bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of radiolabeled racemic and enantiomeric antiarrhythmic agents
Authors: Stolle Wayne T. , Stelzer Lindsay S. , Hester Jackson B. , Perricone Salvatore C. , Hsi Richard S. P.
Published: October 2006

Ibutilide fumarate, racemic N‐[4‐[4‐(ethyl‐n‐heptylamino)‐1‐hydroxybutyl]phenyl]methanesulfonamide hemifumarate, and artilide, the R‐(+)‐enantiomer of N‐[4‐[4‐(di‐n‐butylalmino)‐1‐hydroxybutyl]‐phenyl]methanesulfonamide hemifumerate, are under clinical investigation as Class III antiarrhythmic agents. For conducting drug disposition studies, we synthesized carbon‐14 labeled ibutilide, as well as its two enantiomeric forms. In addition, high specific activity tritium labeled ibutilide was also prepared to facilitate development of a radioimmunoassay and for studying receptor site characteristics of this agent. Results of metabolism studies with [14C]ibutilide led us to prepare tritium labeled artilide, which is more readily accessible than the C‐14 labeled drug. The optical antipode of artilide was also labeled with tritium for comparative drug disposition investigations on the two enantiomers.
Alternative labelling of the cocaine analogue isomers α‐CIT and β‐CIT by direct iodination with no‐carrier‐added Na125I
Authors: Müller Lars , Halldin Christer , Swahn Carl‐Gunnar , Foged Christian
Published: October 2006

An alternative labelling of the cocaine analogue isomers α‐CIT and β‐CIT with no‐carrier‐added 125I by direct iodination of 2α‐ and 2β‐carbomethoxy‐3β‐phenyltropane with Na125I / sulfuric acid / nitric acid / acetic acid and peracetic acid under different reaction conditions, is described. The maximum radiolabelling yield obtained with the two isomers was 48% for [125I]α‐CIT and 28% for [125I]β‐CIT.
Radiolabeled biotin amides from triazenyl precursors: Synthesis, binding, and in‐vivo properties
Authors: Kortylewicz Zbigniew P. , Baranowska‐Kortylewicz Janina , Adelstein S. James , Carmel Alice D. , Kassis Amin I.
Published: October 2006

The synthesis of N‐(4‐[127/125/123l]iodobenzyl)biotin amides 4a — 4c performed by the direct decomposition of N‐[4‐(3′,3′‐dimethyltriazenyl)benzyl)biotin amide with sodium iodide in the presence of CF3COOH is described. Iodinated in this way biotin formed a stable complex with avidin (Kd = 2.84 ± 0.45 × 10−15 M, n = 3.9 ± 0.6) which dissociated in the presence of excess native biotin with a rate constant of 0.034 ± 0.006 hr1. Blood clearance studies and the lack of thyroid uptake indicated that this compound was not deiodinated in vivo and behaved in circulation much like native biotin. This aryltriazene precursor method is suitable for labeling with short‐lived radiohalides. It can be used to produce no‐carrier‐added derivatives of biotin for use in biologic studies and assays involving avidin or streptavidin.
Two‐three steps syntheses of [14C‐ring]o‐xylene, [14C‐ring]o‐toluic acid; [14C‐ring]‐phthalic acid from [14C] barium carbonate
Authors: De Suzzoni S. , Noel J. P.
Published: October 2006

Cocyclotrimerisation of [14C2] acetylene with 2‐butyne gave [3,4,5,6‐14C4] ortho‐xylene (50% overall yield based on [14C] BaCO3). 93% Nitric acid oxidation at 145°C for 55 hours of the crude [3,4,5,6‐14C4] ortho‐xylene gave [3,4,5,6‐14C4] ortho‐toluic acid (25% overall yield based on [14C] BaCO3). Under the same set of reaction conditions but in a sealed tube the crude [3,4,5,6‐14C4] ortho‐xylene gave [3,4,5,6‐14C4] phthalic acid (50% overall yield based on [14C] BaCO3) (specific activity 1.6 GBq. mmol−1).
A practical method for the synthesis and purification of 14C labeled oligonucleotides
Authors: Sasmor Henri M. , Dellinger Douglas J. , Zenk Paul C. , Lee Larry P.
Published: October 2006

