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1.
J Exp Bot ; 69(19): 4633-4649, 2018 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-30053161

RESUMO

Post-translational modifiers such as the small ubiquitin-like modifier (SUMO) peptide act as fast and reversible protein regulators. Functional characterization of the sumoylation machinery has determined the key regulatory role that SUMO plays in plant development. Unlike components of the SUMO conjugation pathway, SUMO proteases (ULPs) are encoded by a relatively large gene family and are potential sources of specificity within the pathway. This study reports a thorough comparative genomics and phylogenetic characterization of plant ULPs, revealing the presence of one ULP1-like and three ULP2-like SUMO protease subgroups within plant genomes. As representatives of an under-studied subgroup, Arabidopsis SPF1 and SPF2 were subjected to functional characterization. Loss-of-function mutants implicated both proteins with vegetative growth, flowering time, and seed size and yield. Mutants constitutively accumulated SUMO conjugates, and yeast complementation assays associated these proteins with the function of ScUlp2 but not ScUlp1. Fluorescence imaging placed both proteins in the plant cell nucleoplasm. Transcriptomics analysis indicated strong regulatory involvement in secondary metabolism, cell wall remodelling, and nitrate assimilation. Furthermore, developmental defects of the spf1-1 spf2-2 (spf1/2) double-mutant opposed those of the major E3 ligase siz1 mutant and, most significantly, developmental and transcriptomic characterization of the siz1 spf1/2 triple-mutant placed SIZ1 as epistatic to SPF1 and SPF2.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Cisteína Endopeptidases/genética , Ligases/genética , Sequência de Aminoácidos , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Cisteína Endopeptidases/química , Cisteína Endopeptidases/metabolismo , Ligases/metabolismo , Filogenia , Alinhamento de Sequência , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/genética , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo
2.
J Oral Maxillofac Surg ; 76(4): 785-792, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29102600

RESUMO

PURPOSE: The aim of this study was to describe the relation between occlusal features and enzyme replacement therapy in patients with mucopolysaccharidoses. MATERIALS AND METHODS: A cross-sectional study was conducted. The sample consisted of 20 patients with mucopolysaccharidoses, 10 of whom were undergoing treatment at a hospital in northeast Brazil. Occlusal features were evaluated by clinical examination and panoramic radiography. A structured questionnaire was administered to evaluate the dental care of each patient. Pearson χ2, Fisher exact, and Mann-Whitney tests were used for data analysis, with a level of significance of 5%. RESULTS: Marked overjet (75%) and anterior open bite (70%) were the most frequent occlusal alterations, and 15% had Class III disorders. Radiography visualized the presence of impacted teeth (75%) and prolonged retention of deciduous teeth (65%). Patients with enzyme replacement therapy had a lower average maximum protrusion (P = .033). A total of 75% of mothers said they had not been advised to take their children to the dentist and 10% of children had never been to the dentist. CONCLUSION: Patients with mucopolysaccharidoses exhibited notable occlusal alterations, especially marked overjet and anterior open bite. Enzyme replacement therapy seems to influence the maximum protrusion of patients.


Assuntos
Terapia de Reposição de Enzimas , Má Oclusão/etiologia , Mucopolissacaridoses/patologia , Adolescente , Adulto , Criança , Pré-Escolar , Estudos Transversais , Terapia de Reposição de Enzimas/efeitos adversos , Terapia de Reposição de Enzimas/métodos , Feminino , Humanos , Masculino , Má Oclusão/diagnóstico por imagem , Má Oclusão Classe III de Angle/diagnóstico por imagem , Má Oclusão Classe III de Angle/etiologia , Mucopolissacaridoses/complicações , Mucopolissacaridoses/tratamento farmacológico , Mordida Aberta/diagnóstico por imagem , Mordida Aberta/etiologia , Sobremordida/diagnóstico por imagem , Sobremordida/etiologia , Radiografia Dentária , Radiografia Panorâmica , Dente Impactado/diagnóstico por imagem , Dente Impactado/etiologia , Adulto Jovem
3.
J Craniofac Surg ; 27(2): 345-9, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26967072

