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4,7-Bis-(4′-cyano-biphenyl-4-yl)-[1, 10]phenanthroline (cnbiphphen) is synthesized from a palladium mediated coupling between 4-cyanobenzene boronic acid and 4,7-bis(4-bromophenyl)-[1,10]phenanthroline. The single crystal X-ray structure was measured at both 298(2) K and 90(2) K. The asymmetric unit of the title compound in the low-temperature phase, C38H22N 4 . HCCl3, is comprised of one cnbiphphen molecule and one chloroform solvent molecule. Additional symmetry relates one-half of the title compound to the other at room temperature. During cooling the structure experiences a reversible phase transition from A 2/a (different setting of C 2/c, with a and c exchanged to match lattice parameters of both phases) to P 21/c. The crystal packing of the title compound consists primarily of electronic charge interactions between the cyano moieties. The chloroform solvent mol-ecule is situated in a cavity between two phenanthroline systems and is associated to two nitrogen atoms of cnbiphphen molecules through Cl3C–H⋯Nphen van der Waals interaction. The synthesis, structure, analytical characterization, optical properties and phase transition of 4,7-bis-(4’-cyano-biphenyl-4-yl)-[1,10]phenanthroline is presented.

Relaxation calorimetry experiments were used to measure the temperature dependence of the heat capacity near the order–disorder phase transition (ODT) in a low molecular weight volumetrically symmetric, lamellae-forming, poly(1,4-isoprene-b-dl-lactide) diblock copolymer with Mn = 2750 g/mol (dispersity Đ = 1.10) and composition fPLA = 0.51. This enabled accurate determination of the latent heat of the ODT, yielding a value ΔHODT = 0.26 ± 0.02 J/g, consistent with previous measurements using differential scanning calorimetry. The relatively small magnitude of the latent heat supports the results of recent molecular simulations and reinforces the importance of composition fluctuations on the thermodynamics of block polymers near the ODT. These thermal measurements reveal no signature of the fluctuation effects in the disordered state as T → TODT, consistent with previous experimental and theoretical work, which supports the notion that composition fluctuations develop over a relatively wide range of tempera...

We report the synthesis, X-ray structure, absorbance and emission of 4,7-bis(4-fluorophenyl)-1,10-phenanthroline and 4,7-bis-(4′-trifluoromethyl-biphenyl-4-yl)-1,10-phenanthroline. 4,7-Bis(4-fluorophenyl)-1,10-phenanthroline was synthesized by successive Skraup reactions and features absorbance at 274 nm and emission at 384 nm while 4,7-bis-(4′-trifluoromethyl-biphenyl-4-yl)-1,10-phenanthroline featured absorbance at 283 nm and emission at 400 nm. The structures show molecular stacking of the phenyl groups and phenanthroline structure and longitudinal fluorine atom associations within the crystal lattice. Furthermore, the three fused rings of the 4,7-bis(4-fluorophenyl)-1,10-phenanthroline unit exhibited torsion up to 13.97(9)° throughout the crystal lattice whereas no torsion is observed for 4,7-bis-(4′-trifluoromethyl-biphenyl-4-yl)-1,10-phenanthroline.

The importance of understanding the genetic and biochemical basis of B cell receptor (BCR) survival signaling in diffuse large B cell lymphoma (DLBCL) is underscored by the recent clinical success of agents that target the BCR pathway. DLBCL is composed of multiple distinct molecular subtypes with divergent clinical outcomes. The activated B cell-like (ABC) subtype is the most aggressive form of DLBCL and is often resistant to standard chemotherapies. ABC DLBCL expresses numerous genes found in antigen-activated B cells, and genetic and pharmacologic studies have demonstrated that ABC DLBCL tumors are addicted to NF-κB activity. The origins of this NF-κB activity remained obscure until RNA interference screens established that the majority of ABC DLBCL cell lines rely on expression of BCR components and downstream signaling effectors for NF-κB activation. Pharmacological inhibition with ibrutinib of Bruton’s tyrosine kinase (Btk), a kinase that is required for BCR signaling to engage NF-κB, is selectively toxic for ABC DLBCL tumors; a finding that has now been translated to the clinic. These novel targets not only offer a promising new therapy options for ABC DLBCL, but also demonstrate the value of a deep molecular understanding of oncogenic signaling pathways.

Summary Adult T cell leukemia/lymphoma (ATLL) is a frequently incurable disease associated with the human lymphotropic virus type I (HTLV-I). RNAi screening of ATLL lines revealed that their proliferation depends on BATF3 and IRF4, which cooperatively drive ATLL-specific gene expression. HBZ, the only HTLV-I encoded transcription factor that is expressed in all ATLL cases, binds to an ATLL-specific BATF3 super-enhancer and thereby regulates the expression of BATF3 and its downstream targets, including MYC. Inhibitors of bromodomain-and-extra-terminal-domain (BET) chromatin proteins collapsed the transcriptional network directed by HBZ and BATF3, and were consequently toxic for ATLL cell lines, patient samples, and xenografts. Our study demonstrates that the HTLV-I oncogenic retrovirus exploits a regulatory module that can be attacked therapeutically with BET inhibitors.

Correlating electromechanical and dielectric properties with nanometre-scale order is the defining challenge for the development of piezoelectric oxides. Current lead (Pb)-based relaxor ferroelectrics can serve as model systems with which to unravel these correlations, but the nature of the local order and its relation to material properties remains controversial. Here we employ recent advances in diffuse scattering instrumentation to investigate crystals that span the phase diagram of PbMg1/3Nb2/3O3-xPbTiO3 (PMN-xPT) and identify four forms of local order. From the compositional dependence, we resolve the coupling of each form to the dielectric and electromechanical properties observed. We show that relaxor behaviour does not correlate simply with ferroic diffuse scattering; instead, it results from a competition between local antiferroelectric correlations, seeded by chemical short-range order, and local ferroic order. The ferroic diffuse scattering is strongest where piezoelectricity is maximal and displays previously unrecognized modulations caused by anion displacements. Our observations provide new guidelines for evaluating displacive models and hence the piezoelectric properties of environmentally friendly next-generation materials.

Canonical transient receptor potential (TRPC) channels are a family of polymodal cation channels with some degree of Ca2+ permeability. Although initially thought to be channels mediating store-operated Ca2+ influx, TRPC channels can be activated by stimulation of Gq-coupled G-protein coupled receptors, or by an increase in intracellular free Ca2+ concentration. Thus, activation of TRPC channels could be a common downstream event of many signaling pathways that contribute to seizure and excitotoxicity, such as N-methyl-D-aspartate (NMDA) receptor-mediated Ca2+ influx, or metabotropic glutamate receptor activation. Recent studies with genetic ablation of various TRPC family members have demonstrated that TRPC channels, in particular heteromeric TRPC1/4 channels and homomeric TRPC5 channels, play a critical role in both pilocarpine-induced acute seizures and neuronal cell death. However, exact underlying mechanisms remain to be fully elucidated, and selective TRPC modulators and antibodies with better specificity are urgently needed for future research.

