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Phosphonates are produced across all domains of life and used widely in medicine and agriculture. Biosynthesis almost universally originates from the enzyme phosphoenolpyruvate mutase (Ppm), EC 5.4.2.9, which catalyzes O-P bond cleavage in phosphoenolpyruvate (PEP) and forms a high energy C-P bond in phosphonopyruvate (PnPy). Mechanistic scrutiny of this unusual intramolecular O-to-C phosphoryl transfer began with the discovery of Ppm in 1988 and concluded in 2008 with computational evidence supporting a concerted phosphoryl transfer via a dissociative metaphosphate-like transition state. This mechanism deviates from the standard 'in-line attack' paradigm for enzymatic phosphoryl transfer that typically involves a phosphoryl-enzyme intermediate, but definitive evidence is sparse. Here we review the experimental evidence leading to our current mechanistic understanding and highlight the roles of previously underappreciated conserved active site residues. We then identify remaining opportunities to evaluate overlooked residues and unexamined substrates/inhibitors.

We quickly approach a future where Internet of Things (IoT) devices are the norm. In this scenario, humans are surrounded by a multitude of heterogeneous devices that assist them in almost every aspect of their daily routines. The realization of this future demands strong authentication guarantees to ensure that these devices are not abused and that their users are not endangered. However, providing authentication for these systems is challenging due to the high heterogeneity of IoT applications. In this paper, we first review several IoT application scenarios and promising authentication methods for each. We identify th e key characteristics of each IoT application scenario, present the strengths and weaknesses of prominent authentication methods from the literature, and review which authentication methods have been proposed in the literature for each application. Then, we present a novel authentication method for IoT based on electromagnetic noise. The key advantage of electromagnetic noise is that any electronic device intrinsically generates electromagnetic noise during normal operation. We extract features from these electromagnetic emanations and use machine learning algorithms to identify devices based on these features. Our method achieves 77% accuracy when identifying devices among a set of seven devices.

Background In the context of the demand for more efficient herbicides, the aim of the present work was to synthesize anilides via simple methods, and evaluate their herbicidal activities through seed germination assays. In silico studies were carried out to identify the enzyme target sites in plants for the most active anilides. Results A total of 18 anilides were prepared via one-pot reaction in yields that varied from 36 to 98% through reactions of anilines with sorbic chloride and hexanoic anhydride. According to seed germination assays in three dicotyledonous and one monocotyledonous plant species, the most active anilides showed root and shoot growth inhibition superior to that of Dual (S-metolachlor). In silico studies indicated that histone deacetylase was the probable enzyme target site in plants for these substances. The affinities of the most active anilides for the binding sites of this enzyme were equal to or higher than those calculated for its inhibitors. Conclusion Anilides 4d, 4e, 4 g, and 4 h are promising candidates for the development of novel herbicides. According to in silico studies, they inhibit histone deacetylase in plants, which can be exploited for the development of new weed control methods. © 2018 Society of Chemical Industry.

Analyzing the structure of hybrid zones is important for inferring their origin, dynamics and evolutionary significance. We examined the geographic structure of phenotypic and genetic variation in the contact zone between two Mexican red oaks, Quercus affinis and Q. laurina. A total of 105 individuals from seven populations were sampled along a 600-km latitudinal gradient representing the distribution area of the two species and their contact zone. Individuals were genotyped for nine nuclear and four chloroplast DNA microsatellite loci (ncSSR and cpSSR, respectively), and characterized for several leaf and acorn traits. The cpSSR data revealed extensive haplotype sharing among populations of the two species, while a Bayesian assignment analysis based on ncSSRs identified two main genetic groups, each corresponding to one of the species, and two populations in the contact zone showing evidence of admixture. The proportion of genetic ancestry in the populations was strongly associated with latitude and showed a pattern of variation with the shape of a narrow sigmoidal cline. The variation in three of the seven phenotypic traits was partially congruent with molecular variation, while the other traits did not conform to a geographic cline but instead were correlated with environmental variables. In conclusion, the hybrid zone between the two oak species has some of the characteristics of a tension zone, but heterogeneous variation across traits suggests differential introgression and the action of extrinsic selection.

Multiple myeloma (MM) is a common and deadly cancer of blood plasma cells. A unique feature of MM is the extremely low somatic mutation rate of the p53 tumor suppressor gene, in sharp contrast with about half of all human cancers where this gene is frequently mutated. Eleven miRNAs have been reported to repress p53 through direct interaction with the 3' untranslated region. The expression of nine of them is higher in MM plasma cells than in healthy donor counterparts, suggesting that miRNA overexpression is responsible for p53 inactivation in MM. Here, we report that the environmental carcinogen benzo[a]pyrene (BaP) upregulated the expression of seven p53-targeting miRNAs (miR-25, miR-15a, miR-16, miR-92, miR-125b, miR-141, and miR-200a), while 2,3,7,8-tetrachlorodibenzo-ρ-dioxin (TCDD) upregulated two of them (miR-25 and miR-92) in MM cells. The miR-25 promoter was activated by both BaP and TCDD, and this response was mediated by the aryl hydrocarbon receptor (AhR). We screened 727 compounds that inhibit MM cell survival and down-regulate the expression of p53-targeting miRNAs. We found that (-)-epigallocatechin-3-gallate (EGCG), a constituent of green tea and a major component of the botanical drug Polyphenon® E, reduced the expression of four p53-targeting miRNAs, including miR-25, miR-92, miR-141, and miR-200a. Collectively, these data implicate polycyclic aromatic hydrocarbons and AhR in the regulation of p53-targeting miRNAs in MM and identify a potential therapeutic and preventive agent to combat this deadly disease.

