Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 43
Filtrar
Mais filtros










Intervalo de ano de publicação
1.
J Wildl Dis ; 60(2): 413-420, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38294761

RESUMO

The order Procellariiformes includes several species of seabirds that perform long-distance migrations crossing all the oceans. These movements may contribute to the dispersal and exchange of hemoparasites, such as haemosporidians. There is a lack of studies regarding the order Haemosporida in Procellariiformes, and, to date, only the genus Plasmodium has been reported. This survey investigated the occurrence of the three genera of haemosporidians, Plasmodium, Haemoproteus, and Leucocytozoon, in samples collected between 2013 and 2022 from 95 individuals of 14 species of Procellariiformes from southern Brazil, including live animals in rehabilitation centers, individuals caught as incidental bycatch, and carcasses found along the coast. A total of 171 samples of blood and fragments of liver and spleen were analyzed, with extracted DNA being subjected to a nested PCR followed by phylogeny analysis. All animals were negative for Plasmodium spp. and Leucocytozoon spp., but one Black-browed Albatross (Thalassarche melanophris) and one Manx Shearwater (Puffinus puffinus) specimen were positive for Haemoproteus spp. The sequences obtained from positive seabirds did not show 100% similarity with other known lineages available in the MalAvi database and thus were probably novel lineages. However, one sequence clustered together with Haemoproteus noctuae, a parasite from Strigiformes, while the other was grouped with Haemoproteus columbae, which is classically related to Columbiformes. These results suggest that both positive animals may have become infected when beached or in rehabilitation centers by a spillover of vectors from local birds. This highlights the importance of surveillance of the health of Procellariiformes regarding the possibility of dissemination of new pathogens in different bird populations.


Assuntos
Doenças das Aves , Haemosporida , Parasitos , Plasmodium , Infecções Protozoárias em Animais , Estrigiformes , Humanos , Animais , Brasil/epidemiologia , Doenças das Aves/epidemiologia , Doenças das Aves/parasitologia , Haemosporida/genética , Plasmodium/genética , Filogenia , Infecções Protozoárias em Animais/epidemiologia , Infecções Protozoárias em Animais/parasitologia
2.
Aesthetic Plast Surg ; 2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38286899

RESUMO

INTRODUCTION: Division of the suspensory ligament of the penis has emerged as a popular surgical approach for penile lengthening, but accurate preoperative predictions of lengthening outcomes remain elusive. This study aimed to identify readily measurable anatomical parameters associated with post-ligamentolysis penile length gain, facilitating more reliable preoperative estimations. METHODS: An experimental cross-sectional study was performed on 16 adult cadavers. Data collected before dissection included: age at death, ethnicity, height, length of the penis before dissection and width of the suspensory ligament of penis. Following the complete dissection of the suspensory ligament of penis, the depth of the pubic symphysis and the penile length after the procedure were measured. The absolute and relative length differences pre- and post-ligamentolysis were calculated. Correlation coefficients were used to study relations between these variables. RESULTS: Penile length increased uniformly after complete division of the suspensory ligament (average gain: 26.38 mm, SD = 14.83 mm; range 4-60 mm). Pearson correlation revealed a significant negative correlation between pre-ligamentolysis penile length and post-ligamentolysis increase (r = - 0.601; p = 0.014), suggesting greater gains in individuals with shorter pre-ligamentolysis lengths. Age, ligament width, and pubic arch depth showed no significant correlations. Ethnicity did not impact post-ligamentolysis length increase (t = - 0.135; p = 0.894). CONCLUSIONS: This study highlights the potential to predict penile length gain post-ligamentolysis through measurable anatomical parameters. The ability to anticipate the outcome of this procedure could empower surgeons to provide informed counseling, potentially elevating patient satisfaction. An experimental cross-sectional study was performed to investigate the outcomes of penile lengthening surgery Penile lengthening was achieved in all subjects via complete dissection of the suspensory ligament of the penis Penile length increase may be predicted preoperatively using easily measurable anatomical parameters NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

