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Background: Patients in intensive care units with traumatic brain injuries (TBI) frequently present acid-base abnormalities and coagulability disorders, which complicate their condition.Objective: To identify protonation through in silico simulations of molecules involved in the process of coagulation in standard laboratory tests.Materials and methods: Ten patients with TBI were selected from the intensive care unit in addition to ten "healthy control subjects", and another nine patients as "disease control subjects"; the latter being a comparative group, corresponding to subjects with diabetes mellitus 2 (DM2). Fibrinogen, FVII, FVIII, FIX, FX, and D-dimer in the presence of acidification were evaluated in 20 healthy subjects in order to compare clinical results with molecular dynamics (MD), and to explain proton interactions and coagulation molecules.Results: The TBI group presented a slight, non-significant increase in D-dimer; but this was not present in "disease control subjects". Levels of fibrinogen, FVII, FIX, FX, and D-dimer were affected in the presence of acidification. We observed that various specific residues of coagulation factors "trap" ions.Conclusion: Protonation of tissue factor and factor VIIa may favor anticoagulant mechanisms, and protonation does not affect ligand binding sites of GPIIb/IIIa (PAC1) suggesting other causes for the low affinity to PAC1.
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Lesões Encefálicas Traumáticas , Prótons , Coagulação Sanguínea , Lesões Encefálicas Traumáticas/complicações , Humanos , Simulação de Dinâmica MolecularRESUMO
Obesity is a serious medical condition worldwide, which needs new approaches and recognized international consensus in treating diseases leading to morbidity. The aim of this review was to examine heterogeneous links among the various phenotypes of obesity in adults. Proteins and associated genes in each group were analysed to differentiate between biomarkers. A variety of terms for classification and characterization within this pathology are currently in use; however, there is no clear consensus in terminology. The most significant groups reviewed include metabolically healthy obese, metabolically abnormal obese, metabolically abnormal, normal weight and sarcopenic obese. These phenotypes do not define particular genotypes or epigenetic gene regulation, or proteins related to inflammation. There are many other genes linked to obesity, though the value of screening all of those for diagnosis has low predictive results, as there are no significant biomarkers. It is important to establish a consensus in the terminology used and the characteristics attributed to obesity subtypes. The identification of specific molecular biomarkers is also required for better diagnosis in subtypes of obesity.
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Biomarcadores , Obesidade/diagnóstico , Obesidade/genética , Proteínas/genética , Adulto , Genótipo , Humanos , Obesidade/classificação , Obesidade/epidemiologia , FenótipoRESUMO
ß-Glucosidase (EC 3.2.1.21) is a prominent member of the GH1 family of glycoside hydrolases. The properties of this ß-glucosidase appear to include resistance to temperature, urea, and iodoacetamide, and it is activated by 2-ME, similar to other members. ß-Glucosidase from chayote (Sechium edule) was purified by ionic-interchange chromatography and molecular exclusion chromatography. Peptides detected by LC-ESI-MS/MS were compared with other ß-glucosidases using the BLAST program. This enzyme is a 116 kDa protein composed of two sub-units of 58 kDa and shows homology with Cucumis sativus ß-glucosidase (NCBI reference sequence XP_004154617.1), in which seven peptides were found with relative masses ranging from 874.3643 to 1587.8297. The stability of ß-glucosidase depends on an initial concentration of 0.2 mg/mL of protein at pH 5.0 which decreases by 33% in a period of 30 h, and then stabilizes and is active for the next 5 days (pH 4.0 gives similar results). One hundred µg/mL ß-D-glucose inhibited ß-glucosidase activity by more than 50%. The enzyme had a Km of 4.88 mM with p-NPG and a Kcat of 10,000 min(-1). The optimal conditions for the enzyme require a pH of 4.0 and a temperature of 50 °C.