RESUMO

The objective of this study was to identify the epidemiological profile of facial fractures and to associate with clinical-epidemiological variables. An epidemiological survey was conducted in which data retrieved from the records of patients with facial fractures were analyzed. Pearson χ(2) test, Student t-test, one-way analysis of variance, Bonferroni post-hoc test, and logistic regression were used for statistical analysis, adopting a level of significance of 5%. The sample consisted of 718 patients ranging in age from 21 to 30 years (56.4%), including 480 (66.9%) men. Traffic accidents were the main etiological agent (62.7%) and the nasal bones were the most affected (41.8%). An association was observed between age and etiology (P < 0.001), fractured bone (P < 0.001), type of treatment (P < 0.001), hospital length of stay (P < 0.001), and complications (P = 0.001). Patients with traffic accident [odds ratio (OR) = 3.08, P < 0.001] and fall (OR = 2.50, P = 0.049) as etiology had more risk of complications. The type of fractured bone was associated with etiology (P < 0.001), type of treatment (P < 0.001), hospital length of stay (P < 0.001), and complications (maxilla--OR = 1.90, P = 0.004). In conclusion, facial fractures predominantly affect men aged 21 to 30 years. Traffic accidents are the main etiology and the nasal bone is the site most affected. It was observed that the older the patient, the greater the susceptibility to falls, surgical treatment, hospital stay, and complications. Patients with maxillary or mandibular fractures remain hospitalized for longer periods of time. Fractures by traffic accident and fall had more risk of complications. The risk of complications was also higher in patients with maxilla fracture.


Assuntos
Ossos Faciais/lesões , Fraturas Cranianas/epidemiologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Brasil , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osso Nasal/lesões , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Fatores de Risco , Fatores Sexuais , Fraturas Cranianas/etiologia , Adulto Jovem
4.
J Oral Maxillofac Surg ; 72(12): 2539-46, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25262398

RESUMO

PURPOSE: Studies addressing the changes in craniofacial morphology of patients with mucopolysaccharidosis (MPS) are important for a better understanding of the progression of this disease. The present objective was to identify major cephalometric abnormalities in these patients. MATERIALS AND METHODS: This was a cross-sectional case-and-control study. The sample was composed of 2 types of study subjects (patients with MPS and normal subjects). The outcome variables were numerous cephalometric elements that measure facial height, dental positioning, facial growth pattern, and mandibular and maxillary positioning. The Student t test was used to compare the mean angular and linear measurements of the case and control groups and the level of significance was set at a P value less than .05. The Bonferroni method was used for adjustment of the P value (P<.003 was deemed significant). RESULTS: The control group consisted of randomly selected subjects matched to the patients with MPS for the demographic variables of gender and age. Seventeen patients with MPS were evaluated (64.7% female). The mean age of the sample was 13.29 years. One patient had MPS I, 8 had MPS IV, and 8 had MPS VI. The results showed important differences between groups in angular measurements (angle formed by the line between the sella and nasion [SN] and the plane of the gonion and gnathion, angle formed by the intersection of the gnathion point and the SN line, angle formed by the intersection of the long axis of the lower incisor with the line between the nasion and the B point, angle formed by the intersection of the long axes of the upper and lower incisors, and angle formed by the Frankfort horizontal plane with the mandibular plane) and linear measurements (condyle to A point, condyle to gnathion, pro-nasal ( Pn) point to the line between the pogonion and upper incisor, and the nearest point of the anterior half of the soft palate to the posterior pharyngeal wall). The angle formed by the SN line with the plane between the gonion and the gnathion and the distance from the condyle to the A point showed meaningful differences after Bonferroni adjustment. CONCLUSION: Patients with MPS present a tendency toward vertical growth that results in a dolichocephalic facial pattern. In addition, a smaller nasopharyngeal space was observed, a factor that might be responsible for the mouth breathing observed in these patients.


Assuntos
Face , Mucopolissacaridoses/patologia , Estudos Transversais , Humanos , Mucopolissacaridoses/diagnóstico por imagem , Radiografia
5.
Adv Med Sci ; 68(2): 402-408, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37837799

RESUMO

PURPOSE: Dipeptidyl peptidase 4 (DPP4) inactivates a range of bioactive peptides. The cleavage of insulinotropic peptides and glucagon-like peptide 1 (GLP1) by DPP4 directly influences glucose homeostasis. This study aimed to describe the mode of interaction between sitagliptin (an antidiabetic drug) and human DPP4 using in silico approaches. MATERIALS AND METHODS: Docking studies were conducted using AutoDock Vina, 2D and 3D schematic drawings were obtained using PoseView and PLIP servers, and the DPP4-sitagliptin complex was visualized with Pymol software. RESULTS: The best affinity energy to form the DPP4-sitagliptin complex was E-value â€‹= â€‹- 8.1 â€‹kcal â€‹mol-1, as indicated by docking simulations. This result suggests a strong interaction. According to our observations, hydrophobic interactions involving the amino acids residues Tyr663 and Val712, hydrogen bonds (Glu203, Glu204, Tyr663, and Tyr667), π-Stacking interactions (Phe355 and Tyr667), and halogenic bonds (Arg123, Glu204, and Arg356) were prevalent in the DPP4-sitagliptin complex. Root Mean Square Deviation prediction also demonstrated that the global structure of the human DPP4 did not have a significant change in its topology, even after the formation of the DPP4-sitagliptin complex. CONCLUSION: The stable interaction between the sitagliptin ligand and the DPP4 enzyme was demonstrated through molecular docking simulations. The findings presented in this work enhance the understanding of the physicochemical properties of the sitagliptin interaction site, supporting the design of more efficient gliptin-like iDPP4 inhibitors.


