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Cytomegalovirus (CMV) is widespread in humans and has been implicated in glioblastoma (GBM) and other tumors. However, the role of CMV in GBM remains poorly understood and the mechanisms involved are not well-defined. The goal of this study was to identify candidate pathways relevant to GBM that may be modulated by CMV. Analysis of RNAseq data after CMV infection of patient-derived GBM cells showed significant upregulation of GBM-associated transcripts including the MET oncogene, which is known to play a role in a subset of GBM patients. These findings were validated in vitro in both mouse and human GBM cells. Using immunostaining and RT-PCR in vivo, we confirmed c-MET upregulation in a mouse model of CMV-driven GBM progression and in human GBM. siRNA knockdown showed that MET upregulation was dependent on CMV-induced upregulation of NF-κB signaling. Finally, proneural GBM xenografts overexpressing c-MET grew much faster in vivo than controls, suggesting a mechanism by which CMV infection of tumor cells could induce a more aggressive mesenchymal phenotype. These studies implicate the CMV-induced upregulation of c-MET as a potential mechanism involved in the effects of CMV on GBM growth.

Cytomegalovirus (CMV) has been implicated in glioblastoma (GBM); however, a mechanistic connection in vivo has not been established. The purpose of this study is to characterize the effects of murine CMV (MCMV) on GBM growth in murine models. Syngeneic GBM models were established in mice perinatally infected with MCMV. We found that tumor growth was markedly enhanced in MCMV(+) mice, with a significant reduction in overall survival compared with that of controls (P < 0.001). We observed increased angiogenesis and tumor blood flow in MCMV(+) mice. MCMV reactivation was observed in intratumoral perivascular pericytes and tumor cells in mouse and human GBM specimens, and pericyte coverage of tumor vasculature was strikingly augmented in MCMV(+) mice. We identified PDGF-D as a CMV-induced factor essential for pericyte recruitment, angiogenesis, and tumor growth. The antiviral drug cidofovir improved survival in MCMV(+) mice, inhibiting MCMV reactivation, PDGF-D expression, pericyte recruitment, and tumor angiogenesis. These data show that MCMV potentiates GBM growth in vivo by increased pericyte recruitment and angiogenesis due to alterations in the secretome of CMV-infected cells. Our model provides evidence for a role of CMV in GBM growth and supports the application of antiviral approaches for GBM therapy.

INTRODUCTION Cytomegalovirus (CMV), a member of the betaherpesvirinae subfamily widespread in human populations has been reported to be detectable in glioblastoma (GBM). We have recently shown that CMV latent infection promotes GBM growth in a mouse model. The mechanisms involved are not well-defined and treatment responses have not been evaluated. METHODS To investigate the effects of CMV on GBM cells we performed an RNAseq study. Differentially expressed genes were identified and validated in vitro and in vivo. RESULTS RNAseq analysis showed multiple alterations induced by CMV infection of GBM cells including significant upregulation of c-MET (8.2 log2 fold change) and MGMT (3.9 log2 fold change) transcripts after CMV infection. c-MET is a tyrosine kinase receptor known to promote GBM growth, and MGMT expression leads to resistance to the alkylating agent temozolomide (TMZ). These findings were validated in vitro in both mouse and human GBM cells, and shown to be dependent on CMV-induced upregulation of NF-kB by siRNA knockdown of the essential RelA NF-kB subunit. The c-MET co-receptor CD44 was also upregulated suggesting a role for the c-MET signalling axis in CMV induced tumor progression. CMV-infected tumor cells were resistant to TMZ treatment as expected due to MGMT upregulation. Treatment of our murine GBM model with TMZ led to a significant increase in survival in control mock-infected mice (median survival 38 days control, 48 days TMZ), whereas the CMV infected mice did not respond (median survival 35 days control, 35 days TMZ; p value between TMZ treated groups = 10–4). CONCLUSIONS Our results define novel and relevant mechanisms by which CMV may influence both GBM growth and resistance to treatment. Further therapeutic and mechanistic studies are underway to investigate these pathways in the context of CMV in GBM. These data support further study of CMV targeting in GBM.

We describe an efficient and convergent synthesis of a series of (1'S,2R,4'S)-3H-4'-azaspiro[benzo[4,5]imidazo[2,1-b]oxazole-2,2'-bicyclo[2.2.2]octanes] displaying potency for the α7 nicotinic acetylcholine receptor (nAChR) and good selectivity vs. the related 5-HT3A receptor.

The motility of blood monocytes is orchestrated by the activity of cell-surface integrins, which translate extracellular signals into cytoskeletal changes to mediate adhesion and migration. Toxoplasma gondii is an intracellular parasite that infects migratory cells and enhances their motility, but the mechanisms underlying T. gondii–induced hypermotility are incompletely understood. We investigated the molecular basis for the hypermotility of primary human peripheral blood monocytes and THP-1 cells infected with T. gondii. Compared with uninfected monocytes, T. gondii infection of monocytes reduced cell spreading and the number of activated β1 integrin clusters in contact with fibronectin during settling, an effect not observed in monocytes treated with lipopolysaccharide (LPS) or Escherichia coli. Furthermore, T. gondii infection disrupted the phosphorylation of focal adhesion kinase (FAK) at tyrosine 397 (Tyr-397) and Tyr-925 and of the related protein proline-rich tyrosine kinase (Pyk2) at Tyr-402. The localization of paxillin, FAK, and vinculin to focal adhesions and the colocalization of these proteins with activated β1 integrins were also impaired in T. gondii–infected monocytes. Using time-lapse confocal microscopy of THP-1 cells expressing enhanced GFP (eGFP)–FAK during settling on fibronectin, we found that T. gondii–induced monocyte hypermotility was characterized by a reduced number of enhanced GFP-FAK–containing clusters over time compared with uninfected cells. This study demonstrates an integrin conformation–independent regulation of the β1 integrin adhesion pathway, providing further insight into the molecular mechanism of T. gondii–induced monocyte hypermotility.