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Cucurbitaceae/enzimologia , beta-Glucosidase/isolamento & purificação , Sequência de Aminoácidos , Precipitação Química , Cromatografia por Troca Iônica , Estabilidade Enzimática , Glucose/química , Concentração de Íons de Hidrogênio , Cinética , Dados de Sequência Molecular , Proteínas de Plantas , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , beta-Glucosidase/químicaRESUMO
BACKGROUND: Dysbiosis during childhood impacts the configuration and maturation of the microbiota. The immaturity of the infant microbiota is linked with the development of inflammatory, allergic, and dysmetabolic diseases. AIMS: To identify taxonomic changes associated with age and GDM and classify the maturity of the intestinal microbiota of children of mothers with GDM and children without GDM (n-GDM). METHODS: Next-generation sequencing was used to analyze the V3-V4 region of 16S rRNA gene. QIIME2 and Picrust2 were used to determine the difference in the relative abundance of bacterial genera between the study groups and to predict the functional profile of the intestinal microbiota. RESULTS: According to age, the older GDM groups showed a lower alpha diversity and different abundance of Enterobacteriaceae, Veillonella, Clostridiales, and Bacteroides. Regarding the functional profile, PWY-7377 and K05895 associated with Vitamin B12 metabolism were reduced in GDM groups. Compared to n-GDM group, GDM offspring had microbiota immaturity as age-discriminatory taxa in random forest failed to classify GDM offspring according to developmental age (OOB error 81%). Conclusion. Offspring from mothers with GDM have a distinctive taxonomic profile related to taxa associated with gut microbiota immaturity.
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Bacteroides , Diabetes Gestacional , Microbioma Gastrointestinal , RNA Ribossômico 16S , Veillonella , Humanos , Diabetes Gestacional/microbiologia , Feminino , Gravidez , Bacteroides/genética , RNA Ribossômico 16S/genética , Veillonella/genética , Lactente , Adulto , Masculino , Disbiose/microbiologia , Fezes/microbiologia , Pré-Escolar , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
Diabetes mellitus is a chronic metabolic disorder, being globally one of the most deadly diseases. This disease requires continually monitoring of the body's glucose levels. There are different types of sensors for measuring glucose, most of them invasive to the patient. Fiber optic sensors have been proven to have advantages compared to conventional sensors and they have great potential for various applications, especially in the biomedical area. Compared to other sensors, they are smaller, easy to handle, mostly non-invasive, thus leading to a lower risk of infection, high precision, well correlated and inexpensive. The objective of this review article is to compare different types of fiber optic sensors made with different experimental techniques applied to biomedicine, especially for glucose sensing. Observations are made on the way of elaboration, as well as the advantages and disadvantages that each one could have in real applications.
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Técnicas Biossensoriais , Glucose/análise , Tecnologia de Fibra Óptica , Humanos , Fibras ÓpticasRESUMO
The human coagulation factor VIII (FVIII) is essential in the intrinsic pathway of blood coagulation and circulates mainly as a non-covalently bound complex with the von Willebrand factor (VWF). This complex (FVIII/VWF) protects FVIII from degradation and cellular uptake, although no biological role has been identified yet for this complex. The FVIII/VWF complex was purified from a healthy donor's plasma by affinity chromatography on a Sepharose 4B-Concanavalin A column and was used to determine its capability to interact with erythrocytes and platelets. The purified FVIII/VWF complex at 6.0 and 12 microg/ml agglutinates rabbit and bovine erythrocytes, and showed negative agglutination with erythrocytes from other species including human ABO. Treatment of erythrocytes with Clostridium perfringens sialidase or trypsin increased four-fold the activity toward rabbit erythrocytes and positive agglutination for human A and B erythrocytes, suggesting the presence of FVIII/VWF-cryptic receptors in these erythrocytes. Goat, pig, or human O erythrocytes were not agglutinated even after enzymatic treatment. Fucose or N-acetyl-glucosamine (GlcNAc), at 10 mM, inhibited agglutinating activity of the complex with rabbit, human A and B erythrocytes, whereas galactose and N-acetyl-galactosamine, even at 200 mM, showed no effect on the complex activity. The FVIII/VWF complex, at 1.5 microg/200,000 platelets, significantly decreased platelet aggregation (p < 0.001) when compared with the effect of platelet-rich plasma; this effect was inhibited with 15 mM GlcNAc or fucose. ELISA assays on FVIII/VWF coated polystyrene plates confirmed specific binding to fucose- or biotinylated GlcNAc-dextran derivatives. We therefore propose that the FVIII/VWF complex possesses lectin activity.
