CHOPIN: a web resource for the structural and functional proteome of Mycobacterium tuberculosis.

Tuberculosis kills more than a million people annually and presents increasingly high levels of resistance against current first line drugs. Structural information about Mycobacterium tuberculosis (Mtb) proteins is a valuable asset for the development of novel drugs and for understanding the biology of the bacterium; however, only about 10% of the ∼4000 proteins have had their structures determined experimentally. The CHOPIN database assigns structural domains and generates homology models for 2911 sequences, corresponding to ∼73% of the proteome. A sophisticated pipeline allows multiple models to be created using conformational states characteristic of different oligomeric states and ligand binding, such that the models reflect various functional states of the proteins. Additionally, CHOPIN includes structural analyses of mutations potentially associated with drug resistance. Results are made available at the web interface, which also serves as an automatically updated repository of all published Mtb experimental structures. Its RESTful interface allows direct and flexible access to structures and metadata via intuitive URLs, enabling easy programmatic use of the models.This work was supported by the Bill & Melinda Gates Foundation (RG60453). University of Cambridge for facilities and support [to TLB]. Funding for open access charge: Bill & Melinda Gates Foundation.This is the final published version. It first appeared at http://database.oxfordjournals.org/content/2015/bav026.long

Hyaloid Vasculature and mmp2 Activity Play a Role during Optic Fissure Fusion in Zebrafish

Vertebrate retinal development requires timely and precise fusion of the optic fissure (OF). Failure of this event leads to congenital vision impairment in the form of coloboma. Recent studies have suggested hyaloid vasculature to be involved in OF fusion. In order to examine this link, we analyzed OF fusion and hyaloid vasculogenesis in the zebrafish pax2a noi mutant line. We first determined that pax2a−/− embryos fail to accumulate F-actin in the OF prior to basement membrane (BM) degradation. Furthermore, using 3D and live imaging we observed reduced OF hyaloid vascularization in pax2a−/− embryos. When examining the connection between pax2a loss of function and hyaloid vasculature, we observed significant reduction of talin1 expression, a regulator of hyaloid vasculature. In addition, cranial VEGF expression was found to be reduced in pax2a−/− embryos. Pharmacological inhibition of VEGF signaling phenocopied the pax2a−/− vasculature, F-actin and BM degradation phenotypes. Lastly, we determined that OF associated hyaloid vasculature is a source of mmp2, mmp14a and mmp14b expression and showed that mmp2 is functionally necessary for degradation of OF BM. Taken together we propose a pax2a driven mechanism that ensures proper and timely hyaloid vasculature invasion of the OF in order to facilitate availability of the BM remodeler mmp2

Spatiotemporal Characterization of Anterior Segment Mesenchyme Heterogeneity during Zebrafish Ocular Anterior Segment Development

Assembly of the ocular anterior segment (AS) is a critical event during development of the vertebrate visual system. Failure in this process leads to anterior segment dysgenesis (ASD), which is characterized by congenital blindness and predisposition to glaucoma. The anterior segment is largely formed via a neural crest-derived population, the Periocular Mesenchyme (POM). In this study, we aimed to characterize POM behaviors and transcriptional identities during early establishment of the zebrafish AS. Two-color fluorescent in situ hybridization suggested that early AS associated POM comprise of a heterogenous population. In vivo and time-course imaging analysis of POM distribution and migratory dynamics analyzed using transgenic zebrafish embryos (Tg[foxc1b:GFP], Tg[foxd3:GFP], Tg[pitx2:GFP], Tg[lmx1b.1:GFP], and Tg[sox10:GFP]) revealed unique AS distribution and migratory behavior among the reporter lines. Based on fixed timepoint and real-time analysis of POM cell behavior a comprehensive model for colonization of the zebrafish AS was assembled. Furthermore, we generated single cell transcriptomic profiles (scRNA) from our POM reporter lines and characterized unique subpopulation expression patterns. Based on scRNA clustering analysis we observed cluster overlap between neural crest associated (sox10/foxd3), POM (pitx2) and finally AS specified cells (lmx1b, and foxc1b). scRNA clustering also revealed several novel markers potentially associated with AS development and/or function including lum, fmoda, adcyap1b, tgfbi, and hmng2. Taken together, our data indicates that AS-associated POM, or Anterior Segment Mesenchyme (ASM), is not homogeneous but rather comprised of several subpopulations with differing colonization patterns, migration behavior, and transcriptomic profiles

A real-time ppg peak detection method for accurate determination of heart rate during sinus rhythm and cardiac arrhythmia

