bluuebunny/crossref_metadata_2025 huggingface dataset:
Vaccinia Virus Telomeres: Interaction with the Viral I1, I6, and K4 Proteins
Authors: DeMasi Joseph , Du Shan , Lennon David , Traktman Paula
Published: July 2002

The 192-kb linear DNA genome of vaccinia virus has covalently closed hairpin termini that are extremely AT rich and contain 12 extrahelical bases. Vaccinia virus telomeres have previously been implicated in the initiation of viral genome replication; therefore, we sought to determine whether the telomeres form specific protein-DNA complexes. Using an electrophoretic mobility shift assay, we found that extracts prepared from virions and from the cytoplasm of infected cells contain telomere binding activity. Four shifted complexes were detected using hairpin probes representing the viral termini, two of which represent an interaction with the “flip” isoform and two with the “flop” isoform. All of the specificity for protein binding lies within the terminal 65-bp hairpin sequence. Viral hairpins lacking extrahelical bases cannot form the shifted complexes, suggesting that DNA structure is crucial for complex formation. Using an affinity purification protocol, we purified the proteins responsible for hairpin-protein complex formation. The vaccinia virus I1 protein was identified as being necessary and sufficient for the formation of the upper doublet of shifted complexes, and the vaccinia virus I6 protein was shown to form the lower doublet of shifted complexes. Competition and challenge experiments confirmed that the previously uncharacterized I6 protein binds tightly and with great specificity to the hairpin form of the viral telomeric sequence. Incubation of viral hairpins with extracts from infected cells also generates a smaller DNA fragment that is likely to reflect specific nicking at the apex of the hairpin; we show that the vaccinia virus K4 protein is necessary and sufficient for this reaction. We hypothesize that these telomere binding proteins may play a role in the initiation of vaccinia virus genome replication and/or genome encapsidation.
Role of Tachykinins in the Host Response to Murine Gammaherpesvirus Infection
Authors: Payne Catherine M. , Heggie Caroline J. , Brownstein David G. , Stewart James P. , Quinn John P.
Published: July 2002

Tachykinins function not only as neurotransmitters but also as immunological mediators. We used infection of tachykinin-deficient (PPT-A−/−) mice and wild-type controls with murine gammaherpesvirus to assess the role of tachykinins in the host response to a virus infection. Although infection was ultimately controlled inPPT-A−/−mice, there were higher titers of infectious virus in the lungs, accompanied by a more rapid influx of inflammatory cells. Clearance of latently infected cells from the spleen was also delayed. This is the first report of the direct influence of tachykinins in the host response to a virus infection.
A Chimeric Human-Bovine Parainfluenza Virus Type 3 Expressing Measles Virus Hemagglutinin Is Attenuated for Replication but Is Still Immunogenic in Rhesus Monkeys
Authors: Skiadopoulos Mario H. , Surman Sonja R. , Riggs Jeffrey M. , Collins Peter L. , Murphy Brian R.
Published: July 2002

The chimeric recombinant virus rHPIV3-N
B
, a version of human parainfluenza virus type 3 (HPIV3) that is attenuated due to the presence of the bovine PIV3 nucleocapsid (N) protein open reading frame (ORF) in place of the HPIV3 ORF, was modified to encode the measles virus hemagglutinin (HA) inserted as an additional, supernumerary gene between the HPIV3 P and M genes. This recombinant, designated rHPIV3-N
B
HA, replicated like its attenuated rHPIV3-N
B
parent virus in vitro and in the upper and lower respiratory tracts of rhesus monkeys, indicating that the insertion of the measles virus HA did not further attenuate rHPIV3-N
B
in vitro or in vivo. Monkeys immunized with rHPIV3-N
B
HA developed a vigorous immune response to both measles virus and HPIV3, with serum antibody titers to both measles virus (neutralizing antibody) and HPIV3 (hemagglutination inhibiting antibody) of over 1:500. An attenuated HPIV3 expressing a major protective antigen of measles virus provides a method for immunization against measles by the intranasal route, a route that has been shown with HPIV3 and respiratory syncytial virus vaccines to be relatively refractory to the neutralizing and immunosuppressive effects of maternally derived virus-specific serum antibodies. It should now be possible to induce a protective immune response against measles virus in 6-month-old infants, an age group that in developing areas of the world is not responsive to the current measles virus vaccine.
Glycoprotein K Specified by Herpes Simplex Virus Type 1 Is Expressed on Virions as a Golgi Complex-Dependent Glycosylated Species and Functions in Virion Entry
Authors: Foster Timothy P. , Rybachuk Galena V. , Kousoulas Konstantin G.
Published: July 2002

To facilitate detection of glycoprotein K (gK) specified by herpes simplex virus, a 12-amino-acid epitope tag was inserted within gK domain III. Recombinant virus gKprotC-DIII, expressing the tagged gK, was isolated. This virus formed wild-type plaques and replicated as efficiently as the wild-type KOS virus in Vero cells. Anti-protein C MAb detected high-mannose and Golgi complex-dependent glycosylated gK within cells as well as on purified virions. The gK-null virus ΔgK (gK
−/−
) entered Vero cells substantially more slowly than the wild-type KOS (gK
+/+
), while ΔgK virus grown in complementing VK302 cells (gK
−/+
) entered with entry kinetics similar to those of the KOS virus.
Improved Adenovirus Vectors for Infection of Cardiovascular Tissues
Authors: Havenga M. J. E. , Lemckert A. A. C. , Grimbergen J. M. , Vogels R. , Huisman L. G. M. , Valerio D. , Bout A. , Quax P. H. A.
Published: July 2002