SUMMARY The timing and magnitude of exposure to preovulatory estradiol followed by post-ovulatory progesterone (periovulatory endocrine milieu) in cattle modulate endometrial gene expression, histotroph composition, and conceptus development, but the mechanisms underlying this regulation remain unknown. Using an experimental model based on the modulation of follicle growth, this work aimed to evaluate if the polyamine metabolic pathway is regulated by the periovulatory endocrine milieu. Nelore cows were manipulated to ovulate small (n = 15) or large (n = 15) follicles, then the profiles of polyamines and their synthetic enzymes were compared between groups. Transcripts for the enzymes of this pathway, ornithine decarboxylase 1 (ODC1; the rate-limiting enzyme in polyamine biosynthesis) protein quantification, adenosylmethionine decarboxylase 1 (AMD1) protein immunolocalization, and concentrations of the different polyamines (putrescine, spermidine, and spermine) were respectively quantified by quantitative reverse-transcriptase PCR, immunoblotting, immunohistochemistry, and gas chromatography-mass spectrometry in both the endometrium and uterine flushing. No differences in gene and protein expression or concentration of polyamines were observed between groups. There were significant correlations between the relative abundance of ODC1 and spermidine/spermine N1-acetyltransferase 1 (SAT1) transcripts as well as between antizyme inhibitor 1 (AZIN1) and adenosylmethionine decarboxylase 1 (AMD1) transcripts. In conclusion, our results show that the polyamine metabolic pathway is present and functional, but not regulated by the periovulatory endocrine milieu in the bovine endometrium. Mol. Reprod. Dev. 81: 584–594, 2014. © 2014 Wiley Periodicals, Inc.

RpoN is a σ(54) factor regulating essential virulence gene expression in several plant pathogenic bacteria, including Pseudomonas syringae and Pectobacterium carotovorum. In this study, we found that mutation of rpoN in the fire blight pathogen Erwinia amylovora caused a nonpathogenic phenotype. The E. amylovora rpoN Tn5 transposon mutant rpoN1250::Tn5 did not cause fire blight disease symptoms on shoots of mature apple trees. In detached immature apple fruits, the rpoN1250::Tn5 mutant failed to cause fire blight disease symptoms and grew to population levels 12 orders of magnitude lower than the wild-type. In addition, the rpoN1250::Tn5 mutant failed to elicit a hypersensitive response when infiltrated into nonhost tobacco plant leaves, and rpoN1250::Tn5 cells failed to express HrpN protein when grown in hrp (hypersensitive response and pathogenicity)-inducing liquid medium. A plasmid-borne copy of the wild-type rpoN gene complemented all the rpoN1250::Tn5 mutant phenotypes tested. The rpoN1250::Tn5 mutant was prototrophic on minimal solid and liquid media, indicating that the rpoN1250::Tn5 nonpathogenic phenotype was not caused by a defect in basic metabolism or growth. This study provides clear genetic evidence that rpoN is an essential virulence gene of E. amylovora, suggesting that rpoN has the same function in E. amylovora as in P. syringae and Pe. carotovorum.

A simple, rapid, and sensitive method for the analysis of paroxetine, in tablets as well as the pure drug, by circular dichroism is described. The method was validated for repeatability, linearity, limit of detection, limit of quantification, and recovery according to the International Conference on Harmonization guidelines. Excellent results were obtained, within the globally accepted validation reference values, particularly taking into account the low concentration levels investigated. This is the first report of the quantitation of paroxetine, a chiral drug, without previous separation of the analyte. Additionally, the solid state CD spectrum of PXT was obtained. Chirality 25:211–214, 2013. © 2013 Wiley Periodicals, Inc.