3.
Int J Mol Sci ; 24(12)2023 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-37373403

RESUMO

Dps proteins (DNA-binding proteins from starved cells) are multifunctional stress defense proteins from the Ferritin family expressed in Prokarya during starvation and/or acute oxidative stress. Besides shielding bacterial DNA through binding and condensation, Dps proteins protect the cell from reactive oxygen species by oxidizing and storing ferrous ions within their cavity, using either hydrogen peroxide or molecular oxygen as the co-substrate, thus reducing the toxic effects of Fenton reactions. Interestingly, the interaction between Dps and transition metals (other than iron) is a known but relatively uncharacterized phenomenon. The impact of non-iron metals on the structure and function of Dps proteins is a current topic of research. This work focuses on the interaction between the Dps from Marinobacter nauticus (a marine facultative anaerobe bacterium capable of degrading petroleum hydrocarbons) and the cupric ion (Cu2+), one of the transition metals of greater biological relevance. Results obtained using electron paramagnetic resonance (EPR), Mössbauer and UV/Visible spectroscopies revealed that Cu2+ ions bind to specific binding sites in Dps, exerting a rate-enhancing effect on the ferroxidation reaction in the presence of molecular oxygen and directly oxidizing ferrous ions when no other co-substrate is present, in a yet uncharacterized redox reaction. This prompts additional research on the catalytic properties of Dps proteins.


Assuntos
Proteínas de Bactérias , Marinobacter , Proteínas de Bactérias/metabolismo , Proteínas de Ligação a DNA/metabolismo , Marinobacter/metabolismo , Oxirredução , Íons , Oxigênio
4.
Parasitol Res ; 122(8): 1935-1941, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37314510

RESUMO

Procellariiformes includes pelagic seabirds that only use land for breeding; and also, these sites mostly occur in insular habitats. These peculiar habits make the investigation of hemoparasites a challenging issue. Thus, the data on the blood parasites of Procellariiformes are still scarce. In the order Piroplasmida, 16 species of Babesia have been described in terrestrial birds and seabirds. However, there is no register for Babesia spp. in procellariiform seabirds. Hence, the objective of this survey was to investigate the occurrence of Babesia spp. in these seabirds. A total of 220 tissue samples from 18 different seabird species were analyzed; the samples comprised blood and fragments of liver and spleen. The samples were obtained from live rescued animals and carcasses found along the southern coast of Brazil. Polymerase chain reaction (PCR) was conducted, followed by phylogenetic analysis. Only one blood sample yielded a positive result, from an adult female Thalassarche chlororhynchos (Atlantic yellow-nosed albatross). The sequence obtained showed the highest identity with sequences of Babesia spp. of birds from the South Pacific, and the isolate was named Babesia sp. strain Albatross. In the phylogenetic analysis, the sequence was grouped within the Babesia sensu stricto group, and further still into a subgroup including Babesia spp. of the Kiwiensis clade (parasites from birds). The phylogenetic analysis also showed that Babesia sp. strain Albatross clustered apart from the Peircei group, a clade that includes Babesia spp. from seabirds. As far as it is known, this is the first report of Babesia sp. in procellariiform seabirds. Babesia sp. strain Albatross may constitute a novel variant of tick-borne piroplasmids associated with the Procellariiformes order.


Assuntos
Babesia , Babesiose , Piroplasmida , Carrapatos , Animais , Feminino , Filogenia , Carrapatos/parasitologia , Aves , Babesiose/parasitologia
5.
Protein Sci ; 32(2): e4567, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36658780

RESUMO

DNA-binding proteins from starved cells (Dps) are small multifunctional nanocages expressed by prokaryotes in acute oxidative stress conditions or during the starvation-induced stationary phase, as a bacterial defense mechanism. Dps proteins protect bacterial DNA from damage by either direct binding or by removing precursors of reactive oxygen species from solution. The DNA-binding properties of most Dps proteins studied so far are related to their unordered, flexible, N- and C-terminal extensions. In a previous work, we revealed that the N-terminal tails of Deinoccocus grandis Dps shift from an extended to a compact conformation depending on the ionic strength of the buffer and detected a novel high-spin ferrous iron center in the proximal ends of those tails. In this work, we further explore the conformational dynamics of the protein by probing the effect of divalent metals binding to the tail by comparing the metal-binding properties of the wild-type protein with a binding site-impaired D34A variant using size exclusion chromatography, dynamic light scattering, synchrotron radiation circular dichroism, and small-angle X-ray scattering. The N-terminal ferrous species was also characterized by Mössbauer spectroscopy. The results herein presented reveal that the conformation of the N-terminal tails is altered upon metal binding in a gradual, reversible, and specific manner. These observations may point towards the existence of a regulatory process for the DNA-binding properties of Dps proteins through metal binding to their N- and/or C-terminal extensions.


