Regulation of kinetochore localization of the Spindle checkpoint kinase Bub1

Le point de contrôle d’assemblage du fuseau mitotique (SAC) est un système de surveillance conservé chez les eucaryotes permettant un attachement précis entre les kinétochores et les microtubules. Le SAC empêche la progression mitotique jusqu'à ce que soit généré un attachement et une tension correcte entre les kinétochores et les microtubules. La dérégulation du SAC a des conséquences graves avec de l'aneuploïdie retrouvé dans la plupart des tumeurs solides. BUB1 est une kinase sérine/thréonine requise pour le fonctionnement du SAC. Elle possède à la fois des rôles dépendants et indépendants de sa fonction kinase. Ce projet définit plusieurs fonctions associées à BUB1 lors de la mitose. L'utilisation d’outils in vivo et in vitro ont permis d’identifier plusieurs sites d'autophosphorylation sur Bub1. Nous avons testé et confirmé le site T589 de BUB1 comme un site d'autophosphorylation. Un mutant de ce site (BUB1-T589A) a été exprimé de manière stable et un anticorps phosphospécifique a été généré pour étudier ce site. Le rôle structural des domaines de BUB1 a été rapporté précédemment. Nous montrons que quand le domaine d'extension du domaine kinase (aa 724-780) située en N-terminal du domaine kinase est nécessaire pour l’autophosphorylation de BUB1-T589 et l'activité de la kinase BUB1, le TPR à l’extrémité N-terminale est localisée normalement kinétochores et n’est pas requis pour l'activité kinase. BUB1-T589A a modifié le taux de renouvellement au kinétochores. Cela conduit à la propagation des signaux de SGO1 et de H2ApT120 au niveau des bras des chromosomes. Enfin, l’autophosphorylation en T589 régule le congression des chromosomes mais pas la fonction de BUB1 pour le SAC. De plus nous montrons que l'inhibition de PLK1, une autre kinase sérine/thréonine, augmente la localisation de BUB1 aux kinétochores après la suppression BUB3 dans les cellules humaines. Ainsi, PLK1 peut réguler la localisation de BUB1 aux kinétochores. Nous montrons également que cette régulation se produit à travers KNL1, une protéine d'échafaudage du SAC. PLK1 pourrait réglementer BUB1 kinétochore localisation pour influencer la progression mitotique. Des futures études se concentreront sur l’élucidation de mécanismes derrière ces interactions.The Spindle assembly checkpoint (SAC) is a monitoring system conserved in eukaryotes for accurate attachments between kinetochores and microtubules. The SAC precludes mitotic progression until correct attachments and tension between kinetochores and microtubules is generated. Deregulation of the SAC has the severe consequence of aneuploidy found in most solid tumors. BUB1 is a serine/threonine kinase required for the SAC function. It has both kinase-dependent and kinase-independent roles. This project defines several BUB1 associated functions during mitosis. Using in vivo and in vitro tools several autophosphorylation sites on BUB1 were identified. We tested and confirmed BUB1 T589 as an autophosphorylation site. A mutant of this site (BUB1-T589A) was stably expressed in cells and a phosphospecific antibody was generated to study this site. The role of structural domains of BUB1 has been studied earlier. We show that while the kinase extension domain (aa 724-780) located N-terminal to the kinase domain is required for BUB1-T589 autophosphorylation and BUB1 kinase activity, the TPR at the N-terminus localizes normally to kinetochores and is not required for kinase activity. BUB1-T589A has altered turnover at kinetochores. This leads to the spread of SGO1 and H2ApT120 signal to chromosome arms. Finally, autophosphorylation at T589 regulates chromosome congression but not the SAC function of BUB1. We further show that inhibition of PLK1, another serine/threonine kinase, increases BUB1 kinetochore localization after BUB3 depletion in human cells. Thus, PLK1 can regulate BUB1 kinetochore localization. We also show that this regulation occurs through KNL1, a scaffold protein of the SAC. It is possible that PLK1 could regulate BUB1 kinetochore localization to influence mitotic progression. Future studies will focus on elucidation of mechanism behind these interactions