Expression of the activating transcription factor 3 (ATF3) gene is induced by Toll-like receptor (TLR) signaling. In turn, ATF3 protein inhibits the expression of various TLR-driven proinflammatory genes. Given its counter-regulatory role in diverse innate immune responses, we defined the effects of ATF3 on neutrophilic airway inflammation in mice. ATF3 deletion was associated with increased lipopolysaccharide (LPS)-driven airway epithelia production of CXCL1, but not CXCL2, findings concordant with a consensus ATF3-binding site identified solely in the Cxcl1 promoter. Unexpectedly, ATF3-deficient mice did not exhibit increased airway neutrophilia after LPS challenge. Bone marrow chimeras revealed a specific reduction in ATF3−/− neutrophil recruitment to wild-type lungs. In vitro, ATF3−/− neutrophils exhibited a profound chemotaxis defect. Global gene expression analysis identified ablated Tiam2 expression in ATF3−/− neutrophils. TIAM2 regulates cellular motility by activating Rac1-mediated focal adhesion disassembly. Notably, ATF3−/− and ATF3-sufficient TIAM2 knockdown neutrophils, both lacking TIAM2, exhibited increased focal complex area, along with excessive CD11b-mediated F-actin polymerization. Together, our data describe a dichotomous role for ATF3-mediated regulation of neutrophilic responses: inhibition of neutrophil chemokine production but promotion of neutrophil chemotaxis.

Diffuse large B-cell lymphoma (DLBCL) is an aggressive cancer of aberrant B-lymphocytes. Although a portion of DLBCL is curable with standard immunochemotherapy, patients who fail this treatment have a poor prognosis. Recently, cancer genomics has paved the way for better understanding of the genetic basis of lymphoma pathogenesis. Characterization of point mutations and structural alterations has uncovered novel molecular targets for lymphoma therapy and provided a comprehensive view of lymphoma development. By performing multiplatform genomic analysis of DLBCL biopsy samples, we have identified KLHL14 as a recurrent target of somatic mutations in activated B-cell-like (ABC) DLBCL biopsies (10.8% of patients). KLHL14 contains a BTB (broad complex, tramtrack, and bric a brac) domain that can potentially mediate dimerization and binding to Cullin3 (CUL3)-a essential scaffold component of the Cullin-RING-based E3 ubiquitin ligase complexes. KLHL14 also contains kelch repeats that can form a B-propeller tertiary structure that can serve as a substrate-binding domain. KLHL14 is highly expressed in B-cells but is found at low levels in non-immune tissues. Deficiency of KLHL14 in mice leads to embryonic lethality while KLHL14 heterozygous mice show reduction of B-1a cells, suggesting a role for KLHL14 in B-cell homeostasis. Importantly, KLHL14 mutations are highly enriched in tumors belonging to the recently defined MCD (MYD88L265P/CD79B mutation) genetic subtype of DLBCL, the subset of ABC DLBCLs. Somatic mutations primarily localize to the N-terminus of the protein in the BTB domain and BACK (BTB and C-terminal Kelch) domain. However, the impact of these mutations as well as the molecular function of KLHL14 is largely unknown. To investigate the biological effect of KLHL14 loss of function, we used an inducible CRISPR/Cas9 system to delete KLHL14 in ABC DLBCL cell lines and monitored cell growth. Ablation of KLHL14 resulted in an increase in cell proliferation and survival, supporting a role for KLHL14 as a tumor suppressor. Next, we performed a multiplatform -omic analysis (proteomics, phosphoproteomics, ubiquitinomics, high-throughput sequencing) to explore the signaling networks and interactome of KLHL14. Whereas ectopic expression of wild-type KLHL14 altered the dynamics of tyrosine phosphorylation and ubiquitylation events in ABC DLBCL lines, KLHL14 lymphoma-associated mutant alleles had little if any effect, suggesting that they are loss-of-function variants. Gene expression profiling by RNA-sequencing revealed that KLHL14-inactivated cells have a higher NF-kB target gene expression than wild-type cells. Thus, tumor-associated inactivating mutations of KLHL14 depend on a subset of essential NF-kB-related oncoproteins for their survival and this might contribute to the proliferative advantage of DLBCL. In summary, we have uncovered a tumor suppressive function of KLHL14 and found that KLHL14 mutants promote ABC DLBCL survival by increasing NF-kB activity. These findings suggest that tumors with KLHL14 inactivating mutations may serve as a marker of resistance to anti-NF-kB treatment and provide the basis for treating MCD subtype patients with downstream NF-kB pathway inhibitors in the clinical settings. Disclosures Staudt: Nanostring: Patents & Royalties.

Although fragile X syndrome (FXS) is caused by a hypermethylated full mutation (FM) expansion with ≥200 cytosine-guanine-guanine (CGG) repeats, and a decrease in FMR1 mRNA and its protein (FMRP), incomplete silencing has been associated with more severe autism features in FXS males. This study reports on brothers (B1 and B2), aged 5 and 2 years, with autistic features and language delay, but a higher non-verbal IQ in comparison to typical FXS. CGG sizing using AmplideX PCR only identified premutation (PM: 55−199 CGGs) alleles in blood. Similarly, follow-up in B1 only revealed PM alleles in saliva and skin fibroblasts; whereas, an FM expansion was detected in both saliva and buccal DNA of B2. While Southern blot analysis of blood detected an unmethylated FM, methylation analysis with a more sensitive methodology showed that B1 had partially methylated PM alleles in blood and fibroblasts, which were completely unmethylated in buccal and saliva cells. In contrast, B2 was partially methylated in all tested tissues. Moreover, both brothers had FMR1 mRNA ~5 fold higher values than those of controls, FXS and PM cohorts. In conclusion, the presence of unmethylated FM and/or PM in both brothers may lead to an overexpression of toxic expanded mRNA in some cells, which may contribute to neurodevelopmental problems, including elevated autism features.

Acute myelogenous leukemia (AML) subtypes that result from oncogenic activation of homeobox (HOX) transcription factors are associated with poor prognosis. The HOXA9 transcription activator and growth factor independent 1 (GFI1) transcriptional repressor compete for occupancy at DNA-binding sites for the regulation of common target genes. We exploited this HOXA9 versus GFI1 antagonism to identify the genes encoding microRNA-21 and microRNA-196b as transcriptional targets of HOX-based leukemia oncoproteins. Therapeutic inhibition of microRNA-21 and microRNA-196b inhibited in vitro leukemic colony forming activity and depleted in vivo leukemia-initiating cell activity of HOX-based leukemias, which led to leukemia-free survival in a murine AML model and delayed disease onset in xenograft models. These data establish microRNA as functional effectors of endogenous HOXA9 and HOX-based leukemia oncoproteins, provide a concise in vivo platform to test RNA therapeutics, and suggest therapeutic value for microRNA antagonists in AML.