Assuntos
Proteínas de Bactérias , Deinococcus , Sequência de Aminoácidos , Proteínas de Bactérias/química , Deinococcus/química , Deinococcus/genética , Deinococcus/metabolismo , DNA Bacteriano/metabolismo
6.
J Zoo Wildl Med ; 53(4): 733-743, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36640075

RESUMO

Plastic ingestion is a problem for seabirds worldwide. In addition to direct health effects such as obstruction or perforation of the gastrointestinal tract, plastic ingestion can also lead to indirect health effects through the release of chemicals that may be absorbed and cause systemic and chronic toxicity. Among chemicals that can be released by plastics are phthalate esters, a group of chemicals widely used as plasticizers or additives to change the physical characteristics of plastics. In this study, three phthalate esters, dimethyl phthalate (DMP), dibuthyl phthalate (DBP), and diethylhexyl phthalate (DEHP), were quantified in the uropygial gland of 48 seabirds from 16 species collected ashore in a tropical region, the coast of Espírito Santo, Eastern Brazil. Including trace levels, DMP was detected in 16 birds (33%) from 10 species, with an average concentration of 0.014 ± 0.005 ng/µl (mean ± SD for individuals with concentrations above the practical level of detection of 0.01 ng/µl). DBP was detected in 15 birds (31%) from 11 species, with an average concentration of 0.049 ± 0.032 ng/µl. DEHP was detected in 21 birds (44%) from 11 species, with an average concentration of 0.115 ± 0.105 ng/µl. DMP concentration in the uropygial gland was positively associated with the presence, number, and mass of plastic items in the upper digestive tract. However, no such relationship was noted for DBP nor DEHP, suggesting the concentration of phthalate compounds in the uropygial gland might not always serve as a reliable proxy for plastic ingestion. In spite of relatively high frequencies of detection, the low concentrations of phthalates detected in this study suggest levels of exposure below known toxicity thresholds. Further studies on the potential adverse effects of phthalate exposure in seabirds are necessary, especially on the reproductive development of embryos and chicks.


Assuntos
Dietilexilftalato , Plásticos , Animais , Plastificantes , Ésteres , Brasil , Galinhas , Ingestão de Alimentos
7.
Int J Mol Sci ; 23(14)2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35887179

RESUMO

Encapsulins are protein nanocages capable of harboring smaller proteins (cargo proteins) within their cavity. The function of the encapsulin systems is related to the encapsulated cargo proteins. The Myxococcus xanthus encapsulin (EncA) naturally encapsulates ferritin-like proteins EncB and EncC as cargo, resulting in a large iron storage nanocompartment, able to accommodate up to 30,000 iron atoms per shell. In the present manuscript we describe the binding and protection of circular double stranded DNA (pUC19) by EncA using electrophoretic mobility shift assays (EMSA), atomic force microscopy (AFM), and DNase protection assays. EncA binds pUC19 with an apparent dissociation constant of 0.3 ± 0.1 µM and a Hill coefficient of 1.4 ± 0.1, while EncC alone showed no interaction with DNA. Accordingly, the EncAC complex displayed a similar DNA binding capacity as the EncA protein. The data suggest that initially, EncA converts the plasmid DNA from a supercoiled to a more relaxed form with a beads-on-a-string morphology. At higher concentrations, EncA self-aggregates, condensing the DNA. This process physically protects DNA from enzymatic digestion by DNase I. The secondary structure and thermal stability of EncA and the EncA-pUC19 complex were evaluated using synchrotron radiation circular dichroism (SRCD) spectroscopy. The overall secondary structure of EncA is maintained upon interaction with pUC19 while the melting temperature of the protein (Tm) slightly increased from 76 ± 1 °C to 79 ± 1 °C. Our work reports, for the first time, the in vitro capacity of an encapsulin shell to interact and protect plasmid DNA similarly to other protein nanocages that may be relevant in vivo.