A practical method for the solid‐phase synthesis and purification of 14C labeled oligonucleotides is described. A streamlined procedure for the synthesis of 5′‐O‐dimethoxytrityl‐2‐(14C)‐thymidine 3′‐(2‐cyanoethyl‐N,N‐diisopropyl) phosphoramidite from commercially available 2‐(14C) labeled thymidine nucleoside is detailed. Methodology for the manual synthesis, isolation, and purification of a high specific activity oligonucleotide phosphorothioate 20‐mer (ISIS 2105) are elaborated.
Synthesis of N1′‐([11C]methyl)naltrindole ([11C]MeNTI): A radioligand for positron emission tomographic studies of delta opioid receptors
Authors: Lever John R. , Kinter Chris M. , Ravert Hayden T. , Musachio John L. , Mathews William B. , Dannals Robert F.
Published: October 2006

A delta opioid receptor antagonist, N1′‐methylnaltrindole (MeNTI), has been labeled with carbon‐11. The precursor for radiolabeling was prepared in 71% yield by benzylation of the phenolic moiety of naltrindole. Alkylation of the indole nitrogen using [11C]iodomethane and aqueous tetra(n‐butyl)ammonium hydroxide at 80 °C in dimethylformamide followed by hydrogenolysis (H2, 10% Pd‐C) of the benzyl protecting group gave [11C]MeNTI. The average (n = 10) time for radiosynthesis, HPLC purification and formulation was 24 min from end‐of‐bombardment. [11C]MeNTI of high radiochemical purity was obtained at end‐of‐synthesis with an average specific activity of 2050 mCi/μmol and radiochemical yield, based on [11C]iodomethane, of 6%.
Preparation of [125I]‐3H2‐iopiperidol‐A by exchange radioiodination
Authors: Ogan Marc , Tomasella Frank , Tu Jan‐I
Published: October 2006

Preparation of radiolabeled [125I]‐N,N′‐bis(2,3‐dihydroxypropyl)‐5‐(3‐hydroxy‐2‐oxo‐1‐piperidinyl)‐2,4,6‐ triiodo‐1,3‐benzenedicarboxamide, ([125I]‐3H2‐iopiperidol‐A), a nonionic radiographic contrast agent, was achieved by HAuCl4 mediated exchange radioiodination. [125I]‐Iopiperidol‐A was isolated by reversed‐ phase HPLC in 83% radiochemical yield. The radiochemical purity of isolated [125I]‐3H2‐iopiperidol‐A was 98.7% and the specific activity was 6.6 mCi/μmol. The role of HAuCl4 in facilitating exchange radioiodination of 3H2‐iopiperidol‐A, however, appears to be a complex process not limited to the production of electrophilic iodine ion.
Deiodination reactions using tributyltin hydride for potential labelling experiments
Authors: Zippi Elizabeth M. , Plourde Gary W. , Satyamurthy N.
Published: October 2006

2,6‐Dinitro‐1‐iodobenzene and 2,4‐dinitro‐1‐iodobenzene were deiodinated with tributyltin hydride at different temperatures using various addition modes. The product ratios of 1,3‐dinitrobenzene and the corresponding tributylstannyldinitrobenzene compounds were determined by NMR in order to evaluate the optimum conditions for impending tritiation experiments.
Synthesis of 4‐hydroxy[4‐3H]‐2(E)‐nonen‐1‐al‐diethylacetal
Authors: Bravais Fabienne , Rao Dinesh , Alary Jacques , Rao Renée C. , Debrauwer Laurent , Bories Georges
Published: October 2006

4‐Hydroxy‐2(E)‐nonen‐1‐al‐diethylacetal 3 (HNE‐DEA) was prepared by condensation of the Grignard compound derived from propiolaldehyde diethylacetal and n‐hexanal followed by reduction of the resulting 4‐hydroxy‐2‐nonyn‐1‐al‐diethylacetal 2 with LiAlH4 according to the literature procedure with minor modifications. Swern oxidation [DMSO + (COCl2)] of 3 gave 30% yield of 4‐oxo‐2(E)‐nonen‐1‐al‐diethylacetal 5. [3H]NaBH4 and [2H]NaBH4 reduction of 5 gave rise respectively to [4‐3H]HNE‐DEA (specific activity 222 GBq/mmol) and [4‐2H]HNE‐DEA.
Synthesis of [2H]5‐epitestosterone
Authors: Wähälä Kristiina , Väänänen Tiina , Hase Tapio , Leinonen Antti
Published: October 2006

Isotopically pure d5‐epitestosterone has been synthesized for the first time. This polydeuterated 17α‐hydroxyandrosterone was prepared by H/D exchange reaction with MeOD‐D2O under basic conditions. Its characteristics are described in detail, including its enolization into a d4‐heterodienic silyl ether derivative upon silylation.
Radioiodinated analogs of xylamine: N‐(2‐chloroethyl)‐N‐ethyl‐2‐[125I]iodobenzylamine and N,N‐diethyl‐2‐[125I]iodobenzylamine as potential tools for monoamine uptake exploration by spect
Authors: Branger C. , Garreau L. , Frangin Y. , Chalon S. , Dubois P. , Dognon A.‐M. , Ombetta‐Goka J.‐E. , Besnard J.‐C. , Guilloteau D.
Published: October 2006