Background Alcohol use occurs across the life span beginning in adolescence and continuing through adulthood. Ethanol (EtOH)-induced pathology varies with age and includes changes in neurogenesis, neurodegeneration, and glial cell activation. EtOH-induced changes in glial activation and immune activity are believed to contribute to EtOH-induced neuropathology. Recent studies indicate an emerging role of glial-derived neuroimmune molecules in alcohol abuse and addiction. Methods Adolescent and adult C57BL/6 mice were treated via gavage with 6 g/kg EtOH for 10 days, and tissue was harvested 1 day post treatment. We compared the effects of EtOH on chemokine and cytokine expression and astrocyte glial fibrillary acidic protein (GFAP) immunostaining and morphology in the hippocampus, cerebellum, and cerebral cortex. Results EtOH increased mRNA levels of the chemokine CCL2/MCP-1 in all 3 regions of adult mice relative to controls. The cytokine interleukin-6 (IL-6) was selectively increased only in the adult cerebellum. EtOH did not affect mRNA levels of the cytokine tumor necrosis factor-alpha (TNF-α) in any of these brain regions in adult animals. Interestingly, CCL2, IL-6, and TNF-α mRNA levels were not increased in the hippocampus, cerebellum, or cortex of adolescent mice. EtOH treatment of adult and adolescent mice resulted in increased GFAP immunostaining. Conclusions Collectively, these data indicate an age- and region-specific susceptibility to EtOH regulation of neuroinflammatory and addiction-related molecules as well as astrocyte phenotype. These studies may have important implications concerning differential alcohol-induced neuropathology and alcohol addiction across the life span.

Dilute magnetic ion doping in LaCo1−y Ni y O3 with y≤1 % leads to the formation of isotropic magnetic clusters that exhibit intracluster interactions which are ferromagnetic in nature. The clusters are comprised of Ni ions surrounded by six magnetically polarized Co ions. The Ni spin is delocalized from the Ni3+ ion but is confined in the vicinity of the six Co coordinated environment forming small magneto-polarons. The cluster ground state is estimated from bulk magnetization to be about gS∼10, in contrast to LaCoO3, which is not magnetic. Using neutron spectroscopy, transitions are observed between the lowest energy levels indicating that the cluster ground state is split. Under a magnetic field of 1 Tesla, the transitions are suppressed while with increasing temperature, the intracluster transitions are overshadowed by the activation of the Co3+ ions to the intermediate spin state.

SummaryMost patients with acute lymphoblastic leukemia (ALL) fail current treatments highlighting the need for better therapies. Because oncogenic signaling activates a p53-dependent DNA damage response and apoptosis, leukemic cells must devise appropriate countermeasures. We show here that growth factor independence 1 (Gfi1) can serve such a function because Gfi1 ablation exacerbates p53 responses and lowers the threshold for p53-induced cell death. Specifically, Gfi1 restricts p53 activity and expression of proapoptotic p53 targets such as Bax, Noxa (Pmaip1), and Puma (Bbc3). Subsequently, Gfi1 ablation cures mice from leukemia and limits the expansion of primary human T-ALL xenografts in mice. This suggests that targeting Gfi1 could improve the prognosis of patients with T-ALL or other lymphoid leukemias.

Objective: Canonical transient receptor potential (TRPC) channels constitute a family of cation channels that exhibit a regional and cell-specific expression pattern throughout the brain. It has been reported previously that TRPC3 channels are effectors of the brain-derived neurotrophic factor (BDNF)/trkB signaling pathway. Given the long postulated role of BDNF in epileptogenesis, TRPC3 channels may be a critical component in the underlying pathophysiology of seizure and epilepsy. In this study, we investigated the precise role of TRPC3 channels in pilocarpine-induced status epilepticus (SE). Methods: The role of TRPC3 channels was investigated using TRPC3 knockout (KO) mice and TRPC3-selective inhibitor Pyr3. Video and electroencephalography (EEG) recording of pilocarpine-induced seizures were performed. Results: We found that genetic ablation of TRPC3 channels reduces behavioral manifestations of seizures and the root-mean-square (RMS) power of SE, indicating a significant contribution of TRPC3 channels to pilocarpine-induced SE. Furthermore, the reduction in SE in TRPC3KO mice is caused by a selective attenuation of pilocarpine-induced theta activity, which dominates both the preictal phase and SE phase. Pyr3 also caused a reduction in the overall RMS power of pilocarpine-induced SE and a selective reduction in the theta activity during SE. Significance: Our results demonstrate that TRPC3 channels unequivocally contribute to pilocarpine-induced SE and could be a novel molecular target for new anticonvulsive drugs. Fil: Phelan, Kevin D.. University of Arkansas for Medical Sciences; Estados Unidos Fil: Shwe, U Thaung. University of Arkansas for Medical Sciences; Estados Unidos Fil: Cozart, Michael A.. University of Arkansas for Medical Sciences; Estados Unidos Fil: Wu, Hong. University of Arkansas for Medical Sciences; Estados Unidos Fil: Mock, Matthew M.. University of Arkansas for Medical Sciences; Estados Unidos Fil: Abramowitz, Joel. National Institute of Environmental Health Sciences; Estados Unidos Fil: Birnbaumer, Lutz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina Fil: Zheng, Fang. University of Arkansas for Medical Sciences; Estados Unidos

The quasi-2D nickelate La4Ni3O8 (La-438), consisting of trilayer networks of square planar Ni ions, is a member of the so-called T' family, which is derived from the Ruddlesden-Popper (R-P) parent compound La4Ni3O10-x by removing two oxygen atoms and rearranging the rock salt layers to fluorite-type layers. Although previous studies on polycrystalline samples have identified a 105-K phase transition with a pronounced electronic and magnetic response but weak lattice character, no consensus on the origin of this transition has been reached. Here, we show using synchrotron X-ray diffraction on high-pO2 floating zone-grown single crystals that this transition is associated with a real space ordering of charge into a quasi-2D charge stripe ground state. The charge stripe superlattice propagation vector, q = (2/3, 0, 1), corresponds with that found in the related 1/3-hole doped single-layer R-P nickelate, La5/3Sr1/3NiO4 (LSNO-1/3; Ni(2.33+)), with orientation at 45° to the Ni-O bonds. The charge stripes in La-438 are weakly correlated along c to form a staggered ABAB stacking that reduces the Coulomb repulsion among the stripes. Surprisingly, however, we find that the charge stripes within each trilayer of La-438 are stacked in phase from one layer to the next, at odds with any simple Coulomb repulsion argument.

