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51.
Keisuke Ishihara Franziska Decker Paulo Caldas James F. Pelletier Martin Loose Jan Brugus Timothy J. Mitchison 《Molecular biology of the cell》2021,32(9):869
Microtubule plus-end depolymerization rate is a potentially important target of physiological regulation, but it has been challenging to measure, so its role in spatial organization is poorly understood. Here we apply a method for tracking plus ends based on time difference imaging to measure depolymerization rates in large interphase asters growing in Xenopus egg extract. We observed strong spatial regulation of depolymerization rates, which were higher in the aster interior compared with the periphery, and much less regulation of polymerization or catastrophe rates. We interpret these data in terms of a limiting component model, where aster growth results in lower levels of soluble tubulin and microtubule-associated proteins (MAPs) in the interior cytosol compared with that at the periphery. The steady-state polymer fraction of tubulin was ∼30%, so tubulin is not strongly depleted in the aster interior. We propose that the limiting component for microtubule assembly is a MAP that inhibits depolymerization, and that egg asters are tuned to low microtubule density. 相似文献
52.
Paulo S. Monzani Samuel Guemra Paulo R. Adona Otavio M. Ohashi Flávio V. Meirelles 《Animal biotechnology》2013,24(2):136-142
Prior to generating transgenic animals for bioreactors, it is important to evaluate the vector constructed to avoid poor protein expression. Mammary epithelial cells cultured in vitro have been proposed as a model to reproduce the biology of the mammary gland. In the present work, three lentiviral vectors were constructed for the human growth hormone (GH), interleukin 2 (IL2), and granulocyte colony-stimulating factor 3 (CSF3) genes driven by the bovine β-casein promoter. The lentiviruses were used to transduce mammary epithelial cells (MAC-T), and the transformed cells were cultured on polystyrene in culture medium with and without prolactin. The gene expression of transgenes was evaluated by PCR using cDNA, and recombinant protein expression was evaluated by Western-blotting using concentrated medium and cellular extracts. The gene expression, of the three introduced genes, was detected in both induced and non induced MAC-T cells. The human GH protein was detected in the concentrated medium, whereas CSF3 was detected in the cellular extract. Apparently, the cellular extract is more appropriate than the concentrated medium to detect recombinant protein, principally because concentrated medium has a high concentration of bovine serum albumin. The results suggest that MAC-T cells may be a good system to evaluate vector construction targeting recombinant protein expression in milk. 相似文献
53.
J. Paulo Macedo Jorge Pereira João Faria J. Luis Alves José López-López 《Computer methods in biomechanics and biomedical engineering》2013,16(12):655-662
AbstractObjectives: The purpose of the present study was to evaluate the distribution and magnitude of stresses through the bone tissue surrounding Morse taper dental implants at different positioning relative to the bone crest. Materials and Methods: A mandibular bone model was obtained from a computed tomography scan. A three-dimensional (3D) model of Morse taper implant-abutment systems placed at the bone crest (equicrestal) and 2?mm bellow the bone crest (subcrestal) were assessed by finite element analysis (FEA). FEA was carried out on axial and oblique (45°) loading at 150 N relatively to the central axis of the implant. The von Mises stresses were analysed considering magnitude and volume of affected peri-implant bone. Results: On vertical loading, maximum von Mises stresses were recorded at 6-7?MPa for trabecular bone while values ranging from 73 up to 118?MPa were recorded for cortical bone. On oblique loading at the equiquestral or subcrestal positioning, the maximum von Mises stresses ranged from 15 to 21?MPa for trabecular bone while values at 150?MPa were recorded for the cortical bone. On vertical loading, >99.9vol.% cortical bone volume was subjected to a maximum of 2?MPa while von Mises stress values at 15?MPa were recorded for trabecular bone. On oblique loading, >99.9vol.% trabecular bone volume was subjected to maximum stress values at 5?MPa, while von Mises stress values at 35?MPa were recorded for >99.4vol.% cortical bone. Conclusions: Bone volume-based stress analysis revealed that most of the bone volume (>99% by vol) was subjected to significantly lower stress values around Morse taper implants placed at equicrestal or subcrestal positioning. Such analysis is commentary to the ordinary biomechanical assessment of dental implants concerning the stress distribution through peri-implant sites. 相似文献
54.