To identify improved adenovirus vectors for cardiovascular gene therapy, a library of adenovirus vectors based on adenovirus serotype 5 (Ad5) but carrying fiber molecules of other human serotypes, was generated. This library was tested for efficiency of infection of human primary vascular endothelial cells (ECs) and smooth muscle cells (SMCs). Based on luciferase, LacZ, or green fluorescent protein (GFP) marker gene expression, several fiber chimeric vectors were identified that displayed improved infection of these cell types. One of the viruses that performed particularly well is an Ad5 carrying the fiber of Ad16 (Ad5.Fib16), a subgroup B virus. This virus showed, on average, 8- and 64-fold-increased luciferase activities on umbilical vein ECs and SMCs, respectively, compared to the parent vector. GFP and
lacZ
markers showed that approximately 3-fold (ECs) and 10-fold (SMCs) more cells were transduced. Experiments performed with both cultured SMCs and organ cultures derived from different vascular origins (saphenous vein, iliac artery, left interior mammary artery, and aorta) and from different species demonstrated that Ad5.Fib16 consistently displays improved infection in primates (humans and rhesus monkeys). SMCs of the same vessels of rodents and pigs were less infectable with Ad5.Fib16 than with Ad5. This suggests that either the receptor for human Ad16 is not conserved between different species or that differences in the expression levels of the putative receptor exist. In conclusion, our results show that an Ad5-based virus carrying the fiber of Ad16 is a potent vector for the transduction of primate cardiovascular cells and tissues.
Essential Roles for CD8+T Cells and Gamma Interferon in Protection of Mice against Retrovirus-Induced Immunosuppression
Authors: Dittmer Ulf , Race Brent , Peterson Karin E. , Stromnes Ingunn M. , Messer Ronald J. , Hasenkrug Kim J.
Published: July 2002

It is known that both animal and human retroviruses typically cause immunosuppression in their respective hosts, but the mechanisms by which this occurs are poorly understood. The present study uses Friend virus (FV) infections of mice as a model to determine how major histocompatibility complex (MHC) genes influence immunosuppression. Previously, MHC-I genes were shown to influence antibody responses to potent antigenic challenges given during acute FV infection. The mapping of an immune response to an MHC-I gene implicated CD8+T cells in the mechanism, so we directly tested for their role by using in vivo CD8+T-cell depletions. Mice resistant to FV-induced immunosuppression became susceptible when they were depleted of CD8+T cells. Resistance also required gamma interferon (IFN-γ), as in vivo neutralization of IFN-γ converted mice from a resistant to susceptible phenotype. On the other hand, susceptibility to FV-induced immunosuppression was dependent on the immunosuppressive cytokine, interleukin-10 (IL-10), as antibody responses were restored in susceptible mice when IL-10 function was blocked in vivo. Thus, FV-induced immunosuppression of antibody responses involves complex mechanisms controlled at least in part by CD8+T cells.
Pivotal Role of the Non-
hr
Origin of DNA Replication in the Genesis of Defective Interfering Baculoviruses
Authors: Pijlman Gorben P. , Dortmans Jos C. F. M. , Vermeesch Angela M. G. , Yang Kai , Martens Dirk E. , Goldbach Rob W. , Vlak Just M.
Published: July 2002

The generation of deletion mutants, including defective interfering viruses, upon serial passage of
Spodoptera exigua
multicapsid nucleopolyhedrovirus (Se
M
NPV) in insect cell culture has been studied. Sequences containing the non-homologous region origin of DNA replication (non-
hr ori
) became hypermolar in intracellular viral DNA within 10 passages in Se301 insect cells, concurrent with a dramatic drop in budded virus and polyhedron production. These predominant non-
hr ori
-containing sequences accumulated in larger concatenated forms and were generated de novo as demonstrated by their appearance and accumulation upon infection with a genetically homogenous bacterial clone of Se
M
NPV (bacmid). Sequences were identified at the junctions of the non-
hr ori
units within the concatemers, which may be potentially involved in recombination events. Deletion of the Se
M
NPV non-
hr ori
using RecE/RecT-mediated homologous ET recombination in
Escherichia coli
resulted in a recombinant bacmid with strongly enhanced stability of virus and polyhedron production upon serial passage in insect cells. This suggests that the accumulation of non-
hr ori
s upon passage is due to the replication advantage of these sequences. The non-
hr ori
deletion mutant Se
M
NPV bacmid can be exploited as a stable eukaryotic heterologous protein expression vector in insect cells.
Binding of FADD and Caspase-8 to Molluscum Contagiosum Virus MC159 v-FLIP Is Not Sufficient for Its Antiapoptotic Function
Authors: Garvey Tara L. , Bertin John , Siegel Richard M. , Wang Guang-hua , Lenardo Michael J. , Cohen Jeffrey I.
Published: July 2002

Molluscum contagiosum virus (MCV), a member of the human poxvirus family, encodes the MC159 protein that inhibits Fas-, tumor necrosis factor (TNF)-, and TNF-related apoptosis-inducing ligant (TRAIL)-induced apoptosis. We used site-directed mutagenesis to change charged or hydrophobic amino acid residues to alanines to identify regions of MC159 that are critical for protection from apoptosis and for protein-protein interactions. Surprisingly, while MC159 is thought to block apoptosis by binding to Fas-associated death domain (FADD) or caspase-8, several mutants that lost apoptosis blocking activity still bound to both FADD and caspase-8. Mutations in the predicted hydrophobic patch 1 and α2 regions of both death effector domains (DEDs) within MC159 resulted in loss of the ability to bind to FADD or caspase-8 and to block apoptosis. Amino acid substitutions in the RXDL motif located in the α6 region of either DED resulted in loss of protection from apoptosis induced by Fas, TNF, and TRAIL and abolished the ability of MC159 to block death effector filament formation. Thus, charged or hydrophobic amino acids in three regions of the MC159 DEDs (hydrophobic patch 1, α2, and α6) are critical for the protein’s ability to interact with cellular proteins and to block apoptosis.
High Prevalence of Simian T-Lymphotropic Virus Type L in Wild Ethiopian Baboons
Authors: Takemura Taichiro , Yamashita Masahiro , Shimada Makoto K. , Ohkura Sadayuki , Shotake Takayoshi , Ikeda Mikio , Miura Tomoyuki , Hayami Masanori
Published: July 2002