The requirement for the B-cell transcription factor OCT2 (octamer-binding protein 2, encoded by Pou2f2) in germinal center B cells has proved controversial. Here, we report that germinal center B cells are formed normally after depletion of OCT2 in a conditional knockout mouse, but their proliferation is reduced and in vivo differentiation to antibody-secreting plasma cells is blocked. This finding led us to examine the role of OCT2 in germinal center-derived lymphomas. shRNA knockdown showed that almost all diffuse large B-cell lymphoma (DLBCL) cell lines are addicted to the expression of OCT2 and its coactivator OCA-B. Genome-wide chromatin immunoprecipitation (ChIP) analysis and gene-expression profiling revealed the broad transcriptional program regulated by OCT2 that includes the expression of STAT3, IL-10, ELL2, XBP1, MYC, TERT, and ADA. Importantly, genetic alteration of OCT2 is not a requirement for cellular addiction in DLBCL. However, we detected amplifications of the POU2F2 locus in DLBCL tumor biopsies and a recurrent mutation of threonine 223 in the DNA-binding domain of OCT2. This neomorphic mutation subtly alters the DNA-binding preference of OCT2, leading to the transactivation of noncanonical target genes including HIF1a and FCRL3 Finally, by introducing mutations designed to disrupt the OCT2-OCA-B interface, we reveal a requirement for this protein-protein interface that ultimately might be exploited therapeutically. Our findings, combined with the predominantly B-cell-restricted expression of OCT2 and the absence of a systemic phenotype in our knockout mice, suggest that an OCT2-targeted therapeutic strategy would be efficacious in both major subtypes of DLBCL while avoiding systemic toxicity.

Lost circulation is a continual problem associated while drilling and cementing during well construction. The failure to provide adequate cement coverage over loss zones can result in costly remediation and/or fines from regulatory agencies. Traditional losses prior to, or during, cement operations are addressed by loss circulation material (LCM) laden pills pumped ahead of the cement treatment as sweeps or spacers. The LCM pills are limited in volume to the size of the mixing tank and can be an insufficient volume compared to the thief zones. The high concentration of LCM in the pill results in higher rheology and can be difficult to properly displace up the annulus by the cement treatment due to inverse friction hierarchy between fluids. The solution being presented consists of using the cement slurry as the carrying fluid along with an inert engineered solid package. A fiber added on-the-fly improves the plugging efficiency of the engineered solid package and the ability to tailor the treatment design to the well conditions up until the treatment is pumped. The results are tangible, with the solution applied successfully in over 300 treatments for multiple operators in the Permian Basin. The Permian Basin is one of the most developed basins in the world with a complex lithology and layered, highly depleted reservoirs. The solution successful performance has been unprecedented in allowing operators to comply with local regulations, avoid remedial work, and extend the life of the completion across corrosive formations. Application of the engineered solid package with fiber is for loss circulation cases in highly depleted zones, permeable formations, and naturally fracture formations for other basins. The progress thus far has been encouraging with additional optimization of the engineered solid package with fiber application could potentially improve the performance further. The final result will be an improved fluid placement and understanding of the effect LCM has on the carrying fluid.

SUNY Cortland and key stakeholders in the Cortland community have developed relationships and leveraged their resources to proactively pursue economic development activity through the creation of an entrepreneurship program. The goal of the college/community partnership is to utilize the expertise in the business community to help foster new business generation from SUNY Cortland students. Additionally, the small businesses would receive help from SUNY Cortland students through internships, conducting business research and providing other services that the business may not have capacity to provide for themselves.

Fetal alcohol spectrum disorder (FASD) is a major health concern worldwide and results from maternal consumption of alcohol during pregnancy. It produces tremendous individual, social, and economic losses. This review will first summarize the structural, functional, and behavior changes seen in FASD. The development of the neuroimmune system will be then be described with particular emphasis on the role of microglial cells in the normal regulation of homeostatic function in the central nervous system (CNS) including synaptic transmission. The impact of alcohol on the neuroimmune system in the developing CNS will be discussed in the context of several key immune molecules and signaling pathways involved in neuroimmune mechanisms that contribute to FASD. This review concludes with a summary of the development of early therapeutic approaches utilizing immunosuppressive drugs to target alcohol-induced pathologies. The significant role played by neuroimmune mechanisms in alcohol addiction and pathology provides a focus for future research aimed at understanding and treating the consequences of FASD.

Canonical transient receptor potential channels (TRPCs) are receptor-operated cation channels that are activated in response to phospholipase C signaling. Although TRPC1 is ubiquitously expressed in the brain, TRPC4 expression is the most restrictive, with the highest expression level limited to the lateral septum. The subunit composition of neuronal TRPC channels remains uncertain because of conflicting data from recombinant expression systems. Here we report that the large depolarizing plateau potential that underlies the epileptiform burst firing induced by metabotropic glutamate receptor agonists in lateral septal neurons was completely abolished in TRPC1/4 double-knockout mice, and was abolished in 74% of lateral septal neurons in TRPC1 knockout mice. Furthermore, neuronal cell death in the lateral septum and the cornu ammonis 1 region of hippocampus after pilocarpine-induced severe seizures was significantly ameliorated in TRPC1/4 double-knockout mice. Our data suggest that both TRPC1 and TRPC4 are essential for an intrinsic membrane conductance mediating the plateau potential in lateral septal neurons, possibly as heteromeric channels. Moreover, excitotoxic neuronal cell death, an underlying process for many neurological diseases, is not mediated merely by ionotropic glutamate receptors but also by heteromeric TRPC channels activated by metabotropic glutamate receptors. TRPC channels could be an unsuspected but critical molecular target for clinical intervention for excitotoxicity.

Summary Total cholesterol and low-density lipoprotein (LDL) cholesterol exhibit an independent, strong, continuous correlation with cardiovascular events. The effectiveness of hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) in the treatment and prevention of atherosclerosis is well-established. However, despite the lowering of LDL targets and the increased use of statins, patients with type 2 diabetes mellitus (DM) continue to experience a higher proportion of adverse coronary artery disease events. This is as a result of an atherogenic dyslipidaemia, characterized by low levels of high-density lipoprotein and elevated plasma triglyceride concentrations, often with high levels of cholesterol-rich remnant particles. This article will review dyslipidaemia and its role in DM, and will discuss available treatment modalities that address residual cardiovascular risk in this disease.

Pressure sensitive paints (PSP) that measure changes in barometric pressure through oxygen quenching are an important analytical tool in wind tunnel and other research. PSPs are often used during the development phase of new aircraft, cars, and trucks where the performance of the proposed vehicle is researched. Previously, many PSPs have been based upon charged phosphorescent metal complex salts that may have poor solubility in the oxygen permeable host material. Charge neutral phosphorescent dyes should have better solubility, and to demonstrate this, neutral osmium and iridium complexes are presented in this report. X-ray structures are measured and DFT calculations are used to explain many of their properties. The complexes are dissolved into poly(1,1,1,3,3,3-hexafluoroisopropylmethacrylate-co-1H,1H-dihydroperfluorobutyl-methacrylate) (FIB), an oxygen permeable host polymer. The complexes exhibit good solubility and compatibility with the FIB polymer. The photo-degradation, temperature sensitivity, and Stern–Volmer curves of PSPs based on these complexes are presented. The complexes show great promise for use in PSPs.

