The carrier frequency in the province was found to be 3.9%. The most common variant is Hb AS. Sixty-two percent of spouses were screened. Molecular diagnosis was performed in 70.3%, of which 21.3% were positive and 69% opted for voluntary abortion. Conclusions: The Cuban Sickle Cell Disease Prevention Program has had a great impact on halting the number of carriers of the disease, the frequency of prenatal molecular diagnoses, and the number of patients with sickle cell anemia in the period 1987-2021.


Introdução: A anemia falciforme é uma doença genética autossómica recessiva, sendo considerada a doença hereditária mais comum em Cuba. O programa cubano de prevenção da anemia falciforme baseia-se no rastreio de todas as mulheres grávidas através da eletroforese da hemoglobina, que tem sido aplicado desde 1983. Objetivo: Descrever o impacto do Programa Cubano de Prevenção da Anemia Falciforme durante o período 1987-2021. Métodos: Foi realizado um estudo observacional de uma série de casos de mulheres grávidas pertencentes à província de Granma no período de 1987 a 2021. Foram revistas estatísticas municipais e provinciais. Foram analisadas variáveis como frequência de portadores, variantes de hemoglobina, cobertura de cônjuges estudados, diagnóstico pré-natal molecular e casos positivos. Para a análise estatística foram utilizadas medidas de síntese como frequências absolutas e relativas. Resultados: A frequência de portadores na província foi de 3,9%. A variante mais comum é a Hb AS. Sessenta e dois por cento dos cônjuges foram rastreados. O diagnóstico molecular foi efectuado em 70,3%, dos quais 21,3% foram positivos e 69% optaram pelo aborto voluntário. Conclusões: O Programa Cubano de Prevenção da Doença Falciforme teve um grande impacto na redução do número de portadores da doença, na frequência dos diagnósticos moleculares pré-natais e no número de pacientes com anemia falciforme no período 1987-2021.

10.
Ciênc. rural (Online) ; 54(1): e20220317, 2024. tab, ilus
Artigo em Inglês | VETINDEX | ID: biblio-1447955

RESUMO

The cheese after processing, generates whey, which is a residue that has potential benefit to human health. However, there are still few studies that seek to evaluate the composition and potential benefits of goat milk whey, even though it is a promising dairy source. Despite this, the large-scale generation of this waste can favor the environmental impact when improperly disposed of. Given this, the use of all content of dairy production can reverse this impact and increase the income of the dairy industry. Thus, this research prepared a powdered product based on goat's milk whey from the spray drying technique and evaluate its physicochemical composition by chemical analytical methods and protein profile through polyacrylamide gel electrophoresis. The elaborated product had a higher carbohydrate content than protein, and the identification of protein fractions showed that the most expressive bands represented beta-lactoglobulin and alpha-lactoalbumin. Therefore, the elaborated product has the potential to generate novel products for human consumption and with potential health benefits.


O queijo após processado gera o soro de leite, sendo este um resíduo que possui potencial benefício à saúde humana. No entanto, ainda são poucos os estudos que buscam avaliar a composição e os potenciais benefícios do soro de leite de cabra, mesmo esta sendo uma fonte láctea promissora. Apesar disso, a geração em larga escala deste resíduo pode favorecer o impacto ambiental quando descartado inadequadamente. Em vista disso, a utilização de todo conteúdo gerado na produção de lácteos pode reverter este impacto e aumentar o rendimento da indústria de laticínios. Diante disso, é importante buscar o desenvolvimento de novos produtos para favorecer a redução do impacto gerado. Assim, este trabalho teve como objetivo elaborar um produto em pó, à base de soro de leite de cabra a partir da secagem pela técnica de spray drying e avaliar sua composição físico-química por métodos químicos analíticos e perfil proteico através da eletroforese em gel de poliacrilamida. O produto elaborado apresentou maior teor de carboidratos do que proteína, e a identificação das frações proteicas mostrou que as bandas mais expressivas representavam beta-lactoglobulina e alfa-lactoalbumina. Portanto, o produto elaborado apresenta potencial para geração de novos produtos para consumo humano e com prováveis benefícios à saúde.


