Diffusion and formation of microtubule asters: physical processes versus biochemical regulation.

Microtubule asters forming the mitotic spindle are assembled around two centrosomes through the process of dynamic instability in which microtubules alternate between growing and shrinking states. By modifying the dynamics of this assembly process, cell cycle enzymes, such as cdc2 cyclin kinases, regulate length distributions in the asters. It is believed that the same enzymes control the number of assembled microtubules by changing the "nucleating activity" of the centrosomes. Here we show that assembly of microtubule asters may be strongly altered by effects connected with diffusion of tubulin monomers. Theoretical analysis of a simple model describing assembly of microtubule asters clearly shows the existence of a region surrounding the centrosome depleted in GTP tubulin. The number of assembled microtubules may in some cases be limited by this depletion effect rather than by the number of available nucleation sites on the centrosome.

Angiogenic properties of human immunodeficiency virus type 1 Tat protein.

Extracellular human immunodeficiency virus type 1 (HIV-1) Tat protein promotes growth of spindle cells derived from AIDS-associated Kaposi sarcoma (AIDS-KS), an angioproliferative disease very frequent in HIV-1-infected individuals. Normal vascular cells, progenitors of AIDS-KS cells, proliferate in response to Tat after exposure to inflammatory cytokines, whose levels are augmented in HIV-1-infected individuals and in KS lesions. Here we show that Tat also promotes AIDS-KS and normal vascular cells to migrate and to degrade the basement membrane and stimulates endothelial cell morphogenesis on a matrix substrate. These effects are obtained at picomolar concentrations of exogenous Tat and are promoted by the treatment of the cells with the same inflammatory cytokines stimulating expression of the receptors for Tat, the integrins alpha 5 beta 1 and alpha v beta 3. Thus, under specific circumstances, Tat has angiogenic properties. As Tat and its receptors are present in AIDS-KS lesions, these data may explain some of the mechanisms by which Tat can induce angiogenesis and cooperate in the development of AIDS-KS.

Estudo da geometria da aresta de corte de ferramentas aplicadas ao torneamento de superligas à base de níquel com alta velocidade de corte

Pesquisadores e indústrias de todo o mundo estão firmemente comprometidos com o propósito de fazer o processo de usinagem ser precisamente veloz e produtivo. A forte concorrência mundial gerou a procura por processos de usinagem econômicos, com grande capacidade de produção de cavacos e que produzam peças com elevada qualidade. Dentre as novas tecnologias que começaram a ser empregadas, e deve tornar-se o caminho certo a ser trilhado na busca da competitividade em curto espaço de tempo, está a tecnologia de usinagem com altas velocidades (HSM de High Speed Machining). A tecnologia HSM surge como componente essencial na otimização dos esforços para manutenção e aumento da competitividade global das empresas. Durante os últimos anos a usinagem com alta velocidade tem ganhado grande importância, sendo dada uma maior atenção ao desenvolvimento e à disponibilização no mercado de máquinas-ferramentas com rotações muito elevadas (20.000 - 100.000 rpm). O processo de usinagem com alta velocidade está sendo usado não apenas para ligas de alumínio e cobre, mas também para materiais de difícil usinabilidade, como os aços temperados e superligas à base de níquel. Porém, quando se trata de materiais de difícil corte, têm-se observado poucas publicações, principalmente no processo de torneamento. Mas, antes que a tecnologia HSM possa ser empregada de uma forma econômica, todos os componentes envolvidos no processo de usinagem, incluindo a máquina, o eixo-árvore, a ferramenta e o pessoal, precisam estar afinados com as peculiaridades deste novo processo. No que diz respeito às máquinas-ferramenta, isto significa que elas têm que satisfazer requisitos particulares de segurança. As ferramentas, devido à otimização de suas geometrias, substratos e revestimentos, contribuem para o sucesso deste processo. O presente trabalho objetiva estudar o comportamento de diversas geometrias ) de insertos de cerâmica (Al2O3 + SiCw e Al2O3 + TIC) e PCBN com duas concentrações de CBN na forma padrão, assim como modificações na geometria das arestas de corte empregadas em torneamento com alta velocidade em superligas à base de níquel (Inconel 718 e Waspaloy). Os materiais foram tratados termicamente para dureza de 44 e 40 HRC respectivamente, e usinados sob condição de corte a seco e com utilização da técnica de mínima quantidade de lubrificante (minimal quantity lubricant - MQL) visando atender requisitos ambientais. As superligas à base de níquel são conhecidas como materiais de difícil usinabilidade devido à alta dureza, alta resistência mecânica em alta temperatura, afinidade para reagir com materiais da ferramenta e baixa condutividade térmica. A usinagem de superligas afeta negativamente a integridade da peça. Por essa razão, cuidados especiais devem ser tomados para assegurar a vida da ferramenta e a integridade superficial de componentes usinados por intermédio de controle dos principais parâmetros de usinagem. Experimentos foram realizados sob diversas condições de corte e geometrias de ferramentas para avaliação dos parâmetros: força de corte, temperatura, emissão acústica e integridade superficial (rugosidade superficial, tensão residual, microdureza e microestrutura) e mecanismos de desgaste. Mediante os resultados apresentados, recomenda-se à geometria de melhor desempenho nos parâmetros citados e confirma-se a eficiência da técnica MQL. Dentre as ferramentas e geometrias testadas, a que apresentou melhor desempenho foi a ferramenta cerâmica CC650 seguida da ferramenta cerâmica CC670 ambas com formato redondo e geometria 2 (chanfro em T de 0,15 x 15º com raio de aresta de 0,03 mm).