Simian T-cell leukemia viruses (STLVs) are the simian counterparts of human T-cell leukemia viruses (HTLVs). A novel, divergent type of STLV (STLV-L) from captive baboons was reported in 1994, but its natural prevalence remained unclear. We investigated the prevalence of STLV-L in 519 blood samples from wild-living nonhuman primates in Ethiopia. Seropositive monkeys having cross-reactive antibodies against HTLV were found among 22 out of 40 hamadryas baboons, 8 of 96 anubis baboons, 24 of 50 baboons that are hybrids between hamadryas and anubis baboons, and 41 of 177 grivet monkeys, but not in 156 gelada baboons. A Western blotting assay showed that sera obtained from seropositive hamadryas and hybrid baboons exhibited STLV-L-like reactivity. A PCR assay successfully amplified STLV sequences, which were subsequently sequenced and confirmed as being closely related to STLV-L. Surprisingly, further PCR showed that nearly half of the hamadryas (20 out of 40) and hybrid (19 out of 50) baboons had STLV-L DNA sequences. In contrast, most of the seropositive anubis baboons and grivet monkeys carried typical STLV-1 but not STLV-L. These observations demonstrate that STLV-L naturally prevails among hamadryas and hybrid baboons at significantly high rates. STLV-1 and -2, the close relative of STLV-L, are believed to have jumped across simian-human barriers, which resulted in widespread infection of HTLV-1 and -2. Further studies are required to know if STLV-L is spreading into human populations.
De Novo Infection and Serial Transmission of Kaposi's Sarcoma-Associated Herpesvirus in Cultured Endothelial Cells
Authors: Lagunoff Michael , Bechtel Jill , Venetsanakos Eleni , Roy Anne-Marie , Abbey Nancy , Herndier Brian , McMahon Martin , Ganem Don
Published: July 2002

Infection by Kaposi's sarcoma-associated herpesvirus (KSHV) is central to the pathogenesis of the endothelial neoplasm Kaposi's sarcoma (KS) and is also linked to the rare B-cell tumor known as primary effusion lymphoma (PEL). Latently infected PEL cell lines can be induced to enter the lytic cycle and produce KSHV virions. However, such cells do not support de novo infection or serial propagation of KSHV. These limitations have prevented the development of systems for the genetic analysis of KSHV and have impeded a deeper understanding of KS pathogenesis. Here we show that human dermal microvascular endothelial cells immortalized by expression of telomerase can be readily infected by KSHV virions produced by PEL cells. Infection is predominantly latent, but a small subpopulation enters the lytic cycle spontaneously. Phorbol ester (tetradecanoyl phorbol acetate [TPA]) treatment of latently infected cells leads to enhanced induction of lytic KSHV replication, resulting in foci of cytopathic effect. There is no cytopathic effect or viral DNA expansion when infected TIME cells (telomerase-immortalized microvascular endothelial cells) are TPA induced in the presence of phosphonoacetic acid (PAA), an inhibitor of herpesvirus replication. Supernatants from phorbol-induced cultures transfer latent KSHV infection to uninfected cells, which can likewise be induced to undergo lytic replication by TPA treatment, and the virus can be further serially transmitted. Serial passage of the virus in TIME cells is completely inhibited when TPA treatment is done in the presence of PAA. Latently infected endothelial cells do not undergo major morphological changes or growth transformation, and infection is lost from the culture upon serial passage. This behavior faithfully recapitulates the behavior of spindle cells explanted from primary KS biopsies, strongly supporting the biological relevance of this culture system. These findings suggest that either the stability or the growth-deregulatory potential of the KSHV latency program in endothelial cells is more limited than might be predicted by analogy with other oncogenic viruses.
Mapping the Rubella Virus Subgenomic Promoter
Authors: Tzeng Wen-Pin , Frey Teryl K.
Published: July 2002

Rubella virus
(RUB), the sole member of the
Rubivirus
genus in the
Togaviridae
family of positive-strand RNA viruses, synthesizes a single subgenomic (SG) RNA containing sequences from the 3′ end of the genomic RNA including the open reading frame (ORF) that encodes the virion proteins. The synthesis of SG RNA is initiated internally on a negative-strand, genome-length template at a site known as the SG promoter (SGP). Mapping the RUB SGP was initiated by using an infectious cDNA vector, dsRobo402/GFP, in which the region containing the SGP was duplicated (K. V. Pugachev, W.-P. Tzeng, and T. K. Frey, J. Virol. 74:10811-10815, 2000). In dsRobo402/GFP, the 5′-proximal nonstructural protein ORF (NS-ORF) is followed by the first SGP (SGP-1), the green fluorescent protein (GFP) gene, the second SGP (SGP-2), and the structural protein ORF. The duplicated SGP, SGP-2, contained nucleotides (nt) −175 to +76 relative to the SG start site, including the 3′ 127 nt of the NS-ORF and 47 nt between the NS-ORF and the SG start site. 5′ Deletions of SGP-2 to nt −40 (9 nt beyond the 3′ end of the NS-ORF) resulted in a wild-type (wt) phenotype in terms of virus replication and RNA synthesis. Deletions beyond this point impaired viability; however, the analysis was complicated by homologous recombination between SGP-1 and SGP-2 that resulted in deletion of the GFP gene and resurrection of viable virus with one SGP. Since the NS-ORF region was not necessary for SGP activity, subsequent mapping was done by using both replicon vectors, RUBrep/GFP and RUBrep/CAT, in which the SP-ORF is replaced with the reporter GFP and chloramphenical acetyltransferase genes, respectively, and the wt infectious clone, Robo402. In the replicon vectors, 5′ deletions to nt −26 resulted in the synthesis of SG RNA. In the infectious clone, deletions through nt −28 gave rise to viable virus. A series of short internal deletions confirmed that the region between nt −28 and the SG start site was essential for viability and showed that the repeated UCA triplet at the 5′ end of SG RNA was also required. Thus, the minimal SGP maps from nt −26 through the SG start site and appears to extend to at least nt +6, although a larger region is required for the generation of virus with a wt phenotype. Interestingly, while the positioning of the RUB SGP immediately adjacent the SG start site is thus similar to that of members of the genus
Alphavirus
, the other genus in the
Togaviridae
family, it does not include a region of nucleotide sequence homology with the alphavirus SGP that is located between nt −48 and nt −23 with respect to the SG start site in the RUB genome.
Ab initio Study of the Potential Curves for CO (X1Σ+), CH (X2Π) and OH (X2Π)
Authors: Bittererová Martina , Biskupič Stanislav , Lischka Hans , Klimo Viliam
Published: July 2002