Neonatal lupus (NL) is a syndrome whose clinical manifestations include transient cutaneous rash, hematologic and hepatic laboratory abnormalities, and irreversible cardiac damage. The cardiac manifestations of NL (cardiac-NL), which include atrioventricular conduction defects (heart block) and life-threatening cardiomyopathy, are associated with substantial mortality and morbidity (1). While maternal antibodies to components of the SSA/Ro-SSB/La ribonucleoprotein complex are necessary for the development of disease, the rarity of advanced injury at 2–5% suggests that fetal factors are highly contributory (2) to a complex pathogenic cascade which links autoantibody to scar. A fetal genetic contribution to the development of cardiac-NL is supported by a high sibling risk ratio (λs= 10–3000), recurrence rate of approximately 18%, concordance rates in monozygotic twins of 33% (reviewed in 3), as well as the results from our published genome-wide association study (GWAS) in cardiac-NL (4). While the precise pathogenic cascade is yet to be fully defined, one scenario posits that injury is initiated by increased apoptosis of the cardiocytes which exposes Ro/SSA antigens and associated ssRNA generating an immune complex phagocytosed by macrophages, which sustain both an inflammatory response and a fibrotic response, resulting in the scarring of the heart (5). This hypothesis is supported by the results from our GWAS (4), where several genome-wide significant variants were identified in the MHC region, as well as at 21q22.3, near the erythroblast transformation-specific transcription factor ERG, which is a regulator of embryonic development, cell proliferation, differentiation, angiogenesis, inflammation, and apoptosis. Despite the significant associations identified in the GWAS (4), we hypothesize that there may be a global enrichment of specific pathways and genetic variation potentially overlooked if only the most significant statistical associations are considered. A judicious and efficient way to identify genetic variation predisposing to complex diseases is the application of a candidate gene framework that incorporates prior biological knowledge. Unlike agnostic GWAS, this approach narrows the hypothesis space to provide a more focused and powerful examination of the data. Indeed, several studies have shown that an essentially Bayesian approach of selecting candidate genes serves to increase the reliability and likelihood of finding genes that are truly associated with disease (6, 7). In accord with these considerations, we tested for a potential enrichment of significant associations in genes with candidate biological functions by mining the cardiac-NL GWAS. Given their potential contribution to inflammation, fibrosis, and conduction abnormalities in the heart, we selected genes with functions contributing to apoptosis, calcium electrogenesis, immune function, and fibrosis. Figure 1 illustrates our approach for the identification of cardiac-NL risk loci. Several associations within genes in candidate pathways relevant to the pathogenesis of cardiac-NL were identified supporting the potential of this approach for gene discovery. Figure 1 Approach for the identification of genes predisposing to cardiac manifestations of neonatal lupus (cardiac-NL). An approach that focuses the hypothesis space based on biological knowledge provides a more powerful and efficient way to identify disease-causing ...

Objective Neonatal lupus (NL) occurs in fetuses exposed to maternal anti-SSA/Ro and/or anti-SSB/La antibodies, although the mothers themselves may not manifest any clinical disease. A focus on transmission of risk factors for NL from maternal grandparents to mothers of children with NL may yield dividends toward understanding the aggregation of autoantibodies and genetic factors in affected families. This study was perforned to determine the role of maternal grandparents in the development of the autoimmune phenotype of mothers of children with NL. Methods Fifty-one mothers of children with cardiac and/or cutaneous NL, 48 maternal grandmothers, and 35 maternal grandfathers in the Research Registry for Neonatal Lupus were interrogated for clinical symptoms by questionnaire and underwent laboratory assessments, including determination of anti-SSA/Ro and anti-SSB/La antibody status (by enzyme-linked immunosorbent assay) and genotype at rs1800629 (TNFα) and rs7775397 (C6orf10) (allelic discrimination). The transmission disequilibrium test (TDT) was computed to test for nonrandom transmission from maternal grandparents to mothers of children with NL. Results The common phenotypic feature in mothers of children with NL was the autoantibody and not the clinical profile; 7 had lupus, 14 had Sjogren's syndrome, 7 had both, and 23 were asymptomatic. Mothers of children with NL were significantly enriched for the risk alleles at both TNFα and C6orf10. The grandparents of children with NL carried minimal burden for autoimmune disease or abnormal antibody production and were not enriched in the genetic risk factors. However, the TDT analysis showed significant excess transmission of the risk alleles at both TNFα (odds ratio [OR] 6.67, P = 3.93 × 10−4) and C6orf10 (OR 35.0, P = 3.74 × 10−5) to mothers of children with NL. Conclusion Mothers of children with NL are enriched for the TNFα and C6orf10 risk alleles, which are preferentially inherited from the asymptomatic maternal grandparents. These findings support the hypothesis that the development of NL and genetic etiology are multigenerational.

BACKGROUND AND PURPOSE Independent studies in experimental models of Trypanosoma cruzi appointed different roles for endothelin-1 (ET-1) and bradykinin (BK) in the immunopathogenesis of Chagas disease. Here, we addressed the hypothesis that pathogenic outcome is influenced by functional interplay between endothelin receptors (ETAR and ETBR) and bradykinin B2 receptors (B2R).