In order to improve the scintigraphy and radiotherapy of neuroendocrine tumors we synthesized two radioiodinated benzylamines [N‐(2‐chloroethyl)‐N‐ethyl‐2‐[125I]iodobenzylamine and N,N‐diethyl‐2‐[125I]iodobenzylamine], analogs of xylamine [N‐(2‐chloroethyl)‐N‐ethyl‐2‐methylbenzylamine]. Xylamine is an irreversible inhibitor of uptake and accumulation of noradrenaline. The two unlabelled iodinated derivatives [N‐(2‐chloroethyl)‐N‐ethyl‐2‐iodobenzylamine and N,N‐diethyl‐2‐iodobenzylamine] were synthesized, purified and checked by HPLC, NMR and mass spectrography. Their affinity for the noradrenaline transporter was determined in vitro on rat brain membrane homogenates with [3H]nisoxetine. Radioiodination was performed by iodide for bromide nucleophilic exchange from brominated precursors. The N,N‐diethyl‐2‐[125I]iodobenzylamine was obtained directly from N,N‐diethyl‐2‐bromobenzylamine. Radiosynthesis of N‐(2‐chloroethyl)‐N‐ethyl‐2‐[125I]iodobenzylamine required three steps. A new brominated precursor [N‐ethyl‐N‐(2‐bromobenzyl)glycine ethyl ester] which was stable for radiolabelling and suitable for reduction to N‐(2‐hydroxyethyl)‐N‐cthyl‐2‐[125I]iodobenzylamine was synthesized. N‐(2‐Hydroxyethyl)‐N‐ethyl‐2‐[125I]iodobenzylamine was converted to N‐(2‐chloroethyl)‐N‐ethyl‐2‐[125I]iodobenzylamine in the presence of an excess of thionyl chloride. Radioiodinated derivatives were purified and checked by HPLC.
N. C. A. radioiodination of idoxifene
Authors: Trivedi Mahendra , Potter Gerrard , Hammersley Peter , Carnochan Paul , Young Helen , Jarman Michael , Ott Robert
Published: October 2006

We describe a method for n.c.a. radioiodination of pyrrolidino tamoxifen (Idoxifene) based on the electrophilic destannylation of tributylstannyl pyrrolidino tamoxifen. The methods of separation using preparative tlc and HPLC give >95% radiochemical purity and >70% recovery.
Synthesis of 1,2[3H]‐1,2‐epoxy analogue of fructose‐6P, an affinity label of escherichia coli glucosamine‐6P synthase
Authors: Leriche Caroline , René Loïc , Derouet Florence , Rousseau Bernard , Badet Bernard
Published: October 2006

1,2‐anhydroglucitol‐6P, a known inhibitor of glucose‐6P isomerase, behaved as a fructose‐6P site‐directed irreversible inhibitor of bacterial glucosamine‐6P synthase. The lack of reproducibility of the aldolase‐mediated condensation of dihydroxyacetone phosphate and glycidaldehyde followed by borohydride reduction previously described prompted us to develop a chemical route to this compound and its radiolabelled counterpart. The compound was synthesized in 13 steps from D‐arabinose with a 6% overall yield. Tritium introduction was performed at step 11 (3 → 4) allowing isolation of the title compound of high specific radioactivity.
Synthesis of Δ4‐3‐oxo‐steroids labelled specifically at the 6β ‐position
Authors: Toft P. , Liston A. J. , Viau A. Y.
Published: October 2006

[6β‐2H] Testosterone and [6β‐3H] Testosterone were each synthesized in good yield in three steps: Treatment of 5a, 6a‐ epoxy‐3, 3‐ethylene‐dioxyandrostan ‐17β‐ol with labelled lithium aluminum hydride is followed by deacetalation and dehydration under carefully controlled basic conditions.
The biosynthesis of 13C labeled starch granules as a source of uniformly labeled 13C D‐glucose
Authors: Buchholz J. R. , Christenson C. W. , Eakin R. T. , Fowler E.
Published: October 2006