Assuntos
Cabras , Laticínios , Soro do Leite , Resíduos de Alimentos
11.
Crit Rev Anal Chem ; : 1-13, 2023 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-38147303

RESUMO

Lipid peroxidation occurs when substances, such as reactive oxygen species, attack lipids. Polyunsaturated fatty acids are the main targets. Several products are formed, including primary products such as lipid hydroperoxides and secondary products such as malondialdehyde (MDA), the most used lipid peroxidation biomarker. As MDA levels are elevated in several diseases, MDA is an essential indicator for assessing pathological states. The thiobarbituric acid reactive substances assay is the most widely used method for MDA determination. However, it lacks specificity. Capillary Electrophoresis (CE) is a separation technique that has been used to quantify MDA in biological samples. This technique has advantages such as the low amount of biological sample required, absence or low volume of organic solvent, short analysis time, separation of interferents, sample preparation step with only protein precipitation, and the possibility of direct detection of the MDA, without derivatization. To our knowledge, this review article is the first to show the CE background for analyzing MDA in biological samples. Therefore, given the potential of MDA in evaluating pathological states, this article demonstrates the potential of CE to become a reference method for MDA determination in clinical analysis laboratories, which will play a significant role in diagnosing and monitoring diseases.

12.
Rev. biol. trop ; Rev. biol. trop;71(1)dic. 2023.
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1449500

RESUMO

Introduction: The Clown anemonefish (Amphiprion ocellaris) is the most popular fish species in the marine aquarium trade; however, there is a lack of information on their digestive physiology during larval ontogeny, valuable information needed for diet design and management protocols. Objective: To characterize the early digestive enzymes of A. ocellaris larvae. Methods: We used three pools (10 larvae each) and extracted 10 samples per tank, from just before hatching to the 38th day. We analyzed the specific activity of acid and alkaline proteases, trypsin, chymotrypsin, leucine aminopeptidase and lipase; and did acid and alkaline protease zymograms. Results: We detected all measured enzymes at hatching. Acid proteases increased in activity until the 38th day. Alkaline proteases, trypsin, chymotrypsin, and leucine aminopeptidase had the same pattern, and maximum activity on the 8th day, decreasing at the 38th day. Lipase activity peaked on the 8th and 30th day. The acid zymogram had a single band, appearing on the 8th day. A total of eight alkaline proteases were revealed (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 and 25.1 KDa), with seven bands on the 1st day and all bands from the 3rd to 8th day, decreasing at two bands (41.9 and 25.1 KDa) in the 38th day. Conclusion: A. ocellaris has a functional stomach on the 8th day, and, on the 38th day, a digestive omnivore pattern with a tendency to carnivory.


Introducción: El pez payaso (Amphiprion ocellaris) es la especie de pez más popular en el comercio de acuarios marinos; sin embargo, falta información sobre su fisiología digestiva durante la ontogenia larval, información valiosa necesaria para protocolos de diseño y manejo dietético. Objetivo: Caracterizar las enzimas digestivas tempranas de larvas de A. ocellaris. Métodos: Usamos tres homogenados (con 10 larvas cada uno) y extrajimos 10 muestras por tanque, justo antes de la eclosión hasta el día 38. Analizamos la actividad específica de proteasas ácidas y alcalinas, tripsina, quimotripsina, leucina aminopeptidasa y lipasa; e hicimos zimogramas de proteasas ácidas y alcalinas. Resultados: Detectamos todas las enzimas medidas en la eclosión. La actividad de proteasas ácidas incrementó hasta el día 38. Proteasas alcalinas, tripsina, quimotripsina, y leucina aminopeptidasa tuvieron el mismo patrón, con actividad máxima en el octavo día, decreciendo en el día 38. Hubo picos en la actividad lipasa a los ocho y 30 días. El zimograma ácido tuvo una banda única, apareciendo al octavo día. Se hallaron ocho proteasas alcalinas (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 y 25.1 KDa), con siete bandas al primer día, y todas las bandas entre el tercer y octavo día, bajando a dos bandas (41.9 y 25.1 KDa) al día 38. Conclusión: A. ocellaris tiene un estómago funcional al octavo día, y, al día 38, un patrón digestivo omnívoro con tendencias carnívoras.