Gangliosides have been considered as potential targets for immunotherapy because they are overexpressed on the surface of melanoma cells. However, immunization with purified gangliosides results in a very poor immune response, usually mediated by IgM antibodies. To overcome this limitation, we immunized mice with R24, a monoclonal antibody (mAb) that recognizes the most tumor-restricted ganglioside (GD3); our goal was to obtain anti-idiotype (Id) antibodies bearing the internal image of GD3. Animals produced anti-Id and anti-anti-Id antibodies. Both anti-Id and anti-anti-Id antibodies were able to inhibit mAb R24 binding to GD3. In addition, the anti-anti-Id antibodies were shown to recognize GD3 directly. Anti-Id and anti-anti-Id mAb were then selected from two fusion experiments for evaluation. The most interesting finding emerged from the characterization of the anti-anti-Id mAb 5.G8. It was shown to recognize two different GD3-expressing human melanoma cell lines in vitro and to mediate tumor cell cytotoxicity by complement activation and antibody-dependent cellular cytotoxicity. The biological activity of the anti-anti-Id mAb was also tested in a mouse tumor model, in which it was shown to be a powerful growth inhibitor of melanoma cells. Thus, activity of the anti-anti-Id mAb 5.G8 matched that of the prototypic anti-GD3 mAb R24 both in vitro and in vivo. Altogether, our results indicate that the idiotype approach might produce high affinity, specific and very efficient antitumor immune responses.

Mindfulness meditation is an ancient and simple form of meditation that has been said to induce several important physical and psychological benefits. The present study was designed with the aim of investigating the psychobiological effects of mindfulness meditation practice in a clinical population. Sixteen patients with depression and anxiety symptoms were submitted to a mindfulness meditation programme for the period of 2 months. Psychological well-being [5-item Mental Health (MH-5)] and several endocrine parameters [adrenocorticotropic hormone (ACTH), cortisol, dehydroepiandrosterone-sulphate (DHEAS), thyroid-stimulating hormone, triiodothyronine, thyroxine and parathyroid hormone (PTH)] were assessed before and after the meditation programme. At post-test, the MH-5 score improved, while the levels of DHEAS and ACTH were raised, and those of PTH decreased. Our results suggest that mindfulness meditation can exert a significant psychobiological modulation by enhancing psychological well-being, as well as by regulating the levels of several hormonal parameters on different axes. Copyright © 2010 John Wiley & Sons, Ltd.

The bee genus Parapsaenythia Friese (Apidae, Protandrenini) is restricted to South America and characterized by the strongly punctate integument and hairy eyes. In the present work, the species of Parapsaenythia are revised, with seven species recognized, of which three are described as new: P. carinulatasp.n. (Argentina, Brazil, Paraguay), P. inornata Moure (Brazil), P. lanatasp.n. (Brazil), P. paspali (Schrottky) (Argentina, Brazil, Paraguay), P. puncticutis (Vachal) (Argentina, Bolivia), P. scutellarissp.n. (Argentina, Bolivia, Brazil, Paraguay) and P. serripes (Ducke) (Argentina, Brazil). Lectotypes are designated for Psaenythia (Parapsaenythia) argentina Friese, Anthrenoides paspali Schrottky and Parapsaenythia paraguayae Brethes. Floral records, distribution maps, illustrations and an identification key are provided. The cladistic analysis of 11 terminal species (all species of Parapsaenythia plus four outgroup species), based on 45 characters of external morphology and male terminalia, produced a single most parsimonious tree. Parapsaenythia was recovered as monophyletic, with the relationships among its species as follows: (P. inornata (P. carinulatasp.n. (P. lanatasp.n. (P. scutellarissp.n. (P. serripes, P. paspali, P. puncticutis))))). Based on these relationships plus the available distributional data, a cladistic biogeographic analysis was performed, and area cladograms for the Chacoan subregion are presented and discussed.

Recent studies have identified AHR as a master regulator of Wilms' tumor suppressor gene (WT1) signaling in the developing kidney. Activation of AHR signaling by environmental chemical is associated with proteasome-mediated degradation of AHR protein, disruption of WT1 alternative splicing, and marked alterations in the regulation of genetic programs of developmental progression in the developing kidney. The complexity of genetic regulatory networks of nephrogenesis controlled by AHR-WT1 interactions will be discussed here with particular emphasis given to the biological and medical consequences that may result from deficits in nephrogenesis that compromise reserve capacity and renal function later in life. Understanding the impact of early-life environmental exposures to chemicals that disrupt AHR signaling can help minimize negative health consequences to pregnant women and their offspring. Birth Defects Research (Part C) 87:192–197, 2009. © 2009 Wiley-Liss, Inc.

The present study was conducted to evaluate the contextual specificity of long interspersed nuclear element-1 (LINE-1 or L1) activation by cellular stress and the role of the aryl hydrocarbon receptor (AHR) transcription factor and oxidative stress in the gene activation response. Activation of the AHR by the genotoxic carcinogen benzo(a)pyrene (BaP) increased L1 expression in human cervical carcinoma (HeLa) cells, human microvascular endothelial cells (HMEC), mouse vascular smooth muscle cells (mVSMC) and mouse embryonic kidney cells (mK4). In contrast, challenge with a different AHR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or UV irradiation (10-20 J/m(2)), induced L1 only in HeLa cells. Transactivation of the mouse L1Md-A5 promoter was observed in all cell types challenged with BaP, while TCDD was without effect, and UV only activated L1 in HeLa cells. Genetic and pharmacological experiments implicated the AHR and oxidative stress as contextual determinants of L1 inducibility by cellular stress.