Silicon-based carbonaceous electrocatalysts for oxygen reduction and evolution properties in alkaline conditions

In this contribution, new electrocatalyst materials, namely silicon-multiwalled carbon nanotubes (Si/MWCNTs), nitrogen-doped multiwalled carbon nano-tubes (Si/NCNTs), and silicon–carbon black (Si/CB), were developed and characterized in an effort to investigate less expensive and more efficient alternatives to Pt-based catalysis for energy storage cells applications. The role of structural behavior of obtained specimens and corresponding electrochemical performances were characterized through X-ray diffraction and scanning electron microscopy, while cyclic voltammetry and electrochemical impedance spectroscopy were analyzed for electrochemical measurements and evaluation of oxygen evolution reaction (OER) along with oxygen reduction reaction (ORR). The electrochemical studies have shown that these materials exhibit reasonable performance for both the ORR and the OER. The findings concluded that the Si/CB base catalyst has shown both OER and ORR activities in comparison to Si/MWCNTs and Si/NCNTs where only ORR performance was monitored. However, Si/NCNTs have shown much higher ORR activity compared to the others. This work highlights the comparison of three possible alternative materials as a potential catalyst to develop optimum alternatives of Pt-free catalysts for fuel cell and lithium-based battery systems

Sorption of As(V) from aqueous solution using in situ growth MgAl–NO3 layered double hydroxide thin film developed on AA6082

In this work, the MgAl–NO3 layered double hydroxide (LDH) developed by the single-step in situ growth method is used as a robust sorbent to remove arsenic from aqueous solution. The MgAl-LDH exhibiting two different distinct morphologies (platelet structure and cauliflower-shaped structure) was developed on the AA6082 substrate with the variation in synthesis parameters, where AA6082 specimen acts as both the reactant and support. The structural characterizations were investigated through scanning electron microscopy, X-ray diffraction analysis, and energy dispersion spectroscopy, while the adsorption of arsenic on MgAl-LDH was studied through Langmuir and Freundlich models. The Langmuir isotherms have shown a maximum adsorption capacity of around 213 and 239 mg/g for platelet and cauliflower-like MgAl-LDH, respectively. The pseudo-first-order and pseudo-second-order Lagergren kinetic models were studied for the understanding of the adsorption kinetics. The results depicted that anion exchange and the electrostatic interaction are the possible reasons of arsenic sorption on MgAl-LDH, but the ion exchange mechanism is found to be the dominant mechanism. The maximum adsorption capacity of cauliflower-shaped MgAl-LDH was found to be slightly higher than platelet structure, but overall maximum arsenic adsorption uptake values of both in situ growth structures have found to be exceeded the mostly reported MgAl-LDH maximum adsorption capacities

Cd20 Expression and Effects on Outcome of Relapsed/ Refractory Diffuse Large B Cell Lymphoma after Treatment with Rituximab