Prenatal smoking cessation has been described as an empathic action “for the baby,” but this has not been empirically demonstrated. We capitalized on a genetically-characterized extant dataset with outstanding measurement of prenatal smoking patterns and maternal face processing data (as an indicator of empathy) to test this hypothesis, and explore how empathy and smoking patterns may be moderated by a genetic substrate of empathy, the oxytocin receptor gene (OXTR). Participants were 143Caucasian women from the East Boston family study with repeated prospective reports of smoking level, adjusted based on repeated cotinine bioassays. Salivary DNA and face processing (Diagnostic Analysis of Nonverbal Accuracy-2) were assessed 14 years later at an adolescent follow-up of offspring. Two-thirds of participants reported smoking prior to pregnancy recognition. Of these, 21% quit during pregnancy; 56% reduced smoking, and 22% smoked persistently at the same level. A significant interaction between face processing and OXTR variants previously associated with increased sensitivity to social context, rs53576GG and rs2254298A, was found (β = −.181; p = .015); greater ability to identify distress in others was associated with lower levels of smoking during pregnancy for rs53576(GG)/rs2254298(A) individuals (p = .013), but not for other genotypes (p = .892). Testing this “empathy hypothesis of prenatal smoking cessation” in larger studies designed to examine this question can elucidate whether interventions to enhance empathy can improve prenatal smoking cessation rates.

A method for the preparation of AB-type monomers with complimentary fluoro-olefin and phenol functionalities is described. The three-step process is amenable to commercial scale up and uses widely-available, commercial reagents including bisphenols and perfluorocycloalkene (PFCA) compounds. Title compounds and intermediates were characterized via multi-nuclear NMR and FT-IR spectroscopy providing structural and mechanistic elucidation of PFCA step-growth polymerizations. The formation of vinyl and allyl substituted products were quantified and shown to be dependent on PFCA ring size and reaction medium. Nearly equal amounts of vinyl- and allyl-substituted products were observed with perfluorocyclohexene (PFCH) while 2–11% allyl-substitution was observed with perfluorocyclopentene (PFCP), depending on reaction medium polarity. Previous PFCA research and characterization of title compounds suggests that fluoride-catalyzed rearrangement is primarily responsible the formation of two substitution products. Polymerization of an AB-type monomer derived from bisphenol A is demonstrated and was shown to reproducibly result in film-forming polymers with higher molecular weights than previously described methods.

A new series of deep blue/blue emitting co-polymers are reported. Poly(9,9-dihexylfluorene-3,6-diyl and 2,7-diyl-co-2,8-dihexyldibenzothiophene-S,S-dioxide-3,7-diyl) derivatives p(F-S) of varying composition have been synthesised. The effects of two different S derivatives with dialkoxy sidechains, the F : S monomer feed ratio, and meta versus para conjugation with respect to the F units have all been investigated in terms of photophysics and polymer light-emitting diode (PLED) device performance in the architecture ITO/PEDOT:PSS/polymer/TPBi/LiF/Al. The meta polymers poly(9,9-dihexylfluorene-3,6-diyl-co-2,8-di(O-methylenecyclohexyl)dibenzothiophene-S,S-dioxide-3,7-diyl) p(Fm-SOCy) in three different co-monomer ratios, P1–3, give deep blue electroluminescence peaking at 415 nm, with the ratio of 70 : 30 p(Fm : SOCy) producing a maximum external quantum efficiency (EQE) ηext, max 2.7%, whilst the ratio of 85 : 15 gave the highest maximum brightness Lmax of 81 cd m−2, with CIE coordinates (0.17, 0.12) The analogous para series poly(9,9-dihexylfluorene-2,7-diyl-co-2,8-di(O-methylenecyclohexyl)dibenzothiophene-S,S-dioxide-3,7-diyl) p(Fp-SOCy) and poly(9,9-dihexylfluorene-2,7-diyl-co-2,8-dihexyloxydibenzothiophene-S,S-dioxide-3,7-diyl) p(Fp-SO6) in two different ratios, P4–7, produced blue emission peaking at ca. 450 nm. The ratio of 70 : 30 F : S units consistently gave better devices than the corresponding 50 : 50 co-polymers. It was also observed that co-polymers incorporating the bulkier SOCy derivatives gave more efficient and brighter devices, with polymer P5 attaining a remarkable ηext, max 3.2%, 4.4 cd A−1, 3.4 lm W−1 and maximum brightness 2500 cd m−2 with CIE (0.16, 0.18).

The design and synthesis of a series of quinuclidine-containing spirooxazolidines (“spiroimidates”) and their utility as α7 nicotinic acetylcholine receptor partial agonists are described. Selected members of the series demonstrated excellent selectivity for α7 over the highly homologous 5-HT3A receptor. Modification of the N-spiroimidate heterocycle substituent led to (1S,2R,4S)-N-isoquinolin-3-yl)-4′H-4-azaspiro[bicyclo[2.2.2]octane-2,5′oxazol]-2′-amine (BMS-902483), a potent α7 partial agonist, which improved cognition in preclinical rodent models.

We describe the synthesis of quinuclidine-containing spiroimidates and their utility as α7 nicotinic acetylcholine receptor (nAChR) partial agonists. A convergent synthetic route allowed for rapid SAR investigation and provided a diverse set of fused 6,5-heteroaryl analogs. Two potent and selective α7 nAChR partial agonists, (1′S,3′R,4′S)-N-(7-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-4H-1′-azaspiro[oxazole-5,3′-bicyclo[2.2.2]octan]-2-amine (20) and (1′S,3′R,4′S)-N-(7-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)-4H-1′-azaspiro[oxazole-5,3′-bicyclo[2.2.2]octan]-2-amine (21), were identified. Both agonists improved cognition in a preclinical rodent model of learning and memory. Additionally, 5-HT3A receptor SAR suggested the presence of a steric site that when engaged led to significant loss of affinity at that receptor.

