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1.
J Mol Biol ; 435(8): 168024, 2023 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-36828271

RESUMO

The biological function of macromolecular complexes depends not only on large-scale transitions between conformations, but also on small-scale conformational fluctuations at equilibrium. Information on the equilibrium dynamics of biomolecular complexes could, in principle, be obtained from local resolution (LR) data in cryo-electron microscopy (cryo-EM) maps. However, this possibility had not been validated by comparing, for a same biomolecular complex, LR data with quantitative information on equilibrium dynamics obtained by an established solution technique. In this study we determined the cryo-EM structure of the minute virus of mice (MVM) capsid as a model biomolecular complex. The LR values obtained correlated with crystallographic B factors and with hydrogen/deuterium exchange (HDX) rates obtained by mass spectrometry (HDX-MS), a gold standard for determining equilibrium dynamics in solution. This result validated a LR-based cryo-EM approach to investigate, with high spatial resolution, the equilibrium dynamics of biomolecular complexes. As an application of this approach, we determined the cryo-EM structure of two mutant MVM capsids and compared their equilibrium dynamics with that of the wild-type MVM capsid. The results supported a previously suggested linkage between mechanical stiffening and impaired equilibrium dynamics of a virus particle. Cryo-EM is emerging as a powerful approach for simultaneously acquiring information on the atomic structure and local equilibrium dynamics of biomolecular complexes.


Assuntos
Aminoácidos , Capsídeo , Microscopia Crioeletrônica , Substâncias Macromoleculares , Aminoácidos/química , Capsídeo/química , Microscopia Crioeletrônica/métodos , Conformação Proteica , Substâncias Macromoleculares/química , Vírus Miúdo do Camundongo/química , Vírus Miúdo do Camundongo/genética
2.
Int. j. morphol ; 39(2): 455-462, abr. 2021. ilus, tab, graf
Artigo em Inglês | LILACS | ID: biblio-1385363

RESUMO

SUMMARY: The objective of this study was to evaluate the bilaterality and symmetry in C-shaped mandibular second molars in Mexican Maya and non-Maya populations using cone-beam computed tomography (CBCT). Five-hundred-twenty-five CBCT scans of patients with left and right mandibular second molars were analyzed to determine the prevalence and bilaterality. The teeth were assessed for the presence of C-shaped root canals, according to Fan et al. (2004) criteria. The sample was subdivided by ethnicity, sex, unilateral/bilateral presence, side of the longitudinal groove, and the C-shaped root canal configuration and symmetry, at the cervical, middle, and apical cross- sections of the root. C-shaped mandibular second molars were present in 24.95% of the individuals. Most (60.30%) of those showing this trait had it bilaterally. When comparing by ethnicity, sex, and side, we detected no significant differences. The vast majority (97.33%) presented the groove along the lingual side. The C3 was the most prevalent configuration in the overall sample, while in the Maya group, the C1 was the most common configuration. When analyzed by sex and ethnic group, the non-Maya females tended to deviate from the other groups in terms of bilaterality. Overall, 55.70% of bilateral C-shaped mandibular second molars were also symmetric in the three radicular thirds. The prevalence of C-shaped mandibular second molars was similar to that reported for northernAsian populations, which is consistent with the evolutionary origins of Native American populations. Most of the sample showed bilaterality and half were symmetric. Clinicians must be aware of the ethnic background of their patients and consider the possible variations to do more predictable root canal treatments.


