bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on organophosphorus compounds: Synthesis and reactions of [1,2,4,3]triaza‐phospholo[4,5‐a]quinoxaline derivative
Authors: Moustafa Hassan M. , Mohamed Mounir A. A.
Published: July 2008

2,4‐Bis‐(4‐methoxyphenyl)‐1,3,2,4‐dithiadiphosphetane‐2,4‐disulfide (Lawesson's reagent) (1) reacted with 2‐hydrazino‐3‐methyl‐quinoxaline (2) to give [1,2,4,3]‐triazaphospholo[4,5‐a]quinoxaline derivative 3. The Mannich reaction using different amines on compound 3 gave Mannich bases 4a–d. Also, compound 3 reacted with formaldehyde to give the corresponding 2‐hydroxymethyl derivative 5, which upon reaction with thionyl chloride gave the corresponding chloromethyl derivative 6. Treatment of compound 6 with some thiols yielded the corresponding sulfides 7a–d. Acylation of compound 3 gave acylated compounds 8a,b. Compound 9, which was prepared through the reaction of compound 3 with ethyl cyanoacetate, was investigated as a starting material for the synthesis of some new heterocyclic systems 10–13. Also, reaction of compound 9 with carbon disulfide and 2 equivalents of methyl iodide in a one‐pot reaction yielded the corresponding ketene‐S,S‐acetal 14, which in turn reacted with bidentates to give some new heterocycles 15–17. © 2008 Wiley Periodicals, Inc. Heteroatom Chem 19:520–529, 2008; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20473
Oxidation of diethyl N‐sulfinylphos‐phoramidates
Authors: Mielniczak Grażyna , Łopusiński Andrzej
Published: July 2008

It is reported that the diethyl N‐sulfinylphosphoramidate (1a) is oxidized with iodoso‐ and/or iodoxybenzene as well as with organic peroxides, to give diethyl N‐sulfonylphosphoramidate (7a). The latter was generated in situ at low temperature and trapped with dialkyl phosphoramidates, affording the N,N′ bis(dialkyl phosphor)sulfamides 3a–c. The oxidation of 1a with cumene peroxide in the presence of 2,2‐dimethyl‐propan‐1‐ol (9) produced the diethylphosphor‐N‐[2,2‐dimethylpropyl]sulfamate (10a). © 2008 Wiley Periodicals, Inc. Heteroatom Chem 19:530–536, 2008; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20474
The impact of using different costing methods on the results of an economic evaluation of cardiac care: microcosting vs gross‐costing approaches
Authors: Clement (Nee Shrive) Fiona M. , Ghali William A. , Donaldson Cam , Manns Braden J.
Published: July 2008

Background: Published guidelines on the conduct of economic evaluations provide little guidance regarding the use and potential bias of the different costing methods.

Objectives: Using microcosting and two gross‐costing methods, we (1) compared the cost estimates within and across subjects, and (2) determined the impact on the results of an economic evaluation.

Methods: Microcosting estimates were obtained from the local health region and gross‐costing estimates were obtained from two government bodies (one provincial and one national). Total inpatient costs were described for each method. Using an economic evaluation of sirolimus‐eluting stents, we compared the incremental cost–utility ratios that resulted from applying each method.

Results: Microcosting, Case‐Mix‐Grouper (CMG) gross‐costing, and Refined‐Diagnosis‐Related grouper (rDRG) gross‐costing resulted in 4‐year mean cost estimates of $16 684, $16 232, and $10 474, respectively. Using Monte Carlo simulation, the cost per QALY gained was $41 764 (95% CI: $41 182–$42 346), $42 538 (95% CI: $42 167–$42 907), and $36 566 (95% CI: $36 172–$36 960) for microcosting, rDRG‐derived and CMG‐derived estimates, respectively (P<0.001).

Conclusions: Within subject, the three costing methods produced markedly different cost estimates. The difference in cost–utility values produced by each method is modest but of a magnitude that could influence a decision to fund a new intervention. Copyright © 2008 John Wiley & Sons, Ltd.
Genetic information, obesity, and labor market outcomes
Authors: Norton Edward C. , Han Euna
Published: July 2008

Economists have argued that obesity may lead to worse labor market outcomes, especially for women. Empirical methods to test this hypothesis have not thus far adequately controlled for the endogeneity of obesity. We use variation in genotype to predict variation in phenotype (obesity). Genetic information from specific genes linked to obesity in the biomedical literature provides strong exogenous variation in the body mass index and thus can be used as instrumental variables. These genes predict swings in weight of between 5 and 20 pounds for persons between five and six feet tall. We use additional genetic information to control for omitted variables correlated with both obesity and labor market outcomes. We analyzed data from the third wave of the Add Health data set, when respondents are in their mid‐twenties. Results from our preferred models show no effect of lagged obesity on the probability of employment or on wages, for either men or women. This paper shows the potential of using genetic information in social sciences. Copyright © 2008 John Wiley & Sons, Ltd.
Is there a meaningful serum hepatitis B virus DNA cutoff level for therapeutic decisions in hepatitis B e antigen–negative chronic hepatitis B virus infection?
Authors: Papatheodoridis George V. , Manesis Emanuel K. , Manolakopoulos Spilios , Elefsiniotis Ioannis S. , Goulis John , Giannousis John , Bilalis Antonios , Kafiri Georgia , Tzourmakliotis Dimitrios , Archimandritis Athanasios J.
Published: July 2008

The diagnosis of hepatitis B e antigen (HBeAg)-negative chronic hepatitis B indicating therapeutic intervention currently requires serum hepatitis B virus (HBV) DNA ≥2,000 IU/mL. We evaluated the severity of liver histology and the presence of histological indication for treatment in patients with HBeAg-negative chronic HBV infection focusing on those with low viremia and/or normal alanine aminotransferase (ALT). In total, 399 patients with increased ALT and detectable serum HBV DNA (chronic hepatitis B patients) and 35 cases with persistently normal ALT and HBV DNA >2,000 IU/mL (inactive carriers) were included. Histological indication for treatment (grading score ≥7 and/or stage ≥2 in Ishak's classification) was found in 91% (185/203), 82% (75/91), 75% (47/63), and 62% (26/42) of chronic hepatitis B patients with HBV DNA ≥200,000, 20,000-199,999, 2,000-19,999, and <2,000 IU/mL, respectively (

P

< 0.001). Histological indication for treatment was more frequent in chronic hepatitis B patients with persistently elevated ALT (86% or 275/321), but it was also found in 74% (58/78) of those with transiently normal ALT (

P

= 0.025). All inactive carriers had HBV DNA <20,000 IU/mL. Histological indication for treatment was present in 17% (6/35) of inactive carriers always due to moderate (stage 2) fibrosis without active necroinflammation.