Assuntos
Myxococcus xanthus , Proteínas de Bactérias/metabolismo , DNA/metabolismo , Ferritinas/metabolismo , Ferro/metabolismo , Myxococcus xanthus/genética , Myxococcus xanthus/metabolismo
8.
Int J Mol Sci ; 23(9)2022 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-35563263

RESUMO

DNA-binding proteins from starved cells (Dps) are homododecameric nanocages, with N- and C-terminal tail extensions of variable length and amino acid composition. They accumulate iron in the form of a ferrihydrite mineral core and are capable of binding to and compacting DNA, forming low- and high-order condensates. This dual activity is designed to protect DNA from oxidative stress, resulting from Fenton chemistry or radiation exposure. In most Dps proteins, the DNA-binding properties stem from the N-terminal tail extensions. We explored the structural characteristics of a Dps from Deinococcus grandis that exhibits an atypically long N-terminal tail composed of 52 residues and probed the impact of the ionic strength on protein conformation using size exclusion chromatography, dynamic light scattering, synchrotron radiation circular dichroism and small-angle X-ray scattering. A novel high-spin ferrous iron-binding site was identified in the N-terminal tails, using Mössbauer spectroscopy. Our data reveals that the N-terminal tails are structurally dynamic and alter between compact and extended conformations, depending on the ionic strength of the buffer. This prompts the search for other physiologically relevant modulators of tail conformation and hints that the DNA-binding properties of Dps proteins may be affected by external factors.


Assuntos
Proteínas de Bactérias , Deinococcus , Sequência de Aminoácidos , Proteínas de Bactérias/metabolismo , DNA/metabolismo , Deinococcus/genética , Ferro/metabolismo , Modelos Moleculares , Concentração Osmolar
9.
Materials (Basel) ; 15(3)2022 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-35161125

RESUMO

Two eco-friendly healing bioproducts generated from microbial mixed cultures (MMC) for the production of polyhydroxyalkanoates (PHA) were used as surface treatments, with two residual materials used as the substrates, namely crude glycerol and pinewood bio-oil. Their ability to improve the durability of concrete samples containing recycled aggregates was assessed. To determine this protective capacity, 180 samples were analyzed using different tests, such as water penetration under pressure, capillary absorption, freeze-thaw and water droplet absorption test. Three types of conditions were used: outdoor-indoor exposure, re-application of biopolymers and application in vertical exposure conditions. The results showed reductions of up to 50% in the water penetration test and a delay in the water droplet absorption test of up to 150 times relative to the reference. The surface application of these bioproducts significantly reduced the degree of water penetration in recycled concrete, increasing its useful lifespan and proving to be a promising treatment for protecting concrete surfaces.

10.
Materials (Basel) ; 14(21)2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34772095

RESUMO

One approach to tackle the problems created by the vast amounts of construction and demolition waste (CDW) generated worldwide while at the same time lengthening concrete durability and service life is to foster the use of recycled aggregate (RA) rather than natural aggregate (NA). This article discusses the use of polyhydroxyalkanoates (PHAs)-producing mixed microbial cultures (MMCs) to treat the surface of recycled concrete with a view to increase its resistance to water-mediated deterioration. The microorganisms were cultured in a minimal medium using waste pinewood bio-oil as a carbon source. Post-application variations in substrate permeability were determined with the water drop absorption and penetration by water under pressure tests. The significant reduction in water absorption recorded reveals that this bioproduct is a promising surface treatment for recycled concrete.