RESUMEN: El objetivo de este estudio fue evaluar la bilateralidad y simetría de los segundos molares mandibulares en forma de C en una población Mexicana Maya y no-Maya mediante tomografía computarizada de haz cónico (CBCT). Material and Métodos: Se analizaron quinientos veinticinco tomografías de pacientes con segundos molares mandibularesderecho e izquierdo para determinar la prevalencia y bilateralidad. Los dientes fueron evaluados según la presencia de conductos en forma de C de acuerdo a los criterios de Fan et al. La muestra fue subdividida por etnicidad, sexo, presencia bilateral/unilateral, lado del surco y configuración y simetría de los conductos en los cortes transversales cervical, medio y apical. Los segundos molares mandibulares en forma de C estuvieron presentes en el 24.95 % de los individuos. La mayoría (60.30%) de los que mostraron este rasgo lo hicieron de forma bilateral. El comparar por etnicidad, sexo y lado, no se encontraron diferencias estadísticamente significativas. La gran mayoría (97.33%) presentó el surco por lingual. C3 fue la configuración más prevalente en la muestra general, mientras que en el grupo Maya el C1 fue la más común. El análisis por sexo y grupo étnico arrojó que las mujeres no Mayas tendieron a desviarse de los otros grupos en términos de bilateralidad. El 55.70% de los segundos molares mandibulares bilaterales fueron simétricos en sus tres tercios radiculares. La prevalencia de los segundos molars mandibulares en forma de C fue similar a la reportada para las poblaciones del norte de Asia, lo cual es consistente con los orígenes evolutivos de las poblaciones nativas americanas. La mayor parte de la muestra presentó el rasgo de forma bilateral y la mitad de éstos fueron simétricos. Los odontólogos deben tomar en cuenta el origen étnico de sus pacientes y considerar las posibles variaciones para realizar tratamientos endodónticos más predecibles.


Assuntos
Humanos , Masculino , Feminino , Cavidade Pulpar/diagnóstico por imagem , Tomografia Computadorizada de Feixe Cônico , Mandíbula/diagnóstico por imagem , Dente Molar/diagnóstico por imagem , Etnicidade , Fatores Etários , Cavidade Pulpar/anatomia & histologia , Mandíbula/anatomia & histologia , México , Dente Molar/anatomia & histologia
3.
Acta Biomater ; 122: 263-277, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33359294

RESUMO

We developed the Fluctuating Nonlinear Spring (FNS) model to describe the dynamics of mechanical deformation of biological particles, such as virus capsids. The theory interprets the force-deformation spectra in terms of the "Hertzian stiffness" (non-linear regime of a particle's small-amplitude deformations), elastic constant (large-amplitude elastic deformations), and force range in which the particle's fracture occurs. The FNS theory enables one to quantify the particles' elasticity (Young's moduli for Hertzian and bending deformations), and the limits of their strength (critical forces, fracture toughness) and deformability (critical deformations) as well as the probability distributions of these properties, and to calculate the free energy changes for the particle's Hertzian, elastic, and plastic deformations, and eventual fracture. We applied the FNS theory to describe the protein capsids of bacteriophage P22, Human Adenovirus, and Herpes Simplex virus characterized by deformations before fracture that did not exceed 10-19% of their size. These nanoshells are soft (~1-10-GPa elastic modulus), with low ~50-480-kPa toughness - a regime of material behavior that is not well understood, and with the strength increasing while toughness decreases with their size. The particles' fracture is stochastic, with the average values of critical forces, critical deformations, and fracture toughness comparable with their standard deviations. The FNS theory predicts 0.7-MJ/mol free energy for P22 capsid maturation, and it could be extended to describe uniaxial deformation of cylindrical microtubules and ellipsoidal cellular organelles.


Assuntos
Fenômenos Mecânicos , Nanopartículas , Módulo de Elasticidade , Elasticidade , Humanos , Dinâmica não Linear
4.
J Vet Sci ; 21(6): e87, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33263234