Assuntos
Diabetes Mellitus Tipo 2 , Inibidores da Dipeptidil Peptidase IV , Humanos , Fosfato de Sitagliptina/farmacologia , Simulação de Acoplamento Molecular , Inibidores da Dipeptidil Peptidase IV/farmacologia , Inibidores da Dipeptidil Peptidase IV/química , Hipoglicemiantes/farmacologia , Hipoglicemiantes/química , Peptídeos
6.
J Am Chem Soc ; 134(44): 18275-85, 2012 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-23030300

RESUMO

Carbapenems are the most potent ß-lactam antibiotics and key drugs for treating infections by Gram-negative bacteria. In such organisms, ß-lactam resistance arises principally from ß-lactamase production. Although carbapenems escape the activity of most ß-lactamases, due in the class A enzymes to slow deacylation of the covalent acylenzyme intermediate, carbapenem-hydrolyzing class A ß-lactamases are now disseminating in clinically relevant bacteria. The reasons why carbapenems are substrates for these enzymes, but inhibit other class A ß-lactamases, remain to be fully established. Here, we present crystal structures of the class A carbapenemase SFC-1 from Serratia fonticola and of complexes of its Ser70 Ala (Michaelis) and Glu166 Ala (acylenzyme) mutants with the carbapenem meropenem. These are the first crystal structures of carbapenem complexes of a class A carbapenemase. Our data reveal that, in the SFC-1 acylenzyme complex, the meropenem 6α-1R-hydroxyethyl group interacts with Asn132, but not with the deacylating water molecule. Molecular dynamics simulations indicate that this mode of binding occurs in both the Michaelis and acylenzyme complexes of wild-type SFC-1. In carbapenem-inhibited class A ß-lactamases, it is proposed that the deacylating water molecule is deactivated by interaction with the carbapenem 6α-1R-hydroxyethyl substituent. Structural comparisons with such enzymes suggest that in SFC-1 subtle repositioning of key residues (Ser70, Ser130, Asn132 and Asn170) enlarges the active site, permitting rotation of the carbapenem 6α-1R-hydroxyethyl group and abolishing this contact. Our data show that SFC-1, and by implication other such carbapenem-hydrolyzing enzymes, uses Asn132 to orient bound carbapenems for efficient deacylation and prevent their interaction with the deacylating water molecule.


Assuntos
Carbapenêmicos/metabolismo , Serratia/enzimologia , beta-Lactamases/metabolismo , Carbapenêmicos/química , Domínio Catalítico , Cristalografia por Raios X , Hidrólise , Simulação de Dinâmica Molecular , Mutação Puntual , Serratia/química , Serratia/genética , Infecções por Serratia/microbiologia , beta-Lactamases/química , beta-Lactamases/genética
7.
Antimicrob Agents Chemother ; 55(11): 5392-5, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21876065

RESUMO

The subclass B2 metallo-ß-lactamase (MBL) Sfh-I from Serratia fonticola UTAD54 was cloned and overexpressed in Escherichia coli. The recombinant protein binds one equivalent of zinc, as shown by mass spectrometry, and preferentially hydrolyzes carbapenem substrates. However, compared to other B2 MBLs, Sfh-I also shows limited hydrolytic activity against some additional substrates and is not inhibited by a second equivalent of zinc. These data confirm Sfh-I to be a subclass B2 metallo-ß-lactamase with some distinctive properties.


Assuntos
Serratia/enzimologia , beta-Lactamases/metabolismo , Carbapenêmicos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , beta-Lactamases/genética
8.
Curr Res Microb Sci ; 2: 100042, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34841333

RESUMO

Nowadays, clinical and scientific interest in antibiotics, as polymyxin, has increased due to the large number of reports of multiresistant Gram-negative bacteria, as Pseudomonas aeruginosa. The aim of this study was to investigate a related group of proteins for resistance to polymyxins, encoded by P. aeruginosa genome, through in silico analysis. The mobilized colistin resistance 1 (MCR1) protein from Escherichia coli was used for comparison. Similar sequences to the protein MCR1 in P. aeruginosa were analysed for physicochemical properties. 31 protein isoforms in P. aeruginosa (EptA) were found able to confer resistance to polymyxin showing protein lengths between 551 and 572 amino acids, with molecular mass values between 61.36 - 62. 80 kDa, isoelectric point between 6.10 to 7.17, instability index between 33.76 to 41.87, aliphatic index between 98.67 to 102.63 and the hydropathyindex between - 0.008 to 0.094. These proteins belong to the DUF1705 superfamily with bit-score values between 559.81 and 629.78. A high degree of similarity between EpTAs in P. aeruginosa was observed in relation to other proteins that confer resistance to polymyxins, present in Gram-negative bacteria species of clinical interest. Although, further studies are needed to identify the actual contribution of EptAs in P. aeruginosa species.