After neonatal HTLV-I infection through breast feeding, approximately 5% of HTLV-I carriers eventually develop Adult T-Cell Leukemia/Lymphoma (ATLL) with a latency of ~50 years, suggesting that acquired genetic and epigenetic changes in cellular genes act in concert with HTLV-I to initiate and maintain oncogenic transformation. We and others have recently utilized next generation sequencing technology to identify mutated genes that could be pivotal in the pathogenesis of ATLL. However, due to the complexity of genomic/epigenetic alteration in the ATLL genome, the identification of indispensable genes for proliferation and/or survival of ATLL cells remains a formidable challenge. To discover essential regulatory networks that are required for the proliferation and survival of ATLL cells, we performed a pooled shRNA screen in 8 ATLL cell lines using a library enriched for shRNAs targeting lymphoid regulatory factors and discovered that two BATF3 shRNAs and one IRF4 shRNA were highly toxic for all ATLL lines, but had little if any effect in other T cell and B cell lines. It is recently shown that a transcriptional complex of Irf4 and Batf binds to AP1-IRF composite (AICE) DNA motifs and plays key roles in the differentiation and function of certain mouse helper T cell subsets. A close paralogue of Batf, Batf3, is an indispensable transcription factor in a mouse dendritic cell subset, but also appears to play a redundant role with Batf in the differentiation of TH2 cells and can substitute for Batf in Batf knockout T cells. Our observations from shRNA screening suggested that IRF4 and BATF3 may cooperate to drive a transcriptional program that is essential for ATLL viability. We next used genome-wide chromatin precipitation (ChIP-seq) to identify the loci that are bound by BATF3 and IRF4. The set of binding peaks and the associated genes in IRF4 and BATF3 ChIP-seq intersected significantly. By integrating the ChIP-seq and gene expression profiling data of shBATF3- and shIRF4-ATLL cells, we defined a set of 68 BATF3-IRF4 direct target genes. Gene set enrichment analysis using gene expression profiling data from primary T cell lymphomas demonstrated that BATF3-IRF4 direct target genes were significantly enriched among genes that are more highly expressed in ATLL than in peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS), suggesting that the BATF3 and IRF4 cooperatively regulate transcription in primary ATLL cells. HBZ is unique among HTLV-I viral proteins in being maintained in expression in all ATLL cases, suggesting that it may help maintain the malignant phenotype. Given that BATF3 and IRF4 are essential regulators in ATLL, we hypothesized possible relationship between HBZ and BATF3-IRF4 complex. We defined HBZ direct target genes by integrating the ChIP-seq and gene expression profiling data of HBZ-knockout ATLL cell lines by CRISPR/Cas9. Notably we discovered that BATF3 was among these. BATF3 mRNA and protein expression decreased following HBZ inactivation. The above considerations suggested that pharmacologic inhibition of the BATF3-IRF4 regulatory network might be a means to attack the HBZ oncogenic program therapeutically. ChIP-seq analysis of two enhancer marks, H3K27ac and BRD4, identified super-enhancers at the BATF3 locus in two ATLL cell lines. The small molecule JQ1 prevents the BET-protein BRD4 from interacting with chromatin, which is required for the function of super-enhancers. JQ1 treatment reduced BATF3 mRNA and protein levels in all ATLL lines tested, correlating with the eviction of BRD4 from the BATF3 super-enhancer. MYC mRNA and protein expression was also broadly downmodulated by JQ1. JQ1 treatment was consistently toxic for all ATLL cell lines tested at dose ranges that killed cell line models of T-ALL and DLBCL, which are known to rely on BET-proteins. In a dose-dependent manner, JQ1 also reduced the viability of primary ATLL samples and downregulated their expression BATF3 and MYC mRNA. Finally, we treated mouse xenograft models of ATLL with the BET-protein inhibitor CPI-203, a JQ1 analog with superior bioavailability in mice. In two different xenograft models, we observed significant tumor regression or growth inhibition, without evidence of systemic toxicity. Our study demonstrates that the HTLV-I virus exploits a regulatory module that can potentially be attacked therapeutically with BET protein inhibitors. Disclosures Yu: Celgene Corporation: Employment.

We report the synthesis of phosphorescent divalent osmium complexes of the form [Os(N–N) 2 (L–L) or Os(L–L) 2 (N–N)] 2+ (PF 6 ) 2 where N–N is a derivative of 1,10-phenanthroline, and L–L is a diarsine or diphosphine ligand: 1,2-bis(dimethylphosphino)ethane, 1,2-bis(dicyclohexylphosphino)ethane, or 1,2-bis(dimethylarseno)benzene. X-ray structures have been determined, luminescent and electrochemical properties have been measured and DFT calculations have been performed on the complexes. The emission lifetime of complexes of structure Os(II)(L–L) 2 (N–N) are longer than the those of Os(II)(N–N) 2 (L–L). The DFT calculations show that there is significant mixing of the π−π ∗ into the dπ−π ∗ charge-transfer state for the complexes of the form Os(II)(L–L) 2 (N–N) resulting in a longer lived excited state. Through DFT calculations we were able to conclude that the HOMO of the complexes is a d orbital on the osmium while the LUMO is the b 1 ( ψ ) π ∗ system of the phenanthroline. However, we found that the HOMO did not have the correct symmetry to enable strong charge transfer to the phenanthroline to be observed, and the strong MLCT transition observed in the spectra is the metal d HOMO(−1) to the b1 π ∗ LUMO of the phenanthroline.

We report new osmium complexes based upon the 2-[(diphenylphosphanyl)-methyl]-pyridine ligand. Hexafluorophosphate salts of bis(3,4,7,8-tetramethyl-1,10-phenantholine)(2-[(diphenylphosphanyl)-methyl]-pyridine)osmium(II) (1), and bis(2-[(diphenylphosphanyl)-methyl]-pyridine)(4,7-diphenyl-1,10-phenantholine)osmium(II) (2) have been synthesized, and X-ray structures have been obtained. Indexing and unit cell refinement of both 1 and 2 indicated a monoclinic P lattice. The space groups were found to be P1 (No. 2) and P21/c, respectively. Both 1 and 2 were found to have the same emission at 645 nm despite the differences in structure. Through density functional theory (DFT) calculations we were able to conclude that the highest occupied molecular orbital (HOMO) of the complexes is the dxy orbital on the osmium, while the lowest unoccupied molecular orbital (LUMO) is the b1(Ψ)π* system of the phenanthroline. However, we found that dxy did not have the correct orientation to enable charge transfer to the phenan...