Conclusion:

HBeAg-negative chronic HBV patients with persistently or transiently increased ALT and HBV DNA ≥20,000 IU/mL almost always require therapeutic intervention, but histological indications for treatment are also present in the majority of such cases with HBV DNA <20,000 and even <2,000 IU/mL. In contrast, minimal histological lesions are observed in the majority of HBeAg-negative patients with persistently normal ALT and HBV DNA >2,000 IU/mL, who may not require immediate liver biopsy and treatment but only close follow-up. (Hepatology 2008.)
Suppressor of cytokine signaling-3 (SOCS-3) and a deficit of serine/threonine (Ser/Thr) phosphoproteins involved in leptin transduction mediate the effect of fructose on rat liver lipid metabolism
Authors: Vilà Laia , Roglans Núria , Alegret Marta , Sánchez Rosa María , Vázquez-Carrera Manuel , Laguna Juan Carlos
Published: July 2008

There is controversy regarding whether fructose in liquid beverages constitutes another dietary ingredient of high caloric density or introduces qualitative changes in energy metabolism that further facilitate the appearance of metabolic diseases. Central to this issue is the elucidation of the molecular mechanism responsible for the metabolic alterations induced by fructose ingestion. Fructose administration (10% wt/vol) in the drinking water of Sprague-Dawley male rats for 14 days induced hyperleptinemia and hepatic leptin resistance. This was caused by impairment of the leptin-signal transduction mediated by both janus-activated kinase-2 and the mitogen-activated protein kinase pathway. The subsequent increase in activity in the liver of the unphosphorylated and active form of the forkhead box O1 nuclear factor, which transrepresses peroxisome proliferator-activated receptor α activity, and a lack of activation of the adenosine monophosphate-activated protein kinase, led to hypertriglyceridemia and hepatic steatosis. These alterations are attributable to two key events: (1) an increase in the amount of suppressor of cytokine signaling-3 protein, which blocks the phosphorylation and activation of janus-activated kinase-2 and Tyr985 on the long form of the leptin receptor; and (2) a common deficit of phosphorylation in serine/threonine residues of key proteins in leptin-signal transduction pathways. The latter is probably produced by the early activation of protein phosphatase 2A, and further sustained by the accumulation in liver tissue of ceramide, an activator of protein phosphatase 2A, due to incomplete oxidation of fatty acids.

Conclusion:

Our data indicate that fructose ingestion as a liquid solution induces qualitative changes in liver metabolism that lead to metabolic diseases. (Hepatology 2008.)
Lignanamides and Sesquiterpenoids from Stems of Mitrephora thorelii
Authors: Ge Fan , Tang Chun‐Ping , Ye Yang
Published: July 2008

The first phytochemical investigation on stems of Mitrephora thorelii led to the isolation of three new lignanamides, thoreliamides A–C (1–3), and a new sesquiterpenoid, thorelinin (4), together with ten known compounds. The structures of the new compounds were established on the basis of extensive spectroscopic analyses. Thoreliamide C is the first trimer derived from cinnamic acid amide units.
Eremophilane‐Type Sesquiterpenes from the Roots of Ligularia virgaurea
Authors: Zhang Zhan‐Xin , Fei Dong‐Qing , Jia Zhong‐Jian
Published: July 2008

From the roots of Ligularia virgaurea, five new eremophilane‐type sesquiterpenes were isolated, including three new eremophilenolides, 6β‐(angeloyloxy)‐1α,8β,10β‐trihydroxyeremophil‐7(11)‐en‐12,8α‐olide (1), 6β‐(angeloyloxy)‐1β,10β‐epoxy‐8β‐ethoxyeremophil‐7(11)‐en‐12,8α‐olide (2), and 1β,10β‐epoxy‐8β‐ethoxy‐6β‐[(2‐methylacryloyl)oxy]eremophil‐7(11)‐en‐12,8α‐olide (3), two new noreremophilanes, 3β‐[(2‐methylacryloyl)oxy]‐8‐oxo‐12‐noreremophil‐6‐en‐11‐one (9), and 9β‐hydroxy‐8‐oxo‐12‐noreremophil‐6‐en‐11‐one (10), and six known eremophilanes, namely 4–8, and 11. Their structures were elucidated by means of spectral methods, such as IR, HR‐ESI‐MS, and 1D‐ and 2D‐NMR, and by comparison of the spectral data with those reported for structurally related compounds.
Four New Dibenzocyclooctadiene Lignans from Schisandra rubriflora
Authors: Li Hong‐Mei , Luo Yong‐Ming , Pu Jian‐Xin , Li Xiao‐Nian , Lei Chun , Wang Rui‐Rui , Zheng Yong‐Tang , Sun Han‐Dong , Li Rong‐Tao
Published: July 2008

From the aerial parts of Schisandra rubriflora (Franch) Rehd. et Wils., four new dibenzocyclooctadiene lignans, methylgomisin R (1), (+)‐14‐tigloylgomisin K3 (2), 12‐demethylwuweilignan I (3), schisandrene A (4), together with 13 known lignans, were isolated. The structures of four new compounds were elucidated by spectroscopic methods including extensive 1D‐ and 2D‐NMR techniques.
Ligand‐Exchange Processes on Solvated Beryllium Cations. Part III
Authors: Puchta Ralph , van Eldik Rudi
Published: July 2008