Quinuclidine-containing spirooxazolines, as described in the previous report in this series, were demonstrated to have utility as α7 nicotinic acetylcholine receptor (α7 nAChR) partial agonists. In this work, the SAR of this chemotype was expanded to include an array of diazine heterocyclic substitutions. Many of the heterocyclic analogs were potent partial agonists of the α7 receptor, selective against other nicotinic receptors and the serotinergic 5HT3A receptor. (1'S,3'R,4'S)-N-(6-phenylpyrimidin-4-yl)-4H-1'-azaspiro[oxazole-5,3'-bicyclo[2.2.2]octan]-2-amine, a potent and selective α7 nAChR partial agonist, was demonstrated to improve cognition in the mouse novel object recognition (NOR) model of episodic memory.

The development of alpha7 nicotinic acetylcholine receptor agonists is considered a promising approach for the treatment of cognitive symptoms in schizophrenia patients. In the present studies we characterized the novel agent, (2R)-N-(6-(1H-imidazol-1-yl)-4-pyrimidinyl)-4'H-spiro[4-azabicyclo[2.2.2]octane-2,5'-[1,3]oxazol]-2'-amine (BMS-933043), in vitro and in rodent models of schizophrenia-like deficits in cognition and sensory processing. BMS-933043 showed potent binding affinity to native rat (Ki = 3.3 nM) and recombinant human alpha7 nicotinic acetylcholine receptors (Ki = 8.1 nM) and agonist activity in a calcium fluorescence assay (EC50 = 23.4 nM) and whole cell voltage clamp electrophysiology (EC50 = 0.14 micromolar (rat) and 0.29 micromolar (human)). BMS-933043 exhibited a partial agonist profile relative to acetylcholine; the relative efficacy for net charge crossing the cell membrane was 67% and 78% at rat and human alpha7 nicotinic acetylcholine receptors respectively. BMS-933043 showed no agonist or antagonist activity at other nicotinic acetylcholine receptor subtypes and was at least 300 fold weaker at binding to and antagonizing human 5-HT3A receptors (Ki = 2,451 nM; IC50 = 8,066 nM). BMS-933043 treatment i) improved 24 hour novel object recognition memory in mice (0.1-10 mg/kg, sc), ii) reversed MK-801-induced deficits in Y maze performance in mice (1-10 mg/kg, sc) and set shift performance in rats (1-10 mg/kg, po) and iii) reduced the number of trials required to complete the extradimensional shift discrimination in neonatal PCP treated rats performing the intra-dimensional/extradimensional set shifting task (0.1-3 mg/kg, po). BMS-933043 also improved auditory gating (0.56-3 mg/kg, sc) and mismatch negativity (0.03-3 mg/kg, sc) in rats treated with S(+)ketamine or neonatal phencyclidine respectively. Given this favorable preclinical profile BMS-933043 was selected for further development to support clinical evaluation in humans.

Interleukin-11 (IL-11) up-regulates the proliferative and invasive capacity of many cancers. Coexpression of glycoprotein 130 (GP130) and IL-11 receptor alpha (IL-11R alpha) is necessary for high-affinity binding of IL-11 to IL-11R alpha. This study investigated the expression of IL-11 and role of prostaglandin F-2 alpha-F-prostanoid receptor (FP receptor) signaling in the modulation of IL-11 expression in endometrial adenocarcinoma cells. Localization of IL-11, IL-11R alpha, and GP130 expression was performed by immunohistochemistry. IL-11 and regulator of calcineurin I isoform 4 (RCAN1-4) mRNA and protein expression were determined by real-time RT-PCR and/or enzyme-linked immunosorbent assay/Western blot analysis using Ishikawa endometrial adenocarcinoma cells stably expressing the FP receptor (FPS cells) and endometrial adenocarcinoma explants. IL-11 mRNA expression was significantly elevated in endometrial adenocarcinoma samples compared with normal endometrium and increased with tumor grade. IL-11 protein expression localized with FP receptor, IL-11R alpha, and GP130 in the neoplastic glandular epithelium of endometrial adenocarcinomas. Prostaglandin F-2 alpha-FP receptor signaling significantly elevated the expression of IL-11 mRNA and protein in a Gq-protein kinase C-calcium-calcineurin-nuclear factor of activated T cells-dependent manner in FPS cells. The calcineurin signaling pathway is known to be controlled by the RCAN (RCAN1-4). indeed, RCAN1-4 expression was significantly elevated in well-differentiated endometrial adenocarcinoma compared with normal endometrium and was found to decrease with tumor grade and negatively regulate IL-11 expression in vitro. This study has highlighted a new mechanism regulating IL-11 expression in endometrial adenocarcinoma cells by the FP receptor via the calcium-calcineurin-nuclear factor of activated T cells pathway. (Am J Pathol 2010, 176:435-445; DOI: 10.2353/ajpath.2010.090403)

Two new deep blue emitting co-polymers are synthesised and exploited in highly-efficient solution-processed polymer light-emitting diodes (PLEDs). A key molecular design feature is selective functionalisation of 9,9-diphenylfluorene to disrupt backbone π-conjugation, thereby promoting luminescence from short emissive domains (λELmax 422 and 401 nm for P1 and P2 devices, respectively). PLEDs using P1 and P2 in the hybrid architecture ITO/PEDOT:PSS/polymer/TPBi/LiF/Al show low turn-on voltages and remarkably high EQE values of ηext,max 3.3%, Lmax of 167 cd m−2 with CIExy coordinates (0.17, 0.10) for P1; and 3.9%, 274 cd m−2, CIExy (0.17, 0.07) for P2. These performances are currently among the best for PLEDs with CIEy ≤ 0.10.