Calculations on spectroscopic properties of the diatomic systems CO, CH, and OH have been carried out by the multiconfiguration SCF, single reference and multireference single and double excitation CI, and average coupled pair functional methods. An evaluation of the different theoretical approaches is performed in order to get better insight into the selection of appropriate procedures for the calculation of the potential energy surface of the H + CO system.
Polarographic Reduction and Potential Carcinogenity of Synthetic 1,3,5-Triazine Bases and Nucleosides
Authors: Novotný Ladislav , Vachálková Anna , Pískala Alois
Published: July 2002

DC polarographic parameters were measured for a series of 15 synthetic 5-aza compounds derived from cytosine, cytidine, uracil and uridine in nonaqueous (dimethylformamide) solutions. The substances in aprotic media are reduced in a single two-electron step at the mercury drop electrode, except for 5,6-dihydro derivatives of 5-azauracil and 5-azauridine which are reduced in two steps. α-Lipoic acid was added to the solutions of the substances, and the slopes tg α of the plots of diffusion current of the substances versus a-lipoic acid concentration, which can serve as an index of potential carcinogenic activity of the substances measured, were determined. The tg α values of all the compounds studied are low as compared to related substances whose carcinogenic activity has been proved. 5-Azacytidine and 5-azauracil are exceptions exhibiting tg a values of 0.295 and 0.400, respectively. For the former compound, this is consistent with the WHO classification as "probably carcinogenic to humans".
Metathesis Catalyst WOCl4/Ph4Sn: The Chemistry of Ripening and Transformation to Polymerization Active Species
Authors: Sedláček Jan , Vohlídal Jiří , Kareš Jiří , Pacovská Marta , Máca Bohumil
Published: July 2002

Ripening of WOCl4/Ph4Sn binary catalyst of composition from 1 : 1 to 1 : 4 in benzene and toluene is a complex reaction in which initially homogeneous solution of the catalyst components continually changes to a colloidal solution, and utterly to a suspension of catalytically active particles consisted of lower-valent tungsten species in the solution of biphenyl, Ph3SnCl and, eventually, unreacted Ph4Sn (at composition 1 : 4). The yield of biphenyl is in a simple stoichiometric relation with the initial concentration of WOCl4 and a proof is given that the phenyl groups constituting biphenyl molecules come from Ph4Sn cocatalyst. The mechanism of the overall process is suggested. Above twenty, mostly phenylated derivatives of phenylacetylene (PhA) and oligomers of two degrees of unsaturation were detected as by-products of PhA reaction with the ripened catalyst formed in addition to main product, poly(phenylacetylene). The excess phenyls built in the oligomers are demonstrated to come from the supplied Ph4Sn. The mechanism is suggested for formation of the both oligomeric derivatives and growing carbene centers in which oxo ligands of tungsten centers are proposed to play a central role.
Sterically Crowded Heterocycles. I. Molecular Structure of 3-Chain Substituted 2-Phenylimidazo[1,2-a]pyridinium Salts
Authors: Böhm Stanislav , Kubík Richard , Hradilek Martin , Němeček Jan , Hušák Michal , Pakhomova Svetlana , Ondráček Jan , Kratochvíl Bohumil , Kuthan Josef
Published: July 2002

2-Phenyl-3-[(Z)-1,3-diphenyl-3-oxopropenyl]imidazo[1,2-a]pyridine (I) was converted to corresponding 1-alkyl-2-phenyl-3-[(Z)-1,3-diphenyl-3-oxopropenyl]imidazo[1,2-a]pyridinium salts III - VI and 2-phenyl- 3-[(Z)-1,3-diphenyl-3-oxopropenyl]imidazo[1,2-a]pyridinium perchlorate (VII). The protonation site is discussed in terms of calculated molecular energies of alternative cations. The X-ray structure analyses of enone I and its quaternary periodide III are reported and compared with the PM3 molecular optimizations. 1H NMR and 13C NMR spectra reveal well changes of molecular structures caused by the transformation of the base I into salts III - VII.
Reactions of 2-Acyl-1,3-indandiones with Nitrogen Nucleophiles
Authors: Hrnčiar Pavel , Švanygová Eva
Published: July 2002