On the basis of DFT calculations (B3LYP/6‐311+G**), the possibility to include solvent effects is considered in the investigation of the H2O‐exchange mechanism on [Be(H2O)4]2+ within the widely used cluster approach. The smallest system in the gas phase, [Be(H2O)4(H2O)]2+, shows the highest activation barrier of +15.6 kcal/mol, whereas the explicit addition of five H‐bonded H2O molecules in [{Be(H2O)4(H2O)}(H2O)5]2+ reduces the barrier to +13.5 kcal/mol. Single‐point calculations applying CPCM (B3LYP(CPCM:H2O)/6‐311+G**//B3LYP/6‐311+G**) on [Be(H2O)4(H2O)]2+ lower the barrier to +9.6 kcal/mol. Optimization of the precursor and transition state of [Be(H2O)4(H2O)]2+ within an implicit model (B3LYP(CPCM:H2O)/6‐311+G** or B3LYP(PCM:H2O)/6‐311+G**) reduces the activation energy further to +8.3 kcal/mol but does not lead to any local minimum for the precursor and is, therefore, unfavorable.
7‐Halogenated 7‐Deazapurine 2′‐Deoxyribonucleosides Related to 2′‐Deoxyadenosine, 2′‐Deoxyxanthosine, and 2′‐Deoxyisoguanosine: Syntheses and Properties
Authors: Seela Frank , Xu Kuiying
Published: July 2008

A series of 7‐fluorinated 7‐deazapurine 2′‐deoxyribonucleosides related to 2′‐deoxyadenosine, 2′‐deoxyxanthosine, and 2′‐deoxyisoguanosine as well as intermediates 4b–7b, 8, 9b, 10b, and 17b were synthesized. The 7‐fluoro substituent was introduced in 2,6‐dichloro‐7‐deaza‐9H‐purine (11a) with Selectfluor (Scheme 1). Apart from 2,6‐dichloro‐7‐fluoro‐7‐deaza‐9H‐purine (11b), the 7‐chloro compound 11c was formed as by‐product. The mixture 11b/11c was used for the glycosylation reaction; the separation of the 7‐fluoro from the 7‐chloro compound was performed on the level of the unprotected nucleosides. Other halogen substituents were introduced with N‐halogenosuccinimides (11a→11c–11e). Nucleobase‐anion glycosylation afforded the nucleoside intermediates 13a–13e (Scheme 2). The 7‐fluoro‐ and the 7‐chloro‐7‐deaza‐2′‐deoxyxanthosines, 5b and 5c, respectively, were obtained from the corresponding MeO compounds 17b and 17c, or 18 (Scheme 6). The 2′‐deoxyisoguanosine derivative 4b was prepared from 2‐chloro‐7‐fluoro‐7‐deaza‐2′‐deoxyadenosine 6b via a photochemically induced nucleophilic displacement reaction (Scheme 5). The pKa values of the halogenated nucleosides were determined (Table 3). 13C‐NMR Chemical‐shift dependencies of C(7), C(5), and C(8) were related to the electronegativity of the 7‐halogen substituents (Fig. 3). In aqueous solution, 7‐halogenated 2′‐deoxyribonucleosides show an approximately 70% S population (Fig. 2 and Table 1).
New Components Including Cyclopeptides from Barks of Christiana africana DC. (Tiliaceae)
Authors: Michalet Serge , Payen‐Fattaccioli Laëtitia , Beney Chantal , Cégiéla Pascale , Bayet Christine , Cartier Gilbert , Noungoué‐Tchamo Diderot , Tsamo Etienne , Mariotte Anne‐Marie , Dijoux‐Franca Marie‐Geneviève
Published: July 2008

Phytochemical investigation of barks of Christiana africana led to the identification of cyclopeptide alkaloids, flavonoids, coumarinolignans, iridoids, sesquiterpenoids, and triterpenes. This plant was classified so far in the Tiliaceae family. This study was started while the genomic study of numerous specimens was described in order to establish new criteria for Malvales botanical classification. In the present work, twenty components were identified, belonging to the three major classes of secondary metabolites: alkaloids, phenols, and terpenes. In the first class, cyclopeptides are well‐known compounds in Rhamnaceae and Sterculiaceae. Their presence in Malvaceae (in APG2 sensus) suggests a possible chemical link between the ex‐Tiliaceae and the Malvaceae.
Two New Alkaloids from Flueggea virosa
Authors: Wang Guo‐Cai , Wang Ying , Li Qian , Liang Jie‐Ping , Zhang Xiao‐Qi , Yao Xin‐Sheng , Ye Wen‐Cai
Published: July 2008

Two new securinega‐type alkaloids and four known ones were isolated from the twigs and leaves of Flueggea virosa. The structures of the new compounds were elucidated by means of spectroscopic methods (UV, IR, HR‐ESI‐MS, and 1D‐ and 2D‐NMR), and the absolute configurations were assigned by CD spectra. The structures of the known compounds were identified by comparison of their physical and spectroscopic data with those reported in the literature.
Surprising Formation of Chlorinated Butenolides from Dialkyl Acetylenedicarboxylates and Hexachloro­acetone in the Presence of Triphenyl Phosphite
Authors: Yavari Issa , Sabbaghan Maryam , Hossaini Zinatossadat , Ghazanfarpour‐Darjani Majid
Published: July 2008

Triphenyl phosphite reacts smoothly with dialkyl acetylenedicarboxylates and hexachloroacetone to produce alkyl 2‐(dichloromethylene)‐2,5‐dihydro‐5‐oxo‐4‐(trichloromethyl)furan‐3‐carboxylates in good yields.
Secondary Metabolites from the Stem Bark of Litsea akoensis and Their Cytotoxic Activity
Authors: Chang Su‐Yen , Cheng Ming‐Jen , Kuo Yueh‐Hsiung , Lee Shiow‐Ju , Chang Hsun‐Shuo , Chen Ih‐Sheng
Published: July 2008