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Fator VIII/metabolismo , Lectinas/metabolismo , Complexos Multiproteicos/metabolismo , Fator de von Willebrand/metabolismo , Animais , Cromatografia de Afinidade , Ensaio de Imunoadsorção Enzimática , Eritrócitos/metabolismo , Humanos , Complexos Multiproteicos/farmacologia , Agregação Plaquetária/efeitos dos fármacosRESUMO
The chronic indeterminate phase of Chagas' disease is asymptomatic despite positive test results for antibodies specific to Trypanosoma cruzi. CD62P-APC (P-selectin) and PAC-1 FITC (GpIIb/IIIa) may improve diagnosis as biomarkers of platelet activity. Nine asymptomatic seropositive subjects, previously untreated, were selected from a blood bank within a year of Chagas' disease detection, in addition to a control group of four. All subjects were evaluated by flow cytometry for CD62P, PAC-1 and CD41, and in a complementary study, by Tissue Doppler Echocardiography for isovolumic relaxation times (IVRT) and E/A ratios. The subjects were classified as positive or negative for CD62P and PAC-1 by a cut off obtained from their mean±2SD. For IVRT and E/A ratios, cut offs were obtained from the American Society of Echocardiography and the European Association of Cardiovascular Imaging recommendations. Fisher's exact test was used for associated findings. Pre-test and post-test probability, sensitivity, specificity, positive and negative predictive values and likelihood ratios were calculated. Abnormalities were expressed as platelet hyperactivity and ventricular dysfunction in CD62P, PAC-1, IVRT and E/A ratios. CD62P appears to have greater sensitivity (0.75) and PAC-1, more accurate specificity (0.75), which may explain thrombotic events in Chagas' disease. We recommend the use of CD62P and PAC-1 as biomarkers of platelet hyperactivity in patients in the chronic indeterminate phase of Chagas' disease.
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The association between type 2 diabetes mellitus (T2DM) and systemic inflammation may increase platelet reactivity and the accelerated development of vascular disease. Platelets are able to modulate the function of immune cells via the direct release of growth factors and pro-inflammatory chemokines through the production of microvesicles. The microvesicles trigger a transcellular delivery system of bioactive molecules to other cells acting as vectors in the exchange of biological information. Here, we consider the influence of platelets and platelet-derived microvesicles on cells of the immune system and the implications in the pathogenesis of T2DM.
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Plaquetas/imunologia , Micropartículas Derivadas de Células/imunologia , Diabetes Mellitus Tipo 2/imunologia , Imunidade Adaptativa , Animais , Plaquetas/metabolismo , Micropartículas Derivadas de Células/metabolismo , Diabetes Mellitus Tipo 2/sangue , Humanos , Imunidade Inata , Mediadores da Inflamação/sangue , Mediadores da Inflamação/imunologia , Estresse Oxidativo , Transdução de SinaisRESUMO
OBJECTIVE: To test the hypothesis that the color of meconial fluid is associated with inflammatory biomarkers, by determining C-reactive protein (CRP) and Interleukin-6 (IL-6) in serum from the umbilical cord. METHODS: In this prospective study, the authors selected 30 newborns with meconium-stained amniotic fluid (MSAF): 14 with green/brown 656 R color and 16 with brown/cinnamon 654 R color, and 20 newborns which showed clear amniotic fluid without MSAF (non-MSAF); all newborns were from mothers without risk factors for neonatal sepsis. RESULTS: IL-6 concentration from umbilical cord blood, [median of 12.9 pg/mL (interquartile range {IQR} 8.7-31.0)] of MSAF-green/brown 656 R increased significantly (p < 0.05) when compared with IL-6 concentration, [median of 9.2 pg/mL (IQR 7.2-12.2)] of newborns with clear amniotic fluid and without meconium. CRP from MSAF-green/brown 656 R was median of 0.5 mg/mL (IQR 0.0-2.7), and median of 1.0 mg/mL (IQR 0.0-5.5) from clear amniotic fluid, without meconium. CONCLUSIONS: Significant association was found between MSAF-green/brown 656 R and increase in IL-6, with normal CRP values.