A potato plant has been grown in a closed chamber with CO2 enriched to a level of 31 at. % in 13C as the sole source of carbon. In 80 days the plant produced 1,684 g of tubers labeled with 29 at. % 13C. of the carbon fixed by the potato plant appromixately 70% was found in the tubers while 30% was found in the tops and roots. Recovered starch granules constituted 10% of the fresh weight of these tubers. A 16% yield of glucose was obtained, based on all of the carbon dioxide used. Distribution of the 13C label among the six species of carbon atoms in strach appears to be nearly uniform. The plant and tubers were indistinguishable from those grown on normal isotopic CO2 indicating no gross isotope effect at this level of 13C enrichement.
Substances naturelles marquees au cardopn14: Synthese de l'acide withranilioue (carboxylc 14C), precurseur bio‐sehtioe de l'ellipticine
Authors: Bac Nguyen Van , Herbert Michel , Pichat Louis , Dat‐Xuong Nguyen
Published: October 2006

La carbonisation du lithien du bromo‐2 acétanilide ne donne qu'sun faible rendement en acide anthranilique.

La carbonisation du promure d'o‐chlorphénylmaanésium pur 14CO2 conduit à l'acide o‐chlorobenzoique ( carboxyle 14c ) dont l'a‐mination deshalogénante fournit l'acide anthranilique ( carboxyle 14C. ) avec un rendement radioactif global de 26% par rapport à Ba14CO3 une activité spécifique de 48 mCi/MM.
Arzneipflanzen, die unverstandene Naturapotheke
Authors: Müller‐Limmroth Wolf
Published: October 2006

„Muß ich dies Körbchen bier voll Kraut und Blumen lesen: voll Pflanzen gift'ger Art und diensam zum Genesen. An vielen Tugenden sind viele drunter reich, ganz ohne Wert nicht eins, doch keins dem andern gleich. Oh, groß Kräfte sind's, weiß man sie recht zu pflegen, die Pflanzen, Krauter, Stein' in ihrem Innern hegen.
Thermally responsive polymeric hydrogel brushes: synthesis, physical properties and use for the culture of chondrocytes
Authors: Collett John , Crawford Aileen , Hatton Paul V , Geoghegan Mark , Rimmer Stephen
Published: October 2006

Hydrogel brushes are materials composed of a water-swollen network, which contains polymer chains that are grafted with another polymer. Using a thermally responsive polymer, poly(
N
-isopropyl acrylamide) (polyNIPAM), as the graft component we are able to maintain the critical solution temperature (
T
crit
), independent of the overall composition of the material, at approximately 32°C. The change in swelling at
T
crit
is a function of the amount of polyNIPAM in the system. However, there is a much smaller change in the surface contact angles at
T
crit
. PolyNIPAM-based materials have generated considerable interest, as ‘smart’ substrates for the culture of cells and here, we show the utility of hydrogel brushes in cell culture. Chondrocytes attached to the hydrogel brushes and yielded viable cell cultures. Moreover, the chondrocytes could be released from the hydrogel brushes without the use of proteases by reducing the temperature of the cultures to below
T
crit
to induce a change in the conformation of the polyNIPAM chain at
T
crit
. The importance of the crosslink hydrogel component is illustrated by significant changes in cell attachment/cell viability as the crosslink density is changed.
Statistical analysis of the dynamics of antibody loss to a disease-causing agent: plague in natural populations of great gerbils as an example
Authors: Park Siyun , Chan Kung-Sik , Viljugrein Hildegunn , Nekrassova Larissa , Suleimenov Bakhtiyar , Ageyev Vladimir S , Klassovskiy Nikolay L , Pole Sergey B , Chr. Stenseth Nils
Published: October 2006

We propose a new stochastic framework for analysing the dynamics of the immunity response of wildlife hosts against a disease-causing agent. Our study is motivated by the need to analyse the monitoring time-series data covering the period from 1975 to 1995 on bacteriological and serological tests—samples from great gerbils being the main host of
Yersinia pestis
in Kazakhstan. Based on a four-state continuous-time Markov chain, we derive a generalized nonlinear mixed-effect model for analysing the serological test data. The immune response of a host involves the production of antibodies in response to an antigen. Our analysis shows that great gerbils recovered from a plague infection are more likely to keep their antibodies to plague and survive throughout the summer-to-winter season than throughout the winter-to-summer season. Provided the seasonal mortality rates are similar (which seems to be the case based on a mortality analysis with abundance data), our finding indicates that the immune function of the sampled great gerbils is seasonal.
Mating trials validate the use of DNA barcoding to reveal cryptic speciation of a marine bryozoan taxon
Authors: Gómez Africa , Wright Peter J , Lunt David H , Cancino Juan M , Carvalho Gary R , Hughes Roger N
Published: October 2006