Application of bis(propylenedioxythiophene) (bis(ProDOT)) π-conjugated bridges bearing alkyl or aryl substituents in electro-optic (EO) chromophores is presented. A series of three bis(ProDOT)-based chromophores and a bithiophene-based control chromophore were prepared and fully characterized with regard to EO applications. The highly planar bis(ProDOT) bridge results in slightly larger (∼10%) molecular hyperpolarizability (β) values as compared to the bithiophene bridge, as measured by hyper-Rayleigh scattering at a variety of wavelengths. In amorphous polycarbonate guest–host films, however, the bulky substituents on the bis(ProDOT) bridge result in significantly larger (∼70%) poling-induced EO coefficient (r33) values, as measured by simple reflection ellipsometry at 1310 nm. This can be attributed to a roughly 2-fold enhancement in poling efficiency due to reduced intermolecular dipole–dipole interactions. This chromophore architecture also exhibits excellent temporal alignment stability and photochem...

Agricultural soils are the primary anthropogenic source of atmospheric nitrous oxide (N2O), contributing to global warming and depletion of stratospheric ozone. Biochar addition has shown potential to lower soil N2O emission, with the mechanisms remaining unclear. We incubated eucalypt biochar (550 °C) – 0, 1 and 5% (w/w) in Ferralsol at 3 water regimes (12, 39 and 54% WFPS) – in a soil column, following gamma irradiation. After N2O was injected at the base of the soil column, in the 0% biochar control 100% of expected injected N2O was released into headspace, declining to 67% in the 5% amendment. In a 100% biochar column at 6% WFPS, only 16% of the expected N2O was observed. X-ray photoelectron spectroscopy identified changes in surface functional groups suggesting interactions between N2O and the biochar surfaces. We have shown increases in -O-C = N /pyridine pyrrole/NH3, suggesting reactions between N2O and the carbon (C) matrix upon exposure to N2O. With increasing rates of biochar application, higher pH adjusted redox potentials were observed at the lower water contents. Evidence suggests that biochar has taken part in redox reactions reducing N2O to dinitrogen (N2), in addition to adsorption of N2O.

The activated B-cell-like (ABC) subtype of diffuse large B-cell lymphoma (DLBCL) relies on chronic active B-cell receptor (BCR) signaling. BCR pathway inhibitors induce remissions in a subset of ABC DLBCL patients. BCR microclusters on the surface of ABC cells resemble those generated following antigen engagement of normal B cells. We speculated that binding of lymphoma BCRs to self-antigens initiates and maintains chronic active BCR signaling in ABC DLBCL. To assess whether antigenic engagement of the BCR is required for the ongoing survival of ABC cells, we developed isogenic ABC cells that differed solely with respect to the IgH V region of their BCRs. In competitive assays with wild-type cells, substitution of a heterologous V region impaired the survival of three ABC lines. The viability of one VH4-34(+) ABC line and the ability of its BCR to bind to its own cell surface depended on V region residues that mediate the intrinsic autoreactivity of VH4-34 to self-glycoproteins. The BCR of another ABC line reacted with self-antigens in apoptotic debris, and the survival of a third ABC line was sustained by reactivity of its BCR to an idiotypic epitope in its own V region. Hence, a diverse set of self-antigens is responsible for maintaining the malignant survival of ABC DLBCL cells. IgH V regions used by the BCRs of ABC DLBCL biopsy samples varied in their ability to sustain survival of these ABC lines, suggesting a screening procedure to identify patients who might benefit from BCR pathway inhibition.

For several decades LaCoO${}_{3}$ attracted interest due the unconventional thermally-driven spin-state crossover from a nonmagnetic to paramagnetic state. More recently, it was realized that the ``nonmagnetic'' ground state harbors some form of magnetism that is likely due to local magnetic entities forming at oxygen vacancies. Such entities dubbed ``magnetic excitons'' were predicted in 1996, with some evidence for their existence emerging around 2005 from muon spin relaxation. In this Rapid Communication the authors provide the first scattering-based evidence for spin excitons, direct information on the size and density of the excitons, as well as their interactions. The form of the scattering is also direct proof of a bulk 3D ensemble of these excitons.

The layered compound ${\mathrm{CePd}}_{1\ensuremath{-}x}{\mathrm{Bi}}_{2}$ with the tetragonal ${\mathrm{ZrCuSi}}_{2}$-type structure was obtained from excess Bi flux. Magnetic susceptibility data of ${\mathrm{CePd}}_{1\ensuremath{-}x}{\mathrm{Bi}}_{2}$ show an antiferromagnetic ordering below 6 K and are anisotropic along the $c$ axis and the $ab$ plane. The anisotropy is attributed to crystal-electric-field (CEF) effects and a CEF model which is able to describe the susceptibility data is given. An enhanced Sommerfeld coefficient $\ensuremath{\gamma}$ of 0.191 J mol $\mathrm{Ce}{}^{\ensuremath{-}1} \mathrm{K}{}^{\ensuremath{-}2}$ obtained from specific-heat measurement suggests a moderate Kondo effect in ${\mathrm{CePd}}_{1\ensuremath{-}x}{\mathrm{Bi}}_{2}$. Other than the antiferromagnetic peak at 6 K, the resistivity curve shows a shoulderlike behavior around 75 K which could be attributed to the interplay between Kondo and CEF effects. Magnetoresistance and Hall-effect measurements suggest that the interplay reconstructs the Fermi-surface topology of ${\mathrm{CePd}}_{1\ensuremath{-}x}{\mathrm{Bi}}_{2}$ around 75 K. Electronic structure calculations reveal that the Pd vacancies are important to the magnetic structure and enhance the CEF effects which quench the orbital moment of Ce at low temperatures.

Divalent osmium complexes of the form [ Os ( N – N ) 2 L – L ] ( PF 6 - ) 2 where N–N was a polypyridyl, and L–L was either cis-1,2-bis(diphenylphosphino)ethene (dppene) or cis-1,2-vinylenebis(diphenylarsine) (dpaene) have been synthesized and characterized. X-ray structures were determined for three complexes and for the free dpaene molecule. It was observed that the P–C bond lengths, and C–P–C bond angles do not change significantly when complexed to osmium. It was observed the As–C bond lengths shorten by 2.3 pm and the C–As–C bond angles broaden by 5.6° when dpaene was complexed to osmium. These changes in the arsine structure may indicate a different method of backbonding between arsenic and osmium. It was found that the arsine complexes had absorption and emission that were to the red of analogous phosphine complexes. In violation of “energy gap law”, the dpaene complexes were found to have higher quantum yields. This may be due to the way that the arsenic atoms bond to osmium.

Juvenile rheumatoid arthritis (JRA), also known as juvenile idiopathic arthritis (JIA), includes the most common chronic autoimmune arthropathies of childhood. These two nomenclatures for classification include components representing the major subclasses of disease. The chromosomal regions and the genes involved in these complex genetic traits are being elucidated, with findings often specific for a particular disease subtype. With the advent of new SNP technologies, progress is being made at an ever-increasing pace. This review discusses the difficulties of deciphering the genetic components in complex disorders, while demonstrating the similarities that JRA shares with other autoimmune disorders. Particular emphasis has been placed on positive findings either for candidate genes that have been replicated independently in JRA/JIA, or findings in JRA for which consistent results have been reported in other forms of autoimmunity.

