Systems-level reconstruction of kinase phosphosignaling networks regulating endothelial barrier integrity using temporal data.

Phosphosignaling networks control cellular processes. We built kinase-mediated regulatory networks elicited by thrombin stimulation of brain endothelial cells using two computational strategies: Temporal Pathway Synthesizer (TPS), which uses phosphoproteomics data as input, and Temporally REsolved KInase Network Generation (TREKING), which uses kinase inhibitor screens. TPS and TREKING predicted overlapping barrier-regulatory kinases connected with unique network topology.

Triazenolysis of alkenes as an aza version of ozonolysis.

Alkenes are broadly used in synthetic applications, thanks to their abundance and versatility. Ozonolysis is one of the most canonical transformations that converts alkenes into molecules bearing carbon-oxygen motifs via C=C bond cleavage. Despite its extensive use in both industrial and laboratory settings, the aza version-cleavage of alkenes to form carbon-nitrogen bonds-remains elusive.

Using machine learning to dissect host kinases required for Leishmania internalization and development.

The Leishmania life cycle alternates between promastigotes, found in the sandfly, and amastigotes, found in mammals. When an infected sandfly bites a host, promastigotes are engulfed by phagocytes (i.e.

Systems-level reconstruction of kinase phosphosignaling networks regulating endothelial barrier integrity using temporal data.

Phosphosignaling networks control cellular processes. We built kinase-mediated regulatory networks elicited by thrombin stimulation of brain endothelial cells using two computational strategies: Temporal Pathway Synthesizer (TPS), which uses phosphoproetiomics data as input, and Temporally REsolved KInase Network Generation (TREKING), which uses kinase inhibitor screens. TPS and TREKING predicted overlapping barrier-regulatory kinases connected with unique network topology.

Using machine learning to dissect host kinases required for internalization and development.

The life cycle alternates between promastigotes, found in the sandfly, and amastigotes, found in mammals. When an infected sandfly bites a host, promastigotes are engulfed by phagocytes (., neutrophils, dendritic cells, and macrophages) to establish infection.

Exploring Subsite Selectivity within -Myristoyltransferase Using Pyrazole-Derived Inhibitors.

-myristoyltransferase (NMT) is a promising antimalarial drug target. Despite biochemical similarities between and human NMTs, our recent research demonstrated that high selectivity is achievable. Herein, we report NMT-inhibiting compounds aimed at identifying novel mechanisms of selectivity.

Multiple receptor tyrosine kinases regulate dengue infection of hepatocytes.

Introduction: Dengue is an arboviral disease causing severe illness in over 500,000 people each year. Currently, there is no way to constrain dengue in the clinic. Host kinase regulators of dengue virus (DENV) infection have the potential to be disrupted by existing therapeutics to prevent infection and/or disease progression.

Interrogating endothelial barrier regulation by temporally resolved kinase network generation.

Kinases are key players in endothelial barrier regulation, yet their temporal function and regulatory phosphosignaling networks are incompletely understood. We developed a novel methodology, Temporally REsolved KInase Network Generation (TREKING), which combines a 28-kinase inhibitor screen with machine learning and network reconstruction to build time-resolved, functional phosphosignaling networks. We demonstrated the utility of TREKING for identifying pathways mediating barrier integrity after activation by thrombin with or without TNF preconditioning in brain endothelial cells.

Host kinase regulation of dormant and replicating liver stages.

Upon transmission to the liver, parasites form replicating schizonts, which continue to initiate blood-stage infection, or dormant hypnozoites that reactivate weeks to months after initial infection. phenotypes in the field vary significantly, including the ratio of schizonts to hypnozoites formed and the frequency and timing of relapse. Evidence suggests that both parasite genetics and environmental factors underly this heterogeneity.

Protocol for assembling and implementing a partially automated system for rearing and handling Anopheles stephensi mosquitoes.

Live mosquitoes are required to comprehensively study vector-borne diseases, including transmission. Traditional mosquito-rearing protocols are laborious and time consuming. Here, we present a protocol for assembling and implementing a partially automated system for rearing and handling Anopheles stephensi mosquitoes.

Identification of potent and selective N-myristoyltransferase inhibitors of Plasmodium vivax liver stage hypnozoites and schizonts.

Drugs targeting multiple stages of the Plasmodium vivax life cycle are needed to reduce the health and economic burdens caused by malaria worldwide. N-myristoyltransferase (NMT) is an essential eukaryotic enzyme and a validated drug target for combating malaria. However, previous PvNMT inhibitors have failed due to their low selectivity over human NMTs.

Liver-stage Plasmodium infection tunes clinical outcomes.

Chora and colleagues show that infection of the liver by Plasmodium modulates severity of disease in the experimental cerebral malaria (ECM) model by generating gamma delta (ɣδ) T cells that produce IL-17. This work calls into question the long-standing assumption that liver infection does not modulate severity of malaria.

Facile Synthesis of an Octagermacubane with Two Three-Coordinate Germanium(0) Atoms and its Unique Radical Anion.

Thermolysis of a 1 : 1 mixture of tris(di-tert-butylmethylsilyl)germane 9 and bis(di-tert-butylmethylsilyl)germane 17 at 100 °C produces unexpectedly octagermacubane 18, having two 3-coordinate Ge atoms (40 % yield). 18 was characterized by X-ray crystallography and it is a singlet biradical (according to DFT quantum mechanical calculations and the absence of an EPR signal). Reactions of 18 with CH Cl and H O yield the novel dichloro-octagermacubane 24 and hydroxy-octagermacubane 25, respectively.

Isoindolinium Groups as Stable Anion Conductors for Anion-Exchange Membrane Fuel Cells and Electrolyzers.