Objective: We have developed a peak detection algorithm for accurate determination of heart rate, using photoplethysmographic (PPG) signals from a smartwatch, even in the presence of various cardiac rhythms, including normal sinus rhythm (NSR), premature atrial contraction (PAC), premature ventricle contraction (PVC), and atrial fibrillation (AF). Given the clinical need for accurate heart rate estimation in patients with AF, we developed a novel approach that reduces heart rate estimation errors when compared to peak detection algorithms designed for NSR. Methods: Our peak detection method is composed of a sequential series of algorithms that are combined to discriminate the various arrhythmias described above. Moreover, a novel Poincaré plot scheme is used to discriminate between basal heart rate AF and rapid ventricular response (RVR) AF, and to differentiate PAC/PVC from NSR and AF. Training of the algorithm was performed only with Samsung Simband smartwatch data, whereas independent testing data which had more samples than did the training data were obtained from Samsung’s Gear S3 and Galaxy Watch 3. Results: The new PPG peak detection algorithm provides significantly lower average heart rate and interbeat interval beat-to-beat estimation errors—30% and 66% lower—and mean heart rate and mean interbeat interval estimation errors—60% and 77% lower—when compared to the best of the seven other traditional peak detection algorithms that are known to be accurate for NSR. Our new PPG peak detection algorithm was the overall best performers for other arrhythmias. Conclusion: The proposed method for PPG peak detection automatically detects and discriminates between various arrhythmias among different waveforms of PPG data, delivers significantly lower heart rate estimation errors for participants with AF, and reduces the number of false negative peaks. Significance: By enabling accurate determination of heart rate despite the presence of AF with rapid ventricular response or PAC/PVCs, we enable clinicians to make more accurate recommendations for heart rate control from PPG data

The effect of pregnancy and stress on the onset of placentophagia in Long-Evans rats

Most virgin rats do not eat placenta when it is presented to them; virtually all parturient rats do. This study was designed to examine the role of the duration and termination of pregnancy on the induction of placentophagia. Time-bred rats determined by a pretest not to be attracted to placenta (nonplacentophages), were tested for placentophagia on one of a number of days of pregnancy. A comparable group was tested for placentophagia after surgical termination of pregnancy on Day 21. Groups of virgins were run controlling for the time interval between the pretest and the placentophagia test, and for a time interval that included a "stressful" event. The results were that (a) the incidence of placentophagia rose gradually from Day 7 to Day 15 of pregnancy, then remained stable, at about 0.4 until delivery; (b) pregnancy termination did not produce an effect on placentophagia greater than that of unterminated pregnancy; (c) a pretest-test interval containing a "stressful" event produced significantly more placentophagia than one that did not contain such an event; and (d) the maximum level of placentophagia observed during pregnancy was the same as that produced in virgins by a "stressful" event

Revealing the molecular signatures of host-pathogen interactions.

Advances in sequencing technology and genome-wide association studies are now revealing the complex interactions between hosts and pathogen through genomic variation signatures, which arise from evolutionary co-existence

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2

<p>Abstract</p> <p>Background</p> <p>Wnt signaling is mediated through 1) the beta-catenin dependent canonical pathway and, 2) the beta-catenin independent pathways. Multiple receptors, including Fzds, Lrps, Ror2 and Ryk, are involved in Wnt signaling. Ror2 is a single-span transmembrane receptor-tyrosine kinase (RTK). The functions of Ror2 in mediating the non-canonical Wnt signaling have been well established. The role of Ror2 in canonical Wnt signaling is not fully understood.</p> <p>Results</p> <p>Here we report that Ror2 also positively modulates Wnt3a-activated canonical signaling in a lung carcinoma, H441 cell line. This activity of Ror2 is dependent on cooperative interactions with Fzd2 but not Fzd7. In addition, Ror2-mediated enhancement of canonical signaling requires the extracellular CRD, but not the intracellular PRD domain of Ror2. We further provide evidence that the positive effect of Ror2 on canonical Wnt signaling is inhibited by Dkk1 and Krm1 suggesting that Ror2 enhances an Lrp-dependent STF response.</p> <p>Conclusion</p> <p>The current study demonstrates the function of Ror2 in modulating canonical Wnt signaling. These findings support a functional scheme whereby regulation of Wnt signaling is achieved by cooperative functions of multiple mediators.</p

Proceedings from the 2018 Canadian Association for the Study of the Liver Single Topic Conference—Decompensated Cirrhosis : from clinic to transplant

Actes publiés dans le Canadian Liver Journal; vol. 2, no. 4, Fall 2019, p. 137-170

Transcriptional Control in Cardiac Progenitors: Tbx1 Interacts with the BAF Chromatin Remodeling Complex and Regulates Wnt5a

Mutations of the Wnt5a gene, encoding a ligand of the non-canonical Wnt pathway, and the Ror2 gene, encoding its receptor, have been found in patients with cardiac outflow tract defects. We found that Wnt5a is expressed in the second heart field (SHF), a population of cardiac progenitor cells destined to populate the cardiac outflow tract and the right ventricle. Because of cardiac phenotype similarities between Wnt5a and Tbx1 mutant mice, we tested potential interactions between the two genes. We found a strong genetic interaction in vivo and determined that the loss of both genes caused severe hypoplasia of SHF–dependent segments of the heart. We demonstrated that Wnt5a is a transcriptional target of Tbx1 and explored the mechanisms of gene regulation. Tbx1 occupies T-box binding elements within the Wnt5a gene and interacts with the Baf60a/Smarcd1 subunit of a chromatin remodeling complex. It also interacts with the Setd7 histone H3K4 monomethyltransferase. Tbx1 enhances Baf60a occupation at the Wnt5a gene and enhances its H3K4 monomethylation status. Finally, we show that Baf60a is required for Tbx1–driven regulation of target genes. These data suggest a model in which Tbx1 interacts with, and probably recruits a specific subunit of, the BAF complex as well as histone methylases to activate or enhance transcription. We speculate that this may be a general mechanism of T-box function and that Baf60a is a key component of the transcriptional control in cardiac progenitors