We have previously shown that cytomegalovirus (CMV) can reactivate in lungs of nonimmunosuppressed patients during critical illness. Our recent work has shown that polymicrobial bacterial sepsis can trigger reactivation of latent murine CMV (MCMV). We hypothesize that MCMV reactivation following bacterial sepsis may be caused by inflammatory mediators. To test this hypothesis, BALB/c mice latently infected with Smith strain MCMV received sublethal intraperitoneal doses of lipopolysaccharide (LPS), tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), or saline. Lung tissue homogenates were evaluated for viral reactivation 3 weeks after mediator injection. Because LPS is known to signal via Toll-like receptor 4 (TLR-4) in mice, further studies blocking this signaling mechanism were performed using monoclonal MTS510. Finally, mice were tested with intravenous TNF-α to determine whether this would cause reactivation. All mice receiving sublethal intraperitoneal doses of LPS, TNF-α, or IL-1β had pulmonary reactivation of latent MCMV 3 weeks following injection, and LPS caused MCMV reactivation with kinetics similar to those for sepsis. When TLR-4 signaling was blocked, exogenous LPS did not reactivate latent MCMV. Intravenous TNF-α administration at near-lethal doses did not reactivate MCMV. Exogenous intraperitoneal LPS, TNF-α, and IL-1β are all capable of reactivating CMV from latency in lungs of previously healthy mice. LPS reactivation of MCMV appears dependent on TLR-4 signaling. Interestingly, intravenous TNF-α did not trigger reactivation, suggesting possible mechanistic differences that are discussed. We conclude that inflammatory disease states besides sepsis may be capable of reactivating CMV from latency.

Mitochondrial damage and dysfunction are thought to play an important role in the pathogenesis of sepsis-induced organ failures. Unfortunately, specific markers of mitochondrial damage in vital organs do not currently exist. Recently, carbomyl phosphate synthase (CPS)-1, a protein primarily localized to liver mitochondria, was found to be present in high concentrations in the plasma of septic humans. Thus, we hypothesized that the circulatory release of CPS-1 would correlate with mitochondrial damage or impaired mitochondrial function in the liver in a clinically relevant model of sepsis.Prospective, randomized, controlled animal study.University medical center research laboratory.Male, Balb/C mice, aged 10-12 wks.Animals were assigned to receive cecal ligation and puncture (CLP sepsis) or sham operation and compared with untreated controls. Plasma CPS-1 levels and liver mitochondrial variables, including morphology, respiratory activity, mass (i.e., cardiolipin content), and protein carbonylation, were assessed at various time points (8, 24, and 48 hrs and 6 days) after surgery.Oxidant stress (i.e., carbonylation) was detected within 8 hrs of CLP and persisted through 48 hrs. Plasma CPS-1 levels increased dramatically at 24 hrs, remained significantly elevated at 48 hrs, and normalized by 6 days in the sepsis group. Abnormalities of liver mitochondrial morphology and function coincided with increased plasma CPS-1 levels. Mitochondrial depletion in the liver was not due to cell death but was associated with increased lysosomal clearance. Increased expression of mitochondrial biogenesis factors preceded restoration of mitochondrial variables and normalization of CPS-1 levels by day 6.Circulating CPS-1 is a marker of mitochondrial damage and depletion in the liver during the subacute phase of CLP sepsis. From a mechanistic standpoint, mitochondrial depletion is not due to cell death but is apparently related to the removal of damaged mitochondria by lysosomes (i.e., autophagy), followed by repletion of mitochondrial populations. Further studies are needed to determine the clinical utility of CPS-1 as a marker of sepsis severity.

Background: Identifying sources of variation in the nicotine and nitrosamine metabolic inactivation pathways is important to understanding the relationship between smoking and cancer risk. Numerous UGT1A and UGT2B enzymes are implicated in nicotine and nitrosamine metabolism in vitro; however, little is known about their roles in vivo. Methods: Within UGT1A1, UGT1A4, UGT1A9, UGT2B7, UGT2B10, and UGT2B17, 47 variants were genotyped, including UGT2B10*2 and UGT2B17*2. The association between variation in these UGTs and glucuronidation activity within European and African American current smokers (n = 128), quantified as urinary ratios of the glucuronide over unconjugated compound for nicotine, cotinine, trans-3′-hydroxycotinine, and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), was investigated in regression models assuming a dominant effect of variant alleles. Results: Correcting for multiple testing, three UGT2B10 variants were associated with cotinine glucuronidation, rs2331559 and rs11726322 in European Americans and rs835309 in African Americans (P ≤ 0.0002). Additional variants predominantly in UGT2B10 were nominally associated with nicotine (P = 0.008–0.04) and cotinine (P = Conclusions: Findings from this initial in vivo study support a role for multiple UGTs in the glucuronidation of tobacco-related compounds in vivo, in particular UGT2B10 and cotinine glucuronidation. Impact: Findings also provide insight into ethnic differences in glucuronidation activity, which could be contributing to ethnic disparities in the risk for smoking-related cancers. Cancer Epidemiol Biomarkers Prev; 24(1); 94–104. ©2014 AACR.