Anion-exchange membrane (AEM) fuel cells (AEMFCs) and water electrolyzers (AEMWEs) have gained strong attention of the scientific community as an alternative to expensive mainstream fuel cell and electrolysis technologies. However, in the high pH environment of the AEMFCs and AEMWEs, especially at low hydration levels, the molecular structure of most anion-conducting polymers breaks down because of the strong reactivity of the hydroxide anions with the quaternary ammonium (QA) cation functional groups that are commonly used in the AEMs and ionomers. Therefore, new highly stable QAs are needed to withstand the strong alkaline environment of these electrochemical devices.

Multimodal Reactivity of N-H Bonds in Triazanes and Isolation of a Triazinyl Radical.

The employment of nitrogen Lewis acids based on nitrenium cations has been increasingly featured in the fields of main group chemistry and catalysis. A formally reduced form of nitrenium ─cyclic triazanes ─are intriguing chemical compounds, the chemistry of which is completely unexplored. In this work, we reveal that N-H-triazanes exhibit unusual N-H bond properties; namely, they can serve as protons, hydrides, or hydrogen atom donors.

Viral protein engagement of GBF1 induces host cell vulnerability through synthetic lethality.

Viruses co-opt host proteins to carry out their lifecycle. Repurposed host proteins may thus become functionally compromised; a situation analogous to a loss-of-function mutation. We term such host proteins as viral-induced hypomorphs.

Hydroxide Chemoselectivity Changes with Water Microsolvation.

Solvent molecules are known to affect chemical reactions, especially if they interact with one or more of the reactants or catalysts. In ion microsolvation, i.e.

Dengue activates mTORC2 signaling to counteract apoptosis and maximize viral replication.

The mechanistic target of rapamycin (mTOR) functions in two distinct complexes: mTORC1, and mTORC2. mTORC1 has been implicated in the pathogenesis of flaviviruses including dengue, where it contributes to the establishment of a pro-viral autophagic state. Activation of mTORC2 occurs upon infection with some viruses, but its functional role in viral pathogenesis remains poorly understood.

Germinal center activity and B cell maturation are associated with protective antibody responses against Plasmodium pre-erythrocytic infection.

Blocking Plasmodium, the causative agent of malaria, at the asymptomatic pre-erythrocytic stage would abrogate disease pathology and prevent transmission. However, the lack of well-defined features within vaccine-elicited antibody responses that correlate with protection represents a major roadblock to improving on current generation vaccines. We vaccinated mice (BALB/cJ and C57BL/6J) with Py circumsporozoite protein (CSP), the major surface antigen on the sporozoite, and evaluated vaccine-elicited humoral immunity and identified immunological factors associated with protection after mosquito bite challenge.

A genome-wide CRISPR-Cas9 screen identifies CENPJ as a host regulator of altered microtubule organization during Plasmodium liver infection.

Prior to initiating symptomatic malaria, a single Plasmodium sporozoite infects a hepatocyte and develops into thousands of merozoites, in part by scavenging host resources, likely delivered by vesicles. Here, we demonstrate that host microtubules (MTs) dynamically reorganize around the developing liver stage (LS) parasite to facilitate vesicular transport to the parasite. Using a genome-wide CRISPR-Cas9 screen, we identified host regulators of cytoskeleton organization, vesicle trafficking, and ER/Golgi stress that regulate LS development.

Mechanistic Insights on the Selectivity of the Tandem Heck-Ring-Opening of Cyclopropyldiol Derivatives.

The preparation of a new class of alkenyl cyclopropyl diols, easily available through a copper-catalyzed carbometalation reaction of cyclopropenes, has enabled the study of key mechanistic aspects of the tandem Heck-cyclopropane ring-opening reaction. Utilizing these substrates containing two distinct hydroxyl groups allowed us to examine parameters affecting the reaction outcome and selectivity. The combination of these experimental results with detailed DFT studies shed light on the mechanism governing the regio- and stereoselectivity of the cyclopropane ring-opening.

Elucidating Spatially-Resolved Changes in Host Signaling During Liver-Stage Infection.

Upon transmission to the human host, sporozoites exit the skin, are taken up by the blood stream, and then travel to the liver where they infect and significantly modify a single hepatocyte. Low infection rates within the liver have made proteomic studies of infected hepatocytes challenging, particularly , and existing studies have been largely unable to consider how protein and phosphoprotein differences are altered at different spatial locations within the heterogeneous liver. Using digital spatial profiling, we characterized changes in host signaling during infection without disrupting the liver tissue.

Cyclosporine Induces Fenestra-Associated Injury in Human Renal Microvessels .

The use of cyclosporine A (CsA) in transplantation is frequently associated with nephrotoxicity, characterized by renal vascular injury, thrombotic microangiopathy, and striped interstitial fibrosis. Here, using human kidney-specific microvascular endothelial cells (HKMECs), we showed that CsA inhibited NFAT1 activation and impaired VEGF signaling in these ECs in a dose- and time-dependent manner. Integrated genome regulatory analyses identified key distinctions in the landscapes of HKMECs compared to human umbilical vein endothelial cells, particularly around genes related to the formation and maintenance of fenestrae.

Host-directed therapy, an untapped opportunity for antimalarial intervention.

Host-directed therapy (HDT) is gaining traction as a strategy to combat infectious diseases caused by viruses and intracellular bacteria, but its implementation in the context of parasitic diseases has received less attention. Here, we provide a brief overview of this field and advocate HDT as a promising strategy for antimalarial intervention based on untapped targets. HDT provides a basis from which repurposed drugs could be rapidly deployed and is likely to strongly limit the emergence of resistance.