11.
Eur Biophys J ; 50(3-4): 561-570, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34009405

RESUMO

Iron-sulfur centers are widespread in living organisms, mostly performing electron transfer functions, either in electron transfer chains or as part of multi-enzymatic complexes, while being also present in enzyme active sites, handling substrate catalysis. Rubredoxin is the simplest iron-sulfur containing protein constituted by a single polypeptide chain of 50 to 60 amino acids, of which four cysteine residues are responsible for metal binding in a tetrahedral coordination sphere. In this manuscript we explore the structure and stability of both apo- and holo-forms of a Rubredoxin from Marinobacter hydrocarbonoclasticus using Synchrotron Radiation Circular Dichroism (SRCD) in combination with other biochemical and spectroscopic techniques. The results are consistent with a holo-protein form containing a monomeric iron center with UV-visible maxima at 760, 578, 494, 386, 356 and 279 nm, an intense EPR resonance with a g value around 4.3 and Mössbauer spectroscopy parameters of δ equal to 0.69 mm/s and ΔEQ equal to 3.25 mm/s, for the ferrous reconstituted state. SRCD data, obtained for the first time for the apo-form, show a quite defined structure with ∆ε maximum at 191 nm and minima at 203 and 231 nm. Most significantly, the presence of isosbestic points at 189 and 228 nm made the interconversion between the two stable apo- and holo-form solution structures clear. SRCD temperature dependence data shows that for both forms the denaturation process proceeds through an intermediate species.


Assuntos
Proteínas Ferro-Enxofre/química , Dicroísmo Circular , Espectroscopia de Ressonância de Spin Eletrônica , Ferro/metabolismo , Marinobacter , Rubredoxinas , Enxofre
12.
Eur Biophys J ; 50(3-4): 513-521, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33900431

RESUMO

DNA-binding proteins from starved cells (Dps) are members of the ferritin family of proteins found in prokaryotes, with hollow rounded cube-like structures, composed of 12 equal subunits. These protein nanocages are bifunctional enzymes that protect the cell from the harmful reaction of iron and peroxide (Fenton reaction), thus preventing DNA damage by oxidative stress. Ferrous ions are oxidized at specific iron-binding sites in the presence of the oxidant and stored in its cavity that can accommodate up to ca. 500 iron atoms. DNA-binding properties of Dps are associated with the N-terminal, positive charge rich, extensions that can promote DNA binding and condensation, apparently by a cooperative binding mechanism. Here, we describe the binding and protection activities of Marinobacter hydrocarbonoclasticus Dps using Electrophoretic Mobility Shift Essays (EMSA), and synchrotron radiation circular dichroism (SRCD) spectroscopy. While no DNA condensation was observed in the tested conditions, it was possible to determine a Dps-DNA complex formation with an apparent dissociation constant of 6.0 ± 1.0 µM and a Hill coefficient of 1.2 ± 0.1. This interaction is suppressed by the inclusion of a single negative charge in the N-terminal region by point mutation. In Dps proteins containing a ferric mineral core (above 96 Fe/protein), DNA binding was impaired. SRCD data clearly showed that no significant modification existed either in secondary structure or protein stability of WT, Q14E variant and core containing proteins. It was, however, interesting to note that, in our experimental conditions, thermal denaturation induced protein aggregation that caused artifacts in thermal denaturation curves, which were dependent on radiation flux and vertical arrangement of the CD cell.


Assuntos
Marinobacter , Proteínas de Bactérias/genética , DNA , Ferro , Modelos Moleculares
13.
Materials (Basel) ; 14(8)2021 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-33921807

RESUMO

The large increase in the world population has resulted in a very large amount of construction waste, as well as a large amount of waste glycerol from transesterification reactions of acyl glycerides from oils and fats, in particular from the production of biodiesel. Only a limited percentage of these two residues are recycled, which generates a large management problem worldwide. For that reason, in this study, we used crude glycerol as a carbon source to cultivate polyhydroxyalkanoates (PHA)-producing mixed microbial cultures (MMC). Two bioproducts derived from these cultures were applied on the surface of concrete with recycled aggregate to create a protective layer. To evaluate the effect of the treatments, tests of water absorption by capillarity and under low pressure with Karsten tubes were performed. Furthermore, SEM-EDS analysis showed the physical barrier caused by biotreatments that produced a reduction on capillarity water absorption of up to 20% and improved the impermeability of recycled concrete against the penetration of water under pressure up to 2.7 times relative to the reference. Therefore, this bioproduct shown to be a promising treatment to protect against penetration of water to concrete surfaces increasing its durability and useful life.