RESUMO

BACKGROUND: A new, extended long-acting tilmicosin (TLAe) preparation was tested against intramammary ceftiofur (CEF) using a non-inferiority trial model during dry-cow therapy (DCT) in a farm with high bovine population density and deficient hygiene application. OBJECTIVES: To evaluate the possibility that TLAe administered parenterally can achieve non-inferiority status compared to CEF administered intramammary for DCT. METHODS: Cows were randomly assigned to TLAe (20 mg/kg subcutaneous; n = 53) or CEF (CEF-HCl, 125 mg/quarter; n = 38 cows) treatment groups. California mastitis testing, colony-forming unit assessment (CFU/mL), and number of cases positive for Staphylococcus aureus were quantified before DCT and 7 d after calving. A complete cure was defined as no bacteria isolated; partial cure when CFU/mL ranged from 150 to 700, and cure-failure when CFU/mL was above 700. RESULTS: TLAe and CEF had overall cure rates of 57% and 53% (p > 0.05) and S. aureus cure rates of 77.7% and 25%, respectively (p < 0.05). The pathogens detected at DCT and 7 days after calving were S. aureus (62.71% and 35.55%), Staphylococcus spp. (22.03% and 35.55%), Streptococcus uberis (10.16% and 13.33%), and Escherichia coli (5.08% and 15.55%). Non-inferiority and binary logistic regression analyses revealed a lack of difference in overall efficacies of TLAe and CEF. Apart from S. aureus, S. uberis was the predominant pathogen found in both groups. CONCLUSIONS: This study is the first successful report of parenteral DCT showing comparable efficacy as CEF, the gold-standard. The extended long-term pharmacokinetic activity of TLAe explains these results.


Assuntos
Antibacterianos , Doenças dos Bovinos , Cefalosporinas , Preparações de Ação Retardada , Mastite Bovina , Tilosina , Animais , Bovinos , Feminino , Antibacterianos/administração & dosagem , Doenças dos Bovinos/tratamento farmacológico , Cefalosporinas/administração & dosagem , Preparações de Ação Retardada/administração & dosagem , Escherichia coli/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/veterinária , Injeções Subcutâneas/veterinária , Mastite Bovina/tratamento farmacológico , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/efeitos dos fármacos , Infecções Estreptocócicas/tratamento farmacológico , Infecções Estreptocócicas/veterinária , Streptococcus/efeitos dos fármacos , Tilosina/administração & dosagem , Tilosina/análogos & derivados
5.
J Virol ; 94(24)2020 11 23.
Artigo em Inglês | MEDLINE | ID: mdl-32938763

RESUMO

Despite their diversity, most double-stranded-RNA (dsRNA) viruses share a specialized T=1 capsid built from dimers of a single protein that provides a platform for genome transcription and replication. This ubiquitous capsid remains structurally undisturbed throughout the viral cycle, isolating the genome to avoid triggering host defense mechanisms. Human picobirnavirus (hPBV) is a dsRNA virus frequently associated with gastroenteritis, although its pathogenicity is yet undefined. Here, we report the cryo-electron microscopy (cryo-EM) structure of hPBV at 2.6-Å resolution. The capsid protein (CP) is arranged in a single-shelled, ∼380-Å-diameter T=1 capsid with a rough outer surface similar to that of dsRNA mycoviruses. The hPBV capsid is built of 60 quasisymmetric CP dimers (A and B) stabilized by domain swapping, and only the CP-A N-terminal basic region interacts with the packaged nucleic acids. hPBV CP has an α-helical domain with a fold similar to that of fungal partitivirus CP, with many domain insertions in its C-terminal half. In contrast to dsRNA mycoviruses, hPBV has an extracellular life cycle phase like complex reoviruses, which indicates that its own CP probably participates in cell entry. Using an in vitro reversible assembly/disassembly system of hPBV, we isolated tetramers as possible assembly intermediates. We used atomic force microscopy to characterize the biophysical properties of hPBV capsids with different cargos (host nucleic acids or proteins) and found that the CP N-terminal segment not only is involved in nucleic acid interaction/packaging but also modulates the mechanical behavior of the capsid in conjunction with the cargo.IMPORTANCE Despite intensive study, human virus sampling is still sparse, especially for viruses that cause mild or asymptomatic disease. Human picobirnavirus (hPBV) is a double-stranded-RNA virus, broadly dispersed in the human population, but its pathogenicity is uncertain. Here, we report the hPBV structure derived from cryo-electron microscopy (cryo-EM) and reconstruction methods using three capsid protein variants (of different lengths and N-terminal amino acid compositions) that assemble as virus-like particles with distinct properties. The hPBV near-atomic structure reveals a quasisymmetric dimer as the structural subunit and tetramers as possible assembly intermediates that coassemble with nucleic acids. Our structural studies and atomic force microscopy analyses indicate that hPBV capsids are potentially excellent nanocages for gene therapy and targeted drug delivery in humans.