LGR8 was recently identified as a cognate receptor for insulin-like peptide-3 (INSL3), and INSL3-LGR8 signaling is best known for its role in testis descent during development. LGR8 mRNA has been detected in various human tissues including brain, but the regional and cellular distribution of LGR8 expression in the mammalian central nervous system is unknown. Therefore, in this study we investigated the presence and localization of LGR8 mRNA in rat brain using reverse transcription-polymerase chain reaction and in situ hybridization histochemistry. Results revealed a distinct distribution of LGR8 in forebrain, with transcripts principally restricted to the posterior thalamus and highest densities detected in the parafascicular nucleus of both adult and developing rats. Unexpectedly, INSL3 mRNA was not detected in brain by similar methods, but preliminary electrophysiologic studies of parafascicular neurons revealed that INSL3 altered their activity. These findings suggest that LGR8 signaling may be involved in sensorimotor control in the rat and perhaps other species, particularly via actions on parafascicular neurons that project to basal ganglia and are depleted in Parkinson's disease.

The preparation, photophysics, and solid-state structures of three osmium cored complexes are reported. The osmium complexes take the general form of [OsCl(N–N)(L–L)(CO)] þ hexafluorophosphate where N stands for a derivative of 1,10-phenanthroline and L stands for a phosphine type ligand. The emission of the complexes is shown to be blue shifted to the osmium emission of Os(bpy) 2þ 3 . The emissions of the various complexes range from yellow (560 nm) to yellow-green (550 nm) to green (520 nm). The quantum yields vary between 60% and 75%. The complexes show lifetimes that are much longer than expected with ranges of 6.5–38 ls. Crystallographic results show that the carbonyl is trans to a phenanthroline nitrogen and the chloro ligand is trans to phosphorus. A discussion will be presented as to the nature of the bonding in these complexes based upon the data from the crystallography.

Europium beta diketonates are easily synthesized highly luminescent complexes with high temperature sensitivity. We report on the temperature dependence of the luminescence of recently synthesized europium complexes originally prepared for use as light emitting diodes. It has been discovered that when incorporated in a polymer matrix, their decay lifetime can provide accurate measurement of temperature. Their lifetime as a function of temperature depends on three factors: (i) the type and number of ligands in the complex, (ii) the particular polymer used for the matrix, and (iii) the europium chelate to polymer matrix concentration ratio. Various tris and tetrakis europium chelates are used to study ligand effects, while the polymers FIB, polycarbonate, and Teflon© are used to analyze matrix effects. In all cases studied, higher concentrations give rise to shorter lifetimes and higher temperature sensitivities, with sensitivity defined as ΔI/(IrefΔT). We propose to explain this phenomenon by using the following equation: 1/τobs≡Ktotal=kr+knr(T)+kc([Eu]). Here Ktotal is the observed decay rate, which is the inverse of the observed lifetime, while kr and knr(T) are the radiative and nonradiative decay rates, respectively. As well as being dependent on temperature, knr(T) for these complexes is very dependent on the environment, i.e., solvent or polymer, and can be considered as ken(T). The rate kc([Eu]) is the quenching term dependent on the concentration of the europium complex.

Two newly synthesized luminescent compounds provide a lifetime based dual pressure and temperature sensor when jointly dissolved in a polymer matrix. The temperature sensor is a tris(β-diketonate) phenanthroline europium complex, with a broad absorption peaked at 370 nm and a characteristic but relatively sharp 615 nm europium emission. The lifetime of this compound varies between 260 and 120 μs over the temperature range 5–45 °C for a derivative of −3.50 μs/°C. Our pressure sensor is a porphyrin compound called platinum tetra(pentafluorophenyl)porpholactone (PtTFPL) developed recently in our labs. This compound has a principal absorption band at 390 nm and a red-shifted phosphorescence at 730 μm. In silicone polycarbonate co-polymer, PtTFPL has a lifetime range of 42.7–6.6 μs in going from 0 to 1 atm at 1 atm. The two sensors have maximum absorption in the range 370–400 nm but non-overlapping emission spectra upon excitation with a broadband illumination at 400 nm. As a result, with appropriate band pass optical filters, the two emission lines can be detected separately. In addition to the high photostability of the dual system, our studies show that the luminescence properties of one sensor are independent of the presence of the other sensor. We note that measurement of decay rate is more robust than measuring intensity, since it is unaffected by changes in the light source, the index of refraction of the medium, optical geometry variations and fluctuations in film thickness and dye concentration. Hence the two compounds afford the possibility of a dual luminophor lifetime measurement of pressure and temperature.

Background: In asthma, measurements of airway inflammation correlate poorly with clinical markers and airway hyperresponsiveness. While the relation between determinants of asthma severity is known, that for cough is unknown. We hypothesised that cough sensitivity changes relate to changes in cough scores and objectively measured cough frequency. Aims: To examine the relation between commonly used outcome measurements of cough severity in children. Methods: The concentration of capsaicin causing two and five or more coughs (C2 and C5 respectively), cough frequency objectively measured using an ambulatory cough meter, and parent and child recorded subjective cough scores were determined in 40 children with recurrent cough on two occasions. Results: On occasion one, log cough frequency significantly correlated with parent and child recorded log cough score (rs = 0.32, p = 0.05; and rs = 0.32, p = 0.046 respectively) and significantly negatively correlated with log C2 (rs= -0.5, p = 0.005). Subjective cough scores did not relate to either C2 or C5. On occasion two, the relation between cough frequency and C2 and C5 measures was lost, but C2 had a weak but significant relation to parent recorded cough score (rs = -0.38, p = 0.047). When the changes in the log values were determined, C5 but not C2 significantly related to cough frequency. Conclusion: In children, measures of cough sensitivity have a weak relation with cough frequency. Subjective cough scores have a stronger and consistent relation with cough frequency. These cough severity indices measure different aspects of cough. The choice of indices depends on the reason for performing the measurement.

A new europium (Eu) complex with dendron-substituted diketone ligands was synthesized and found to exhibit a photoluminescence efficiency of 45%. Double-layer light-emitting diodes based on polymer matrices doped with the Eu complex were fabricated. An electroluminescence external quantum efficiency of 0.80% was achieved when a copolymer containing side-chain carbazole and 1,3,4-oxadiazole groups was used as the matrix. The results are analyzed in the context of Forster energy transfer.

Objective: To determine High Dependency Unit (HDU) utilization patterns over time and its impact on Intensive Care Unit (ICU) workload. Design: Retrospective analysis of prospectively collected data. Setting: High Dependency and Intensive Care Units of an Irish University Hospital. Subjects: Data prospectively collected on all HDU admissions was analyzed for the first 6 months of operation and comparable 3-month periods during years 2 and 4. ICU workload was assessed by comparing ICU admissions by speciality, length of ICU stay, and need for ventilatory support before and after HDU opening. Interventions: None Measurements and main results: Demographic data, reason for admission, patient source, admitting speciality, length of HDU stay, and discharge destination were analysed. ICU data included casemix, length of stay, and requirement for ventilatory support. Seventy four percent of HDU admissions were from the cardiothoracic, vascular and general surgical specialities. The mean HDU stay was 2.1 days, wit...