Background Clinical abnormalities associated with drug therapy may lead to injury if untreated. Detection of preceding drug-related hazardous conditions (DRHCs) may provide the opportunity to prevent injury. This study evaluated the frequency of abnormal laboratory values attributed to the effect of drugs. Methods A total of 590 patients consecutively admitted to a surgical intensive care unit during a 3-month period were prospectively studied. Serum electrolyte, platelet, creatinine, glucose, magnesium, and liver enzyme concentrations were classified as either normal or abnormal using standard criteria. DRHCs were abnormal laboratory results determined to be caused by drugs. Results DRHCs were detected in 16.4% of patients, reflecting a rate of 47 events per 1,000 patient days. 66%, 22%, and 12% of DRHCs were categorized as either definite, probable, or possible, respectively. In 97% of suspected DRHCs, clinicians responded by ordering additional laboratory tests or changing drug regimens. Conclusions Monitoring laboratory values and assessing the association to medications may reduce the risk of injury consequent to adverse drug events.

Irrigation (or ‘floating’) of watermeadows in Europe was economically important before the 20th century because it enhanced grazing sward productivity in the spring. Today, few operational systems remain, but interest in floating as a technique is returning as concern grows over floodplain management, the fate of nutrients and the sustainability of agricultural production systems. Investigations into grass productivity, phosphorus and nitrogen relations were undertaken at a surviving watermeadow system, the Britford Site of Special Scientific Interest, Salisbury, England (1995–2001) and at an experimental site at Norwich, England (1998–2001). Irrigation significantly increased late-winter and early-spring grass productivity at Britford in the order of 60–80% over 5 weeks compared with un-irrigated controls. Phosphorus travelled across the system; some included with the clay fraction in suspended sediment, but available phosphorus in the topsoil reduced across the meadows by between one-half and one-third. Nitrate in irrigation water was utilised, but the meadow system did not reduce concentrations of dissolved nitrate. Increased nitrate concentration in soil-water is attributed to mineralisation after drainage. Annual nitrogen cycle process modelling suggests the present levels of production and soil organic matter accumulation are attributable to environmental eutrophication. A conceptual model is presented for enhanced spring growth following floating and it is concluded that, while temperature is the trigger that enhances productivity early in the season, this is achieved by enhancing the mineralisation of soil organic matter and release of available nitrogen. There was no evidence that watermeadows act as a sink for nitrogen mobile in surface waters but there was significant uptake of topsoil available phosphorus.

Mulch was added to sand and sandy loam soil in order to simulate repeated additions of organic matter into soils of the tropics where high temperatures cause rapid decomposition. The experiments used pots (0.003 m3) and lysimeters (0.18 m3) to investigate the effect of farmyard manure (FYM) and municipal waste compost (MWC) incorporation on temperature, soil water retention and evaporation, and MWC on transpiration, photosynthetic rate and yield as performance indicators for cowpea [Vigna unguiculata (L)]. Incorporation of FYM and MWC in un-cropped pots increased early stage evaporation compared with surface mulch treatments and compost mulch conserved soil water at depths between 0.05 and 0.6 m. Repeated incorporation of compost in lysimeters increased crop water availability between 7 and 10% and yield increased 3 to 5 times, despite organic matter content remaining below 1%. Results show cowpea performance indicators in light, low fertility soils of the tropics would benefit substantially from repeated...

A kinase-anchoring proteins (AKAPs) coordinate cAMP-mediated signaling by binding and localizing cAMP-dependent protein kinase (PKA), using an amphipathic helical docking motif. Peptide disruptors of PKA localization that mimic this helix have been used successfully to assess the involvement of PKA in specific signaling pathways. However, these peptides were developed as disruptors for the type II regulatory subunit (RII) even though both RI and RII isoforms can bind to AKAPs and have discrete functions. To evaluate the effects of each localized isoform, we designed peptides that specifically bind to either RI or RII. Using a peptide array, we have defined the minimal binding sequence of dual specific-AKAP 2 ( d -AKAP2), which binds tightly to both RI and RII. Side-chain requirements for affinity and isoform specificity were evaluated by using a peptide substitution array where each position along the A kinase binding domain of d -AKAP2 was substituted by the other 19 l -amino acids. This array comprises 513 single-site substitution analogs of the d -AKAP2 sequence. Peptides containing single and multiple mutations were evaluated in a quantitative fluorescence binding assay and a cell-based colocalization assay. This strategy has allowed us to design peptides with high affinity ( K D = 1–2 nM) and high specificity for RIα versus RIIα. These isoform-specific peptides will be invaluable tools to evaluate functional differences between localized RI and RII PKA and are RIα-specific disruptors. This array-based analysis also provides a foundation for biophysical analysis of this docking motif.

A second form of tryptophan hydroxylase (TPH) is expressed in the brain by the gene Tph2. The presence of the gene was discovered when Tph 1(-/-)mice were found to express normal amounts of serotonin in brain, but not in the periphery. Additionally, Tph1(-/-) mice showed no observed behavioral differences from wild-type littermates. Veenstra-Vanderweele and Cook discuss the ramifications of these findings and what they might mean for designing drugs that target the expression and activity of TPH in differing tissues.

Emissions of N2O were measured following combined applications of inorganic N fertiliser and crop residues to a silt loam soil in S.E. England, UK. Effects of cultivation technique and residue application on N2O emissions were examined over 2 years. N2O emissions were increased in the presence of residues and were further increased where NH4NO3 fertiliser (200 kg N ha−1) was applied. Large fluxes of N2O were measured from the zero till treatments after residue and fertiliser application, with 2.5 kg N2O-N ha−1 measured over the first 23 days after application of fertiliser in combination with rye (Secale cereale) residues under zero tillage. CO2 emissions were larger in the zero till than in the conventional till treatments. A significant tillage/residue interaction was found. Highest emissions were measured from the conventionally tilled bean (Vicia faba) (1.0 kg N2O-N ha−1 emitted over 65 days) and zero tilled rye (3.5 kg N2O-N ha−1 over 65 days) treatments. This was attributed to rapid release of N following incorporation of bean residues in the conventionally tilled treatments, and availability of readily degradable C from the rye in the presence of anaerobic conditions under the mulch in the zero tilled treatments. Measurement of 15N-N2O emission following application of 15N-labelled fertiliser to microplots indicated that surface mulching of residues in zero till treatments resulted in a greater proportion of fertiliser N being lost as N2O than with incorporation of residues. Combined applications of 15N fertiliser and bean residues resulted in higher or lower emissions, depending on cultivation technique, when compared with the sum of N2O from single applications. Such interactions have important implications for mitigation of N2O from agricultural soils.