13.
Int J Mol Sci ; 24(19)2023 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-37834205

RESUMO

The emergence of hyper-virulent and multidrug-resistant (MDR) strains of Klebsiella pneumoniae isolated from patients with hospital- and community-acquired infections is a serious health problem that increases mortality. The molecular analysis of virulome expression related to antimicrobial-resistant genotype and infection type in K. pneumoniae strains isolated from patients with hospital- and community-acquired infections has been poorly studied. In this study, we analyzed the overall expression of the virulence genotype associated with the antimicrobial resistance genotype and pulse field gel electrophoresis (PFGE) type (PFtype) in K. pneumoniae. We studied 25 strains of K. pneumoniae isolated from patients who developed bacteremia and pneumonia during their hospital stay and 125 strains from outpatients who acquired community-acquired infections. Susceptibility to 12 antimicrobials was determined by Kirby-Bauer. The identification of K. pneumoniae and antibiotic-resistance genes was performed using polymerase chain reaction (PCR). To promote the expression of the virulence genes of K. pneumoniae, an in vitro infection model was used in human epithelial cell lines A549 and A431. Bacterial RNA was extracted with the QIAcube robotic workstation, and reverse transcription to cDNA was performed with the Reverse Transcription QuantiTect kit (Qiagen). The determination of the expression of the virulence genes was performed by real-time PCR. In addition, 57.3% (n = 86) of the strains isolated from patients with hospital- and community-acquired infections were multidrug-resistant (MDR), mainly to beta-lactam antibiotics (CB, AM, CFX, and CF), aminoglycosides (GE), quinolones (CPF and NOF), nitrofurantoin (NF), and sulfamethoxazole/trimethoprim (SXT). The most frequently expressed genes among strains isolated from hospital- and community-acquired infections were adhesion-type, ycfm (80%), mrkD (51.3%), and fimH (30.7%); iron uptake, irp2 (84%), fyuA (68.7%), entB (64.7%), and irp1 (56.7%); and protectins, rpmA (26%), which were related to antibiotic-resistance genes, blaTEM (96%), blaSHV (64%), blaCITM (52.6%), blaCTXM-1 (44.7%), tetA (74%), sul1 (57.3%), aac(3)-IV (40.7%), and aadA1 (36%). The results showed the existence of different patterns of expression of virulome related to the genotype of resistance to antimicrobials and to the PFtypes in the strains of K. pneumoniae that cause hospital- and community-acquired infections. These findings are important and may contribute to improving medical treatment strategies against infections caused by K. pneumoniae.


Assuntos
Infecções Comunitárias Adquiridas , Infecção Hospitalar , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Klebsiella pneumoniae , Infecções Comunitárias Adquiridas/tratamento farmacológico , Infecções Comunitárias Adquiridas/genética , Genótipo , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/genética , Infecção Hospitalar/microbiologia , Hospitais , Testes de Sensibilidade Microbiana , beta-Lactamases/genética , Farmacorresistência Bacteriana Múltipla/genética
14.
Electrophoresis ; 2023 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-37853649

RESUMO

Developing paper-based electrophoretic methods involve dealing with significant uncertainty levels when compared to their capillary counterparts. Critical information for developing these kinds of methods are the electrophoretic mobility of background electrolytes and samples. This work presents the design and characterization of a device for measuring the electrophoretic mobilities of dyes in porous media. The device was developed with the aim of validating a previously presented model and also proposing a protocol for the straightforward determination of electrophoretic mobilities in porous media when open-channel values are already known. Whatman #1 paper was used as a model substrate as far as it is the most common porous medium substrate for paper-based electrophoresis. The device was designed using a numerical simulation-assisted approach, utilizing OpenFOAM® and specific solvers for capillary transport and electromigration, namely porousMicroTransport and electroMicroTransport, respectively. The electrophoretic mobilities of five dyes were analyzed experimentally with the proposed device. To establish appropriate comparative values at different pHs, experiments in fused silica capillaries were also performed. An effective parameter model for describing the electrophoretic behavior of dyes in porous media, that is, the constriction factor, was found consistent with previous reports for the Whatman #1 paper. This consistency was found after considering (via direct measurements) the chromatographic effect of the medium over each dye. Consequently, the recorded values hold significant worth due to their potential for direct application in designing new experiments or devices in Whatman #1 paper. With the validation of the model through the experiments with the proposed device, those researchers interested on developing electrophoretic methods in porous substrates can make use of the open-channel electrophoretic mobilities reported in the literature, or in the well-known software databases, and correct them for the media of interest just by performing two simple characterization steps.