14.
Org Biomol Chem ; 18(45): 9300-9307, 2020 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-33169764

RESUMO

A missense mutant of a Dps protein (DNA-binding protein from starved cells) from Marinobacter hydrocarbonoclasticus was used as a building block to develop a new supramolecular assembly complex which enhances the iron uptake, a physiological function of this mini-ferritin. The missense mutation was conducted in an exposed and flexible region of the N-terminal, wherein a threonine residue in position 10 was replaced by a cysteine residue (DpsT10C). This step enabled a click chemistry approach to the variant DpsT10C, where a thiol-ene coupling occurs. Two methods and two types of linker were used resulting in two different mini-ferritin supramolecular polymers, which have maintained secondary structure and native iron uptake physiological function. Electrophoretic assays and mass spectrometry were utilized to confirm that both functionalization and coupling reactions occured as predicted. The secondary structure has been investigated by circular dichroism and synchrotron radiation circular dichroism. Size and morphology were obtained by dynamic light scattering, size exclusion chromatography and atomic force microscopy, respectively. The iron uptake of the synthesized protein polymers was confirmed by UV-Vis spectroscopy loading assays.

15.
Angew Chem Int Ed Engl ; 58(4): 1013-1018, 2019 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-30481405

RESUMO

Dps proteins (DNA-binding protein from starved cells) are hollow-sphere-shaped, dodecameric enzymes found in bacteria and archaeal species. They can oxidize ferrous iron in a controlled manner using hydrogen peroxide or molecular oxygen as co-substrate, and most of them confer physical protection through DNA binding. Oxidized iron is stored, as a mineral core, in a central cavity. Direct evidence is now provided that, furthermore, Dps proteins containing small mineral cores can oxidize and mineralize toxic ferrous ions in anaerobic conditions and in the absence of any additional aqueous oxidant co-substrate. Dps proteins containing cores of 24 irons per dodecamer can oxidize about 5 ferrous irons per dodecamer, with that number approximately doubling for protein particles containing in average 96 irons per protein. This additional activity carries importance as it can be a detoxification mechanism present during anaerobic or oxygen-limited growth conditions.


Assuntos
Proteínas de Bactérias/química , Proteínas de Ligação a DNA/química , Ferritinas/química , Compostos Ferrosos/química , Marinobacter/química , Sítios de Ligação , Modelos Moleculares , Oxirredução
16.
Plast Reconstr Surg Glob Open ; 6(2): e1635, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29616161

RESUMO

BACKGROUND: Orofacial clefts are among the most common congenital craniofacial malformations and may be associated with other birth defects. However, the proportion and type of additional anomalies vary greatly between studies. This study assessed the prevalence and type of associated congenital malformations in children with orofacial clefts, who attended the largest cleft lip and palate tertiary referral center in Portugal. METHODS: Consecutive children with orofacial clefts who attended at least 1 consultation in our Clefts Unit between 1981 and 2012 were studied. Demographic and clinical data regarding the number and type of associated malformations were retrospectively collected and analyzed. RESULTS: Of the 701 patients studied, 219 (31.2%) had associated congenital malformations. These malformations were more frequent in children with cleft palate (43.4%) than in children with cleft lip and palate (27.5%) or with cleft lip only (19.4%). Within the group with associated anomalies, 73 cases (33.3%) had conditions related with known chromosomal defects, monogenic syndromes or sequences, and 146 cases (66.7%) had multiple congenital anomalies of unknown origin. From those, head and neck malformations were the most common (60.3%), followed by malformations in the cardiovascular (28.3%) and musculoskeletal systems (26%). CONCLUSIONS: The overall prevalence of associated malformations of nearly 1 in 3 children with orofacial clefts stressed the need for a comprehensive evaluation of these patients by a multidisciplinary cleft team. Moreover, one-third of the children had multiple congenital anomalies of known origins. Thus, early routine screening for other malformations and genetic counseling might be valuable for orofacial clefts management.