Introduction: Down regulation of CD20 expression has been reported in diffuse large B cell lymphoma (DLBCL)). Therefore, it is important to determine whether chemotherapy with rituximab induces CD20 down regulation and effects survival. Objectives: To determine the incidence of down regulation of CD20 expression in relapsed DLBCL after treatment with rituximab and to compare outcomes and assess pattern of relapse between CD20 negative and CD20 positive cases. Methodology: We retrospectively reviewed patients with relapsed DLBCL who received rituximab in the first line setting at Aga Khan University Hospital between January 2007 and December 2014. Data were recorded on predesigned questionnaires, with variables including demographics, details regarding date of diagnosis and relapse, histology, staging, international prognostic index, treatment and outcomes at initial diagnosis and at relapse. The Chi square test was applied to determine statistical significance between categorical variables. Survival curves were generated by the Kaplan–Meier method. Results: A total of 54 patients with relapsed DLBCL were included in our study, 38 (70 %) males and 16(30%) females. Some 23 (43%) patients were at stage IV at the time of diagnosis and 34 (63%) had B symptoms. The most frequent R-IPI at diagnosis was II in 24 (44%) patients. Only 6 (11%) did not show CD20 expression on re-biopsy for relapsed/refractory disease, 2 with CD20 negative DLBCL responding to second line chemotherapy. A complete response after salvage chemotherapy was noted in 16 (29.6%) cases with relapsed/refractory DLBCL. Seven (13%) patients underwent an autologous bone marrow transplant as consolidation after second line treatment. Median overall survival was 18 months in CD20 positive vs. 13 months in CD20 negative patients. Conclusion: This study demonstrated that a small percentage of patients treated with rituximab lose their CD20 expression at the time of relapse. However, it is unclear whether this is associated with an inferior outcome

Image Local Features Description through Polynomial Approximation

This work introduces a novel local patch descriptor that remains invariant under varying conditions of orientation, viewpoint, scale, and illumination. The proposed descriptor incorporate polynomials of various degrees to approximate the local patch within the image. Before feature detection and approximation, the image micro-texture is eliminated through a guided image filter with the potential to preserve the edges of the objects. The rotation invariance is achieved by aligning the local patch around the Harris corner through the dominant orientation shift algorithm. Weighted threshold histogram equalization (WTHE) is employed to make the descriptor in-sensitive to illumination changes. The correlation coefficient is used instead of Euclidean distance to improve the matching accuracy. The proposed descriptor has been extensively evaluated on the Oxford's affine covariant regions dataset, and absolute and transition tilt dataset. The experimental results show that our proposed descriptor can categorize the feature with more distinctiveness in comparison to state-of-the-art descriptors. - 2013 IEEE.This work was supported by the Qatar National Library.Scopu

Current Status of Intravesical Therapies for Bladder Pain Syndrome (BPS): A Narrative Review of Emerging Evidence

Bladder pain syndrome (BPS) is a complex condition, which can have debilitating sequelae for patients. Many elements of BPS remain poorly understood including pathophysiology, diagnosis and treatment. Navigating patient care can therefore be challenging for the clinician. Management mandates a multidisciplinary and symptom-based approach. Intravesical treatments such as instillation therapies remain a cornerstone of most treatment algorithms and there are a range of agents that can be selected. This review offers an up-to-date evaluation of the evidence for these intravesical treatments.publishedVersio

Management of Bladder Pain Syndrome (BPS): A Practical Guide

Bladder pain syndrome (BPS) is a prevalent and pervasive disease. The physical and psychological sequelae can be very burdensome for the patient, and the condition represents a real challenge for the clinician as well. With no simple pathognomonic test, finding harmony in navigating patient care can be demanding. Diagnosis and management rely upon a multidisciplinary and holistic approach. Treatment options include conservative measures and pharmacotherapies as well as bladder instillation therapies. Ultimately, surgery may be offered but only in cases of refractory disease. This article offers a pragmatic guide for clinicians managing this challenging disease.publishedVersio

Ensuring food safety with molecularly imprinted polymers: innovative methods for the detection of aflatoxins in food and feed samples

Aflatoxins, a group of mycotoxins, represent a heterogeneous class of secondary metabolites that pose a significant risk to food safety and public health due to their potent toxicity. Aflatoxins are widely distributed in the environment, with high levels frequently observed in hot and humid conditions. There is an ongoing development of various methods for detecting aflatoxins in food and feed samples. Herein, a review of these methods is presented with special emphasis on molecularly imprinted polymers (MIPs) as selective materials for aflatoxins’ detection. The key findings of various methods for real-time analysis of food and feed samples are presented and analyzed, providing a comparative assessment of their performance. Furthermore, the challenges and limitations of these methods are discussed, considering their commercialization prospects and real-world requirements