Three patients are reported with a prior history of alcohol misuse accompanied by abnormal liver toxicity tests and other indices of alcohol misuse. A decreased but persistently raised serum gamma-glutamyl transferase activity during subsequent abstinence or controlled drinking was interpreted incorrectly as due to continued alcohol misuse whereas obesity-related fatty liver was the probable cause. The value of serum carbohydrate deficient transferrin assays in the differential diagnosis of abnormal liver toxicity tests is emphasized.

Systematic evaluation of the structure–activity relationships of a new class of 4-aryl-pyridine glucagon antagonists led to the discovery of potent analogues bearing a key secondary hydroxy moiety as seen in compound 1a . Due to the importance of this new class of compounds, it became necessary to establish an efficient synthesis of the pure enantiomer. A resolution and two chiral syntheses of alcohol 1a were discovered and herein presented.

A series of N-benzylpyroglutamyl-L-phenylalanine derivatives bearing carbamoyl substituents in the 3- or 4-positions was prepared and assayed for inhibition of the interaction between VCAM and VLA-4. Potent inhibition was observed in a number of analogues with substitution in the 4-position favored over the 3-position. A crystal structure of the key intermediate 25 indicates that it accesses a low energy conformation which closely matches key pharmacophores of a structurally characterized cyclic peptide.

This study was conducted to examine the relationship between whole blood serotonin level and behavioral symptoms in 78 subjects with autism. No significant associations were found between serotonin level and the primary behavioral outcome measures. However, a significant inverse relationship between serotonin level and self-injury was demonstrated.

Although it is well known that B-vitamin deficiencies directly affecting the brain are common in alcohol misuse, no concise guidelines on the use of vitamin supplements in alcohol misusers currently exist in the UK. The purpose of this study was to assess current practice and opinion among UK physicians. Questionnaires were completed by a total of 427 physicians comprising Accident and Emergency (A&E) specialists and psychiatrists, with a response rate of 25%. The main findings were that vitamin deficiency was perceived as being uncommon amongst alcohol misusers (

Aviation accidents due to alcohol consumption by aircrew appear to be rare, especially in commercial aviation. However, a small proportion of general aviation accidents are attributable to alcohol use by aircrew, and aircrew are not well informed about the metabolism of alcohol and its effects on performance. Furthermore, there is evidence that aircrew performance may be impaired by alcohol consumption even after their Blood Alcohol Concentration has returned to "zero" (i.e. < 5 mg/dl). Accidents caused by impairment of aircrew performance by alcohol may therefore not be attributed to alcohol use at all. Aviation safety relies upon faultless human performance and is thus highly sensitive to alcohol-related impairment of performance. This paper provides a review of research regarding aircrew alcohol consumption, impairment of aircrew performance by alcohol, incidence of aviation accidents attributable to alcohol use by aircrew, and other related subjects.

Friedel-Crafts acylations of 3-alkyl-1-(phenylsulfonyl)pyrroles appear to be kinetically favored at the adjacent C-2 position but rearrangement to the C-5 position can occur after prolonged reaction times. This reversible Friedel-Crafts methodology has been employed for the synthesis of the ant trail pheromone, methyl 4-methylpyrrole-2-carboxylate.

Acidification of liquid egg prior to drying did not improve subsequent storage life, although pH measurements showed that powder from untreated egg became acid more rapidly during storage. Reduction in the moisture content (total volatiles) from 4.7 to 3.0% doubled, and reduction from 4.7 to 1.7% tripled, the storage life of dried whole egg powder as assessed by fluorescence tests. The maximum storage life predicted for the low moisture powder by this test was only 36 wk. at 27 °C. and 5 wk. at 38 °C. Palatability tests suggested that the product was somewhat less perishable, as a powder of 1.7% moisture was considered fit for use as an egg dish after 64 wk. at 27 °C. Gas-packing low moisture powders in an atmosphere of carbon dioxide appeared to be slightly more effective as a means of retaining palatability than packing in an atmosphere of air or nitrogen, but was particularly effective in preventing loss of solubility (assessed by potassium chloride values) during storage.

Friedel-Crafts acylations of 3-alkyl-1-(phenylsulfonyl)pyrroles appear to be kinetically favored at the adjacent C-2 position but rearrangement to the C-5 position can occur after prolonged reaction times. This reversible Friedel-Crafts methodology has been employed for the synthesis of the ant trail pheromone, methyl 4-methylpyrrole-2-carboxylate.

5-Hydroxyalkyl-4-phenylpyridines have been identified as a novel class of glucagon antagonists with potential utility for the treatment of diabetes. A lead structure with moderate activity was discovered through a high throughput screening assay. Structure–activity relationships led to the discovery of a potent antagonist, IC 50 =0.11 μM, more than 60-fold improvement over the lead structure.

A series of N-benzylpyroglutamyl-L-phenylalanine derivatives bearing carbamoyl substituents in the 3- or 4-positions was prepared and assayed for inhibition of the interaction between VCAM and VLA-4. Potent inhibition was observed in a number of analogues with substitution in the 4-position favored over the 3-position. A crystal structure of the key intermediate 25 indicates that it accesses a low energy conformation which closely matches key pharmacophores of a structurally characterized cyclic peptide.