17.
J Chromatogr A ; 1472: 55-65, 2016 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-27773392

RESUMO

A common strategy for the production and purification of recombinant proteins is to fuse a tag to the protein terminal residues and employ a "tag-specific" ligand for fusion protein capture and purification. In this work, we explored the effect of two tryptophan-based tags, NWNWNW and WFWFWF, on the expression and purification of Green Fluorescence Protein (GFP) used as a model fusion protein. The titers obtained with the expression of these fusion proteins in soluble form were 0.11mgml-1 and 0.48mgml-1 for WFWFWF and NWNWNW, respectively. A combinatorial library comprising 64 ligands based on the Ugi reaction was prepared and screened for binding GFP-tagged and non-tagged proteins. Complementary ligands A2C2 and A3C1 were selected for the effective capture of NWNWNW and WFWFWF tagged proteins, respectively, in soluble forms. These affinity pairs displayed 106M-1 affinity constants and Qmax values of 19.11±2.60ugg-1 and 79.39ugg-1 for the systems WFWFWF AND NWNWNW, respectively. GFP fused to the WFWFWF affinity tag was also produced as inclusion bodies, and a refolding-on column strategy was explored using the ligand A4C8, selected from the combinatorial library of ligands but in presence of denaturant agents.


Assuntos
Cromatografia de Afinidade/métodos , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/isolamento & purificação , Ligantes , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Triptofano/química , Triptofano/isolamento & purificação , Sequência de Aminoácidos , Proteínas de Fluorescência Verde/biossíntese , Corpos de Inclusão/metabolismo , Desnaturação Proteica , Redobramento de Proteína , Proteínas Recombinantes de Fusão/biossíntese , Solubilidade
18.
Appl Microbiol Biotechnol ; 99(20): 8329-36, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26278535

RESUMO

The increasing dependency on fossil fuels has driven researchers to seek for alternative energy sources. Renewable energy sources such as sunlight, wind, or water are the most common. However, since the 1990s, other sources for energy production have been studied. The use of microorganisms such as bacteria or archaea to produce energy is currently in great progress. These present several advantages even when compared with other renewable energy sources. Besides the energy production, they are also involved in bioremediation such as the removal of heavy metal contaminants from soils or wastewaters. Several research groups have demonstrated that these organisms are able to interact with electrodes via heme and non-heme iron proteins. Therefore, the role of iron as well as iron metabolism in these species must be of enormous relevance. Recently, the influence of cellular iron regulation by Fur in the Geobacter sulfurreducens growth and ability to produce energy was demonstrated. In this review, we aim to briefly describe the most relevant proteins involved in the iron metabolism of bacteria and archaea and relate them and their biological function with the ability of selected organisms to produce energy.


Assuntos
Archaea/metabolismo , Bactérias/metabolismo , Fontes de Energia Bioelétrica , Eletricidade , Eletrodos/microbiologia , Ferro/metabolismo , Archaea/crescimento & desenvolvimento , Bactérias/crescimento & desenvolvimento
19.
Radiat Environ Biophys ; 54(1): 111-121, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25249071

RESUMO

Organisms are often exposed to different types of ionizing radiation that, directly or not, will promote damage to DNA molecules and/or other cellular structures. Because of that, organisms developed a wide range of response mechanisms to deal with these threats. Endonuclease III is one of the enzymes responsible to detect and repair oxidized pyrimidine base lesions. However, the effect of radiation on the structure/function of these enzymes is not clear yet. Here, we demonstrate the effect of UV-C radiation on E. coli endonuclease III through several techniques, namely UV-visible, fluorescence and Mössbauer spectroscopies, as well as SDS-PAGE and electrophoretic mobility shift assay. We demonstrate that irradiation with a UV-C source has dramatic consequences on the absorption, fluorescence, structure and functionality of the protein, affecting its [4Fe-4S] cluster and its DNA-binding ability, which results in its inactivation. An UV-C radiation-induced conversion of the [4Fe-4S](2+) into a [2Fe-2S](2+) was observed for the first time and proven by Mössbauer and UV-visible analysis. This work also shows that the DNA-binding capability of endonuclease III is highly dependent of the nuclearity of the endogenous iron-sulfur cluster. Thus, from our point of view, in a cellular context, these results strengthen the argument that cellular sensitivity to radiation can also be due to loss of radiation-induced damage repair ability.