Reactions of 2-acyl-1,3-indandiones with nitrogen nucleophiles were studied rarely. The question, if they react with carbonyl carbon of acyl group or indandione skeleton, has not been answered unambiguously. To make clear the question which carbonyl carbon of 2-acyl-1,3-indandiones enters the reaction with nitrogen nucleophiles we carried out the reactions with 2-acetyl- (Ia), 2-propionyl- (Ib), 2-pivaloyl- (Ic), and 2-benzoyl-1,3-indandione (Id). We used different 2-acyl-1,3-indandiones with the aim to find out if the character of acyl group affects the course of reaction. We used ethoxyamine, primary amines, phenylhydrazine, hydrazine and methylhydrazine as nucleophile reactants. The reactions were carried out in methanol at reflux at 10% excess of nitrogen base. The reactions with phenylhydrazine, hydrazine and methylhydrazine were performed with twofold excess of nitrogen base. The separation of reaction products was carried out by chromatography on silica gel. We found that 2-acyl-1,3-indandiones I react with ethoxyamine both at the acylcarbonyl carbon to produce 2-(1-ethoxyiminoalkyl)-1,3-indandiones II and the carbonyl carbon of indandione skeleton to give rise 3-(ethoxyimino)-2-acyl-1-indanones III. In all cases, the carbonyl carbon of acyl group was preferred (the observed ratio of products II to III was 6 - 8 : 1). From the reaction of 2-acyl-1,3-indandiones with primary amines only the products IV of reaction with the acylcarbonyl carbon were isolated. The hydrazines used reacted with 2-acyl-1,3-indandiones also at carbonyl carbon of acyl group in the first step to produce hydrazones. However, the products isolated in most cases were formed by the attack of hydrazone nitrogen at carbonyl carbon of indandione skeleton giving rise to derivatives of indeno[2,3-d]pyrazole-4-one V. It is interesting that 2-acetyl-1,3-indandione and 2-propionyl-1,3-indandione, reacting with phenylhydrazine and hydrazine, yielded only corresponding hydrazones VI.
Synthesis of 3,7,11-Trimethyl-10-oxododecanoic Acid
Authors: Sharma Madan L. , Chand Tek
Published: July 2002

3,7,11-Trimethyl-10-oxododecanoic acid (I) was prepared from 6-bromo-2-methylhept-2-ene (II) in six steps.
A Short and Convenient Synthesis of (2E,4E)-N-Isobutyl-7-(3,4-methylenedioxyphenyl)hepta-2,4-dienamide
Authors: Kad Goverdhan L. , Arora Ajay K. , Singh Sukhwinder , Singh Jasvinder
Published: July 2002

(2E,4E)-N-Isobutyl-7-(3,4-methylenedioxyphenyl)hepta-2,4-dienamide (I) possessing insecticidal activity against flies and mosquitoes has been synthesized in fine yield from 3-(3,4-methylenedioxyphenyl)prop-1-ene (II) using hydroboration-oxidation with acetoxyborohydride and a modified Wittig reaction as the key steps.
Study of Microwave Irradiation Effect on Condensation of 6-R-3-Formylchromones with Active Methylene Compounds
Authors: Gašparová Renata , Lácová Margita
Published: July 2002

Condensation of 6-R-3-formylchromones (I) with 3,3-dimethyl-1,3-cyclohexanedione (II), 1,3-indandione (III), 1,2e-biindenylidene-3,1e,3e-trione (bindone, IV), 2-oxo-1,4-benzothiazine (V) and 3-oxo-2,3-dihydro-1-thia-3a,8-diazacyclopent[a]indene (VI) by the "classical" method, as well as condensation in a microwave oven, has been studied. Some subsequent reactions of these products are described.
Approximate Calculation of Pressure Drop in Laminar Flow of Generalized Newtonian Fluid Through Channels with Insert
Authors: Dolejš Václav , Doleček Petr , Machač Ivan , Šiška Bedřich
Published: July 2002

An equation of Rabinowitsch-Mooney type has been suggested for approximate calculation of pressure drop in flow of generalized Newtonian fluid through channels with insert both in the region of creeping flow and at higher values of the Reynolds number, and this calculation method has been verified for four types of insert using own numerical solution and experimental results as well as literature data.
Polysaccharides from the Flowers of Malva Mauritiana L.: Structure of an Arabinogalactan
Authors: Capek Peter , Kardašová Alžbeta
Published: July 2002

A water-soluble arabinogalactan composed of D-galactose and L-arabinose in the mole ratio 1 : 1.4 has been isolated from the flowers of Malva mauritiana L. Partial acid hydrolysis, methylation analysis, periodate oxidation, and 13C NMR spectroscopy were employed in structure elucidation. The arabinogalactan was shown to have a highly branched structure. The core consisted of 1,6-linked β-D-galactopyranose units, about 65% of which were substituted in position C-3 by side-chains of mainly 1,5-linked α-L-arabinofuranosyl residues.
Preparation of 7-[O-(Carboxymethyl)oxime] Derivatives of Dehydroepiandrosterone and Pregnenolone
Authors: Pouzar Vladimír , Slavíková Tereza , Černý Ivan
Published: July 2002

Novel simple syntheses of 7-(O-carboxymethyloxime) derivatives of 3β-hydroxyandrost-5-en-17-one (dehydroepiandrosterone) and of 3β-hydroxypregn-5-en-20-one (pregnenolone) are reported. In the first one, 17-oxoandrost-5-en-3β-yl acetate was oxidized to give the 7,17-dione which was then selectively reduced in the position 17. Oximation, reoxidation, and deacetylation yielded the desired oxime. The second synthesis started with (20R)-pregn-5-ene-3β,20-diol 3-acetate which was converted to the 20-nitrate, oxidized to the C-7 ketone, oximated, partially deprotected in position 20, reoxidized, and deacetylated. Both the 7-(O-carboxymethyloxime) derivatives have been devised as immunoassay components.
Synthesis of Juvenile Hormone Bioanalogs Derived from 2-(4-Hydroxybenzyl)-6-methylcyclohexanone
Authors: Rejzek Martin , Zarevúcka Marie , Wimmer Zdeněk , Vaněk Tomáš , Tykva Richard , Kuldová Jelena , Šaman David , Bennettová Blanka
Published: July 2002

