Microtubules do not promote mitotic slippage when the spindle assembly checkpoint cannot be satisfied

© 2008 Brito et al. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Journal of Cell Biology 182 (2009): 623-629, doi:10.1083/jcb.200805072.When the spindle assembly checkpoint (SAC) cannot be satisfied, cells exit mitosis via mitotic slippage. In microtubule (MT) poisons, slippage requires cyclin B proteolysis, and it appears to be accelerated in drug concentrations that allow some MT assembly. To determine if MTs accelerate slippage, we followed mitosis in human RPE-1 cells exposed to various spindle poisons. At 37°C, the duration of mitosis in nocodazole, colcemid, or vinblastine concentrations that inhibit MT assembly varied from 20 to 30 h, revealing that different MT poisons differentially depress the cyclin B destruction rate during slippage. The duration of mitosis in Eg5 inhibitors, which induce monopolar spindles without disrupting MT dynamics, was the same as in cells lacking MTs. Thus, in the presence of numerous unattached kinetochores, MTs do not accelerate slippage. Finally, compared with cells lacking MTs, exit from mitosis is accelerated over a range of spindle poison concentrations that allow MT assembly because the SAC becomes satisfied on abnormal spindles and not because slippage is accelerated.This research was supported by The National Institutes of Health (GMS 40198 to C.L. Rieder) and a doctoral research fellowship (SFRH/ BD/13663/2003) from Fundacao para a Ciencia e a Tecnologia (to D.A. Brito)

Cells satisfy the mitotic checkpoint in Taxol, and do so faster in concentrations that stabilize syntelic attachments

© The Authors, 2009 . This article is distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. The definitive version was published in Journal of Cell Biology 186 (2009): 675-684, doi:10.1083/jcb.200906150.To determine why the duration of mitosis (DM) is less in Taxol than in nocodazole or Eg5 inhibitors we studied the relationship between Taxol concentration, the DM, and the mitotic checkpoint. We found that unlike for other spindle poisons, in Taxol the DM becomes progressively shorter as the concentration surpasses ~0.5 µM. Studies on RPE1 and PtK2 expressing GFP/cyclin B or YFP/Mad2 revealed that cells ultimately satisfy the checkpoint in Taxol and do so faster at concentrations >0.5 µM. Inhibiting the aurora-B kinase in Taxol-treated RPE1 cells accelerates checkpoint satisfaction by stabilizing syntelic kinetochore attachments and reduces the DM to ~1.5 h regardless of drug concentration. A similar stabilization of syntelic attachments by Taxol itself appears responsible for accelerated checkpoint satisfaction at concentrations >0.5 µM. Our results provide a novel conceptual framework for how Taxol prolongs mitosis and caution against using it in checkpoint studies. They also offer an explanation for why some cells are more sensitive to lower versus higher Taxol concentrations.This work was supported by National Institutes of Health GMS grant 40198 to C.L. Rieder

Top-down human pose estimation with depth images and domain adaptation

In this paper, a method for estimation of human pose is proposed, making use of ToF (Time of Flight) cameras. For this, a YOLO based object detection method was used, to develop a top-down method. In the first stage, a network was developed to detect people in the image. In the second stage, a network was developed to estimate the joints of each person, using the image result from the first stage. We show that a deep learning network trained from scratch with ToF images yields better results than taking a deep neural network pretrained on RGB data and retraining it with ToF data. We also show that a top-down detector, with a person detector and a joint detector works better than detecting the body joints over the entire image.This work is supported by: European Structural and Investment Funds in the FEDER component, through the Operational Competitiveness and Internationalization Programme (COMPETE 2020) [Project no 002797; Funding Reference: POCI-01-0247-FEDER-002797]

Automated generation of synthetic in-car dataset for human body pose detection

In this paper, a toolchain for the generation of realistic synthetic images for human body pose detection in an in-car environment is proposed. The toolchain creates a customized synthetic environment, comprising human models, car, and camera. Poses are automatically generated for each human, taking into account a per-joint axis Gaussian distribution, constrained by anthropometric and range of motion measurements. Scene validation is done through collision detection. Rendering is focused on vision data, supporting time-of-flight (ToF) and RGB cameras, generating synthetic images from these sensors. Ground-truth data is then generated, comprising the car occupants' body pose (2D/3D), as well as full body RGB segmentation frames with different body parts' labels. We demonstrate the feasibility of using synthetic data, combined with real data, to train distinct machine learning agorithms, demonstrating the improvement in their algorithmic accuracy for the in-car scenario.This work is supported by: European Structural and Investment Funds in the FEDER component, through the Operational Competitiveness and Internationalization Programme (COMPETE 2020) [Project no 039334; Funding Reference: POCI-01-0247-FEDER-039334]

