Complete genome sequence of Torque teno indri virus 1, a novel anellovirus in blood from a free-living lemur

ABSTRACT We identified Torque teno indri virus 1 (TTIV1), the first anellovirus in a free-living lemur ( Indri indri ). The complete circular 2,572-nucleotide (nt) TTIV1 genome is distantly related to torque teno sus virus. Phylogenetic and sequence analyses support TTIV1 as a putative member of a new genus within the Anelloviridae family. </jats:p

Nanoprodrugs of NSAIDs: Preparation and Characterization of Flufenamic Acid Nanoprodrugs

We demonstrated that hydrophobic derivatives of the nonsteroidal anti-inflammatory drug (NSAID)flufenamic acid (FA), can be formed into stable nanometer-sized prodrugs (nanoprodrugs) that inhibit the growth of glioma cells, suggesting their potential application as anticancer agent. We synthesized highly hydrophobic monomeric and dimeric prodrugs of FA via esterification and prepared nanoprodrugs using spontaneous emulsification mechanism. The nanoprodrugs were in the size range of 120 to 140 nm and physicochemically stable upon long-term storage as aqueous suspension, which is attributed to the strong hydrophobic interaction between prodrug molecules. Importantly, despite the highly hydrophobic nature and water insolubility, nanoprodrugs could be readily activated into the parent drug by porcine liver esterase, presenting a potential new strategy for novel NSAID prodrug design. The nanoprodrug inhibited the growth of U87-MG glioma cells with IC50 of 20 μM, whereas FA showed IC50 of 100 μM, suggesting that more efficient drug delivery was achieved with nanoprodrugs

The linear ubiquitin assembly complex (LUBAC) is essential for NLRP3 inflammasome activation

Independent of its known role in NF-κB transcription, the HOIL-1L containing LUBAC is required for assembly and activation of the NLRP3 inflammasome via linear ubiquitination of ASC.Linear ubiquitination is a newly discovered posttranslational modification that is currently restricted to a small number of known protein substrates. The linear ubiquitination assembly complex (LUBAC), consisting of HOIL-1L, HOIP, and Sharpin, has been reported to activate NF-κB–mediated transcription in response to receptor signaling by ligating linear ubiquitin chains to Nemo and Rip1. Despite recent advances, the detailed roles of LUBAC in immune cells remain elusive. We demonstrate a novel HOIL-1L function as an essential regulator of the activation of the NLRP3/ASC inflammasome in primary bone marrow–derived macrophages (BMDMs) independently of NF-κB activation. Mechanistically, HOIL-1L is required for assembly of the NLRP3/ASC inflammasome and the linear ubiquitination of ASC, which we identify as a novel LUBAC substrate. Consequently, we find that HOIL-1L−/− mice have reduced IL-1β secretion in response to in vivo NLRP3 stimulation and survive lethal challenge with LPS. Together, these data demonstrate that linear ubiquitination is required for NLRP3 inflammasome activation, defining the molecular events of NLRP3 inflammasome activation and expanding the role of LUBAC as an innate immune regulator. Furthermore, our observation is clinically relevant because patients lacking HOIL-1L expression suffer from pyogenic bacterial immunodeficiency, providing a potential new therapeutic target for enhancing inflammation in immunodeficient patients

Afferent Cortical Connectivity to Rodent Posterior Parietal Cortex Along the Mediolateral Axis

The posterior parietal cortex (PPC), which can be found in rodents and primates, is thought to be a multimodal convergence area that is interconnected with important visual, auditory, somatosensory, motor and vestibular regions. The PPC is responsible for complex processes such as forming intentions, directing attention, spatial processing and sensorimotor integration. These processes all contribute to decisionmaking. Though rodents would serve as a good model organism for studying the PPC, the anatomy and connectivity of this area are poorly characterized in rats, especially regarding corticocortical projections to the PPC. This thesis analyzes differences in cortical projections to the PPC using fluorescent retrograde Cholera Toxin B subunit (CTB) neuronal tracers to find patterns of connectivity and to define the border between dysgranular retrosplenial cortex (RSD) and medial PPC (mPPC) as well as between mPPC and lateral PPC (lPPC). The borders we identify are consistent with the Paxinos and Watson atlas delineations that were made previously on the basis of cytoarchitecture. We provide connectivity-based evidence to support the existence of at least two subregions. Finally, we examine frontal orienting field (FOF) projections to the PPC in order to test the reciprocity of connectivity between these two regions. Our results are compared to the afferent connectivity of primate PPC. Finally, we suggest a series of future experiments that may elucidate the functional connectivity of the PPC using the results of this anatomical study. Characterizing the subdivisions and connectivity of the rodent PPC will hopefully permit more accurately targeted future experiments that may give insight into the characteristics of this important component of decision-making