A new simple approach to isomeric 2-(4-alkoxybenzyl)-6-methylcyclohexanones and the corresponding alcohols and a subsequent synthesis of compounds imitating the action of the insect juvenile hormones is described. Biological activity of the isomeric juvenile hormone bioanalogs differed considerably when tested on the flesh fly (Sarcophaga bullata) and the yellow mealworm (Tenebrio molitor).
Sterically Crowded Heterocycles. IV. Diastereoisomeric Products by Ferricyanide Oxidation of Quaternary Pyridinium Salts
Authors: Kubík Richard , Kuthan Josef
Published: July 2002

While ferricyanide oxidation of achiral 4-(4-dimethylaminophenyl)-2,6-diphenyl-1-(pyridin-2-yl)pyridinium perchlorate (2) gave racemic 3-(4-dimethylaminophenyl)-1-phenyl-3-(2-phenylimidazo[1,2-a]pyridin-3-yl)prop-2-en-1-one (7), the same oxidative procedure applied to racemic 1-[5-(1-methylpyrrolidin-2-yl)pyridin-2-yl]-2,4,6-triphenylpyridinium perchlorate (5) or its dimethylamino derivative 6 let to mixtures of diastereoisomeric 3-[6-(1-methylpyrrolidin-2-yl)-2-phenylimidazo[1,2-a]pyridin-3-yl]-1,3-diphenyl-2-en-1-ones (8) or their dimethylamino derivatives 9, respectively.
Synthesis of 5-Amino-5-deoxypentonolactams
Authors: Kefurt Karel , Kefurtová Zdeňka , Marková Věra , Slívová Karla
Published: July 2002

5-Azido-5-deoxy-1,2-O-isopropylidene-α-D-xylofuranose (4) and 5-azido-5-deoxy-1,2-O-isopropylidene-β-D-arabinofuranose (10) were prepared starting from D-xylose and D-arabinose, respectively. Using the oxidation-reduction way for the C-3 epimerization, 5-azido-5-deoxy-1,2-O-isopropylidene-α-D-ribofuranose (15) and 5-azido-5-deoxy-1,2-O-isopropylidene-β-D-lyxofuranose (17) were obtained from 4 and 10, respectively. The derivatives 4, 10, 15 and 17 afforded by acid hydrolysis, oxidation with bromine and catalytic hydrogenation successively the corresponding 5-azido-5-deoxy-D-pentofuranoses 6, 11, 18, 19, 5-azido-5-deoxy-D-pentonolactones 7, 12, 20, 21 and 5-amino-5-deoxy-D-pentonolactams 8, 13, 22, 23.
Synthesis of New L-Proline Amides with Anticonvulsive Effect
Authors: Šilhánková Alexandra , Šindelář Karel , Dobrovský Karel , Krejčí Ivan , Hodková Jarmila , Polívka Zdeněk
Published: July 2002

Series of heterocyclic L-proline amides were prepared from BOC-L-proline and heterocyclic amines (mostly substituted piperazines and morpholines) via active ester with hydroxysuccinimide. 4-(4-Fluorobenzoyl)piperidine afforded L-proline 4-(4-(4-(4-fluorobenzoyl)piperidin-1-yl)benzoyl)piperidine (7b) simultaneously with expected L-proline 4-(4-fluorobenzoyl)piperidide (7a). D-Proline N-(3-(4-(3-chlorophenyl)piperazin-1-yl)propyl)amide (2) was prepared starting from D-proline. The amides were tested by methods of biochemical and behavioural pharmacology.
Catalytic Oxidation of Substituted Phenols in a Trickle Bed Reactor
Authors: Tukač Vratislav , Hanika Jiří
Published: July 2002

The catalytic wet oxidation was studied of phenol, 2-aminophenol, salicylic acid and 5-sulfosalicylic acid performed in a laboratory trickle bed reactor. A three-phase high-pressure catalytic reactor with an inside diameter of 18 mm and length of catalytic bed of 200 mm was operated at temperatures 90-180 °C, pressures 2-7 MPa and liquid space velocity 1-10 h-1. Simultaneously, the catalytic activity and the stability of extruded active carbon black Chezacarb and active carbon Chemviron were tested. At a comparable activity, the active carbon Chemviron exhibited a greater mechanical strength and stability. The influence of phenol substituents on the oxidation conversion corresponded to their inductive effect: The electropositive amino group supported the oxidation, on the contrary, the presence of carboxy and sulfo groups on aromatic ring led to only low conversion. The complications on evaluating the experimental data are caused by the non-isothermal temperature profile along the catalyst bed, the non-ideal oxygen dissolution in aqueous solutions and especially the hydrodynamic effect of flow rate on the degree of catalyst wetting and thus on the entire effectiveness of the oxidation process.
The Use of a Membrane GasLiquid Separator for Flow Injection Hydride Generation Atomic Absorption Spectrometry On-Line Speciation and Determination of As(III) and As(V)
Authors: Krenželok Milan , Rychlovský Petr
Published: July 2002

This method employs a membrane gas-liquid separator and the hydride generation technique in the FIA AAS method for speciation of As(V) and As(III). The gas-liquid membrane separator achieves high measurement sensitivity and shows a very fast response (short delay time) because of its minimum dead volume. Conditions were optimized for the determination of arsenic by the flow injection hydride generation atomic absorption spectrometry method using this membrane separator. An apparatus was proposed for on-line speciation of both As(V) and As(III). Several reduction systems were used for reduction of As(V) to As(III) in the on-line arrangement. The best results were obtained using solution of 15 g KI in 100 ml of 6 M HCl at the reaction coil temperature 60 °C. A detection limit of 150 pg for As(III), 80 pg for As(III + V) and sampling rate of 25 samples per hour were reached.
Chicken Pituitary Glycoproteins: New Isolation Method
Authors: Ryšlavá Helena , Krešlová Jana , Barthová Jana , Barth Tomislav
Published: July 2002