Assuntos
Proteínas do Capsídeo/química , Capsídeo/ultraestrutura , Microscopia Crioeletrônica/métodos , Picobirnavirus/genética , Picobirnavirus/metabolismo , Capsídeo/metabolismo , Proteínas do Capsídeo/genética , Genoma Viral , Humanos , Modelos Moleculares , Conformação Proteica , Conformação Proteica em alfa-Hélice , Domínios Proteicos , RNA de Cadeia Dupla , Vírion/ultraestrutura , Montagem de Vírus
6.
Sci Rep ; 10(1): 3248, 2020 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-32094421

RESUMO

Here we studied HLA blocks and haplotypes in a group of 218 Lacandon Maya Native American using a high-resolution next generation sequencing (NGS) method. We assessed the genetic diversity of HLA class I and class II in this population, and determined the most probable ancestry of Lacandon Maya HLA class I and class II haplotypes. Importantly, this Native American group showed a high degree of both HLA homozygosity and linkage disequilibrium across the HLA region and also lower class II HLA allelic diversity than most previously reported populations (including other Native American groups). Distinctive alleles present in the Lacandon population include HLA-A*24:14 and HLA-B*40:08. Furthermore, in Lacandons we observed a high frequency of haplotypes containing the allele HLA-DRB1*04:11, a relatively frequent allele in comparison with other neighboring indigenous groups. The specific demographic history of the Lacandon population including inbreeding, as well as pathogen selection, may have elevated the frequencies of a small number of HLA class II alleles and DNA blocks. To assess the possible role of different selective pressures in determining Native American HLA diversity, we evaluated the relationship between genetic diversity at HLA-A, HLA-B and HLA-DRB1 and pathogen richness for a global dataset and for Native American populations alone. In keeping with previous studies of such relationships we included distance from Africa as a covariate. After correction for multiple comparisons we did not find any significant relationship between pathogen diversity and HLA genetic diversity (as measured by polymorphism information content) in either our global dataset or the Native American subset of the dataset. We found the expected negative relationship between genetic diversity and distance from Africa in the global dataset, but no relationship between HLA genetic diversity and distance from Africa when Native American populations were considered alone.


Assuntos
Variação Genética , Genética Populacional , Haplótipos , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe I/genética , Desequilíbrio de Ligação , Adolescente , Adulto , África , Alelos , Feminino , Frequência do Gene , Genótipo , Geografia , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Cadeias HLA-DRB1/genética , Homozigoto , Humanos , Masculino , México/etnologia , Pessoa de Meia-Idade , Análise de Componente Principal , Adulto Jovem , Indígena Americano ou Nativo do Alasca
7.
Methods Mol Biol ; 1886: 259-278, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30374873

RESUMO

Microscopes are used to characterize small objects with the help of probes that interact with the specimen, such as photons and electrons in optical and electron microscopies, respectively. In atomic force microscopy (AFM) the probe is a nanometric tip located at the end of a micro cantilever which palpates the specimen under study as a blind person manages a walking stick. In this way AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in liquid milieu. Beyond imaging, AFM also enables not only the manipulation of single protein cages, but also the characterization of every physicochemical property able of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In this chapter we start revising some recipes for adsorbing protein shells on surfaces. Then we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted for extracting physical information, such as mechanical and electrostatic properties. We also explain how a convenient combination of AFM and fluorescence methodologies entails monitoring genome release from individual viral shells during mechanical unpacking.


