High content live cell imaging for the discovery of new antimalarial marine natural products

<p>Abstract</p> <p>Background</p> <p>The human malaria parasite remains a burden in developing nations. It is responsible for up to one million deaths a year, a number that could rise due to increasing multi-drug resistance to all antimalarial drugs currently available. Therefore, there is an urgent need for the discovery of new drug therapies. Recently, our laboratory developed a simple one-step fluorescence-based live cell-imaging assay to integrate the complex biology of the human malaria parasite into drug discovery. Here we used our newly developed live cell-imaging platform to discover novel marine natural products and their cellular phenotypic effects against the most lethal malaria parasite, <it>Plasmodium falciparum</it>.</p> <p>Methods</p> <p>A high content live cell imaging platform was used to screen marine extracts effects on malaria. Parasites were grown <it>in vitro </it>in the presence of extracts, stained with RNA sensitive dye, and imaged at timed intervals with the BD Pathway HT automated confocal microscope.</p> <p>Results</p> <p>Image analysis validated our new methodology at a larger scale level and revealed potential antimalarial activity of selected extracts with a minimal cytotoxic effect on host red blood cells. To further validate our assay, we investigated parasite's phenotypes when incubated with the purified bioactive natural product bromophycolide A. We show that bromophycolide A has a strong and specific morphological effect on parasites, similar to the ones observed from the initial extracts.</p> <p>Conclusion</p> <p>Collectively, our results show that high-content live cell-imaging (HCLCI) can be used to screen chemical libraries and identify parasite specific inhibitors with limited host cytotoxic effects. All together we provide new leads for the discovery of novel antimalarials.</p

Absolute Winding Number Differentiates Mouse Spatial Navigation Strategies With Genetic Risk for Alzheimer’s Disease

Spatial navigation and orientation are emerging as promising markers for altered cognition in prodromal Alzheimer’s disease, and even in cognitively normal individuals at risk for Alzheimer’s disease. The different APOE gene alleles confer various degrees of risk. The APOE2 allele is considered protective, APOE3 is seen as control, while APOE4 carriage is the major known genetic risk for Alzheimer’s disease. We have used mouse models carrying the three humanized APOE alleles and tested them in a spatial memory task in the Morris water maze. We introduce a new metric, the absolute winding number, to characterize the spatial search strategy, through the shape of the swim path. We show that this metric is robust to noise, and works for small group samples. Moreover, the absolute winding number better differentiated APOE3 carriers, through their straighter swim paths relative to both APOE2 and APOE4 genotypes. Finally, this novel metric supported increased vulnerability in APOE4 females. We hypothesized differences in spatial memory and navigation strategies are linked to differences in brain networks, and showed that different genotypes have different reliance on the hippocampal and caudate putamen circuits, pointing to a role for white matter connections. Moreover, differences were most pronounced in females. This departure from a hippocampal centric to a brain network approach may open avenues for identifying regions linked to increased risk for Alzheimer’s disease, before overt disease manifestation. Further exploration of novel biomarkers based on spatial navigation strategies may enlarge the windows of opportunity for interventions. The proposed framework will be significant in dissecting vulnerable circuits associated with cognitive changes in prodromal Alzheimer’s disease

The Golden Age of European Cabaret

Program for the second annual RISD Cabaret held in the Cellar in the Pit. Design and layout by Anne Johnson, Susan Sellers and Georgie Stout.https://digitalcommons.risd.edu/liberalarts_cabaret_programs/1001/thumbnail.jp

Bostonia: The Boston University Alumni Magazine. Volume 23

Founded in 1900, Bostonia magazine is Boston University's main alumni publication, which covers alumni and student life, as well as university activities, events, and programs

Seasonal and Regional Dynamics of M. ulcerans Transmission in Environmental Context: Deciphering the Role of Water Bugs as Hosts and Vectors

Buruli ulcer, caused by Mycobacterium ulcerans, is a devastating skin disease. Most cases of Buruli ulcer occur in poor rural communities. As a result, treatment is frequently sought too late and about 25% of those infected—particularly children—become permanently disabled. Outbreaks of Buruli ulcer have always been associated with swampy areas. However, the route(s) of bacillus transmission is (are) still unclear. This Mycobacterium species resides in water where it colonizes many ecological niches such as aquatic plants, herbivorous animals and predatory/carnivorous insects. For several years the role of water bugs as hosts and vectors of M. ulcerans was suspected and was demonstrated under laboratory conditions. The aim of this work was to further assess the role of water bugs as hosts and vectors of M. ulcerans in environmental context. This work identifies several water bug families as hosts of M. ulcerans in Buruli ulcer endemic area. The detection of bacilli in saliva of human biting insects provides further evidence for their role in M. ulcerans transmission. Interestingly, three of these insects are good flyers, and as such could participate in M. ulcerans dissemination