A new method for isolation of glycoproteins from chicken pituitaries was applied. The procedure consist of chromatography on ConA-Sepharose and by HPLC on S Hyper D and Vydac C4 columns. The hormonal activity of the glycoproteins was tested by determining their stimulatory effect on cAMP or testosterone production. Molecular weights of the products of tryptic cleavage of the hormone were determined using mass spectrometry (MALDI TOF). A comparison of the values obtained with theory shows that the protein is the β-unit of chicken luteinizing hormone.
C3-Symmetric Azaphosphatranes
Authors: Lake Fredrik , Hagberg Lars , Svensson Mats , Moberg Christina
Published: July 2002

C3-Symmetric azaphosphatranes, (3S,7S,10S)-2,3,7,8,9,10-hexamethyl-2,8,9-triaza-5-azonia-1-λ5-phosphabicyclo[3.3.3]undecane chloride and (3S,7S,10S)-3,7,10-triisopropyl-2,8,9-trimethyl-2,8,9-triaza-5-azonia-1-λ5-phosphabicyclo[3.3.3]undecane chloride, have been prepared starting from the corresponding chiral tripodal tetraamines and chlorobis(diethylamino)phosphane. The compounds are weak acids which are not fully deprotonated by potassium tert-butoxide. Density functional calculations of the compounds and their conjugate bases demonstrate that the weak acidity originates in a conformational change upon deprotonation leading to substantial steric repulsion.
Ulcer associated cell lineage glands expressing trefoil peptide genes are induced by chronic ulceration in ileal pouch mucosa
Authors: Pera M , Heppell J , Poulsom R , Teixeira F V , Williams J
Published: July 2002

BACKGROUND

Chronic ulcerative conditions in the gastrointestinal tract result in the appearance of the ulcer associated cell lineage (UACL). The glands of this new cell lineage secrete epidermal growth factor, transforming growth factor α, and the trefoil factor family (TFF) peptides, which are known to participate in repair processes. Pouchitis is the most frequent complication of ileal pouch-anal anastomosis.

AIM

Our aim was to determine whether the mucosal ulceration present in pouchitis can induce the development of UACL glands.

METHODS

Biopsies from ileal pouches with pouchitis (n=10), healthy pouches (n=5), and normal terminal ileum (n=5) were studied. Expression of TFF mRNA was assessed by in situ hybridisation. TFF1 and TFF2 proteins were localised by immunochemistry.

RESULTS

UACL glands containing TFF1 and TFF2 were observed in six patients with pouchitis. In some glands, there was TFF3 mRNA as has been reported for Crohn's UACL. None of the biopsies from ileal reservoirs without pouchitis showed UACL glands (p<0.05). Neither TFF1 nor TFF2 expression was detected in ileal reservoirs without pouchitis.

CONCLUSION

UACL glands arise de novo in ileal pouch mucosa of patients with pouchitis and express all three TFF peptide genes. Chronic inflammation alone, present in healthy pouches, is not enough to stimulate the growth of the UACL, and additional stimuli consequent on ulceration may be needed.
Metaphors, models and organisational ethics in health care
Authors: McCrickerd Jennifer
Published: July 2002

Crucial to discussions in organisational ethics is an evaluation of the metaphors and models we use to understand the organisations we are discussing. I briefly defend this contention and evaluate three possible models: the current corporate model, an orchestrator model which puts hospitals in the same class as malls and airports, and a community model. I argue that the corporate and orchestrator model push to the background some important organisational ethics issues and bias us inappropriately towards certain solutions. Furthermore, I argue that the community model allows these to be more easily brought up. I also respond to the likely challenge that hospitals really are corporations by arguing that this is not relevant to evaluations of the appropriateness of the corporate model.
Words of Tohkaku Wada: medical heritage in Japan
Authors: Matsumoto Masatoshi , Inoue Kazuo , Kajii Eiji
Published: July 2002

The origins of Japan's medical ideas, which are deeply rooted in its religion, culture and history, are not widely understood in medical societies of other countries. We have taken up the task of summarising this tradition here so that some insight can be gained into the unique issues that characterise the practice of medicine in Japan. We borrow from the sayings of Tohkaku Wada, a medical philosopher of late eighteenth-century Japan, for a look at Japanese medical tradition. Wada's medical thought was very much reflective of the Buddhism, Zen, and swordsmanship that informed eighteenth-century philosophy in Japan. His central concepts were “chu” and “sei”, that is, complete and selfless dedication to the patient and the practice of medicine. This paper explores Wada's thought, explaining it mainly from the standpoint of Japanese traditional culture.
The value of taking an ‘ethics history’
Authors: Sayers Gwen M , Barratt David , Gothard Catherine , Onnie Clive , Perera Sunimalee , Schulman Daniel
Published: July 2002

Objectives—To study the value of taking an ethics history as a means of assessing patients' preferences for decision making and for their relatives' involvement.

Design—Questionnaire administered by six junior doctors to 56 mentally competent patients, admitted into general and geriatric medical beds.

Setting—A large district general hospital in the United Kingdom.

Main measures—To establish whether patients were adequately informed about their illness and whether they minded the information being communicated to their relatives. To establish their preference regarding truthful disclosure and participation in decision making with risk attached. To establish whether they wished to be involved in CPR decision making, and if not, who should make the decision. To establish whether they knew of living wills and whether they had any advance directives.

Results—Twenty-four (43%) were inadequately informed of their illness.Forty-six (82%) said they would want to know were something serious to be found. Twenty-eight (50%) wanted to make their own decision if requiring risky treatment and 11 (20%) wanted family members involved. Thirty-one (55%) wanted to make a cardiopulmonary resuscitation (CPR) decision and five of these decisions differed from those made by the doctors. Twenty-five (45%) preferred the doctors to decide. Eleven (20%) of the patients had heard of living wills but only one had executed such a will. Seven (13%) of the patients wished to provide advance directives. Three (5%) did not find the history taking helpful but none were discomforted.