9.
Carbohydr Polym ; 255: 117432, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33436231

RESUMO

Grape pectic polysaccharides-malvidin-3-O- ß -d-glucoside binding was studied, aiming to unveil the impact of structural diversity of polysaccharides on anthocyanins-polysaccharides interactions. Polysaccharides were extracted with solutions of imidazole (ISP) and carbonate at 4 °C (CSP-4 °C) and room temperature (CSP-RT) and also recovered from the dialysis supernatant of the remaining cellulosic residue after the aqueous NAOH extraction of hemicellulosic polysaccharides (Sn-CR). Polysaccharides richer in homogalacturonan domains, like those present in the CSP-4 °C fraction had approximately 50-fold higher binding affinity to malvidin-3-O- ß-d-glucoside, than polysaccharides with side chains (as ISP and CSP-RT extractable polysaccharides). CSP-4 °C polysaccharides showed a positive effect on malvidin-3-O- ß-d-glucoside colour fading. Hydration equilibrium constant of malvidin-3-O- ß-d-glucoside in the presence of CSP-4 °C polysaccharides was higher, showing the preferential stabilization of the flavylium cation. The results showed that anthocyanins colour stabilization can be promoted by pectic polysaccharide structures such as those extracted by cold carbonate.

10.
Biochim Biophys Acta Biomembr ; 1862(9): 183314, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32304757

RESUMO

α-Synuclein (αsyn) is a cytosolic intrinsically disordered protein (IDP) known to fold into an α-helical structure when binding to membrane lipids, decreasing protein aggregation. Model membrane enable elucidation of factors critically affecting protein folding/aggregation, mostly using either small unilamellar vesicles (SUVs) or nanodiscs surrounded by membrane scaffold proteins (MSPs). Yet SUVs are mechanically strained, while MSP nanodiscs are expensive. To test the impact of lipid particle size on α-syn structuring, while overcoming the limitations associated with the lipid particles used so far, we compared the effects of large unilamellar vesicles (LUVs) and lipid-bilayer nanodiscs encapsulated by diisobutylene/maleic acid copolymer (DIBMA) on αsyn secondary-structure formation, using human-, elephant- and whale -αsyn. Our results confirm that negatively charged lipids induce αsyn folding in h-αsyn and e-αsyn but not in w-αsyn. When a mixture of zwitterionic and negatively charged lipids was used, no increase in the secondary structure was detected at 45 °C. Further, our results show that DIBMA/lipid particles (DIBMALPs) are highly suitable nanoscale membrane mimics for studying αsyn secondary-structure formation and aggregation, as folding was essentially independent of the lipid/protein ratio, in contrast with what we observed for LUVs having the same lipid compositions. This study reveals a new and promising application of polymer-encapsulated lipid-bilayer nanodiscs, due to their excellent efficiency in structuring disordered proteins such as αsyn into nontoxic α-helical structures. This will contribute to the unravelling and modelling aspects concerning protein-lipid interactions and α-helix formation by αsyn, paramount to the proposal of new methods to avoid protein aggregation and disease.


Assuntos
Lipídeos de Membrana/química , Polímeros/farmacologia , Lipossomas Unilamelares/química , alfa-Sinucleína/química , Alcenos/química , Alcenos/farmacologia , Humanos , Proteínas Intrinsicamente Desordenadas/química , Bicamadas Lipídicas/química , Maleatos/química , Maleatos/farmacologia , Proteínas de Membrana/química , Polímeros/química , Agregados Proteicos/efeitos dos fármacos , Conformação Proteica em alfa-Hélice/efeitos dos fármacos , Dobramento de Proteína/efeitos dos fármacos , Estrutura Secundária de Proteína/efeitos dos fármacos
11.
Trop Med Int Health ; 14(9): 1117-21, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19624473

RESUMO

OBJECTIVES: To evaluate performance characteristics and ease of use of the new commercially available Crystal VC Rapid Dipstick (VC) test (Span Diagnostics, India) for Vibrio cholerae O1 and O139. METHODS: Whole stool was collected from patients presenting to a hospital cholera ward during a 2008 epidemic in Guinea-Bissau. The VC test on stool samples was conducted on-site; samples were subsequently stored in Cary-Blair transport media and sent to the Centers for Disease Control and Prevention for diagnostic testing by culture and polymerase chain reaction (PCR). In addition, four local laboratory technicians who were unfamiliar with the test were provided with stool samples, the VC test kit, and simple written instructions and asked to perform the test and interpret results. RESULTS: A total of 101 stool specimens were collected and tested. Compared with PCR, the test was 97% sensitive and 71-76% specific. Laboratory technicians in Bissau performed the test and interpreted results correctly using only simple written instructions. CONCLUSIONS: The VC test may be useful for cholera diagnosis in outbreak situations where laboratory capacity is limited.