As the postgenomic era continues to unfold, a new wave of scientific investigation is upon us focusing on the application of genomic technologies to study the meanings encrypted on the DNA code and the responses of living organisms to changes in their environment. Recent functional genomics studies in this laboratory have focused on the role of the aryl hydrocarbon receptor, a ubiquitous transcription factor, in genetic programming during renal development. Also of interest is the application of genomics investigations to the study of chronic medical conditions associated with early life exposures to environmental contaminants. Molecular evidence is discussed in this review within the framework of human molecular medicine.

F-centers, generated in mixed single crystals KCl1–xBrx:Pb2+ by gamma radiation exposure in the range of 0.5–30 kGy, were studied in order to elucidate the effect of these defects in the thermally stimulated luminescence emission of the doped crystal. The F-centers were identified by its characteristic F absorption band which was located between 520 and 640 nm, depending on the ion composition in the mixed crystal. The thermoluminescence emission was measured before and after room temperature optical bleaching with F-light and significant effects in the TL glow curve were observed. The results suggested that the defects like F-centers play a very important role in the recombination mechanism responsible for the thermoluminescence emission process. (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A multiplex PCR method was developed to identify P. aeruginosa, B. cepacia complex, and S. maltophilia directly in sputum and oropharyngeal samples from CF patients. One hundred and six patients (53 male, and 53 female) attending our pulmonology clinic were studied from September 2000-April 2001. Two hundred and fifty-seven samples were cultured in selective media and submitted to multiplex PCR reactions, using three primer pairs targeting specific genomic sequences of each species, with an additional primer pair targeting a stretch of ribosomal 16S DNA, universal for bacteria, to act as a control. P. aeruginosa was isolated by culture in 56% of samples, B. cepacia complex in 4.3%, and S. maltophilia in 2.7%, while multiplex PCR identified P. aeruginosa in 78.7%, B. cepacia complex in 3.9%, and S. maltophilia in 3.1% of samples. Multiplex PCR results were verified by PCR reactions using different species-specific primers described in the literature and DNA sequencing of amplicons from a few samples. Comparing to culture results, the sensitivity and specificity values of multiplex PCR for bacterial identification were, respectively, 97.2% and 45.5% for P. aeruginosa, 45.5% and 97.9% for B. cepacia complex, and 40% and 97.6% for S. maltophilia. All 10 multiplex PCR-positive results for B. cepacia complex were confirmed using other species-specific primers described in the literature, while this approach confirmed results for S. maltophilia identification in 7/8 samples (87.5%). Sequencing of amplicons from samples culture-negative but multiplex PCR-positive for P. aeruginosa and B. cepacia complex confirmed their identity, while minor nucleotide differences among amplicons ruled out the hypothesis of PCR contamination.

This study assessed in vitro marginal leakage of class V cavities prepared by turbine and Er:YAG laser and restored with different materials. Sixty cavities with enamel and dentine margins were prepared and assigned to six groups: I, II, III by turbine and IV, V, VI by Er:YAG laser. The following restorative systems were used: groups I and IV: Bond 1 + Alert; II and V: Fuji II LC; III and VI: SBMP + Dispersalloy. After finishing, specimens were thermocycled for 8 h and 45 min (500 cycles), isolated, immersed in a 0.2% Rhodamine B solution, sectioned oro-facially and analysed for leakage. The dye penetration means (%) were: occlusal I: 10.09 (+/- 21.28), II: 3.25 (+/- 10.27), III: 0, IV: 41.77 (+/- 42.48), V: 23.37 (+/- 33.79), VI: 12.66 (+/- 24.06); cervical I: 16.49 (+/- 26.67), II: 4.34 (+/- 13.71), III: 0, IV: 37.71 (+/- 30.47), V: 39.56 (+/- 43.35) and VI: 72.53 (+/- 37.79). The use of Er:YAG laser for cavity preparation yielded higher degree of marginal leakage, as compared with the use of conventional air-turbine. The enamel interface provided better marginal sealing, comparing with dentine/cementum margin. As to the cavity preparation device (i.e. laser or bur), the analysis of the results showed that bonded amalgam and Fuji II LC provided less infiltration, than Alert. On the other hand, for lased cavities, Alert provided the best results, similar to those of Fuji II LC and superior to those reached by bonded amalgam.