Six new butanolides, litseadioxanins A and B (1 and 2, resp.) bearing a 1,2‐dioxane moiety, litseatrinolides A and B (3 and 4, resp.), and litseakolides D1 and D2 (5 and 6, resp.), were isolated from the stem bark of Litsea akoensis, together with six known compounds. The structures of the new compounds were characterized by in‐depth NMR‐spectroscopic and mass‐spectrometric analyses. Butanolides 1–4, and a mixture of 6 and litsenolide E2, and litsenolide B1 were tested against human tumor cells, including MCF‐7 (human breast adenocarcinoma), NCI‐H460 (non‐small‐cell lung cancer), and SF‐268 (glioblastoma) cell lines. Among the tested compounds, litsenolide B1 exhibited marginal cytotoxic activity against MCF‐7, NCI‐H460, and SF‐268 cell lines in vitro.
Fluorescence Switches Triggered by Metal Ions
Authors: Wessig Pablo , Czapla Sylvia , Schedler Uwe
Published: July 2008

Molecular switches 1 with a modular architecture whose functional principle is based on conformational changes around two acetylene axes were developed. By systematic screening of various metal ions, it was found that especially Pb2+ ions effectively trigger the switching event. The reporting of this event takes place by the change of the ratio between excimer and monomer fluorescence emission of two pyrene moieties.
Efficient Synthesis of Functionalized Thiazoles from Acid Chlorides, Tetramethylthiourea, Ethyl Bromopyruvate, and Ammonium Thiocyanate
Authors: Yavari Issa , Hossaini Zinatossadat , Seyfi Samereh , Shirgahi‐Talari Faezeh
Published: July 2008

An efficient synthesis of ethyl 2‐(dimethylamino)‐1,3‐thiazole‐4‐carboxylates is described via a four‐component reaction between acid chlorides, tetramethylthiourea, ethyl bromopyruvate, and ammonium thiocyanate.
The persistence in hedge fund performance: extended analysis
Authors: Capocci Daniel P. J.
Published: July 2008

This study analyses and decomposes hedge fund returns to detect a systematic hedge fund selection criterion that enables investors to consistently and significantly outperform classical equities and bond indices over a full market cycle and over bullish and bearish market periods. The methodology used is adapted from Capocci and Hübner. The measures used include the returns, the volatility, the Sharpe score, the alpha, the beta, the skewness and the kurtosis. Measures incorporating the volatility display very strong ability to assist investors in creating alpha and consistently and significantly outperform classical indices. A sub‐period analysis is performed to check the robustness of the results. Copyright © 2008 John Wiley & Sons, Ltd.
Exchange rate pass‐through to prices in macrodata: a comparative sensitivity analysis
Authors: Mihailov Alexander
Published: July 2008

This paper compares exchange rate pass‐through to aggregate prices in the US, Germany and Japan across a number of dimensions. Building on the empirical approaches in the recent literature, our contribution is to perform a thorough sensitivity analysis of pass‐through estimates. We find that the econometric method, data frequency and variable proxy employed matter for the precision of details, yet they often agree on some general trends. Thus, pass‐through to import prices has declined in the 1990s relative to the 1980s, pass‐through to export prices remains country‐specific and pass‐through to consumer prices is nowadays negligible in all three economies we considered. Copyright © 2008 John Wiley & Sons, Ltd.
Cloning and characterization of E‐dlg, a novel splice variant of mouse homologue of the Drosophila discs large tumor suppressor binds preferentially to SAP102
Authors: Mao Peizhong , Tao Yuan‐Xiang , Fukaya Masahiro , Tao Feng , Li Dechun , Watanabe Masahiko , Johns Roger A.
Published: July 2008

Membrane‐associated guanylate kinases (MAGUKs) act as scaffolds to coordinate signaling events through their multiple domains at the plasma membrane. The MAGUK SH3 domain is noncanonical and its function remains unclear. To identify potential binding partners of MAGUK SH3, the synapse‐associated protein 102 (SAP102) SH3 domain was used as bait in a yeast two‐hybrid screen of a mouse embryonic cDNA library. A mouse homologue of the Drosophila discs large tumor suppressor (Dlg, also known as SAP97) bound preferentially to SAP102 SH3. The 4347bp cDNA sequence encoded an 893 amino acid protein with 94% identity to mouse SAP97. A deleted region (33‐aa) strongly suggests this is a novel splice variant, which we call Embryonic‐dlg/SAP97 (E‐dlg). The interaction of SAP102 and E‐dlg was confirmed in mammalian cells. E‐dlg can also bind to potassium channel Kv1.4 in a pull‐down assay. E‐dlg was highly expressed in embryonic and some adult mouse tissues, such as brain, kidney, and ovary. Furthermore, in situ hybridization showed that E‐dlg was mostly expressed in olfactory bulb and cerebellum. © 2008 IUBMB IUBMB Life, 60(10): 684–692, 2008
Caveolin‐1 promotes the transformation and anti‐apoptotic ability of mouse hepatoma cells
Authors: Wang Shujing , Jia Li , Zhou Huimin , Wang Xiaomin , Zhang Jianing
Published: July 2008