Assuntos
Cólera/diagnóstico , Vibrio cholerae O1/isolamento & purificação , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Cólera/epidemiologia , Fezes/microbiologia , Feminino , Guiné-Bissau/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade , Adulto Jovem
12.
Elife ; 82019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31868587

RESUMO

RCK domains regulate the activity of K+ channels and transporters in eukaryotic and prokaryotic organisms by responding to ions or nucleotides. The mechanisms of RCK activation by Ca2+ in the eukaryotic BK and bacterial MthK K+ channels are well understood. However, the molecular details of activation in nucleotide-dependent RCK domains are not clear. Through a functional and structural analysis of the mechanism of ATP activation in KtrA, a RCK domain from the B. subtilis KtrAB cation channel, we have found that activation by nucleotide requires binding of cations to an intra-dimer interface site in the RCK dimer. In particular, divalent cations are coordinated by the γ-phosphates of bound-ATP, tethering the two subunits and stabilizing the active state conformation. Strikingly, the binding site residues are highly conserved in many different nucleotide-dependent RCK domains, indicating that divalent cations are a general cofactor in the regulatory mechanism of many nucleotide-dependent RCK domains.


Assuntos
Proteínas de Bactérias/química , Proteínas de Transporte de Cátions/química , Nucleotídeos/química , Conformação Proteica , Trifosfato de Adenosina/química , Bacillus subtilis/química , Bacillus subtilis/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/ultraestrutura , Sítios de Ligação/genética , Cálcio/metabolismo , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/ultraestrutura , Cátions/química , Cristalografia por Raios X , Canal de Potássio Kv1.6/química , Canal de Potássio Kv1.6/ultraestrutura , Nucleotídeos/genética , Potássio/química , Potássio/metabolismo , Canais de Potássio/química , Canais de Potássio/genética , Canais de Potássio/ultraestrutura , Domínios Proteicos/genética , Estrutura Terciária de Proteína , Proteínas Ribossômicas
13.
J Agric Food Chem ; 67(34): 9579-9590, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31381329

RESUMO

The first contact of tannins with the human body occurs in the mouth, where some of these tannins are known to interact with salivary proteins, in particular with proline-rich proteins (PRPs). These interactions are important at a sensory level, especially for astringency development, but could also affect the biological activities of the tannins. This study gathers information on the relative affinity of the interaction, complex stoichiometry, and tannin molecular epitopes of binding for the interactions between the families of PRPs (bPRPs, gPRPs, and aPRPs) and three representative ellagitannins (castalagin, vescalagin, and punicalagin). These interactions were studied by saturation-tranfer difference NMR and microcalorimetry. The effect of the PRP-ellagitannin interaction on their antioxidant ability was also assessed by ferric reduction antioxidant power (FRAP) assays. The results support a significant interaction between the studied tannins and PRPs with binding affinities in the micromolar range. Punicalagin was always the ellagitannin with higher affinity. aPRPs were the salivary PRPs with higher affinity. Moreover, it was observed that when ellagitannins are present in low concentrations (5-50 µM), as occurs in food, the antioxidant ability of these tannins when complexed with salivary PRPs could be significantly impaired.


Assuntos
Taninos Hidrolisáveis/química , Proteínas Salivares Ricas em Prolina/química , Antioxidantes/química , Antioxidantes/metabolismo , Adstringentes/química , Adstringentes/metabolismo , Humanos , Taninos Hidrolisáveis/metabolismo , Cinética , Ligação Proteica , Saliva/química , Saliva/metabolismo , Proteínas Salivares Ricas em Prolina/metabolismo , Paladar
14.
Food Chem ; 276: 33-42, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30409602

RESUMO

At red wine pH, malvidin-3-glucoside (mv-3-glc), the major anthocyanin of red wine, is expected to be present mainly in its non-colored hemiketal form. However, due to copigmentation with flavanols (e.g. epicatechin), the stabilization of the colored forms of mv-3-glc occurs. Some flavanols have been linked to astringency, due to their ability to interact/precipitate salivary proteins, namely proline-rich proteins (PRPs). So, a major question is if this copigmentation interaction could affect the ability of flavanols to interact with SP. To answer this, the effect of the interaction between mv-3-glc and epicatechin with basic and acidic PRPs, was investigated by saturation-tranfer difference (STD)-NMR and isothermal titration calorimetry (ITC). The most relevant result was that epicatechin:mv-3-glc mixture presents a synergic effect toward the interaction with both PRPs when compared to individual polyphenols. Furthermore, was observed that epicatechin interaction was driven by hydrophobic and hydrophilic interactions while mv-3-glc interaction was driven by electrostatic interactions.