Conclusion—Taking an ethics history is a simple means of obtaining useful information about patients' preferences.
Is current practice around late termination of pregnancy eugenic and discriminatory? Maternal interests and abortion
Authors: Savulescu Julian
Published: July 2002

The attitudes of Australian practitioners working in clinical genetics and obstetrical ultrasound were surveyed on whether termination of pregnancy (TOP) should be available for conditions ranging from mild to severe fetal abnormality and for non-medical reasons.These were compared for terminations at 13 weeks and 24 weeks. It was found that some practitioners would not facilitate TOP at 24 weeks even for lethal or major abnormalities, fewer practitioners support TOP at 24 weeks compared with 13 weeks for any condition, and the difference in attitudes to TOP between 13 weeks and 24 weeks is most marked for pregnancies which are normal or involve a mild disorder.

It is argued that a fetal abnormality criterion for late TOP is inconsistently applied, discriminatory and eugenic. Four possible moral justifications for current practice are examined, each of which would require significant changes to current practice. I argue in favour of a maternal interests criterion for any TOP.
Economics and ethics in health care
Authors: Culyer Anthony J
Published: July 2002

This editorial provides a review of the current ways in which health economics is impacting on policy and reviews some of the key ethical and value-judgmental issues that commonly arise in and as a result of the work of economists. It also briefly highlights the contributions of the authors of this special issue of the journal, all of which illustrate how economists have approached ethical issues in health service policy (both in its financing and its delivery), and some of which explore the major methodological matters that arise and go on to discuss their potential as sources of conflict or harmony with other approaches to the same questions.
Two challenges to the double effect doctrine: euthanasia and abortion
Authors: Shaw A B
Published: July 2002

The validity of the double effect doctrine is examined in euthanasia and abortion. In these two situations killing is a method of treatment. It is argued that the doctrine cannot apply to the care of the dying. Firstly, doctors are obliged to harm patients in order to do good to them. Secondly, patients should make their own value judgments about being mutilated or killed. Thirdly, there is little intuitive moral difference between direct and indirect killing. Nor can the doctrine apply to abortion. Doctors kill fetuses as a means of treating the mother. They also kill them as an inevitable side effect of other treatment. Drawing a moral distinction between the direct and the indirect killing gives counterintuitive results. It is suggested that pragmatic rules, not ethics, govern practices around euthanasia and cause it to be more restricted than abortion.
Informed consent doesn't exist in AMI trials
Authors: Verdú-Pascual F , Castelló-Ponce A
Published: July 2002

Hilden and Gammelgaard's (H&G) comments on the evaluation and application of statistical trials and on informed consent lead to the same conclusion that there are special circumstances when clinical trials are performed in emergency situations.

The decision to comment on the work that H&G responded to was motivated by the wish to manifest the need to reach a consensus on modifying the conditions in which clinical trials should be performed in special circumstances.

One unquestionable fact must first be addressed: clinical trials are necessary for acquiring new knowledge and consequent improvements in health care services. At the same time, patient participation should be encouraged under conditions that respect patients' decision making capability.

Are these conditions being met at present? An answer to this is given below.
A future like ours revisited
Authors: Brown M T
Published: July 2002

It is claimed by the future like ours anti-abortion argument that since killing adult humans is wrong because it deprives them of a future of value and the fetus has a future of value, killing fetuses is wrong in the same way that killing adult human beings is wrong. In The morality of abortion and the deprivation of futures (this journal, April 2000) I argued that the persuasive power of this argument rests upon an equivocation on the term “future of value”. If the expression means “a potential future of value” then the moral claim is implausible because people do not in general have rights to what they need to fulfill their potential; if the expression means “self-represented future of value” then the argument fails because the fetus does not represent its future. Under no interpretation is the argument sound. In Deprivations, futures and the wrongness of killing (this journal, December 2001) Donald Marquis, author of the future like ours argument, responds at length to this objection. In the present essay the focus of the debate shifts to the proper interpretation of the right not to be killed. Donald Marquis argues that this liberty right entails the welfare right to the means necessary to sustain life; I argue that the right not to be killed does not entail unlimited welfare rights. On Marquis's view, the right not to be killed confers upon the fetus the right to whatever it takes to sustain life; on the view I defend, the right not to be killed does not confer upon the fetus or anyone else the right to another person's body. On Marquis's view, abortion is almost never permissible; on my view abortion is almost always permissible.
Present self-represented futures of value are a reason for the wrongness of killing
Authors: Parsons S J
Published: July 2002

In Marquis's recent paper he has not satisfactorily shown that killing does not adversely affect the victim's present self-represented desires for their future. Marquis is correct in believing life and death are distinct, but living and dying are not. In fact, to use a well-known saying, “the second we are born we start to die”. During the process of dying, whether it be long as in over our lifetime or short as in as we are being killed, there comes a point when the present realistic desires we have we know will never be satisfied. This is why killing can be wrong. This would imply killing an unconscious person, infant, or fetus cannot be wrong. But such killing can be wrong, despite the person killed not experiencing the desire not to be killed as he was dying. Killing can be wrong because others can have a present self-represented desire for that person not to be killed to have been killed. If this line of reasoning is correct, then the “best interests” principle often applied to life and death considerations regarding unconscious persons, infants, and fetuses, is invalid, as such human beings do not have present desires. All that matters is whatrelevantothers rationally desire, after being informed of the facts and the consequences, for that unconscious person, infant or fetus.
One principle and three fallacies of disability studies
Authors: Koch T
Published: July 2002

A question between John Harris and I is the degree to which lessons may be learned, and insights gained, from a life distinguished by physical differences. He argues it as the “aborting Beethoven fallacy”, I insist on the evidence that what we learn from physical differences may be critical and life enhancing.