Caveolin‐1 is a major structural protein of caveolae and plays important roles in signal transduction, cellular transformation and tumor metastasis. Our previous study demonstrated that caveolin‐1 expression level was positively correlated with the invasive ability of mouse hepatoma Hepa1‐6 and Hca‐F cells. However, the role of caveolin‐1 in cellular transformation and apoptosis remains undetermined. We found that exogenous expression of caveolin‐1 in Hepa1‐6 cells enhanced cell transformation capability both in vitro and in vivo and prevented actinomycin D‐induced apoptosis via the activation of survivin‐mediated survival pathway. Conversely, downregulation of caveolin‐1 in Hca‐F cells significantly attenuated cell transformation ability in vitro and in vivo and increased cell sensitivity to actinomycin D by inhibiting survivin‐mediated survival pathway. These results indicate that caveolin‐1 could play an active role in mediating the transformation and survival of mouse hepatoma cells and might be a potential target for gene and antitumor drugs therapy. © 2008 IUBMB IUBMB Life, 60(10): 693–699, 2008
Extra‐nuclear signaling of estrogen receptors
Authors: Fu Xiao‐Dong , Simoncini Tommaso
Published: July 2008

Estrogen controls multiple biological functions through binding to estrogen receptors (ERs). Traditionally, ERs have been regarded as transcription factors regulating the expression of target genes. However, growing evidence of rapid estrogen's actions in a number of tissues has been accumulating and alternative mechanisms of signal transduction have been proposed. These so called “extra‐nuclear actions” do not require gene expression or protein synthesis and are independent of the nuclear localization of ERs. Indeed, some of these actions are elicited by ERs residing at or near the plasma membrane. Membrane‐associated molecules such as ion channels, G proteins, the tyrosine kinase c‐Src as well as growth factor receptors are modulated by liganded ERs within the membrane, leading to the activation of downstream cascades such as mitogen‐activated protein kinase, phosphatidylinositol 3‐OH kinase, protein kinase A, and protein kinase C. These cascades mediate some important rapid actions of estrogen, such as the activation of nitric oxide synthesis or the remodeling of actin cytoskeleton. In addition, these pathways are critical for the regulation of the expression of a number of target proteins implicated in cell proliferation, apoptosis, differentiation, movement, and homeostasis. In this manner, the extra‐nuclear pathways are tightly integrated with the genomic pathways to orchestrate the full spectrum of estrogen's biological functions. The recent advancements in the characterization of the molecular basis of the extra‐nuclear signaling of estrogen helps to understand the role of estrogen on human cells, and may in future turn out to be of relevance for clinical purposes. © 2008 IUBMB IUBMB Life, 60(8): 502–510, 2008
Molecular characterization of nitrite reductase gene (aniA) and gene product in Neisseria meningitidis isolates: Is aniA essential for meningococcal survival?
Authors: Stefanelli Paola , Colotti Gianni , Neri Arianna , Salucci Maria Luisa , Miccoli Roberto , Di Leandro Luana , Ippoliti Rodolfo
Published: July 2008

The present study evaluates sequence conservation in the gene coding for nitrite reductase (aniA) and AniA expression from a panel of Neisseria meningitidis isolates. Sequence analysis of the coding region in 19 disease‐associated and 4 carrier strains notwithstanding a high degree of sequence similarity showed a number of nucleotide changes, some of which possibly resulted in premature translation termination or function loss. In particular, in one disease‐associated strain a 9‐residues insertion was found to be located close to the type I Cu‐site and a catalytic histidine at position 280 was mutated into a leucine. In two strains from carriers, a sequence corresponding to a portion of a transposase gene within the aniA was also found. The AniA protein was always expressed, except for these two carriers strains and for other two strains in which the presence of the premature stop codons was recognized. The biochemical properties of the cloned soluble domain of the enzyme (sAniA) from N. meningitidis reference MC58 strain and from a clinical invasive isolate were studied. In particular, biochemical analysis of sAniA from MC58 demonstrated a clear dependence of its catalytic activity upon acidification, while the clinical isolate‐derived sAniA was not functional. Thus, the results obtained suggest that the presence of a conserved and functional aniA gene is not essential for meningococcal survival. © 2008 IUBMB IUBMB Life, 60(9): 629–636, 2008
Comparison of two extraction procedures for determination of drugs of abuse in human saliva by high‐performance liquid chromatography
Authors: Fernández P. , Morales L. , Vázquez C. , Lago M. , Bermejo A. M.
Published: July 2008

High performance liquid chromatography in combination with diode array detection (HPLC‐DAD) was used to determine morphine, 6‐acetylmorphine, cocaine, benzoylecgonine, cocaethylene, methadone and 2‐ethylene‐1,5‐dimethyl‐3,3,‐diphenylpyrrolidine in human saliva. For comparison, samples were prepared by either liquid–liquid extraction in Toxitubes A® or microwave‐assisted extraction (MAE), by mixing 1 ml of saliva with 10 ml of chloroform and operating at 100 ˚C for 10 min. Acetonitrile and 0.02 m phosphate buffer at pH 6.5 were used as mobile phase in HPLC in gradient mode. The detector response was linear over the drug concentration range of 0.05–2.0 µg ml−1 in human saliva. The analytical method was validated by determining its precision and accuracy (n = 5), which were lower than 5% as relative standard deviation and 6% as relative error. Limits of detection ranged from 10 to 35 ng ml−1; mean recoveries of drugs were from 53 to 95% with Toxitubes A and from 83 to 100% with MAE at two different concentrations (0.1 and 1.0 µg ml−1). The proposed method was applied to 24 saliva samples from individuals poisoned with opiates and/or cocaine. Copyright © 2008 John Wiley & Sons, Ltd.
Self‐aggregation of water‐soluble chitosan and solubilization of thymol as an antimicrobial agent
Authors: Hu Ying , Du Yumin , Wang Xiaoying , Feng Tao
Published: July 2008