Assuntos
Antocianinas/metabolismo , Catequina/metabolismo , Glucosídeos/metabolismo , Proteínas Salivares Ricas em Prolina/metabolismo , Ligação Proteica , Vinho/análise
15.
J Mol Biol ; 430(24): 5029-5049, 2018 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-30381148

RESUMO

The Drosophila EAG (dEAG) potassium channel is the founding member of the superfamily of KNCH channels, which are involved in cardiac repolarization, neuronal excitability and cellular proliferation. In flies, dEAG is involved in regulation of neuron firing and assembles with CaMKII to form a complex implicated in memory formation. We have characterized the interaction between the kinase domain of CaMKII and a 53-residue fragment of the dEAG channel that includes a canonical CaMKII recognition sequence. Crystal structures together with biochemical/biophysical analysis show a substrate-kinase complex with an unusually tight and extensive interface that appears to be strengthened by phosphorylation of the channel fragment. Electrophysiological recordings show that catalytically active CaMKII is required to observe active dEAG channels. A previously identified phosphorylation site in the recognition sequence is not the substrate for this crucial kinase activity, but rather contributes importantly to the tight interaction of the kinase with the channel. The available data suggest that the dEAG channel is a docking platform for the kinase and that phosphorylation of the channel's kinase recognition sequence modulates the strength of the interaction between the channel and the kinase.


Assuntos
Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/química , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Proteínas de Drosophila/química , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Canais de Potássio Éter-A-Go-Go/química , Canais de Potássio Éter-A-Go-Go/metabolismo , Animais , Sítios de Ligação , Domínio Catalítico , Cristalografia por Raios X , Drosophila melanogaster/química , Fenômenos Eletrofisiológicos , Modelos Moleculares , Simulação de Acoplamento Molecular , Fosforilação , Ligação Proteica , Conformação Proteica
16.
Food Chem ; 243: 175-185, 2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29146325

RESUMO

In this work, saturation transfer difference-NMR, isothermal microcalorimetry and molecular dynamics simulations have been used to study the individual interactions between basic, glycosylated and acidic proline-rich proteins (bPRPS, gPRPs, aPRPs) and P-B peptide with some representative food tannins [procyanidin B2, procyanidin B2 3'-O-gallate (B2g) and procyanidin trimer (catechin-4-8-catechin-4-8-catechin)]. Results showed that P-B peptide was in general the salivary protein (SP) with higher affinity whereas aPRPs showed lower affinity to the studied procyanidins. Moreover, B2g was the procyanidin with higher affinity for all SP. Hydrophobic and hydrogen bonds were present in all interactions but the major driving force depended on the procyanidin-SP pair. Furthermore, proline clusters or residues in their vicinity were identified as the probable sites of proteins for interaction with procyanidins. For bPRP and aPRP a significant change to less extended conformations was observed, while P-B peptide did not display any structural rearrangement upon procyanidins binding.


Assuntos
Proteínas Salivares Ricas em Prolina/metabolismo , Taninos/metabolismo , Glicosilação , Humanos , Simulação de Dinâmica Molecular , Ligação Proteica , Conformação Proteica , Proteínas Salivares Ricas em Prolina/química
17.
FEMS Microbiol Ecol ; 59(3): 638-50, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17381519

RESUMO

In order to investigate the bacterial diversity in a number of rivers, reservoirs and lakes in northern and central Portugal during the winter of 2004/5 (atypically dry), we applied molecular methodologies, namely denaturing gradient gel electrophoresis with primers targeting fractions of the bacterial 16S rRNA gene. Environmental parameters such as pH, conductivity, inorganic nutrients, total suspended solids and chlorophyll a were determined in order to characterize the trophic status of the studied water bodies. We found water bodies with oligotrophic to hypereutrophic characteristics. Organisms belonging to the Bacteroidetes and Alphaproteobacteria were found at the highest pH environment. Bacteroidetes were also related to high nutrient concentrations. Verrucomicrobia were associated with the most oligotrophic reservoir and low pH values. Actinobacteria were present in all samples from lakes and reservoirs, indicating its preference for lentic water bodies. Cyanobacteria dominance was related to high pH and conductivity levels. In general the conductivity values recorded in winter 2005 were the highest over recent years and chlorophyll a also reached very high levels. The data emphasize an enhanced risk of eutrophication for the studied water bodies, especially in the subsequent months when the temperature rises.