Archives, Memory, and Interfaces with the Past

Archival interfaces are criticalnodes in archival systems where archivistsnegotiate and exercise power over theconstitution and representation of archives. Drawing on notions of interfaces from physical,technological, and computer systems, archivalinterfaces are both a metaphor for archivists'roles as intermediaries between documentaryevidence and its readers and a tangible set ofstructures and tools that place archivaldocuments in a context and provide aninterpretative framework. Interfaces in moderninstitutions and technological systems areneither natural nor neutral. In probingarchival interfaces, what may appear as neutraland objective processes are revealed as placeswhere archivists determine what constituteslegitimate evidence of the past and shapesocial memories. The emergence of computerinterfaces as an increasingly common mode ofuser interaction with archives demands thatarchivists confront the interpretative natureof their work and exploit opportunities toplace themselves visibly in the interfaces theyconstruct.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41817/1/10502_2004_Article_5096450.pd

Widespread white matter microstructural abnormalities in bipolar disorder: Evidence from mega- and meta-analyses across 3,033 individuals

Fronto-limbic white matter (WM) abnormalities are assumed to lie at the heart of the pathophysiology of bipolar disorder (BD); however, diffusion tensor imaging (DTI) studies have reported heterogeneous results and it is not clear how the clinical heterogeneity is related to the observed differences. This study aimed to identify WM abnormalities that differentiate patients with BD from healthy controls (HC) in the largest DTI dataset of patients with BD to date, collected via the ENIGMA network. We gathered individual tensor-derived regional metrics from 26 cohorts leading to a sample size of N = 3033 (1482 BD and 1551 HC). Mean fractional anisotropy (FA) from 43 regions of interest (ROI) and average whole-brain FA were entered into univariate mega- and meta-analyses to differentiate patients with BD from HC. Mega-analysis revealed significantly lower FA in patients with BD compared with HC in 29 regions, with the highest effect sizes observed within the corpus callosum (R2 = 0.041, Pcorr < 0.001) and cingulum (right: R2 = 0.041, left: R2 = 0.040, Pcorr < 0.001). Lithium medication, later onset and short disease duration were related to higher FA along multiple ROIs. Results of the meta-analysis showed similar effects. We demonstrated widespread WM abnormalities in BD and highlighted that altered WM connectivity within the corpus callosum and the cingulum are strongly associated with BD. These brain abnormalities could represent a biomarker for use in the diagnosis of BD. Interactive three-dimensional visualization of the results is available at www.enigma-viewer.org

Widespread white matter microstructural abnormalities in bipolar disorder: evidence from mega- and meta-analyses across 3033 individuals

Fronto-limbic white matter (WM) abnormalities are assumed to lie at the heart of the pathophysiology of bipolar disorder (BD); however, diffusion tensor imaging (DTI) studies have reported heterogeneous results and it is not clear how the clinical heterogeneity is related to the observed differences. This study aimed to identify WM abnormalities that differentiate patients with BD from healthy controls (HC) in the largest DTI dataset of patients with BD to date, collected via the ENIGMA network. We gathered individual tensor-derived regional metrics from 26 cohorts leading to a sample size of N = 3033 (1482 BD and 1551 HC). Mean fractional anisotropy (FA) from 43 regions of interest (ROI) and average whole-brain FA were entered into univariate mega- and meta-analyses to differentiate patients with BD from HC. Mega-analysis revealed significantly lower FA in patients with BD compared with HC in 29 regions, with the highest effect sizes observed within the corpus callosum (R2 = 0.041, Pcorr < 0.001) and cingulum (right: R2 = 0.041, left: R2 = 0.040, Pcorr < 0.001). Lithium medication, later onset and short disease duration were related to higher FA along multiple ROIs. Results of the meta-analysis showed similar effects. We demonstrated widespread WM abnormalities in BD and highlighted that altered WM connectivity within the corpus callosum and the cingulum are strongly associated with BD. These brain abnormalities could represent a biomarker for use in the diagnosis of BD. Interactive three-dimensional visualization of the results is available at www.enigma-viewer.org