Lesão hiperemiada do bordo palpebral

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Reductionism at the vertebrate kinetochore

The kinetochore forms the site of attachment for mitotic spindle microtubules driving chromosome segregation. The interdependent protein interactions in this large structure have made it difficult to dissect the function of its components. In this issue, Hori et al. (2013. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201210106) present a novel and powerful methodology to address the sufficiency of individual proteins for the creation of a functional de novo centromere.

Temporal control of epigenetic centromere specification

All living organisms require accurate mechanisms to faithfully inherit their genetic material during cell division. The centromere is a unique locus on each chromosome that supports a multiprotein structure called the kinetochore. During mitosis, the kinetochore is responsible for connecting chromosomes to spindle microtubules, allowing faithful segregation of the duplicated genome. In most organisms, centromere position and function is not defined by the local DNA sequence context but rather by an epigenetic chromatin-based mechanism. Centromere protein A (CENP-A) is central to this process, as chromatin assembled from this histone H3 variant is essential for assembly of the centromere complex, as well as for its epigenetic maintenance. As a major determinant of centromere function, CENP-A assembly requires tight control, both in its specificity for the centromere and in timing of assembly. In the last few years, there have been several new insights into the molecular mechanism that allow this process to occur. We will review these here and discuss the general implications of the mechanism of cell cycle coupling of centromere inheritance.

Polo-like kinase 4 controls centriole duplication but does not directly regulate cytokinesis

Centrioles organize the centrosome, and accurate control of their number is critical for the maintenance of genomic integrity. Centrioles duplicate once per cell cycle, and duplication is coordinated by Polo-like kinase 4 (Plk4). We previously demonstrated that Plk4 accumulation is autoregulated by its own kinase activity. However, loss of heterozygosity of Plk4 in mouse embryonic fibroblasts has been proposed to cause cytokinesis failure as a primary event, leading to centrosome amplification and gross chromosomal abnormalities. Using targeted gene disruption, we show that human epithelial cells with one inactivated Plk4 allele undergo neither cytokinesis failure nor increase in centrosome amplification. Plk4 is shown to localize exclusively at the centrosome, with none in the spindle midbody. Substantial depletion of Plk4 by small interfering RNA leads to loss of centrioles and subsequent spindle defects that lead to a modest increase in the rate of cytokinesis failure. Therefore, Plk4 is a centriole-localized kinase that does not directly regulate cytokinesis.

Distinct mechanisms eliminate mother and daughter centrioles in meiosis of starfish oocytes

Centriole elimination is an essential process that occurs in female meiosis of metazoa to reset centriole number in the zygote at fertilization. How centrioles are eliminated remains poorly understood. Here we visualize the entire elimination process live in starfish oocytes. Using specific fluorescent markers, we demonstrate that the two older, mother centrioles are selectively removed from the oocyte by extrusion into polar bodies. We show that this requires specific positioning of the second meiotic spindle, achieved by dynein-driven transport, and anchorage of the mother centriole to the plasma membrane via mother-specific appendages. In contrast, the single daughter centriole remaining in the egg is eliminated before the first embryonic cleavage. We demonstrate that these distinct elimination mechanisms are necessary because if mother centrioles are artificially retained, they cannot be inactivated, resulting in multipolar zygotic spindles. Thus, our findings reveal a dual mechanism to eliminate centrioles: mothers are physically removed, whereas daughters are eliminated in the cytoplasm, preparing the egg for fertilization.