Assuntos
Microscopia de Força Atômica , Vírus/ultraestrutura , Capsídeo , Análise de Dados , Humanos , Processamento de Imagem Assistida por Computador/métodos , Fenômenos Mecânicos , Microscopia de Força Atômica/métodos , Eletricidade Estática , Proteínas Virais/química
8.
Methods Mol Biol ; 1665: 281-296, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28940075

RESUMO

In Atomic Force Microscopy (AFM) the probe is a nanometric tip located at the end of a microcantilever which palpates the specimen under study as a blind person uses a white cane. In this way AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in liquid milieu. Beyond imaging, AFM also enables the manipulation of single protein cages, and the characterization a variety physicochemical properties able of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In this chapter we start revising some recipes for adsorbing protein shells on surfaces. Then we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted to extracting physical information, such as mechanical and electrostatic properties.


Assuntos
Capsídeo/química , Microscopia de Força Atômica/métodos , Nanotecnologia/métodos , Proteínas/química
9.
Biochem Soc Trans ; 45(2): 499-511, 2017 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-28408490

RESUMO

Microscopes are used to characterize small objects with the help of probes that interact with the specimen, such as photons and electrons in optical and electron microscopies, respectively. In atomic force microscopy (AFM), the probe is a nanometric tip located at the end of a microcantilever which palpates the specimen under study just as a blind person manages a walking stick. In this way, AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in a liquid milieu. Beyond imaging, AFM also enables not only the manipulation of single protein cages, but also the characterization of every physicochemical property capable of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In the present revision, we start revising some recipes for adsorbing protein shells on surfaces. Then, we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted to extracting physical information, such as mechanical and electrostatic properties. We also explain how a convenient combination of AFM and fluorescence methodologies entails monitoring genome release from individual viral shells during mechanical unpacking.


Assuntos
Proteínas Virais/metabolismo , Vírus/ultraestrutura , Adsorção , Fenômenos Biomecânicos , Genoma Viral , Microscopia de Força Atômica/métodos , Vírus/genética
10.
ACS Nano ; 9(11): 10826-33, 2015 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-26491879

RESUMO

Tight confinement of naked genomes within some viruses results in high internal pressure that facilitates their translocation into the host. Adenovirus, however, encodes histone-like proteins that associate with its genome resulting in a confined DNA-protein condensate (core). Cleavage of these proteins during maturation decreases core condensation and primes the virion for proper uncoating via unidentified mechanisms. Here we open individual, mature and immature adenovirus cages to directly probe the mechanics of their chromatin-like cores. We find that immature cores are more rigid than the mature ones, unveiling a mechanical signature of their condensation level. Conversely, intact mature particles demonstrate more rigidity than immature or empty ones. DNA-condensing polyamines revert the mechanics of mature capsid and cores to near-immature values. The combination of these experiments reveals the pressurization of adenovirus particles induced by maturation. We estimate a pressure of ∼30 atm by continuous elasticity, which is corroborated by modeling the adenovirus mini-chromosome as a confined compact polymer. We propose this pressurization as a mechanism that facilitates initiating the stepwise disassembly of the mature particle, enabling its escape from the endosome and final genome release at the nuclear pore.


Assuntos
Adenovírus Humanos/química , Capsídeo/química , Cromatina/química , Pressão , Vírion/química , Entropia , Células HEK293 , Células HeLa , Humanos , Espermidina/farmacologia
11.
ACS Nano ; 9(11): 10571-9, 2015 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-26381363

RESUMO

Viruses package their genome in a robust protein coat to protect it during transmission between cells and organisms. In a reaction termed uncoating, the virus is progressively weakened during entry into cells. At the end of the uncoating process the genome separates, becomes transcriptionally active, and initiates the production of progeny. Here, we triggered the disruption of single human adenovirus capsids with atomic force microscopy and followed genome exposure by single-molecule fluorescence microscopy. This method allowed the comparison of immature (noninfectious) and mature (infectious) adenovirus particles. We observed two condensation states of the fluorescently labeled genome, a feature of the virus that may be related to infectivity. Beyond tracking the unpacking of virus genomes, this approach may find application in testing the cargo release of bioinspired delivery vehicles.