Amilcar Chagas Freitas Júnior Paulo Henrique dos Santos Ching-Chang Ko Manoel Martín Júnior Erika Oliveira de Almeida 《Computer methods in biomechanics and biomedical engineering》2013,16(5):515-521
This study aimed to evaluate the influence of loading on a maxillary central incisor with the periodontal ligament (PDL) represented by 2D elastic beam elements using a 2D finite element analysis. Two models (M) were built varying the PDL representation: Mh (homogeneous PDL) and Mht (heterogeneous PDL with beam3 elements). Stress and displacements were determined for three loading conditions (L): Ll, lingual face loading at 45° with the tooth long axis; Li, perpendicular to the incisal edge; and Lip, on the incisal edge, parallel to the tooth long axis. Evaluation was performed on ANSYS software. Lip provided lower stress variation on the tooth and support structures when compared to Ll and Li. PDL's influence on stress values was lower for Lip. Oblique loading showed stress and displacement not observed in parallel loading condition through PDL's heterogeneous representation and it is probably incompatible with the in vivo condition. 相似文献
55.
Diego Ribeiro Migueis Ortega Roberto Hiroaki Nagahama Paulo Cesar Motta Rogério Bertani 《ZooKeys》2013,(352):93-116
The mygalomorph neotropical genus Fufius Simon, 1888 comprises ten species, distributed from Guatemala in Central America to southeastern Brazil, in South America. Most of the species were described from northern South America, in the Amazonian region. Only F. funebris Vellard, 1924 and F. lucasae Guadanucci & Indicatti, 2004 are known from regions more to the south of the continent. Herein we describe three new Brazilian species, Fufius minusculus
sp. n. and F. jalapensis
sp. n. from the state of Tocantins, and F. candango
sp. n. from Distrito Federal. The female of F. lucasae is described for first time and the male and female of F. funebris are redescribed based on specimens collected at the type locality. 相似文献
56.
Lionete Nunes de Lima Caio C. Aragon Cesar Mateo Jose M. Palomo Raquel L.C. Giordano Paulo W. Tardioli Jose M. Guisan Gloria Fernandez-Lorente 《Process Biochemistry》2013,48(1):118-123
The soluble lipase from Pseudomonas fluorescens (PFL) forms bimolecular aggregates in which the hydrophobic active centers of the enzyme monomers are in close contact. This bimolecular aggregate could be immobilized by multipoint covalent linkages on glyoxyl supports at pH 8.5. The monomer of PFL obtained by incubation of the soluble enzyme in the presence of detergent (0.5% TRITON X-100) could not be immobilized under these conditions. The bimolecular aggregate has two amino terminal residues in the same plane. A further incubation of the immobilized derivative under more alkaline conditions (e.g., pH 10.5) allows a further multipoint attachment of lysine (Lys) residues located in the same plane as the amino terminal residues. Monomeric PFL was immobilized at pH 10.5 in the presence of 0.5% TRITON X-100. The properties of both PFL derivatives were compared. In general, the bimolecular derivatives were more active, more selective and more stable both in water and in organic solvents than the monomolecular ones. The bimolecular derivative showed twice the activity and a much higher selectivity (100 versus 20) for the hydrolysis of R,S-2-hydroxy-4-phenylbutyric acid ethyl ester (HPBEt) in aqueous media at pH 5.0 compared to the monomeric derivative. In experiments measuring thermal inactivation at 75 °C, the bimolecular derivative was 5-fold more stable than the monomeric derivative (and 50-fold more stable than a one-point covalently immobilized PFL derivative), and it had a half-life greater than 4 h. In organic solvents (cyclohexane and tert-amyl alcohol), the bimolecular derivative was much more stable and more active than the monomeric derivative in catalyzing the transesterification of olive oil with benzyl alcohol. 相似文献
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