Assuntos
Desoxirribonuclease (Dímero de Pirimidina)/efeitos da radiação , Proteínas de Escherichia coli/efeitos da radiação , Proteínas Ferro-Enxofre/efeitos da radiação , Raios Ultravioleta , DNA/metabolismo , Reparo do DNA , Desoxirribonuclease (Dímero de Pirimidina)/química , Desoxirribonuclease (Dímero de Pirimidina)/genética , Desoxirribonuclease (Dímero de Pirimidina)/metabolismo , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Proteínas Ferro-Enxofre/química , Proteínas Ferro-Enxofre/metabolismo , Plasmídeos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/efeitos da radiação , Análise Espectral
20.
Aval. psicol ; 13(3): 447-455, dez. 2014. tab
Artigo em Português | Index Psicologia - Periódicos | ID: psi-62725

RESUMO

Este estudo tem como objetivo validar, em uma população portuguesa jovem, a Escala de Percepção de Funcionamento Parental (EPFP) desenvolvida com base na versão espanhola Estilos Educativos Materno y Paterno. Seiscentos e cinco participantes, equitativamente divididos pelos dois sexos, responderam à EPFP. A estrutura fatorial original composta por seis dimensões foi adaptada, com a remoção de alguns itens, tendo apresentado bons índices de ajustamento (análise fatorial confirmatória) e valores de consistência variando entre α=0,81 e 0,95. Apesar de significativamente correlacionadas, verificaram-se diferenças importantes entre as percepções relativas às dimensões do funcionamento materno e paterno. As mães são consideradas mais afetuosas e comunicativas, promotoras da autonomia, confidentes, bem-humoradas e menos controladoras. A EPFP se revelou um instrumento válido e fidedigno na avaliação do funcionamento parental, através da percepção dos filhos.(AU)


This study aimed to validate the Parental Functioning Perception Scale – Mother and Father (EPFP), as the Portuguese language version of the Spanish scale Estilos Educativos Materno y Paterno (Maternal and Paternal Educational Styles). A sample of 605 college students participated, equally divided by gender, who responded to the EPFP regarding the two parental figures. The original factorial structure (which was composed of six dimensions, adapted, with some items removed), showed good adjustment indexes (confirmatory factor analysis) and acceptable internal consistency values ranging from α=0.81 to 0.95. Despite being significantly correlated, statistically significant differences were found between the perceptions of the dimensions of maternal and paternal parental functioning. Mothers were considered to be more affectionate, communicative, good-humored and trustworthy, while encouraging autonomy, and less controlling than fathers. The EPFP proved to be a valid and reliable instrument in the assessment of the adolescents’ perception of maternal and paternal parental functioning.(AU)


Este estudio tuvo como objetivo validar la Escala de Percepción del Funcionamiento Parental (EPFP) en una populación portuguesa joven, desarrollada a partir de la versión española de Estilos Educativos Materno y Paterno. Participaron en el estudio 605 jóvenes, de los dos sexos, que contestaron a la EFPF para las dos figuras parentales. La estructura factorial original (seis dimensiones) fue adaptada con la retirada de algunos ítems, habiendo presentado buenos índices de ajustamiento (análisis fatorial confirmatória), con apropiados valores de consistencia interna (α=0,81 α=0,95). A pesar de significativamente correlacionadas, se han comprobado diferencias significativas entre las percepciones relativas a las dimensiones del funcionamiento materno y paterno. Las madres fueran consideradas más afectuosas y comunicativas, promotoras de autonomía, confidentes, de buen humor y menos controladoras. La EPFP se mostró un instrumento válido y confiable en la evaluación de la percepción que los adolescentes tienen del funcionamiento parental.(AU)


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto Jovem , Adulto , Relações Pais-Filho , Pais/psicologia , Relações Familiares , Reprodutibilidade dos Testes
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...