This work was carried out to determine the effect of overload cycles on the fatigue life of a structural steel used for offshore applications. Single and multiple overloads were adopted and the corresponding fatigue crack growth retardation was evaluated. Residual stress fields were measured in the vicinity of the crack tip using an X-ray diffraction technique and their size compared with that of the overload cyclic plastic zone. In regard to crack growth retardation, the results indicated that the extension in fatigue life increases with an increase in overload, as a consequence of the generation of higher compressive residual stress levels over a larger distance ahead of the crack tip. The effect of two equal and consecutive overloads, with the second one applied at different intervals of crack propagation from the first, was also considered. Larger intervals were shown to lead to a longer residual fatigue life.

Introduction The phytochemistry of species of the genus Piper has been studied extensively, including Piper solmsianum. However, no studies have addressed the phytochemistry of the sap content of Piper species. Objective To evaluate the transferring of secondary compounds from the saps of P. solmsianum to the honeydew of Edessa meditabunda. Methodology The honeydew of E. meditabunda and saps of P. solmsianum were analysed by GC-MS, 1H-NMR and LC-MS. Results The lignan (−)-grandisin and the phenylpropanoid (E)-isoelemicin were detected in both saps of P. solmsianum and honeydew of E. meditabunda. Conclusion Analysis of honeydew secreted by the sap-sucking insect E. meditabunda indicated that (−)-grandisin and (E)-isoelemicin are absorbed from the phloem of Piper solmsianum. Copyright © 2012 John Wiley & Sons, Ltd.

Self-diffusion in model Al and Ni has been studied by molecular static and molecular dynamic techniques. The structure of defect-lattice configurations has been obtained with the former technique. With the latter, the vacancy diffusion mechanism has been analysed over a wide temperature range, and particular attention has been paid to multiple jumps in the high temperature region. The possible contribution of divacancies, within the limits imposed by the interatomic potentials used, has also been considered.

Although monomethine cyanines and reactive dyes, used mainly in dyeing of textile goods, are well studied, the development of cyanines with groups capable of being bound covalently to a solid support has been largely unexplored. The scope of this study was the synthesis and characterisation of aminocyanines capable of bind to microcrystalline cellulose, through the polyfunctional cross-linking agent cyanuric chloride. In a second approach, the cyanines bound to microcrystalline cellulose in varying amounts were used as specific ligands in affinity chromatography. Two different methods to determine the amount of cyanine linked to the dyed cellulose were used, based on Fourier transformed infrared spectroscopy and elemental analysis. The fixation yields of the two different cyanines bound to cellulose were determined.

Ethyl and methyl esters of commercial rhodamines B, 19, 101 and 110 and propyl and butyl esters of commercial rhodamine B were synthesized and isolated with different counterions (yields 70–98%). The 1H and 13C NMR spectral data for these compounds were fully assigned by a combination of one- and two-dimensional experiments. The Fourier transform IR and UV–visible spectra were also recorded and the main bands were identified. Copyright © 2000 John Wiley & Sons, Ltd.

A collection of 26 cultivars of wheat Triticum aestivum were screened for resistance against the two main aphid pests of cereals, the greenbug Schizaphis graminum Rond. and the Russian wheat aphid (RWA) Diuraphis noxia Mordvilko. Since genetic variability has been found in Argentinean populations of both aphid species, this work was aimed at determining the response of different types of resistance in wheat cultivars when infested with aphids. Antixenosis, antibiosis and tolerance were evaluated with traditional tests in controlled environmental conditions using a clone of greenbug biotype C and a clone of RWA collected on wheat. Genetic resistance was found against one or both aphid species in several wheats. Most of the highest levels of antixenosis, antibiosis and tolerance against the two aphids occurred in different cultivars; as a consequence the resistance mechanisms for both pests appear to be partly independent. Antibiosis against greenbug or RWA appears to be determined by two different sets of genes, one affecting development time and the other reducing fecundity and longevity. The antibiosis against both aphid species in terms of their development time and the intrinsic rate of population increase resulted in a partial cross effect of these aphid traits against the alternative insect species. Nonetheless, the same cultivars affected the total fertility and the longevity of both aphids. Since the highest plant performance levels and the least plant damage were recorded in different wheats, different patterns of tolerance were displayed against the greenbug and the RWA. Consequently, different genes appear to be involved in several traits of the resistance mechanisms against the two aphids. The genes that independently conferred resistance to aphids could be combined in new cultivars of wheat to broaden their genetic base of resistance against the greenbug and the RWA.