A group of children with a past history of wheezing was randomly selected from the Melbourne community at the age of 7 years in 1964, and a further group of children with severe wheezing was selected from the same birth cohort at the age of 10 years. These subjects have been followed prospectively at 7-year intervals, with the last review in 1999, when their average age was 42 years. Eighty-seven percent of the original cohort who were still alive participated in the 1999 review. This study showed that the majority of children who had only a few episodes of wheezing associated with symptoms of a respiratory infection had a benign course, with many ceasing to wheeze by adult life. Most who continued with symptoms into adult life were little troubled by them. Conversely, those children with asthma mostly continued with significant wheezing into adult life, and the more troubled they were in childhood, the more likely symptoms continued. There was a loss in lung function by the age of 14 years in those with severe asthma, but the loss did not progress in adult life. The childhood asthma had been treated before the availability of inhaled steroids. There was no significant loss of lung function in those with milder symptoms.

T follicular helper (Tfh) cells are specialized CD4 helper T cells that signal B cells to produce high affinity antibodies and provide long-term humoral immunity. Identifying genes that govern Tfh differentiation and function could provide targets for improving vaccines and therapies for autoimmunity. However, Tfh differentiation is only partially understood, and not fully recapitulated in vitro. To discover novel Tfh-regulating genes, we developed a CRISPR-based functional genetic system to mutate genes in primary mouse T cells, and measured the effects on Tfh cells in vivo. Transgenic Cas9-expressing mouse CD4 T cells were transduced in vitro with an optimized retroviral vector encoding an sgRNA and GFP. With a guide targeting Tcf7 (encoding a Tfh-essential transcription factor Tcf1), >90% cells could be transduced; up to >80% of GFP+ cells were Tcf1neg by flow and mutational analyses. To evaluate the functional consequence of CRISPR editing, we adoptively transferred virus-specific CD4 T cells that were transduced with either a control or Tcf7 sgRNA construct into wild-type hosts. After viral challenge, control T cells differentiated into both Tfh and non-Tfh cells, while very few Tcf7-edited T cells became Tfh cells. Similarly, guides against the Tfh master transcription factor Bcl6 abrogated Tfh differentiation in vivo. Next, we constructed sgRNA libraries of up to 400 guides, targeting up to 80 genes, including ones implicated in primary immunodeficiencies and druggable targets. NGS analysis of T cells post-viral challenge revealed depletion of multiple guides targeting known Tfh-required genes in the Tfh vs non-Tfh populations, validating this approach. We are currently following up on potential hits from these screens.

Status epilepticus (SE) is a life-threatening disease that has been recognized since antiquity but still causes over 50,000 deaths annually in the United States. The prevailing view on the pathophysiology of SE is that it is sustained by a loss of normal inhibitory mechanisms of neuronal activity. However, the early process leading to the initiation of SE is not well understood. Here, we show that, as seen in electroencephalograms, SE induced by the muscarinic agonist pilocarpine in mice is preceded by a specific increase in the gamma wave, and genetic ablation of canonical transient receptor potential channel (TRPC) 7 significantly reduces this pilocarpine-induced increase of gamma wave activity, preventing the occurrence of SE. At the cellular level, TRPC7 plays a critical role in the generation of spontaneous epileptiform burst firing in cornu ammonis (CA) 3 pyramidal neurons in brain slices. At the synaptic level, TRPC7 plays a significant role in the long-term potentiation at the CA3 recurrent collateral synapses and Schaffer collateral-CA1 synapses, but not at the mossy fiber-CA3 synapses. Taken together, our data suggest that epileptiform burst firing generated in the CA3 region by activity-dependent enhancement of recurrent collateral synapses may be an early event in the initiation process of SE and that TRPC7 plays a critical role in this cellular event. Our findings reveal that TRPC7 is intimately involved in the initiation of seizures both in vitro and in vivo. To our knowledge, this contribution to initiation of seizures is the first identified functional role for the TRPC7 ion channel. Fil: Phelan, K. D.. University of Arkansas for Medical Sciences; Estados Unidos Fil: Shwe, U. T.. University of Arkansas for Medical Sciences; Estados Unidos Fil: Abramowitz, J.. National Institute of Environmental Health Sciences; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos Fil: Zheng, F.. University of Arkansas for Medical Sciences; Estados Unidos

Fetal alcohol spectrum disorders (FASD) result from fetal exposure to alcohol and are the leading cause of mental retardation in the United States. There is currently no effective treatment that targets the causes of these disorders. Thus, novel therapies are critically needed to limit the neurodevelopmental and neurodegenerative pathologies associated with FASD.A neonatal mouse FASD model was used to examine the role of the neuroimmune system in ethanol (EtOH)-induced neuropathology. Neonatal C57BL/6 mice were treated with EtOH, with or without pioglitazone, on postnatal days 4 through 9, and tissue was harvested 1 day post treatment. Pioglitazone is a peroxisome proliferator-activated receptor (PPAR)-γ agonist that exhibits anti-inflammatory activity and is neuroprotective. We compared the effects of EtOH with or without pioglitazone on cytokine and chemokine expression and microglial morphology in the hippocampus, cerebellum, and cerebral cortex.In EtOH-treated animals compared with controls, cytokines interleukin-1β and tumor necrosis factor-α mRNA levels were increased significantly in the hippocampus, cerebellum, and cerebral cortex. Chemokine CCL2 mRNA was increased significantly in the hippocampus and cerebellum. Pioglitazone effectively blocked the EtOH-induced increase in the cytokines and chemokine in all tissues to the level expressed in handled-only and vehicle-treated control animals. EtOH also produced a change in microglial morphology in all brain regions that was indicative of microglial activation, and pioglitazone blocked this EtOH-induced morphological change.These studies indicate that EtOH activates microglia to a pro-inflammatory stage and also increases the expression of neuroinflammatory cytokines and chemokines in diverse regions of the developing brain. Further, the anti-inflammatory and neuroprotective PPAR-γ agonist pioglitazone blocked these effects. It is proposed that microglial activation and inflammatory molecules expressed as a result of EtOH treatment during brain development contribute to the sequelae associated with FASD. Thus, pioglitazone and anti-inflammatory pharmaceuticals more broadly have potential as novel therapeutics for FASD.

Novel europium complexes containing either symmetrical or unsymmetrical diphenanthryl β-diketone ligands were synthesized and characterized. The ligand conjugation length has a profound effect on the spectral properties of the complexes and the device performance of the complexes as emitting dopants in light-emitting diodes. The unsymmetrical ligand has a longer conjugation, which red-shifts the absorption of the complex. The longer wavelength absorption facilitates the choice of host material that can efficiently transfer energy to the complex. However, the longer conjugation also results in a less efficient energy transfer from the ligand to the central Eu3+ ion.