OF several recent syntheses of dl-methionine, that commencing with y-butyrolactone1 is relatively long, while others2 are additionally inconvenient in that they employ methyl-B-chloroethylsulphide, a vesicant intermediate. It seems, therefore, desirable to state that the following series of reactions has been found to afford an overall yield of dl-methionine of 29 percent (that is, based on acrolein), and to be free from disadvantages such as those mentioned above

Polymorphism of the dopamine transporter genotype (DAT1) confers a small but significant susceptibility to attention deficit hyperactivity disorder (ADHD). We examined whether the volume of the head of caudate, a striatal structure with high DAT expression that is important for inhibitory function, differs by DAT1 in children diagnosed with the disorder relative to age and IQ matched controls. Volume of the head of caudate was delineated in the right and left hemisphere and compared between 7- and 13-year-old children with and without ADHD (combined type) who were carriers of two (10/10) or one (9/10) copy of the 10-repeat DAT1 allele. Caudate volumes were overall smaller in 10/10 than 9/10 children, particularly in the left than right hemisphere. While DAT1 effects did not vary by ADHD diagnosis, overall caudate volumes were smaller in ADHD relative to control children. Altered caudate development associated with 10-repeat homozygosity of DAT1 may contribute susceptibility to ADHD.

Ninety-six crossbred barrows were randomly allotted to five replications of four treatments to determine the effects of diets containing 20% extruded full-fat soybeans (FFS) or 4% butter (B) on the growth, composition, and sensory characteristics of finishing pigs. Pigs were housed five per pen, except for one replication that had four pigs per pen, in environmentally regulated barns. Pigs were given ad libitum access to a corn-soybean meal diet for 11 wk (control); a corn-extruded soybeans and soft wheat midds diet for 11 wk (FFS-11); a corn-soybean meal diet for 6 wk, changed to a corn-extruded soybeans and soft wheat midds diet for 5 wk (FFS-5); or a corn-soybean meal diet for 6 wk, changed to a corn-soybean meal, soft wheat midds, and 4% butter diet for 5 wk (B). Feed intake and weight gain were measured once every 2 wk. Pigs were slaughtered and carcass data were collected. Sensory characteristics (tenderness, juiciness, pork flavor intensity, off-flavor intensity, and overall acceptability), shear force, moisture, and fat content were determined for the longissimus muscle. Sensory characteristics (pork flavor, off-flavor, rubbery, cohesiveness, and juiciness) and 2- thiobarbituric acid values were determined for ground pork (30% fat) after 1, 4, and 7 d for control, FFS-11, and B treatments. No differences (P > .05) in ADG were observed between diets. Feed efficiency of the FFS-11 group was better (P < .05) than that of the control or B groups. No consistent differences were observed for carcass, sensory, or shear-force characteristics of the longissimus muscle or ground pork.(ABSTRACT TRUNCATED AT 250 WORDS)

Previous work from this laboratory has shown that proline isomerization can be used as a kinetic trap for intermediates in the folding of RNase A. There are two classes of unfolded RNase A: a fast-folding class, U/sub F/, and a major (80%) slow folding class, U/sub S/ (1), The U/sub F/ in equilibrium U/sub S/ interconversion reaction in unfolded RNase A has been shown to be acid-catalyzed and to have other specific properties of proline isomerization. Thus, it is probable that U/sub S/ contains one or more proline isomers in wrong cis or trans conformations. We have evidence that, in the folding of RNase A at low temperatures, structural intermediates accumulate before proline isomerization. An assay has been devised for wrong proline isomers which can be used during folding. These data point to the existence of a quasi-native intermediate, I/sub N/, formed late in the folding reaction. I/sub N/ appears to be folded, as measured both by tyrosine absorbance and by 2'CMP binding, but has one or more prolines in the wrong cis-trans conformation, as measured by the assay for wrong proline isomers. Proline isomerization provides a suitable trap for intermediates in folding since: (a) proline isomerization is the finalmore » and slowest step of the folding reaction; (b) folding does occur before proline isomerization takes place; and (c) the kinetic trap is applicable to the major form of the unfolded protein (U/sub S/).« less

This paper reports on ceramic-to-metal seals prepared by sputtering a titanium metallizing layer onto ceramic disks and then brazing to metal tubes. The ceramics used were alumina, MACOR, spinel, AlON, and a mixture of Al{sub 2}O{sub 3} and Si{sub 3}N{sub 4}. Except for the MACOR, which was brazed to a titanium tube, the ceramics were brazed to niobium tubes. The seals were leak tested and then sent to Los Alamos National Laboratory, where they were irradiated using the spallation neutron source at the Los Alamos Meson Physics Facility. Following irradiation for {approx} 90 days to a fluence of 2.8 {times} 10{sup 23} n/m{sup 2}, the samples were moved to hot cells and again leak tested. Only the MACOR samples showed any measurable leaks. One set of samples was then pressurized to 6.9 MPa (1000 psi) and subsequently leak tested. No leaks were found. Bursting the seals required hydrostatic pressures of at least 34 MPa (5000 psi). The high seal strength and few leaks indicate that ceramic-to-metal seals can resist radiation-induced degradation.

We have fabricated polycrystalline diamond hemispheres by hot-filament CVD (HFCVD) in spherical cavities wet-etched into a high temperature glass substrate CTE matched to silicon. Hemispherical resonators 1.4 mm in diameter have a Q of up to 143 000 in the fundamental wineglass mode, for a ringdown time of 2.4 s. Without trimming, resonators have the two degenerate wineglass modes frequency matched as close as 2 Hz, or 0.013% of the resonant frequency (~16 kHz). Laser trimming was used to match resonant modes on hemispheres to 0.3 Hz. Experimental and FEA energy loss studies on cantilevers and hemispheres examine various energy loss mechanisms, showing that surface related losses are dominant. Diamond cantilevers with a Q of 400 000 and a ringdown time of 15.4 s were measured, showing the potential of polycrystalline diamond films for high Q resonators. These resonators show great promise for use as hemispherical resonant gyroscopes (HRGs) on a chip.