In this work, it is shown that the Ca 2+ -transport ATPase found in the microsomal fraction of the cerebellum can use both glucose 6-phosphate/hexokinase and fructose 1,6-bisphosphate/phosphofructokinase as ATP-regenerating systems. The vesicles derived from the cerebellum were able to accumulate Ca 2+ in a medium containing ADP when either glucose 6-phosphate and hexokinase or fructose 1,6-bisphosphate and phosphofructokinase were added to the medium. There was no Ca 2+ uptake if one of these components was omitted from the medium. The transport of Ca 2+ was associated with the cleavage of sugar phosphate. The maximal amount of Ca 2+ accumulated by the vesicles with the fructose 1,6-bisphosphate system was larger than that measured either with glucose 6-phosphate or with a low ATP concentration and phosphoenolpyruvate/pyruvate kinase. The Ca 2+ uptake supported by glucose 6-phosphate was inhibited by glucose, but not by fructose 6-phosphate. In contrast, the Ca 2+ uptake supported by fructose 1,6-bisphosphate was inhibited by fructose 6-phosphate, but not by glucose. Thapsigargin, a specific SERCA inhibitor, impaired the transport of Ca 2+ sustained by either glucose 6-phosphate or fructose 1,6-bisphosphate. It is proposed that the use of glucose 6-phosphate and fructose 1,6-bisphosphate as an ATP-regenerating system by the cerebellum Ca 2+ -ATPase may represent a salvage route used at early stages of ischemia; this could be used to energize the Ca 2+ transport, avoiding the deleterious effects derived from the cellular acidosis promoted by lactic acid.

A collection of tritordeum amphiploids (Hordeum chilense × Triticum turgadum) and their wheat parents were screened for resistance against the two main aphid pesis of cereals, the greenhug. Schizaphis graminum Rond. and ihe Russian wheat aphid (RWA) Diuraphis naxia Mord-vilko. Antixenosis. antibiosis and tolerance were evaluated in controlled environmental conditions using a. clone of greenbug biotypc C and a clone of RWA collected on pasta wheat. Tritordeum amphiploids pos-sess genetic resistance against greenbug and RWA; some of the lines tested were more resistant than the parental wheat line. Four principal components explained the resistance against both aphid species. The antixenosis shown against both pests was mainly contributed by their wheat parents. The antibiosis againsl both aphid species was obviously dependent on diflerent plant traits. The highest levels of antibiosis against the two aphids occurred in different amphiploids. Different genes are involved in the antibiotic reaction against the two aphids. The Tritordeum resistance to RWA is based on anlixenosis and ant-biosis since the tolerance trails were not independent of the other types of resistance. The level of tolerance shown to the greenbug was variable and appears to be controlled by differeni mechanisms. The tolerance to aphids shown by H. chilense is expressed in the amphiploids. but with some genomic interaction. Genes conferring resistance to aphids in H. chilensee could be incorporated into new cultivars of wheat to broaden their genetic base of resistance against greenbug and RWA.

Pattern recognition approaches were developed and applied to the classification of 600 MHz 1 H NMR spectra of urine from rats dosed with compounds that induced organ-specific damage in either the liver or kidney. Male rats were separated into groups (n = 5) and each treated with one of the following compounds; adriamycin, allyl alcohol, 2-bromoethanamine hydrobromide, hexachlorobutadiene, hydrazine, lead acetate, mercury II chloride, puromycin aminonucleoside, sodium chromate, thioacetamide, 1,1,2-trichloro-3,3,3-trifluoro-1-propene or dose vehicle. Urine samples were collected over a 7 day time-course and analysed using 600 MHz 1 H NMR spectroscopy. Each NMR spectrum was data-reduced to provide 256 intensity-related descriptors of the spectra. Data corresponding to the periods 8-24 h, 24-32 h and 32-56 h post-dose were first analysed using principal components analysis (PCA). In addition, samples obtained 120-144 h following the administration of adriamycin and puromycin were included in the analysis in order to compensate for the late onset of glomerular toxicity. Having established that toxin-related clustering behaviour could be detected in the first three principal components (PCs), three-quarters of the data were used to construct a soft independent modelling of class analogy (SIMCA) model. The remainder of the data were used as a test set of the model. Only three out of 61 samples in the test set were misclassified. Finally as a further test of the model, data from the 1 H NMR spectra of urine from rats that had been treated with uranyl nitrate were used. Successful prediction of the toxicity type of the compound was achieved based on NMR urinalysis data confirming the robust nature of the derived model. © 1998 John Wiley & Sons, Ltd.

Repeated cycles of vascular injury by allylamine induce vascular lesions similar to those seen in atherosclerotic vessels, or following balloon catheterization. Vascular (aortic) smooth muscle cells harvested from allylamine-treated animals (i.e., allylamine cells) acquire a proliferative advantage relative to control counterparts that is associated with differential secretion and extracellular matrix sequestration of several proteins. In the present study, we have characterized two of these proteins (M(r) 52 and 36 kDa; pl 5.6 and 5.2, respectively) and their putative role in the expression of a proliferative phenotype. Because the physical properties of these proteins were comparable to those of osteopontin (OPN) and its thrombin-generated fragment(s), initial experiments were conducted to examine the expression and processing of OPN in this cell system. OPN mRNA expression was enhanced during early G1 cell cycle progression in allylamine cells relative to control counterparts. However, comparable amounts of OPN (M(r) 56, 52, and 50 kDa) were detected by Western analysis in media conditioned by both cell types using the OP-199 or B77-Rat1 antibodies to OPN. Allylamine cells, however, produced increased amounts of a 36 kDa protein recognized by the OP-199 antibody. Incubation of conditioned media from [35S]methionine-labeled allylamine cells with thrombin decreased the intensity of the 52 kDa protein, while increasing the intensity of a 36 kDa protein. RT-PCR analysis demonstrated expression of a 1.2 kb OPN band in both cell types consistent with the predicted size of OPN mRNA, suggesting that the 36 kDa fragment recognized by OP-199 in allylamine cells was likely not due to altered splicing of the OPN transcript. To determine if OPN and/or the 36 kDa fragment played a central role in the proliferative capacity of allylamine cells, the effect of an antibody to an alpha v integin subunit was examined. An antibody to the alpha v subunit, but not alpha 4, nullified the proliferative advantage of allylamine cells relative to control counterparts, suggesting that integrin-mediated signaling is a key feature of the proliferative phenotype of allylamine cells. We conclude that enhanced proteolytic cleavage of OPN may characterize the modulation of vascular SMCs to a more proliferative phenotype following chemical injury by allylamine.