Assuntos
Adenoviridae/genética , Adenoviridae/fisiologia , Genoma Viral , Montagem de Vírus , Benzoxazóis/química , Capsídeo/metabolismo , Linhagem Celular Tumoral , Humanos , Microscopia de Força Atômica , Microscopia de Fluorescência , Compostos de Quinolínio/química
12.
Methods Mol Biol ; 1089: 1-24, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24132474

RESUMO

In this chapter we compile a battery of biophysical and imaging methods suitable to investigate adenovirus structural stability, structure, and assembly. Some are standard methods with a long history of use in virology, such as embedding and sectioning of infected cells, negative staining, or immunoelectron microscopy, as well as extrinsic fluorescence. The newer cryo-electron microscopy technique, which combined with advanced image processing tools has recently yielded an atomic resolution picture of the complete virion, is also described. Finally, we detail the procedure for imaging and interacting with single adenovirus virions using the atomic force microscope in liquid conditions. We provide examples of the kind of data obtained with each technique.


Assuntos
Adenoviridae/ultraestrutura , Adenoviridae/fisiologia , Microscopia de Força Atômica/métodos , Microscopia Eletrônica/métodos , Espectrometria de Fluorescência/métodos
13.
Sci Rep ; 3: 1434, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23486377

RESUMO

The standard pathway for virus infection of eukaryotic cells requires disassembly of the viral shell to facilitate release of the viral genome into the host cell. Here we use mechanical fatigue, well below rupture strength, to induce stepwise disruption of individual human adenovirus particles under physiological conditions, and simultaneously monitor disassembly in real time. Our data show the sequence of dismantling events in individual mature (infectious) and immature (noninfectious) virions, starting with consecutive release of vertex structures followed by capsid cracking and core exposure. Further, our experiments demonstrate that vertex resilience depends inextricably on maturation, and establish the relevance of penton vacancies as seeding loci for virus shell disruption. The mechanical fatigue disruption route recapitulates the adenovirus disassembly pathway in vivo, as well as the stability differences between mature and immature virions.


Assuntos
Adenovírus Humanos/fisiologia , Estresse Mecânico , Adenovírus Humanos/química , Capsídeo/química , Capsídeo/fisiologia , Proteínas do Capsídeo/metabolismo , Humanos , Microscopia de Força Atômica , Simulação de Dinâmica Molecular , Montagem de Vírus
14.
J Biol Chem ; 287(37): 31582-95, 2012 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-22791715

RESUMO

Adenovirus assembly concludes with proteolytic processing of several capsid and core proteins. Immature virions containing precursor proteins lack infectivity because they cannot properly uncoat, becoming trapped in early endosomes. Structural studies have shown that precursors increase the network of interactions maintaining virion integrity. Using different biophysical techniques to analyze capsid disruption in vitro, we show that immature virions are more stable than the mature ones under a variety of stress conditions and that maturation primes adenovirus for highly cooperative DNA release. Cryoelectron tomography reveals that under mildly acidic conditions mimicking the early endosome, mature virions release pentons and peripheral core contents. At higher stress levels, both mature and immature capsids crack open. The virus core is completely released from cracked capsids in mature virions, but it remains connected to shell fragments in the immature particle. The extra stability of immature adenovirus does not equate with greater rigidity, because in nanoindentation assays immature virions exhibit greater elasticity than the mature particles. Our results have implications for the role of proteolytic maturation in adenovirus assembly and uncoating. Precursor proteins favor assembly by establishing stable interactions with the appropriate curvature and preventing premature ejection of contents by tightly sealing the capsid vertices. Upon maturation, core organization is looser, particularly at the periphery, and interactions preserving capsid curvature are weakened. The capsid becomes brittle, and pentons are more easily released. Based on these results, we hypothesize that changes in core compaction during maturation may increase capsid internal pressure to trigger proper uncoating of adenovirus.