Water‐soluble chitosan (WCS) with five different molecular weights was synthesized by N‐acetylation of chitosan with acetic anhydride. Degree of deacetylation of sample was determined by potentiometric titration. The structure of WCS was characterized by FTIR, XRD, and gel permeation chromatography. The critical aggregation concentration (cac) of self‐aggregation was determined by measuring the fluorescence intensity of pyrene as a fluorescent probe and the surface tension. With the decrease of weight‐average molecular weight (Mw), the cac values of WCSs in aqueous media were 0.48, 0.41, 0.35, 0.16, and 0.06 mg/mL, respectively. The surface tension–concentration plots of WCS5 had more than one turning point. It showed WCS had tendency to form monomolecular micelles in the very dilute solutions. In addition, WCSs were evaluated for efficacy of nanoparticle formation, which can solubilize and carry thymol, a water‐insoluble antimicrobial agent. The diameter and morphology of thymol‐loaded WCS nanoparticles were characterized by Zeta size nano series and TEM. Mean diameters of these thymol‐loaded WCS nanoparticles in aqueous media were 189, 167, 134, 35, and 21 nm, respectively. TEM photographs exhibited the thymol‐loaded WCS nanoparticle as a bimolecular micelle. Furthermore, thymol‐loaded WCS nanoparticles had stronger antibacterial activity than thymol. With the decrease of the size, the thymol‐loaded nanoparticles showed a stronger antimicrobial effect on gram‐positive bacteria and fungi. The minimum inhibitory concentration value of thymol‐loaded WCS5 nanoparticle against Staphyococcus aureus and Bacillus subtilis was 0.00313–0.00157% (w/v). © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Increasing cell adhesion on plasma deposited fluorocarbon coatings by changing the surface topography
Authors: Gristina Roberto , D'Aloia Erica , Senesi Giorgio S. , Milella Antonella , Nardulli Marina , Sardella Eloisa , Favia Pietro , d'Agostino Riccardo
Published: July 2008

In designing new biomaterials, it is of outstanding importance to consider how cells respond to specific chemical and topographical features on the material surface. The behavior of most cell types in vivo is strictly related to specific chemical and topographical cues that characterize the extra cellular environment. In particular, during their lives cells react to topographical patterns such as those of the extracellular matrix (ECM), of micro and/or nanometric dimensions. The production of micrometric and/or nanometric features on artificial materials usually involves expensive and time‐consuming methods of manufacturing, such as electron beam and colloidal lithography. In this article, different “Teflon‐like” structured surfaces were deposited from tetrafluoroethylene (C2F4)‐fed plasmas, for the study of cell adhesion and growth. The reaction of different cell lines to different topographical features was evaluated and compared with cell behavior on flat samples with the same chemical composition. Cell adhesion was calculated from area covered by cells at different time of culture. Beside this, cell proliferation was determined with the MTT test. Cell morphology and filopodia interaction with the nanofeatures were also estimated by optical and scanning electron microscopy. A dramatic difference both in adhesion and growth was found between cells seeded on flat and rough surfaces with the density and spreading of adhered cells varying as a function of the roughness of coatings. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009
Chitosan‐NAC nanoparticles as a vehicle for nasal absorption enhancement of insulin
Authors: Wang Xin , Zheng Chao , Wu Zhongming , Teng Dayong , Zhang Xinge , Wang Zhen , Li Chaoxing
Published: July 2008

The purpose of this work was to investigate chitosan‐N‐acetyl‐L‐cysteine (chitosan‐NAC) nanoparticles as a potential carrier system for the nasal delivery of insulin. For the study, we used insulin‐loaded chitosan‐NAC nanoparticles (140–210 nm in diameter) prepared by in situ gelation with tripolyphosphate (TPP), with positive zeta potential values of +19.5–31.7 mV and insulin loading capacities of 13–42%. The physicochemical properties of the nanoparticles were affected by the number of thiol groups present. Mucoadhesive properties, which were evaluated by measuring the in vitro absorbed mass of mucin, of chitosan‐NAC nanoparticles were >1.8‐fold that of unmodified chitosan nanoparticles. In aqueous solution, chitosan‐NAC nanoparticles exhibited fast swelling behavior. Insulin was released from chitosan‐NAC nanoparticles in vitro in an initial burst followed by slow release. Intranasal administration of chitosan‐NAC nanoparticles in rats enhanced the absorption of insulin by the nasal mucosa compared with unmodified chitosan nanoparticles and control insulin solution. In light of these observations, the novel thiolated chitosan nanoparticles represent a promising vehicle for nasal insulin administration. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009
HPLC analysis of eluted monomers from two composite resins cured with LED and halogen curing lights
Authors: Tabatabaee Masumeh Hasani , Mahdavi Hosein , Zandi Shiller , Kharrazi M. J.
Published: July 2008

This study investigated the leaching of monomers (Bis‐GMA and TEGDMA) from nano‐hybrid (Filtek Supreme) and flowable (Filtek Flow) dental composite resins cured with LED or conventional halogen curing lights, and immersed in saliva or water for 24 h. Nine disc specimens were made for each experimental group. After the polymerization process, the specimens were immersed in either water or saliva and incubated at 37°C for 24 h. Eluted Bis‐GMA and TEGDMA monomers were detected using high performance liquid chromatography (HPLC). The data were analyzed using three‐way ANOVA (p = 0.05) and the independent samples t test. TEGDMA (53.15–1 μg/L) was leached from the resins at a higher level than Bis‐GMA (28–0.5 μg/L) (p < 0.01), regardless of the affecting factors: composite type, solvent (media) and type of curing light. In general, Filtek Flow resin released more TEGDMA than Filtek Supreme (p < 0.05), but the Supreme resin released more Bis‐GMA than TEGDMA (p < 0.05). Halogen light induced greater monomer elution than LED light immersion in water. Saliva released more TEGDMA than water (p < 0.05). We conclude that (1) total leached TEGDMA was higher than total Bis‐GMA, (2) saliva and halogen light (lower intensity than LED) leached more monomers from the resins, and (3) the flowable composite resin leached more TEGDMA than the nano‐hybrid. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009
Twisted product of cocycles and factorization of semi‐regular relative difference sets
Authors: Chen Yu Qing
Published: July 2008