Despite increasing threats to the marine environment, only a fraction of the biodiversity of the oceans has been described, owing in part to the widespread occurrence of cryptic species. DNA-based barcoding through screening of an orthologous reference gene has been proposed as a powerful tool to uncover biological diversity in the face of dwindling taxonomic expertise and the limitations of traditional species identification. Although DNA barcoding should be particularly useful in the sea, given the prevalence of marine cryptic species, the link between taxa identified through DNA barcodes and reproductively isolated taxa (biological species) has rarely been explicitly tested. Here, we use an integrated framework comparing breeding compatibility, morphology and mitochondrial (cytochromecoxidase 1) and nuclear (elongation factor-1-alpha) DNA sequence variation among globally distributed samples of the cosmopolitan marine bryozoanCelleporella hyalina(L.). Our results reveal thatC. hyalinacomprises numerous deep, mostly allopatric, genetic lineages that are reproductively isolated, yet share very similar morphology, indicating rampant cryptic speciation. The close correspondence between genetic lineages and reproductively isolated taxa in the context of minimal morphological change suggests that DNA barcoding will play a leading role in uncovering the hidden biodiversity of the oceans and that the sole use of morphologically based taxonomy would grossly underestimate the number of marine species.
Allopatric origins of microbial species
Authors: Whitaker Rachel J
Published: October 2006

Although allopatric divergence is a well-accepted mechanism of speciation for eukaryotic macro-organisms, the importance of geographical barriers to divergence in microbial populations is a subject of great debate. Do geographically separated populations of micro-organisms diverge independently, or does their structure fit the often quoted Bass-Becking description ‘everything is everywhere; the environment selects’? Aided by high-resolution genetic and genomic tools, the search for ‘microbial marsupials’ has revealed that in fact both are true; some species of micro-organisms demonstrate allopatric divergence, while others do not. This discovery opens the door for comparative analyses, where questions about the differences in evolutionary and ecological mechanisms that drive divergence and speciation in different microbial species can begin to be explored. Investigating these differences in evolutionary mechanisms will greatly enhance interest in, and understanding of, the dynamic processes that create and maintain the vast diversity of the microbial world.
Serum insulin‐like growth factor II levels in normal adolescents and those with insulin dependent diabetes mellitus
Authors: Acerini C. L. , Clayton K. L. , Hintz R. , Baker B. , Watts A. , Holly J. M. P. , Dunger D. B.
Published: October 2006

OBJECTIVE Unlike IGF‐I and its principal binding proteins, data regarding IGF‐II levels have not been well defined in normal subjects and those with insulin‐dependent diabetes mellitus (IDDM). We have therefore measured IGF‐II, as well as IGF‐I, and IGFBP‐3, levels In a large cohort of subjects with IDDM and in age/sex matched controls.

PATIENTS One hundred and fourteen patients with IDDM (57 males, 57 females) and 89 control subjects (49 males, 40 females).

MEASUREMENTS Random blood samples were obtained from each subject for the measurement of IGF‐II, IGF‐I and IGFBP‐3 levels.

RESULTS Mean values of IGF‐II (±SEM) were 630 (±27.8) μg/l and 646 (±32.3) μg/l in female and male controls, compared to 569 (±23.3) μg/l and 623.3 (±28.1) μg/l in female and male diabetics respectively. IGF‐II levels did not differ significantly between the sexes or show any change with transition through puberty in either control or diabetic groups. In contrast, IGF‐I levels increased through puberty peaking at stages 3–5 in controls (P < 0.001) and G4–5 (P= 0.002) in diabetic males but not females. IGF‐I levels in all diabetics were generally lower than in controls, differences reaching significance at G4–5 in males (P= 0.002) and B5 in females (P= 0.002). IGFBP‐3 levels did not show any variation with puberty stage in diabetics, in contrast to controls where levels increased, peaking at G4–5 in males (P= 0.001) and B3 in females. IGFBP‐3 levels were lower in diabetics of both sexes and at all stages compared to controls (P range 0.047 to < 0.001). Multiple regression analysis revealed significant correlations between IGF‐II and IGFBP‐3 (F= 20.1, P= < 0.001) and reaffirmed previously observed associations for IGF‐I and IGFBP‐3. The sum of IGF‐I and IGF‐II (expressed as nmol/l) correlated with IGFBP3; r= 0.47 in controls and 0.60 in diabetics.

CONCLUSIONS Insulin‐dependent diabetes mellitus is not associated with any significant changes in IGF‐II levels during puberty. The binding of IGFBP‐3 for both IGF‐I and IGF‐II is unaltered by insulin‐dependent diabetes mellitus.
Pharmacokinetics and pharmacodynamics of subcutaneous testosterone implants in hypogonadal men
Authors: Jockenhövel F. , Vogel E. , Kreutzer M. , Relnhardt W. , Lederbogen S. , Reinwein D.
Published: October 2006

OBJECTIVE There are advantages and disadvantages with all of the presently available types of testosterone replacement for hypogonadal men. We performed this investigation to establish detailed data about the pharmacokinetics, pharmacodynamics, feasibility and side‐effects of subcutaneously implanted testosterone (T) pellets.