15.
Trop Life Sci Res ; 34(3): 259-277, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37860088

RESUMO

The sperm interacts with seminal plasma proteins during its transport through the female reproductive tract to reach the oocyte. Seminal plasma proteins have been associated as biomarkers of fertility in bovine males, while two-dimensional electrophoresis in polyacrylamide gels under denaturing conditions (2D-PAGE) is a useful technique for their separation, allowing their subsequent analysis with the aid of specialised software. Brahman bulls are known for their tolerance to tropical conditions such as low-quality pastures, high temperatures, and relative humidity as well as moderate resistance to infestations by parasites and insects. The present study describes the two-dimensional electrophoretic profiles of the seminal plasma proteins in the rainy and dry seasons, associating them with the fertility of Brahman bulls in the Colombian Orinoquía in a 90-days breeding season and a single-sire mating system (1 bull per 50 Brahman cows) with 60 consecutive days of rest. The fertility-related seminal plasma protein spots increased in the dry season. Likewise, a meaningful relationship was found between the protein spots that possibly coincide with the Binder of Sperm Proteins. It was also found that bulls with the highest percentages of pregnancy also had similarities in their 2D seminal plasma maps. We conclude that the seminal plasma protein profile of Brahman bulls raised in the Colombian low tropic changes between rainy and dry seasons, and such changes may influence the reproductive performance of those animals.

16.
Odontol.sanmarquina (Impr.) ; 26(3): e24828, jul.-set.2023.
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1537672

RESUMO

Currently, the FDI recommendation (World Dental Federation) for fluoride concen­tration in toothpastes is that the total fluoride content declared by manufacturers be between 1,000 and 1,500 ppm, and of this amount, at least 800 ppm should be bioavai­lable fluoride. The aim of this study was to describe the market of toothpastes marketed in Medellín and to measure the concentration of bioavailable fluoride by capillary elec­trophoresis to verify their compliance with the FDI recommendation for the prevention of dental caries. After sampling the toothpastes available for sale in the 16 communes of the city of Medellin, a single operator measured in triplicate the concentration of bioavailable fluoride in 36 samples of previously blinded toothpastes using the capi­llary electrophoresis (EC) technique. Most of the evaluated toothpastes that declared a content of 1,000­1,500 ppm fluoride met the recommendation of presenting at least 800 ppm bioavailable fluoride. Measurement of bioavailable fluoride in toothpastes with MFP (sodium monofluorophosphate) is recommended, as it is known that binding to the abrasive can decrease its concentration over time.


Actualmente la recomendación de la FDI (Federación Dental Internacional) sobre la concentración de fluoruro en cremas dentales es que el contenido de fluoruro total de­clarado por los fabricantes sea entre 1.000 y 1.500 ppm, y que de esta cantidad al menos 800 ppm sea fluoruro biodisponible. El objetivo de este estudio fue describir el mercado de las cremas dentales comercializadas en Medellín y medir en estas la concentración de fluoruro biodisponible por electroforesis capilar para verificar su cumplimiento de la recomendación de la FDI para la prevención de la caries dental. Luego de realizar un mues­treo en las 16 comunas de la ciudad de Medellín sobre las cremas dentales disponibles a la venta, un solo operador midió por triplicado la concentración de fluoruro biodisponible en 36 muestras de cremas dentales previamente cegadas, mediante la técnica de electroforesis capilar (EC). La mayoría de las cremas dentales evaluadas que declaraban contenido de 1.000 a 1.500 ppm de fluoruro, cumplieron la recomendación de presentar al menos 800 ppm de fluoruro biodisponible. Se recomienda realizar la medición del fluoruro biodisponible en las cremas con MFP, ya que se conoce que la unión al abrasivo puede disminuir su concentración en el tiempo.

17.
J Sep Sci ; 46(19): e2300442, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37582647

RESUMO

A capillary electrophoresis-mass spectrometry method was used to analyze naphthenic acids in produced water samples. It was possible to detect cyclopentanecarboxylic, benzoic, cyclohexanebutyric, 1-naphthoic, decanoic, 3,5-dimethyladamantane-1-carboxylic, 9-anthracenecarboxylic, and pentadecanoic acids within ca. 13 min using a buffer composed of 40 mmol/L ammonium hydroxide, 32 mmol/L acetic acid and 20% v/v isopropyl alcohol, pH 8.6. The proposed method showed good repeatability, with relative standard deviation (RSD) values of 6.6% for the sum of the peak areas and less than 2% for the analysis time. In the interday analysis, the RSD values for the sum of the peak areas and migration time were 10.3% and 10%, respectively. The developed method demonstrated linear behavior in the concentration range between 5 and 50 mg/L for benzoic, decanoic, 3,5-dimethyladamantane-1-carboxylic and 9-anthracenecarboxylic acids, and between 10 and 50 mg/L for cyclopentanecarboxylic, cyclohexanebutyric, 1- naphthoic, and pentadecanoic acids. The detection limits values ranged from 0.31 to 1.64 mg/L. Six produced water samples were analyzed and it was possible to identify and quantify cyclopentanecarboxylic, benzoic, cyclohexanebutyric, and decanoic acids. The concentrations varied between 4.8 and 98.9 mg/L, proving effective in the application of complex samples.