In this article, we introduce what we call twisted Kronecker products of cocycles of finite groups and show that the twisted Kronecker product of two cocycles is a Hadamard cocycle if and only if the two cocycles themselves are Hadamard cocycles. This enables us to generalize some known results concerning products and factorizations of central semi‐regular relative difference sets. © 2008 Wiley Periodicals, Inc. J Combin Designs 16: 431–441, 2008
Abductive inferences to psychological variables: Steiger's question and best explanations of psychopathy
Authors: Markus Keith A. , Hawes Samuel W. , Thasites Rula J.
Published: July 2008

Abductive inference often involves inference to the best explanation. A focus on the bestness of explanations facilitates a comparative analysis of how abductive inference would differ if approached with four contrasting sets of assumptions about how scientific inference works: positivism, realism, and two kinds of pragmatism. As a thought experiment, one can imagine a situation in which competing models of psychopathy differ in parsimony and fit to the data, but produce a tie when considering both virtues in combination. The thought experiment demonstrates that Steiger's (1990) question about how best to combine competing virtues in scientific inference applies to abductive inference and that the answers depend upon other assumptions about how science works. The comparative analysis helps focus some of the issues that require clarification before abductive inference can enter the Pantheon of standard research methods in psychology. More constructively, the analysis also demonstrates that one need not accept scientific realism to accept and use abductive inference. © 2008 Wiley Periodicals, Inc. J Clin Psychol 64:1–20, 2008.
In situ immunocytochemical detection of altered membrane composition induced by cell–cell contact in cultured mammalian cells
Authors: Du Wei , Cui Zheng , Tsui Zhao‐Chun , Chen Qirui , Willingham Mark C.
Published: July 2008

In cultured mammalian cells, both normal and transformed, cell–cell contact was shown to alter the detergent extractability of nuclear and plasma membranes detected using immunocytochemistry of fixed cells. This alteration occurred in each cell individually, occurred in less than 1 hour after altered cell–cell contact, and did not involve new protein or mRNA synthesis. These results indicate that composition of cellular membranes is highly affected by cell–cell contact and culture density. This suggests a possible role for alterations in membrane composition in the regulation of normal cell behavior, including signaling of contact inhibition of growth and movement, behaviors that are not effectively regulated by cell–cell contact in malignantly transformed cells. Microsc. Res. Tech., 2008. © 2008 Wiley‐Liss, Inc.
Adaptive or maladaptive response to adenoviral adrenomedullin gene transfer is context‐dependent in the heart
Authors: Leskinen Hanna , Rauma‐Pinola Tanja , Szokodi István , Kerkelä Risto , Pikkarainen Sampsa , Uusimaa Paavo , Hautala Timo , Vuolteenaho Olli , Ruskoaho Heikki
Published: July 2008

Background

Adrenomedullin (AM) is a potent vasodilator and natriuretic peptide produced in the heart, but controversy persists regarding its cardiac effects. We explored the potential role of AM on cardiac function and remodeling by direct recombinant adenoviral AM gene delivery into the anterior wall of the left ventricle (LV).

Methods

AM was overexpressed in healthy rat hearts and in hearts during the remodeling process in response to pressure overload and myocardial infarction. The AM effects were analysed with echocardiography and in an isolated perfused rat heart preparation. The expression of AM and the activation of underlying signaling pathways were also investigated.

Results

AM mRNA increased by 20.9‐fold (p < 0.001) in healthy rat heart and improved fractional shortening by 14% (p < 0.05) and ejection fraction by 8% (p < 0.05). In isolated perfused hearts, an increase (p < 0.05) in the first derivative of isovolumic LV pressure rise (dP/dtmax) without alteration in diastolic properties was noted. The overexpression of AM activated protein kinase Cε and Cδ isoforms in the LV, whereas p38 mitogen‐activated protein kinase activity decreased. Angiotensin II‐induced LV hypertrophy was significantly attenuated by AM (p < 0.01) without compromising cardiac contractility. By contrast, AM enhanced LV dilatation (p < 0.01) and anterior wall thinning (p < 0.001) and augmented the deterioration of LV function (p < 0.05) post‐infarction.

Conclusions

The results obtained in the present study show that AM overexpression improves LV systolic function without altering cardiac diastolic properties in the normal heart. Moreover, AM is a potent context‐dependent modulator of LV remodeling because it promotes an adaptive response in pressure overload‐induced LV hypertrophy and triggers a maladaptive process in post‐infarction remodeling. Copyright © 2008 John Wiley & Sons, Ltd.
Clinical‐grade preparation of human natural regulatory T‐cells encoding the thymidine kinase suicide gene as a safety gene
Authors: Guillot‐Delost Maude , Cheraï Mustapha , Hamel Yamina , Rosenzwajg Michelle , Baillou Claude , Simonin Ghislaine , Leclercq Virginie , Mariotti‐Ferrandiz Maria Encarnita , Six Adrien , Bon‐Durand Véronique , Maury Sébastien , Salomon Benoit L. , Cohen José L. , Klatzmann David , Lemoine François M.
Published: July 2008

Background

Human CD4+CD25+FOXP3+ natural regulatory T‐cells (nTreg) have a great therapeutic potential for the induction of tolerance in allo‐transplanted patients or for the control of severe auto‐immune diseases. However, clinical‐grade production of nTreg remains difficult to achieve because of the absence of a truly specific surface marker and of their low frequency that implies a need for their ex vivo expansion. Furthermore, safety issues should be taken into consideration due to the risk of either uncontrolled nTreg‐induced immunosuppression or uncontrolled proliferation of autoreactive contaminating T‐cells particularly in an auto‐immune context.

Methods

We compared different clinical‐grade conditions for immuno‐magnetic selection and ex vivo expansion of nTreg. For safety, expanded cells were genetically modified with retroviral vectors co‐expressing human CD90 and HSV1 thymidine kinase. The CD90 surface marker and thymidine kinase allow for selection and elimination of transduced cells by ganciclovir, respectively.