A system for the generation of in-car human body pose datasets

Published online: 8 October 2020With the advent of autonomous vehicles, detection of the occupants’ posture is crucial to tackle the needs of infotainment interaction or passive safety systems. Generative approaches have been recently proposed for human body pose in-car detection, but this type of approaches requires a large training dataset for a feasible accuracy. This requirement poses a difficulty, given the substantial time required to annotate such large amount of data. In the in-car scenario, this requirement risk increases even further, since a robust human body pose ground-truth system capable of working in it is needed but inexistent. Currently, the gold standard for human body pose capture is based on optical systems, requiring up to 39 visible markers for a plug-in gait model, which in this case are not feasible given the occlusions inside the car. Other solutions, such as inertial suits, also have limitations linked to magnetic sensitivity and global positioning drift. In this paper, a system for the generation of images for human body pose detection in an in-car environment is proposed. To this end, we propose to smartly combine inertial and optical systems to suppress their individual limitations: By combining the global positioning of 3 visible head markers provided by the optical system with the inertial suit’s relative human body pose, we obtain an occlusion-ready, drift-free full-body global positioning system. This system is then spatially and temporally calibrated with a time-of-flight sensor, automatically obtaining in-car image data with (multi-person) pose annotations. Besides quantifying the inertial suit inherent sensitivity and accuracy, the feasibility of the overall system for human body pose capture in the in-car scenario was demonstrated. Our results quantify the errors associated with the inertial suit, pinpoint some sources of the system’s uncertainty and propose how to minimize some of them. Finally, we demonstrate the feasibility of using system generated data (which was made publicly available), independently or mixed with two publicly available generic datasets (not in-car), to train 2 machine learning algorithms, demonstrating the improvement in their algorithmic accuracy for the in-car scenario.This work is supported by: European Structural and Investment Funds in the FEDER component, through the Operational Competitiveness and Internationalization Programme (COMPETE 2020) [Project n.º 002797; Funding Reference: POCI-01-0247-FEDER-002797]

Modulation of the Effects of Lung Immune Response on Bone Marrow by Oral Antigen Exposure

Allergic airway inflammation is attenuated by oral tolerization (oral exposure to allergen, followed by conventional sensitization and challenge with homologous antigen), which decreases airway allergen challenge-induced eosinophilic infiltration of the lungs and bone marrow eosinophilia. We examined its effects on bone marrow eosinophil and neutrophil production. Mice of wild type (BP-2, BALB/c, and C57BL/6) and mutant strains (lacking iNOS or CD95L) were given ovalbumin (OVA) or water (vehicle) orally and subsequently sensitized and challenged with OVA (OVA/OVA/OVA and H2O/OVA/OVA groups, resp.). Anti-OVA IgG and IgE, bone marrow eosinophil and neutrophil numbers, and eosinophil and neutrophil production ex vivo were evaluated. T lymphocytes from OVA/OVA/OVA or control H2O/OVA/OVA donors were transferred into naïve syngeneic recipients, which were subsequently sensitized/challenged with OVA. Alternatively, T lymphocytes were cocultured with bone marrow eosinophil precursors from histocompatible sensitized/challenged mice. OVA/OVA/OVA mice of the BP-2 and BALB/c strains showed, relative to H2O/OVA/OVA controls, significantly decreased bone marrow eosinophil counts and ex vivo eosinopoiesis/neutropoiesis. Full effectiveness in vivo required sequential oral/subcutaneous/intranasal exposures to the same allergen. Transfer of splenic T lymphocytes from OVA/OVA/OVA donors to naive recipients prevented bone marrow eosinophilia and eosinopoiesis in response to recipient sensitization/challenge and supressed eosinopoiesis upon coculture with syngeneic bone marrow precursors from sensitized/challenged donors