DESIGN AND MEASUREMENT In a single‐dose, open‐label, non‐randomized study, 6 T‐pellets, each containing 200 mg of fused crystalline T, were Implanted in the subdermal fat tissue of the lower abdominal wall of 14 hypogonadal men. Blood samples for determination of T, LH, FSH, 5α‐dihydrotestosterone (DHT), sex hormone binding globulin (SHBG) and oestradiol (E2) were obtained at 0, 0.5, 1, 2, 4, 8, 12, 24, 36, 48 hours and on day 21 after implantation and then every 3 weeks until day 189, and on days 246 and 300 during follow‐up. In another 36 hypogonadal men the feasibility and side‐effects of T‐pellets were evaluated.

PATIENTS Fourteen patients participated in the detailed pharmacokinetic study and another 36 patients in the assessment of feasibility and side‐effects. All patients (age range 18–61 years) suffered from primary or secondary hypogonadism (T < 3.6nmol/l).

RESULTS The pharmacokinetic study in 14 hypogonadal men revealed an initial short‐lived burst release of T with a peak concentration of 49.0 ± 3.7 nmol/l at 0–5 ± 0.13 days which was followed by a stable plateau lasting until day 63 (day 2, 35.2 ± 2.3; day 63, 34.8 ± 2.6 nmol/l). Thereafter serum T gradually declined and was close to baseline concentrations on day 300. Apparent terminal elimination half‐life (t1/2) was 70.8 ± 10.7 days and apparent mean residence time 87.0 ± 45 days. On average, serum T was below 10 nmol/l after 180 days. Absorption of T followed a zero‐order release kinetic with an absorption half‐time of 74.7 days (95% confidence Interval: 71.1–78.5) and was almost complete by day 189 (95.9 ± 0.84%). Serum DHT and E2 were significantly elevated from day 21 to day 105 and correlated significantly with T (DHT, r = 0–65, P < 00001, E2, r = 067, P < 0.0001). SHBG was significantly decreased from day 21 to day 168. In 6 men with primary hypogonadism T suppressed LH and FSH to the eugonadal range from day 21 to 126 and 42 to 105, respectively, with nadirs occurring at day 84 (LH) and day 63 (FSH). LH and FSH were highly inversely correlated with T (r=−0.47 and −0.57).

The only side‐effect observed during 112 implantations in the total group of 50 men were 6 local infections (5.4%) leading to extrusion of 5 pellets In 3 men. When given the choice, all patients except one preferred T‐pellets to their previous T medication for permanent substitution therapy.

CONCLUSION T‐pellets are the androgen formulation with the longest biological action and strongest pharmacodynamic efficacy in terms of gonadotrophin suppression. The pharmacokinetic features are advantageous compared to other T preparations and the patient acceptance is high.
Fulminant methicillin‐sensitive Staphylococcus aureus infection in a healthy adolescent, highlighting ‘Panton–Valentine leucocidin syndrome’
Authors: Swaminathan A. , Massasso D. , Gotis‐Graham I. , Gosbell I.
Published: October 2006

A 14‐year‐old immunocompetent adolescent presented systemically unwell with left knee septic arthritis. Within several days, disseminated bone and soft tissue collections became evident, associated with deep venous thrombosis and pulmonary involvement. Methicillin‐sensitive Staphylococcus aureus was isolated, harbouring Panton–Valentine leucocidin genes. Aggressive antibiotic and surgical therapies eventually lead to recovery. Intrafamilial spread of the pathogenic isolate was shown by household screening. This presentation is consistent with ’PVL Syndrome’ and is typical of severe S. aureus infection emerging in young populations globally.
Headspace analysis of volatile flavour compounds of teleme cheese made from sheep and goat milk
Authors: MASSOURAS T , PAPPA E C , MALLATOU H
Published: October 2006

The headspace compounds of teleme cheese made from sheep's milk, goats’ milk or mixture of sheep's and goats’ milk (50:50) were analysed during ripening by static headspace gas chromatography–mass spectrometry. A total of 21 major compounds were identified, including aldehydes (7), alcohols (5), ketones (4), and acids (2). All types of cheeses contained approximately the same volatiles at different concentrations. The total volatile compounds (TVC) increased during ripening. Cheeses made from sheep's milk showed the highest level of TVC, whereas cheeses made from goats’ milk showed the lowest one.
Flow‐mediated dilatation in overweight and obese women with polycystic ovary syndrome
Authors: Brinkworth GD , Noakes M , Moran LJ , Norman R , Clifton PM
Published: October 2006

Objective  There remains a large degree of disagreement about the association of polycystic ovary syndrome (PCOS) with impaired endothelial dysfunction and cardiovascular disease (CVD) risk. The purpose of this study was to determine whether overweight and obese women with PCOS have impaired endothelial function compared with weight‐matched controls without PCOS and whether endothelial function is associated with cardiovascular risk markers and hormonal parameters.