Assuntos
Bactérias/classificação , Biodiversidade , Água Doce/microbiologia , Microbiologia da Água , Bactérias/genética , Bactérias/isolamento & purificação , Clima Frio , Eletroforese em Gel de Poliacrilamida , Água Doce/análise , Genes Bacterianos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Portugal , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Estações do Ano
18.
Res Microbiol ; 157(10): 938-47, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17125975

RESUMO

The aim of the present study was to assess the occurrence and molecular diversity of beta-lactamase genes and integrons among Gram-negative ampicillin-resistant bacteria from Ria de Aveiro. Ampicillin-resistant isolates were selected and subjected to genotyping using REP-PCR. Representatives from each REP pattern were affiliated with the following taxa by sequencing the 16S rRNA gene: Aeromonas caviae, A. hydrophila, A. media, A. molluscorum, A. veronii, A. salmonicida, Aeromonas sp., Pseudomonas putida, Pseudomonas sp., Escherichia coli, Escherichia sp., Shigella sonnei, Shigella sp., Klebsiella pneumoniae, K. oxytoca, Raoultella ornithinolytica, R. terrigena, R. planticola, Citrobacter freundii, Morganella morganii and Enterobacter sp. Isolates affiliated with genera Escherichia or Shigella were identified as Escherichia coli using phenotypic-based tests. PCR was used to assess beta-lactamase encoding sequences (bla(TEM), bla(SHV), bla(CARB), bla(CTX-M), bla(IMP), bla(VIM), bla(CphA/IMIS), bla(OXA-A), bla(OXA-B), bla(OXA-C)), class 1 and class 2 integrases, and integron variable regions. Sequence analysis of PCR products was performed. beta-Lactamase genes were detected in 77.8% of the Enterobacteriaceae and in 10.5% of the Aeromonas. The most frequently detected gene was bla(TEM), followed by bla(SHV,)bla(OXA-B), bla(CphA/IMIS) and bla(CARB). Retrieved sequences shared high homology with previously described beta-lactamases. The intI1 gene was present in 29.6% of the Enterobacteriaceae and in 21% of the Aeromonas isolates. The intI2 gene was present in 4 isolates. A total of 13 cassettes included in 12 different cassette arrays were identified. The most frequently found resistance gene cassettes were aadA variants. Previous investigations based on cultivation-independent approaches revealed higher molecular diversity among beta-lactamase-encoding sequences in this estuary. This fact reinforces the hypothesis that cultivation-dependent approaches may underestimate the prevalence of antibiotic resistance genes in environmental samples and may introduce bias in the recovery of their molecular variants.


Assuntos
Resistência a Ampicilina/genética , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/genética , Integrons/genética , Microbiologia da Água , beta-Lactamases/genética , Ampicilina/farmacologia , Antibacterianos/farmacologia , Genes Bacterianos , Bactérias Gram-Negativas/isolamento & purificação , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
19.
Arq. odontol ; 57: 244-252, jan.-dez. 2021. ilus, tab
Artigo em Português | LILACS, BBO - odontologia (Brasil) | ID: biblio-1348380

RESUMO

Objetivo: Analisar na literatura quais são os hábitos associados à mordida aberta anterior em crianças. Métodos: Realizou-se uma pesquisa bibliográfica referente aos artigos publicados de 2015 a 2020 nos bancos de dados on-line PubMed, Embase, SciELO, LILACS e BBO utilizando descritores e sinônimos MeSH, DeCS e Emtree com as seguintes etapas: identificação, triagem, elegibilidade e inclusão. Foram incluídos estudos observacionais, transversais ou de coorte, e artigos que demonstrassem hábitos associados à mordida aberta anterior em crianças de 2 a 12 anos de idade. Relatos de caso, revisões da literatura e outros tipos de estudos que não estavam de acordo com os critérios foram excluídos. Os artigos foram analisados por dois pesquisadores independentes e os estudos selecionados foram avaliados quanto à qualidade metodológica. Resultados: De um total de 462 artigos encontrados apenas 9 foram selecionados para o estudo. Desses, 8 (88,9%) apresentaram alta ou moderada qualidade metodológica, sendo apenas 1 (11,1%), dentre os artigos, de baixa qualidade. Mediante à análise dos estudos inclusos, observou-se que a manutenção de hábitos orais como sucção digital (66,6% dos artigos) e de chupeta (77,7%), bem como uso de mamadeira (33,3%) e duração do tempo de aleitamento materno (22,2%) pode ocasionar alterações na oclusão, fala, respiração, crescimento craniofacial, afetando diretamente a qualidade de vida da criança. Conclusão:Uma vez que a infância é a fase adequada para a implementação de novos hábitos saudáveis e tratamentos, é imprescindível que o cirurgião-dentista conheça essa associação, e adote medidas terapêuticas e preventivas.