18.
Artigo em Inglês | MEDLINE | ID: mdl-37535992

RESUMO

Through density functional theory calculations was studied theoretically the formation process of a magnetic and mesoporous molecularly imprinted polymer for ractopamine (RAC), evaluating the molecular electrostatic potential map, functional monomers, functional monomer / template stoichiometry and crosslink agents. The results revealed that the best conditions for the synthesis were established with acrylic acid as functional monomer in a 1: 4 stoichiometry using acetonitrile as the solvent and ethylene glycol dimethacrylate as crosslink agent. It was observed that nine hydrogen bonds established between the RAC and acrylic acid play a key role on the pre-polymerization complex. In addition, three analytical methods using HPLC, UHPLC and CE instruments were optimized for rapid analysis. The adsorbent was experimentally synthesized considering the best conditions found at the molecular level and characterized by FTIR, DRX, TGA, SEM, TEM, surface analysis, and wettability. After that, the synthesized material was used in magnetic solid phase extraction combined with capillary electrophoresis in a preliminary RAC recovery study from milk samples. Finally, greenness metric with a score of 0.55 have been obtained for the sample preparation procedure using the online AGREEprep metric.


Assuntos
Impressão Molecular , Polímeros Molecularmente Impressos , Impressão Molecular/métodos , Adsorção , Extração em Fase Sólida/métodos , Cromatografia Líquida de Alta Pressão/métodos , Fenômenos Magnéticos
19.
J Clin Immunol ; 43(8): 1873-1880, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37505322

RESUMO

PURPOSE: This study aimed to investigate the correlation between calculated globulin (CG, total protein level minus albumin level) and the gamma globulin fraction (Gamma), obtained from serum protein electrophoresis with serum IgG levels in adults (≥ 18 years). METHODS: Using linear regression models, analyses of CG and Gamma levels correlation with IgG levels in adults were performed. Receiver-operator curves were created to determine cutoff values and the respective sensitivity and specificity measures. RESULTS: A total of 886 samples were analyzed. CG and Gamma were positively and statistically correlated with IgG levels (r2 = 0.4628 for CG, and = 0.7941 for Gamma, p < 0.0001 for both analyses). For the detection of hypogammaglobulinemia, i.e., IgG level below the reference value (6 g/L), a CG cutoff value of 24 g/L showed a sensitivity of 86.2% (95% CI 69.4-94.5) and a specificity of 92% (90.0-93.6). A Gamma cutoff value of 7.15 g/L yielded a sensitivity of 100% (88.3-100) and a specificity of 96.8 (95.3-97.8). CONCLUSION: Both CG and Gamma levels determined by protein electrophoresis analysis may be used to screen for antibody deficiencies in adults, enabling earlier diagnosis of antibody deficiencies in a routine clinical setting.


Assuntos
Agamaglobulinemia , Doenças da Imunodeficiência Primária , Humanos , Adulto , Eletroforese , Globinas , Imunoglobulina G
20.
Hematol., Transfus. Cell Ther. (Impr.) ; 45(supl.2): S140-S147, July 2023. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1514184

RESUMO

ABSTRACT Introduction: Colombia has been subject to intense genetic and cultural currents due to its geographical location. Hemoglobinopathies are the most common recessive diseases found worldwide and represent an important public health problem, according to the region and ancestry of each country. Objectives: To evaluate the frequency of hemoglobin variants according to the geographical region in a population group adjusted to sex and age in Colombia. Methods: This was a descriptive retrospective study of hemoglobin variants performed by electrophoresis in patients treated at and/or referred to specialized care institutions in Bogota, Colombia between January 2009 and December 2020. Results: A total of 2,224 results were analyzed, 48.4% male and 51.5% female; 63.3% of patients were without alterations, 14.3% presented with thalassemia, 17.3%, HbS, 2.3%, HbS/C, 1.8%, HbC, 0.5%, HbE and 0.5% persistent HbF, with HbS being more prevalent in males (p = 0.005). When assessing the geographical regions of Colombia, a higher prevalence of HbS was found in the Pacific (p = 0.005) and Caribbean regions, while Thalassemia and HbS were more prevalent in the Andean and Orinoquia regions, and it was rare to find any hemoglobinopathies (p = 0.0001) in the Amazonian region. Conclusions: The main hemoglobinopathies found in Colombia are HbS, predominantly in males, and Thalassemia. The distribution of hemoglobinopathies in different geographical regions of Colombia is influenced by ancestry.

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