Design  Cross‐sectional analysis.

Setting  An outpatient trial at the Commonwealth Scientific Industrial Research Organisation Clinical Research Unit.

Population  Overweight and obese women with PCOS (n= 12) and weight‐matched controls without PCOS (n= 10).

Methods  Endothelial function, cardiovascular risk markers and hormonal parameters were assessed in the patients.

Main outcome measures  Endothelial function was assessed by flow‐mediated dilatation (FMD) of the brachial artery using high‐resolution ultrasound. Lipid profile, fasting insulin level, glucose level, insulin resistance, C‐reactive protein level, folate level, Vitamin B12 level and hormonal parameters.

Results  Women with PCOS had significantly higher testosterone levels (P < 0.001) and free androgen index (P= 0.006) compared with the controls without PCOS. Both groups were normoinsulinaemic, and there were no significant differences in any of the markers of CVD between women with and without PCOS. Furthermore, FMD was similar in both groups (PCOS 6.1 ± 1.2% versus control 5.6 ± 1.0%, P= 0.77).

Conclusions  Compared with a group of weight‐matched women with similar metabolic profiles, normoinsulinemic, overweight and obese women with PCOS did not show any greater impairment in endothelial function assessed by FMD. A normoinsulinemic phenotype of PCOS with low metabolic risk factors may reduce the risk of endothelial dysfunction in overweight and obese women with this syndrome. Further studies are required that directly compare FMD in normoinsulinemic and hyperinsulinaemic women with PCOS.
Location of resistance factors in the leaves of potato and wild tuber‐bearing Solanum species to the aphid Myzus persicae
Authors: Alvarez A. E. , Tjallingii W. F. , Garzo E. , Vleeshouwers V. , Dicke M. , Vosman B.
Published: October 2006

Analysis of electrically recorded feeding behaviour of aphids was combined with colony‐development tests to search for sources of resistance to Myzus persicae (Sulzer) (Homoptera: Aphididae) in tuber‐bearing Solanum species (Solanaceae), aiming at a reduction of potato leaf roll virus (PLRV) transmission. Twenty genotypes, originating from 14 gene bank accessions, representing 13 wild tuber‐bearing Solanum spp., three Solanum tuberosum L. (potato) cultivars, and one S. tuberosum breeding line, were selected. Colony‐development tests were carried out in no‐choice experiments by placing adult aphids on plants of each genotype and counting numbers of nymphs and adults on young plants after 8 and 15 days, and on flowering plants after 14 and 30 days. Large differences were observed among genotypes: some developed small colonies and others developed large ones. Also, in a few genotypes, resistance in mature plants was different for leaves of different ages; young leaves were resistant to aphids whereas old senescent leaves were susceptible. The electrical penetration graph (DC‐EPG system) technique was used to study aphid feeding behaviour on each Solanum genotype for 6 h. Electrical penetration graph (EPG) results also showed large differences among the genotypes, indicating resistance at the leaf surface and at three different levels of plant tissue (epidermis, mesophyll, and phloem). Therefore, it was concluded that different mechanisms of resistance to M. persicae exist among the genotypes analysed. EPGs recorded from aphids on Solanum berthaultii Hawkes and Solanum tarijense Hawkes with and without glandular trichomes showed that strong surface resistance can bias EPG parameters associated with resistance located in deeper tissues. Experimental evidence is presented that the resistance to aphids in the genotypes with glandular trichomes strongly depends on these morphological structures.
Characterization of Membrane Association Domains within the
Tomato Ringspot Nepovirus
X2 Protein, an Endoplasmic Reticulum-Targeted Polytopic MembraneProtein
Authors: Zhang Guangzhi , Sanfaçon Hélène
Published: October 2006