Results

We showed that (i) nTreg could be enriched in a one step using CD25 microbeads, were functionally suppressive and mainly FOXP3+; (ii) using anti‐CD28‐ and anti‐CD3‐coated beads, interleukin‐2 and rapamycin, nTreg were expanded 150–200‐fold after 3 weeks. Under these clinical‐grade conditions, they remained suppressive, and no major alteration of the TCR repertoire was observed; (iii) after efficient retroviral transduction and CD90 selection, nTreg maintained their suppressive activity; (iv) transduced nTreg could be eliminated by ganciclovir upon activation.

Conclusions

The efficient procedure reported here for the preparation of nTreg, whose safety has been ensured, is now applicable for further clinical trials. Copyright © 2008 John Wiley & Sons, Ltd.
The administration of an adenoviral thymidine kinase suicide gene to the uterine artery of rabbits does not affect fertility: a safety study of pregnant and nonpregnant rabbits and their offspring
Authors: Laurema Anniina , Riekkinen Mervi , Heikura Tommi , Vähäkangas Elisa , Manninen Hannu , Heinonen Seppo , Ylä‐Herttuala Seppo
Published: July 2008

Background

Adenoviral gene therapy, based on herpes simplex virus thymidine kinase (AdvTK) is being developed for clinical use but no safety data are available with respect to the effects on female germ cells should the virus accidentally be released into systemic circulation. We studied the effects of AdvTK gene therapy on ovaries and germ cells in pregnant and nonpregnant rabbits and also the potential transmission of a transgene to offspring.

Methods

To mimic the severest of conditions, gene transfer was made by direct catheter‐mediated injection into the uterine artery of pregnant and nonpregnant rabbits. AdvTK or AdvLacZ at 1 × 1010 pfu were used for gene transfer. Ganciclovir was administered to AdvTK‐treated rabbits to induce gene therapy. The rabbits were mated 6 and 12 weeks following gene transfer and the surviving young (89 from a total of 114) were analysed.

Results

No change in fertility was observed in the two matings after the gene transfer. In addition, no change was observed in ovarian histology between the AdvTK group, the AdvLacZ group and the nontreated controls. Southern blotting analysis showed no genomic integration of the transgene. However, in PCR analysis, transgene DNA was found in 9.3% of the litter samples. This was not the case for results from the reverse transcription‐PCR assay.

Conclusions

Although AdvTK gene therapy may initially affect ovarian cells, the influence appears to be transient. However, after direct exposure of the ovarian cells in high concentration of adenoviruses, transmission of a transgene in the offspring cannot be excluded. Copyright © 2008 John Wiley & Sons, Ltd.
Development of adenovirus immobilization strategies for in situ gene therapy
Authors: Hu Wei‐Wen , Lang Michael W. , Krebsbach Paul H.
Published: July 2008

Background

Regenerative gene therapy using viral vectors enables transduced cells to express bioactive factors in vivo. Viral delivery with spatial control can enhance transduction efficiency and may limit systemic infection. Consequently, we tethered biotinylated adenovirus via interactions with avidin on chitosan surfaces to gain robust control for in situ transduction.

Methods

Avidin was either directly conjugated to chitosan (virus–biotin–avidin‐material; VBAM) or indirectly docked on biotinylated chitosan surfaces (virus–biotin–avidin–biotin‐material; VBABM) to tether biotinylated adenovirus. Enzyme‐linked immunosorbent assay (ELISA) and spectroscopic analysis were performed to demonstrate the binding profiles. Biotin‐alkaline phosphatase and biotinylated adenovirus were used as different sized particles to evaluate binding efficiencies and were compared by the Sips isotherm adsorption method. Scanning electron microscopy (SEM) examination illustrated virus distribution, and the transduction efficiency was determined by in vitro cell transduction.

Results

ELISA and spectroscopic analysis both demonstrated that the VBAM system led to multilayer avidin formation on biomaterial surfaces, whereas VBABM formed a monolayer of avidin. Sips isotherm adsorption indicated that the VBAM method increased heterogeneity and steric hindrance of binding sites. By contrast, the VBABM method docked avidin on chitosan surfaces and orientated the binding sites to facilitate ligand binding. In addition, SEM images illustrated that the VBABM method led to more even viral distribution. In vitro cell infection experiments also demonstrated that the VBABM system enhanced virus immobilization and thus improved cell transduction efficiency over the VBAM system.

Conclusions

The VBABM strategy is a superior method for in situ transduction from biomaterials. This strategy could be adapted for use with a variety of biomaterials as well as viral vectors, and thus may be an alternative method for in vivo regenerative gene therapy. Copyright © 2008 John Wiley & Sons, Ltd.
Circular chromatic index of type 1 Blanuša snarks
Authors: Mazák Ján
Published: July 2008

We determine the exact value of the circular chromatic index of generalized Blanuša snarks of type 1 introduced by Watkins more than two decades ago. In this case, the index takes infinitely many values and can get arbitrarily close to 3. Generalized Blanuša snarks are the first explicit class with this property; until now only finitely many values of the circular chromatic index of snarks have been known. © 2008 Wiley Periodicals, Inc. J Graph Theory 59: 89–96, 2008
Live single‐cell video‐mass spectrometry for cellular and subcellular molecular detection and cell classification
Authors: Mizuno Hajime , Tsuyama Naohiro , Harada Takanori , Masujima Tsutomu
Published: July 2008

The molecular content from the cytoplasm of a live, single mammalian cell and its organelle were trapped with a nano‐electrospray ionization (ESI) tip acting as a micropipette under a video microscope, and hundreds of small molecular peaks were detected by direct nano‐ESI mass spectrometry (MS). Granule‐ or cytoplasm‐specific peaks in a mast cell (RBL 2H3) model were extracted by paired t‐test to demonstrate their specific localization. Some of the typical and specific molecules were successfully identified by MS/MS analysis. This method was also applied to the cell classification of seven types of cell lines at the single‐cellular level by principal component analysis (PCA), revealing seven clusters in the multivariate score plot. Copyright © 2008 John Wiley & Sons, Ltd.