Combinaciones de aceites esenciales para el control de Sitophilus zeamais

Sitophilus zeamais ataca granos de maíz almacenados y tradicionalmente es controlado con pesticidas sintéticos. Sin embargo, el uso frecuente de estos compuestos causa daños al ambiente y a la salud, además de favorecer el desarrollo de poblaciones resistentes de insectos. El objetivo del trabajo fue evaluar el efecto de aceites esenciales (AEs) y sus combinaciones binarias contra S. zeamais, con el fin de encontrar alternativas menos riesgosas para su control. Los AEs utilizados fueron: Pimenta racemosa var. ozua, Rosmarinus officinalis, Pimenta haitiensis, Citrus sinensis e Illicium verum. Se realizó un ensayo de toxicidad fumigante probando los AEs individuales y sus combinaciones binarias (1:1 v/v). Para los tratamientos binarios se determinó el Índice Combinatorio (IC). Considerando los resultados de los AEs individuales y las combinaciones con mayor efecto fumigante, se realizó un test de atracción/repelencia.Las combinaciones binarias mostraron mayor efecto fumigante y repelente que los AEs individuales. Pimenta racemosa-R. Officinalis, P. racemosa-C. sinensis y R. officinalis-C. sinensis fueron las combinaciones con mayor actividad fumigante (CL95= 43.59, 52.27 y 62.27 μl/l, respectivamente). Los valores IC de estas combinaciones revelan sinergismo entre estos AEs. Además, P. racemosa-R.officinalis fue la combinación más repelente (IR: −62,92, −67,46, −70,62 para 4 μl/l, 0,4 μl/l y 0,2 μl/l, respectivamente). Resumiendo P. racemosa-R. officinalis tiene un mayor efecto insecticida y repelente contra S. zeamais con respecto a los AEs individuales. En conclusión, el efecto sinérgico de las combinaciones binarias disminuye la cantidad de AE requerido, proporcionando una interesante alternativa para el control del gorgojo del maíz.Fil: Brito, Vanessa Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Achimón, Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Pizzolitto, Romina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Ramirez Sanchez, Adriana Ines. Instituto de Innovación en Biotecnología e Industria; República DominicanaFil: Gómez Torres, E. A.. Instituto de Innovación en Biotecnología e Industria; República DominicanaFil: Zygadlo, Julio Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Zunino, María Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina1° Congreso Argentino de SemillasVirtualArgentinaUniversidad Nacional de Córdoba. Facultad de Ciencias AgropecuariasAsociación de Laboratorios Agropecuarios Privado

Primary resistance of HIV to antiretrovirals among individuals recently diagnosed at voluntary counselling and testing centres in the metropolitan region of Recife, Pernambuco

Determining the prevalence and type of antiretroviral (ARV) resistance among ARV-naïve individuals is important to assess the potential responses of these individuals to first-line regimens. The prevalence of primary resistance and the occurrence of recent infections among individuals with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) were identified among recently diagnosed patients at five sexually transmitted disease/AIDS testing and counselling centres in the metropolitan region of Recife (RMR), Pernambuco, Brazil, between 2007-2009. One-hundred and eight samples were analysed using the Calypte® BED assay. Males predominated (56%), as did patients aged 31-50 years. Twenty-three percent presented evidence of a recent HIV infection. The median CD4+ T lymphocyte count was 408 cells/mm³ and the median viral load was 3.683 copies/mL. The prevalence of primary resistance was 4.6% (confidence interval 95% = 1-8.2%) based on criteria that excluded common polymorphisms in accordance with the surveillance drug resistance mutation criteria. The prevalence of resistance to non-nucleoside reverse transcriptase, nucleoside/nucleotide reverse transcriptase and protease inhibitors were 3.8%, 1.5% and 0.8%, respectively. Fifty-seven percent of strains were from clade B, 37.7% were clade F and 3.1% were clade C; there were no statistically significant differences with respect to resistance between clades. Recent infection tended to be more common in men (p = 0.06) and in municipalities in the south of the RMR (Jaboatão dos Guararapes and Cabo de Santo Agostinho) (p = 0.046). The high prevalence of recent infection and the high prevalence of non-B strains in this poor Brazilian region merit further attention.Laboratório Central de Saúde Pública de Pernambuco Setor de VirologiaUniversidade Federal de Pernambuco Programa de Pós-Graduação em Medicina TropicalFiocruz Centro de Pesquisa Aggeu MagalhãesCentro de Testagem e Aconselhamento Herbert de SouzaUniversidade Federal de São Paulo (UNIFESP) Laboratório de RetrovirologiaUNIFESP, Laboratório de RetrovirologiaSciEL