Assuntos
Adenoviridae/fisiologia , Capsídeo/fisiologia , DNA Viral/metabolismo , Internalização do Vírus , Células HEK293 , Humanos
15.
Ultramicroscopy ; 114: 56-61, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22356789

RESUMO

Control and minimization of tip-sample interaction forces are imperative tasks to maximize the performance of atomic force microscopy. In particular, when imaging soft biological matter in liquids, the cantilever dragging force prevents identification of the tip-sample mechanical contact, resulting in deleterious interaction with the specimen. In this work we present an improved jumping mode procedure that allows detecting the tip-sample contact with high accuracy, thus minimizing the scanning forces (-100 pN) during the approach cycles. To illustrate this method we report images of human adenovirus and T7 bacteriophage particles which are prone to uncontrolled modifications when using conventional jumping mode.


Assuntos
Adenoviridae/ultraestrutura , Bacteriófago T7/ultraestrutura , Microscopia de Força Atômica/métodos , Humanos
16.
Chem Commun (Camb) ; (6): 678-9, 2004 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-15010777

RESUMO

The novel dimer [Cp(2)Zr[upper bond start]([minus sign in circle])S([plus sign in circle])CH(Ph)CH[double bond]C[upper bond end]PPh(2)](2), the first example of a structurally characterised sulfur-bridged binuclear zirconathiolane complex, was prepared, characterised by NMR spectroscopy and X-ray crystallography, and some aspects of its solution behaviour were studied.

17.
Obstet Gynecol ; 99(5 Pt 2): 930-2, 2002 May.
Artigo em Inglês | MEDLINE | ID: mdl-11975963

RESUMO

BACKGROUND: As more women with cystic fibrosis are living into their reproductive years, this disease can complicate pregnancy and coexist with other entities. We report a case of cystic fibrosis with hemolytic uremic syndrome. CASE: An 18-year-old primigravida with cystic fibrosis was admitted at term with pulmonary symptoms, hypertension, and thrombocytopenia. She was delivered with the admitting diagnosis of severe preeclampsia. Postpartum, thrombocytopenia, and microangiopathic hemolytic anemia worsened. She developed renal failure and acute respiratory distress syndrome, requiring plasmapheresis, mechanical ventilation, and hemodialysis. Renal biopsy was consistent with the diagnosis of hemolytic uremic syndrome. CONCLUSION: Cystic fibrosis, a disease once managed predominantly by pediatric subspecialists, will be seen increasingly by physicians caring for adults, including obstetrician-gynecologists. We may also expect to see it coexisting with other disorders. The management of such patients may prove challenging.


Assuntos
Fibrose Cística/complicações , Síndrome Hemolítico-Urêmica/complicações , Complicações na Gravidez/fisiopatologia , Adolescente , Feminino , Síndrome Hemolítico-Urêmica/diagnóstico , Humanos , Gravidez , Complicações na Gravidez/diagnóstico
18.
Artigo em Espanhol | MEDLINE | ID: mdl-2773689

RESUMO

The authors study aggression with the "Instrument I to measure aggression of Ledesma Jimeno, Rodrigues Isidoro and Izquierdo de la Torre" in a sample of teachers (N = 48) made up of 15 university professors and 33 non-university professors (primary school, secondary school and professional training). The teacher sample is compared with a control sample of 479 men. The group of non-university professors showed more self-aggression and less aggression toward others than the control sample and university sample did. Both groups of teachers showed high levels of "technical intelligence". The aggression of the university professors was very similar to the control group and showed special peculiarities giving high values of intrapsychic incoherence that made their conduct more unforeseeable, with higher creative dispositions and a higher risk to their mental health than the other two groups studied.


Assuntos
Agressão/psicologia , Personalidade , Ensino , Adulto , Humanos , Masculino , Pessoa de Meia-Idade
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