Spectroscopie et Imagerie RMN multi-noyaux à très haut champ magnétique

Bipolar disorder is a chronic affective disorder affecting 1 to 3% of the adult population worldwide and has a high level of comorbidity with suicide rates, substance abuse and other harmful conditions. The disorder has possible ties to schizophrenia and has been observed to have a strong genetic component. The exact biological underpinnings have not been firmly established, however abnormalities in limbic subcortical and prefrontal areas have been observed.Ever since its discovery more than half a century ago, a daily intake of Lithium salts has arguably become the most reliable treatment of the disorder, despite us possessing little to no understanding of its biochemical action. In order to shed some light on the effect of Lithium in the brain, we have developed Lithium-7 MR imaging at 7 and 17 Tesla in order to assess its cerebral concentration and distribution. Specifically, I worked on developing and validating several acquisition, reconstruction and quantification methods dedicated to 7Li MRI and MRS. Those methods were first applied to study ex vivo the cerebral distribution of lithium in rats. These rats were pretreated for 28 days with Li2CO3, sacrificed and their head fixated with PFA. Using a home-made 1H/7Li radiofrequency surface coil and a 7Li Turbo Spin echo acquisition and a modified phantom replacement method for quantification, we were able to measure Li concentration maps. Regional Li concentration values were then compared with those obtained with mass spectrometry.After this preclinical proof-of-concept study, an in vivo 7Li MRI protocol was designed to map the cerebral Li concentration in euthymic bipolar subjects at 7T. These individuals all followed a regular lithium treatment. For this study, we chose to use an ultra-short echo-time Steady State Free Precession sequence with non-Cartesian k-space sampling. A quantification and analysis pipeline similar to the one used for our preclinical study was applied for this study, with the addition of a correction step for B0 inhomogeneities. After conducting a statistical analysis at the cohort level, it was assessed that the left hippocampus, a major part of the limbic system that has been associated with BD on multiple occasions, exhibited systematically a high level of lithium. Finally, I developed a quantification method accounting for the different relaxation times of 7Li in the CSF and in the brain parenchyma. This method was applied to image lithium at 7T in a subset of bipolar patients reducing drastically the differences initially observed between the SSFP and bSSFP sequences.Le trouble bipolaire est un trouble de l'humeur récurrent affectant de 1 à 3% de la population adulte à travers le monde et ayant une comorbidité importante avec une hausse du taux de suicides, l'abus de drogues et d'autres troubles médicaux. Ce trouble semble avoir plusieurs liens avec la schizophrénie et une vulnérabilité au trouble est souvent héréditaire dans une famille. Même si les causes biologiques n'ont pas encore été établies, de nombreuses anomalies dans le système limbique sous-cortical et la zone préfrontal ont été observés.Depuis sa découverte il y a plus d'un demi-siècle, une prise de sels de Lithium a été le traitement le plus fiable, mais l'action biochimique du Lithium sur le cerveau et le pourquoi de l'efficacité du traitement reste un mystère. Afin de pouvoir mieux comprendre cet effet, nous avons développé des séquences d'imagerie du Lithium-7 via résonance magnétique à 7 et 17 Tesla afin de pouvoir établir sa distribution et concentration cérébrale. Spécifiquement, j'ai travaillé sur le développement et la validation des méthodes d'acquisition, de reconstruction et de quantification de notre protocole. Ces méthodes ont d'abord été appliqués afin d'étudier la distribution cérébrale du Lithium sur des cerveaux de rats ex-vivo. Ces rats étés traités pendant 28 jours avec du Li2CO3, sacrifiés et leurs têtes fixés avec du PFA. En utilisant une antenne surfacique double canaux 1H/7Li fait maison, une acquisition 7Li Turbo Spin echo et une méthode de remplacement par fantôme pour la quantification, nous avons pu mesurer les cartes de concentration du Lithium. Les concentrations moyennes obtenus dans des régions d’intérêt prédéfinis ont été mesurés afin de les comparer avec les résultats obtenus par spectrométrie de masse.Après cette étude préclinique qui a permis de valider notre approche, un protocole similaire fut créé pour une étude in-vivo 7Li d'imagerie par résonance magnétique chez des patients bipolaires euthymiques sur un scanner 7T. Ces individus ont tous suivis un traitement régulier de Lithium. Pour cette étude, nous avons utilisé une séquence Steady State Free Precession à TE ultra-court et avec un échantillonnage du k-space non-cartésien. Un pipeline de quantification et d'analyse similaire à celle utilisé pour notre étude préclinique fut appliqué pour cette étude, avec l'ajout d'une étape de correction pour les inhomogénéités de B0. Après avoir fait une analyse statistique au niveau de toute la cohorte par régions d'intérêt, il a été observé que l'hippocampe gauche, une part majeur du système limbique associé au trouble bipolaire à de multiples occasions, possède systématiquement une haute concentration de Lithium. Finalement, la méthode de quantification fut modifiée en quantification bi-compartimentale afin de prendre en compte les différences dans les taux de relaxation du Lithium dans le CSF et dans le parenchyme du cerveau. Cette méthode fut appliqué afin de pouvoir quantifier le Lithium à 7T dans un sous-groupe des patients bipolaires et radicalement réduire les différences initialement observés entre les séquences SSFP et bSSFP