A series of 4,5-disubstituted cis-pyrrolidinones was investigated as inhibitors of 17beta-HSD II for the treatment of osteoporosis. Biochemical data for several compounds are given. Compound 42 was selected as the lead candidate.

Wernicke's encephalopathy, a disorder with significant mortality and high morbidity, is common amongst alcohol-dependent patients. Thiamine deficiency appears to play a key role in its aetiology, and parenteral high-dose thiamine is effective in prophylaxis and treatment. Unfortunately, reports of rare anaphylactoid reactions have led to a dramatic reduction in the use of parenteral thiamine, and it is possible that this change in treatment has led, or will lead, to an increase in morbidity and mortality. There is a need for education of doctors who treat alcohol-dependent patients, in order to ensure appropriate use of parenteral thiamine in prophylaxis and treatment of this disorder.

Resin Properties Cure Mechanism Applications Keywords: vinyl ester resins; free-radical cure mechanism

17β-Hydroxysteroid dehydrogenase II (17β-HSD II) antagonists may provide an important way into preventing the onset of osteoporosis. The discovery of 4,5-disubstituted cis-pyrrolidinones as 17β-HSD II inhibitors led to the development of an efficient intramolecular Michael addition, followed by catalytic hydrogenation to obtain the desired cis configuration.

4,5-Disubstituted cis-pyrrolidinones were investigated as inhibitors of type II 17beta-hydroxysteroid dehydrogenase (17beta-HSD). Early structure-activity relationship patterns for this class of compounds are discussed.

The tubulin-binding agent combretastatin A-4-phosphate (CA-4-P), rapidly disrupts the vascular network of tumors leading to secondary tumor cell death. In vitro, CA-4-P destabilizes microtubules and causes endothelial cell death. In this study we analyze the mechanisms by which CA-4-P induces the death of proliferating endothelial cells. We demonstrate that at/=7.5 nmol/L, CA-4-P damages mitotic spindles, arrests cells at metaphase, and leads to the death of mitotic cells with characteristic G(2)/M DNA content. Mitotic arrest was associated with elevated levels of cyclin B1 protein and p34(cdc2) activity. Inhibition of p34(cdc2) activity by purvalanol A caused mitotic-arrested cells to rapidly exit mitosis, suggesting that sustained p34(cdc2) activity was responsible for metaphase arrest. Pharmacological prevention of entry into mitosis protected cells from undergoing cell death, further establishing the link between mitosis and cell death induction by CA-4-P. CA-4-P-mediated cell death shared characteristics of apoptosis but was independent of caspase activation suggesting the involvement of a non-caspase pathway(s). These data suggest that induction of apoptosis in endothelial cells by CA-4-P is associated with prolonged mitotic arrest. Therefore, by activating cell death pathways, CA-4-P, in addition to being an effective anti-vascular agent, may also interfere with regrowth of blood vessels in the tumor.

A series of (5-(2H)-isoxazolonyl) ureas were developed as nanomolar inhibitors of hormone-sensitive lipase, an enzyme of potential importance in the treatment of diabetes.

High-pressure freezing is applicable to both morphological and immunocytochemical studies. We are investigating the morphogenesis of foot-and-mouth disease virus and African swine fever virus by the use of high-pressure freezing of infected cells. Foot-and-mouth disease virus particles are not detected in sections of conventionally immersion-fixed infected cells, but when the cells are prepared by high-pressure freezing, newly formed virions are readily seen throughout the cell. We report two methods for high-pressure freezing of virally infected cells: first, two sapphire discs frozen 'face to face' with a narrow spacer to prevent cell damage and, second, a fibrous filter substrate that can be easily cut into discs to fit into the freezing planchettes. Cells readily adhere to the fibres in vitro, and the complete disc can be rapidly transferred to the planchettes for freezing. Immunolabelling studies of the microneme proteins of the parasite Eimeria tenella indicate that high-pressure freezing followed by freeze-substitution in acetone with uranyl acetate allows high-sensitivity immunolabelling for these proteins.

Purpose: Despite its clinical success, methotrexate (MTX) therapy is associated with toxicities such as seizures, the pathogenesis of which remains unclear. It has been suggested that hyperhomocysteinemia is caused by MTX and is responsible for its neurotoxic effects. The purposes of this study were to explore whether hyperhomocysteinemia was related to MTX administration and toxicity and whether homocysteine or MTX toxicity differed by methylenetetrahydrofolate reductase (MTHFR) or reduced folate carrier (RFC) genetic polymorphisms. Patients and Methods: We studied 53 children with newly diagnosed acute lymphoblastic leukemia who were consecutively treated on a single clinical protocol that included two courses of high-dose MTX (high-dose methotrexate [HDMTX]; 2.5 or 5.0 g/m2 per day) as consolidation therapy. Results: The study participants’ median plasma homocysteine concentrations at 23 and 44 hours after HDMTX (9.00 μmol/L and 10.12 μmol/L, respectively) were greater than the concentrations immediately before HDMTX (5.77 μmol/L, P < .0001 for both comparisons). Seven days after HDMTX treatment, their plasma concentration returned to baseline. Nine patients experienced seizures, and five patients experienced thrombosis during the first 15 months of therapy, with a tendency for there to be higher plasma homocysteine in patients with seizures across all time points (P = .063) but not in patients with thrombosis (P = .59). We observed no significant differences in plasma or cerebrospinal fluid homocysteine levels or in toxicity based on the MTHFR 677C/T or RFC 80G/A genotypes. Conclusion: We conclude that homocysteine was transiently elevated after HDMTX and may be related to seizure risk in children with leukemia.