13C NMR chemical shift assignments of five α- and β- epimeric pairs of cholestanes functionalized at C-3 are presented. Empirical increment estimations proved to be a valuable tool for the unequivocal structural elucidation when compared with the chemical shift values of cholestanes derivatized by introduction of N- and S-containing groups at C-3 in equatorial and axial positions. Moreover, the possibility is demonstrated to anticipate the effect of OC(S)R substituents at neighboring carbon atoms of the ring A backbone. Copyright © 2005 John Wiley & Sons, Ltd.

Although the role of several protooncogenes, including sis, myc, and myb in the regulation of growth and differentiation of vascular cells has been examined in some detail, limited information is available on the contribution of ras genes to these processes. In the present studies the influence of oncogenic ras transfection on the phenotypic expression of rat aortic smooth muscle cells (SMCs) was examined. Cultured rat aortic SMCs during early passage (P4) were transfected by lipofection with c-Ha-rasEJ in a pSV2 neo vector or with pSV2 neo vector alone. Stable transfectants were selected in G418 over a 6-week period. Oncogene-transfected cells (ras-LF-1) exhibited differences in morphology and growth pattern relative to vector controls (neo-LF-1), or naive SMCs, including the development of prominent processes and the appearance of focal cellular arrangements giving rise to latticelike structures. Southern analysis revealed multiple integration of oncogenic ras in ras LF-1 cells. Transfection of c-Ha-rasEJ was associated with a twofold increase in p21 levels relative to pSV2 vector controls demonstrating that exogenous ras was expressed in these cells. Overexpression of ras p21 afforded SMCs a lower serum requirement for growth compared to vector controls, anchor-age independent growth on soft agar, and acquisition of epidermal growth factor (EGF) responsiveness. Stimulation of serum-deprived SMCs with 5% fetal bovine serum (FBS) increased steady-state levels of c-Ha-ras mRNA in both ras-LF-1 and neo-LF-1 but ras induction was more pronounced in ras-transfected cells. α-smooth muscle (SM) actin gene expression was markedly reduced in ras-transfected cells relative to vector controls. These results show that transfection of c-Ha-rasEJ into aortic SMCs induces an altered phenotypic state characterized by alterations in growth factor-related signal transduction and tumorigenic potential. © 1994 Wiley-Liss, Inc.

The physico-chemical properties of the intermetallic phases in the Cu–In system have been a matter of considerable theoretical and experimental interest in connection with, i.a., the application of In–Sn alloys as lead-free micro-soldering alloys. Recently, a new binary compound with the chemical formula Cu10In7 has been detected in a study of the η-phase field. The structure of the Cu10In7 phase has been determined as closely related to that of the Cu11In9 compound occurring in the phase diagram, but no experimental or theoretical information on its electronic structure, thermodynamic and equation-of-state properties has yet been reported. In the present work we report the lattice parameters, bulk modulus, energy of formation from the constituent elements and the electronic structure of the new phase, calculated by applying an ab initio density-functional-theory method. Our calculation technique uses the projector augmented wave potentials and the exchange-correlation functions of Perdew and Wang in the generalized gradient approximation. The present results for the Cu10In7 phase are compared with the experimental data available, and with the trends in structural and thermodynamic properties emerging from ab initio calculations also performed in the present study for various structurally related and neighboring compounds in the Cu–In phase diagram, viz., the ideal B82–Cu2In, B81–CuIn, B82–CuIn2 phases and the Cu11In9 compound.

(2R,4R)-N-Acyl-2-phenylthiazolidine-4-carboxylic acids were used to generate 5H,7H-thiazolo[3,4-c]oxazol-4-ium-1-olates with internal dipolarophiles. The intramolecular 1,3-dipolar cycloaddition of these mesoionic species led to the synthesis of new 1H-pyrrolo[1,2-c]thiazole derivatives (8a, 8b and 12) as single enantiomers. The structure of 8a was determined by X-ray crystallography.