Dynamical Scenarios for Chromosome Bi-orientation

Chromosome bi-orientation at the metaphase spindle is essential for precise segregation of the genetic material. The process is error-prone, and error-correction mechanisms exist to switch misaligned chromosomes to the correct, bi-oriented configuration. Here, we analyze several possible dynamical scenarios to explore how cells might achieve correct bi-orientation in an efficient and robust manner. We first illustrate that tension-mediated feedback between the sister kinetochores can give rise to a bistable switch, which allows robust distinction between a loose attachment with low tension and a strong attachment with high tension. However, this mechanism has difficulties in explaining how bi-orientation is initiated starting from unattached kinetochores. We propose four possible mechanisms to overcome this problem (exploiting molecular noise; allowing an efficient attachment of kinetochores already in the absence of tension; a trial-and-error oscillation; and a stochastic bistable switch), and assess their impact on the bi-orientation process. Based on our results and supported by experimental data, we put forward a trial-and-error oscillation and a stochastic bistable switch as two elegant mechanisms with the potential to promote bi-orientation both efficiently and robustly.

UM Men's Swimming Team, 1927

Back Row: Clarence Batter, Richard S. Spindle, Robert W. Wagner, Albert Mayer, Jr., J. B. Allan Seager, Thomas Y. Watson

Middle Row: Carl R. Darnall, Robert L. Halsted, Paul Starrett, captain Paul C. Samson, head coach Matt Mann, J.M. Halsted, Maurice J. Shorr

Front Row: Harold W. Bailey, Clarence A. Horn, George E. Hubbell, George W. Bement

UM Men's Swimming Team, 1928

Back Row: Henry Dinkelspiel, John C. Benedict, Charles F. McCaffree, William F. McDonald, Edward L. Warner, Robert C. Goldsmith, Jack L. Lair,

Third Row: Harold W. Bailey, Ernest C. Reif, Frank W. Walaitis, Walter Chaffee, Robert P. Walker, J.J Thompson, Garnet Ault,

Garfield Hubbell, Thomas Y. Watson, J. B. Allan Seager, captain Carl R. Darnall, head coach Matt Mann, Robert W. Wagner, Richard S. Spindle, Clarence Batter

Front Row: Byron O. Hughes, J.M. Halstead, Clarence A. Horn, Meyer Rosenberg

UM Men's Swimming Team, 1929

Back Row: Harold W. Bailey, Howard Brown, William F. McDonald, Robert C. Goldsmith, Edward L. Warner, Henry Dinkelspiel,

Third Row: head coach Matt Mann, Richard C. Mertz, Charles F. McCaffree, Garnet W. Ault, J.J. Thompson, O. Bruce. Goldsmith, Rawson F. Hosmer, assistant coach John W. MacMahon

Second Row:, Thomas Y. Watson, George E. Hubbell, Robert P. Walker, captain Richard S. Spindle, J.B. Allan Seager, Frank W. Walaitis, Ernest C. Reif

Front Row: Byron O. Hughes, Frederick J. Grimshaw, Harold E. Nimz

Neuromuscular efficiency of the sternocleidomastoid and anterior scalene muscles in patients with chronic neck pain

Purpose: This study compared the neuromuscular efficiency (NME) of the sternocleidomastoid (SCM) and anterior scalene (AS) muscles between 20 chronic neck pain patients and 20 asymptomatic controls. Method: Myoelectric signals were recorded from the sternal head of SCM and the AS muscles as subjects performed sub-maximal isometric cervical flexion contractions at 25 and 50% of the maximum voluntary contraction (MVC). The NME was calculated as the ratio between MVC and the corresponding average rectified value of the EMG signal. Ultrasonography was used to measure subcutaneous tissue thickness over the SCM and AS to ensure that differences did not exist between groups. Results: For both the SCM and AS muscles, NME was shown to be significantly reduced in patients with neck pain at 25% MVC (p < 0.05). Subcutaneous tissue thickness over the SCM and AS muscles was not different between groups. Conclusions: Reduced NME in the superficial cervical flexor muscles in patients with neck pain may be a measurable altered muscle strategy for dysfunction in other muscles. This aberrant pattern of muscle activation appears to be most evident under conditions of low load. NME, when measured at 25% MVC, may be a useful objective measure for future investigation of muscle dysfunction in patients with neck pain.

Lectin-induced haemocyte inactivation in insects

Most multimeric lectins are adhesion molecules, promoting attachment and spreading on surface glycodeterminants. In addition, some lectins have counter-adhesion properties, detaching already spread cells which then acquire round or spindle-formed cell shapes. Since lectin-mediated adhesion and detachment is observed in haemocyte-like Drosophila cells, which have haemomucin as the major lectin-binding glycoprotein, the two opposite cell behaviours may be the result of lectin-mediated receptor rearrangements on the cell surface. To investigate oligomeric lectins as a possible extracellular driving force affecting cell shape changes, we examined lectin-mediated reactions in lepidopteran haemocytes after cytochalasin D-treatment and observed that while cell-spreading was dependent on F-actin, lectin-uptake was less dependent on F-actin. We propose a model of cell shape changes involving a dynamic balance between adhesion and uptake reactions. (C) 2004 Elsevier Ltd. All rights reserved.