Reactive Oxygen Species Responsive Nanoprodrug to Treat Intracranial Glioblastoma

Chemotherapy for intracranial gliomas is hampered by limited delivery of therapeutic agents through the blood brain barrier (BBB). An optimal therapeutic agent for brain tumors would selectively cross the BBB, accumulates in the tumor tissue and be activated from an innocuous prodrug within the tumor. Here we show brain tumor-targeted delivery and therapeutic efficacy of a nanometer-sized prodrug (nanoprodrug) of camptothecin (CPT) to treat experimental glioblastoma multiforme (GBM). The CPT nanoprodrug was prepared using spontaneous nanoemulsification of a biodegradable, antioxidant CPT prodrug and α-tocopherol. The oxidized nanoprodrug was activated more efficiently than nonoxidized nanoprodrug, suggesting enhanced therapeutic efficacy in the oxidative tumor microenvironment. The <i>in vitro</i> imaging of U-87 MG glioma cells revealed an efficient intracellular uptake of the nanoprodrug <i>via</i> direct cell membrane penetration rather than <i>via</i> endocytosis. The <i>in vivo</i> study in mice demonstrated that the CPT nanoprodrug passed through the BBB and specifically accumulated in brain tumor tissue, but not in healthy brain tissue and other organs. The accumulation preferably occurred at the periphery of the tumor where cancer cells are most actively proliferating, suggesting optimal therapeutic efficacy of the nanoprodrug. The nanoprodrug was effective in treating subcutaneous and intracranial tumors. The nanoprodrug inhibited subcutaneous tumor growth more than 80% compared with control. The median survival time of mice implanted with an intracranial tumor increased from 40.5 days for control to 72.5 days for CPT nanoprodrug. This nanoprodrug approach is a versatile method for developing therapeutic nanoparticles enabling tumor-specific targeting and treatment. The nontoxic, tumor-specific targeting properties of the nanoprodrug system make it a safe, low cost, and versatile nanocarrier for pharmaceuticals, imaging agents, and diagnostic agents

The linear ubiquitin assembly complex (LUBAC) is essential for NLRP3 inflammasome activation

Linear ubiquitination is a newly discovered posttranslational modification that is currently restricted to a small number of known protein substrates. The linear ubiquitination assembly complex (LUBAC), consisting of HOIL-1L, HOIP, and Sharpin, has been reported to activate NF-κB–mediated transcription in response to receptor signaling by ligating linear ubiquitin chains to Nemo and Rip1. Despite recent advances, the detailed roles of LUBAC in immune cells remain elusive. We demonstrate a novel HOIL-1L function as an essential regulator of the activation of the NLRP3/ASC inflammasome in primary bone marrow–derived macrophages (BMDMs) independently of NF-κB activation. Mechanistically, HOIL-1L is required for assembly of the NLRP3/ASC inflammasome and the linear ubiquitination of ASC, which we identify as a novel LUBAC substrate. Consequently, we find that HOIL-1L(−/−) mice have reduced IL-1β secretion in response to in vivo NLRP3 stimulation and survive lethal challenge with LPS. Together, these data demonstrate that linear ubiquitination is required for NLRP3 inflammasome activation, defining the molecular events of NLRP3 inflammasome activation and expanding the role of LUBAC as an innate immune regulator. Furthermore, our observation is clinically relevant because patients lacking HOIL-1L expression suffer from pyogenic bacterial immunodeficiency, providing a potential new therapeutic target for enhancing inflammation in immunodeficient patients