Aim: To perform an analysis of in the literature regarding which habits are associated with anterior open bite in children. Methods:This was a bibliographic study conducted using articles published from 2015 to 2020 in the PubMed, Embase, SciELO, LILACS, and BBO online databases, using MeSH, DeCS, and Emtree descriptors and synonyms with the following steps: identification, screening, eligibility, and inclusion. Observational, cross-sectional, or cohort studies, as well as and articles demonstrating habits associated with anterior open bite in children aged 2 to 12 years, were included. Case reports, literature reviews, and other types of studies that were not in accordance with the criteria were excluded. The articles were evaluated by two independent researchers, and the selected studies were evaluated for methodological quality. Results:Of a total of 462 articles found, only nine were selected for the study. Of these, eight (88.9%) presented high or moderate methodological quality, with only 1 (11.1%) of the articles presenting a low quality. Through the analysis of the included studies, it was observed that the maintenance of oral habits, such as finger sucking (66.6% of the articles) and pacifiers (77.7%), as well as the use of a bottle (33.3%) and the duration of breastfeeding time (22.2%), may cause changes in occlusion, speech, breathing, and craniofacial growth, directly affecting the child's quality of life. Conclusion: Since childhood is the appropriate phase for the implementation of new healthy habits and treatments, it is essential for the dentist to understand this association and adopt therapeutic and preventive measures.


Assuntos
Humanos , Pré-Escolar , Criança , Comportamento de Sucção , Mordida Aberta/etiologia , Chupetas/efeitos adversos , Mamadeiras , Sucção de Dedo , Fatores de Risco , Assistência Odontológica para Crianças
20.
Int. j. odontostomatol. (Print) ; 15(1): 132-136, mar. 2021. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1385710

RESUMO

Allergic reactions are host immune responses to endogenous or exogenous antigens, which can result in local and systemic problems. Among the main allergens are the dental materials used in orthodontics, which faces some challenges with regard to biocompatibility with oral tissues.This study aimed to analyze in the literature which materials used in orthodontics are related to the appearance of oral and perioral allergic reactions, as well as to investigate the most prevalen t manifestations.An integrative review was carried out with articles published between 2010 and 2020 on the PubMed, Embase, and ScienceDirect on-line database. For this, MeSH descriptors and synonyms were used following two semantic fields: "Hypersensitivity" in association with "Orthodontic Appliances", the selection of studies counted with the stages of identifica tion, screening, eligibility and inclusion. Randomized clinical trials, cross-sectional or cohort clinical trials; Patients with alle rgic reactions affecting the oral or perioral region due to contact with orthodontic material, were included in the review.From a to tal of 549 articles only 04 were selected for the study. In accordance with what has been analyzed in the evidence of these studies, periodontal changes such as gingival inflammation as well as erythema, edema, papules and blisters were the main reactions described. Furthermore, in all articles there was an association with nickel and in only one with chromium. It is possible to i nfer that inflammatory lesions are frequent manifestations on the use of nickel in orthodontic patients.


Las reacciones alérgicas son respuestas inmunes del huésped a antígenos endógenos o exógenos, los cuales pueden provocar problemas locales y sistémicos. Entre los principales alérgenos se encuentran los materiales dentales utilizados en la ortodoncia, la cual se enfrenta a algunos desafíos respecto a la biocompatibilidad de los tejidos orales. Este estudio tuvo como objetivo analizar en la literatura que materiales utilizados en ortodoncia están relacionados con la aparición de reacciones alérgicas orales y periorales, así como investigar las manifestaciones más prevalentes. Se llevó a cabo una revisión con artículos publicados entre 2010 y 2020 en las bases de datos PubMed, Embase y ScienceDirect. Para ello, se utilizaron descriptores y sinónimos de MeSH siguiendo dos campos semánticos: "Hipersensibilidad" en asociación con "Aparatos de Ortodoncia", la selección de estudios contó con las etapas de identificación, cribado, elegibilidad e inclusión. Ensayos clínicos aleatorizados, ensayos clínicos transversales o de cohortes; en la revisión se incluyeron pacientes con reacciones alérgicas que afectaron a la regiones oral o perioral debido al contacto con material de ortodoncia. De un total de 549 artículos sólo cuatro fueron seleccionados para el estudio. De acuerdo con lo que se analizó en la evidencia, las principales reacciones descritas fueron cambios periodontales, como inflamación gingival, eritema, edema, pápulas y ampollas. Además, en todos los artículos hubo una asociación con níquel y solamente en uno con cromo. Es posible inferir que las lesiones inflamatorias son manifestaciones frecuentes sobre el uso de níquel en pacientes de ortodoncia.


Assuntos
Humanos , Queilite/induzido quimicamente , Dermatite de Contato , Hipersensibilidade/epidemiologia , Ortodontia , Materiais Biomédicos e Odontológicos , Níquel/efeitos adversos
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