Replication
of nepoviruses (family
Comoviridae
) occurs in association with
endoplasmic reticulum (ER)-derived membranes. We have previously shown
that the putative nucleoside triphosphate-binding protein (NTB) of
Tomato ringspot nepovirus
is an integral membrane protein with
two ER-targeting sequences and have suggested that it anchors the viral
replication complex (VRC) to the membranes. A second highly hydrophobic
protein domain (X2) is located immediately upstream of the NTB domain
in the RNA1-encoded polyprotein. X2 shares conserved sequence motifs
with the comovirus 32-kDa protein, an ER-targeted protein implicated in
VRC assembly. In this study, we examined the ability of X2 to associate
with intracellular membranes. The X2 protein was fused to the green
fluorescent protein and expressed in
Nicotiana benthamiana
by
agroinfiltration. Confocal microscopy and membrane flotation
experiments suggested that X2 is targeted to ER membranes. Mutagenesis
studies revealed that X2 contains multiple ER-targeting domains,
including two C-terminal transmembrane helices and a less-well-defined
domain further upstream. To investigate the topology of the protein in
the membrane, in vitro glycosylation assays were conducted using X2
derivatives that contained N-glycosylation sites introduced at the N or
C termini of the protein. The results led us to propose a topological
model for X2 in which the protein traverses the membrane three times,
with the N terminus oriented in the lumen and the C terminus exposed to
the cytoplasmic face. Taken together, our results indicate that X2 is
an ER-targeted polytopic membrane protein and raises the possibility
that it acts as a second membrane anchor for the
VRC.
Pathogenesis of a Genogroup II Human Norovirus in Gnotobiotic Pigs
Authors: Cheetham Sonia , Souza Menira , Meulia Tea , Grimes Sheila , Han Myung Guk , Saif Linda J.
Published: October 2006

We
evaluated the gnotobiotic (Gn) pig as a model to study the pathogenesis
of human norovirus (HuNoV) and to determine the target cells for viral
replication. Sixty-five Gn pigs were inoculated with fecal filtrates of
the NoV/GII/4/HS66/2001/US strain or with pig-passaged intestinal
contents (IC) and euthanized acutely (
n
= 43) or after
convalescence (
n
= 22). Age-matched Gn piglets
(
n
= 14) served as mock-inoculated controls.
Seventy-four percent (48/65) of the inoculated animals developed mild
diarrhea compared to 0 of 14 controls. Pigs from postinoculation days
(PID) 1 to 4 tested positive for HuNoV by reverse
transcription-PCR of rectal swab fluids (29/65) and IC
(9/43) and by antigen (Ag) enzyme-linked immunosorbent assay (ELISA)
using antiserum to virus-like particles of HuNoV GII/4. No
control pigs were positive. Histopathologic examination showed mild
lesions in the proximal small intestine of only one pig (1/7).
Seroconversion after PID 21 was detected by antibody ELISA in 13 of 22
virus-inoculated pigs (titers, 1:20 to 1:200) but not in controls.
Immunofluorescent microscopy using a monoclonal antibody to HuNoV GII
capsid revealed patchy infection of duodenal and jejunal enterocytes of
18 of 31 HuNoV-inoculated pigs with a few stained cells in the ileum
and no immunofluorescence (IF) in mock-inoculated controls.
Immunofluorescent detection of the viral nonstructural N-terminal
protein antigen in enterocytes confirmed translation. Transmission
electron microscopy of intestines from HuNoV-inoculated pigs showed
disrupted enterocytes, with cytoplasmic membrane vesicles containing
calicivirus-like particles of 25 to 40 nm in diameter. In summary,
serial passage of HuNoV in pigs, with occurrence of mild diarrhea and
shedding, and immunofluorescent detection of the HuNoV structural and
nonstructural proteins in enterocytes confirm HuNoV replication in Gn
pigs.
At Least 50% of Human-Specific HERV-K (HML-2) Long Terminal Repeats Serve In Vivo as Active Promoters for Host Nonrepetitive DNA Transcription
Authors: Buzdin Anton , Kovalskaya-Alexandrova Elena , Gogvadze Elena , Sverdlov Eugene
Published: October 2006

We report the first genome-wide comparison of in vivo promoter activities of a group of human-specific endogenous retroviruses in healthy and cancerous germ line tissues. To this end, we employed a recently developed technique termed genomic repeat expression monitoring. We found that at least 50% of human-specific long terminal repeats (LTRs) possessed promoter activity, and many of them were up- or downregulated in a seminoma. Individual LTRs were expressed at markedly different levels, ranging from ∼0.001 to ∼3% of the housekeeping beta-actin gene transcript level. We demonstrated that the main factors affecting the LTR promoter activity were the LTR type (5′-proviral, 3′ proviral, or solitary) and position with regard to genes. The averaged promoter strengths of solitary and 3′-proviral LTRs were almost identical in both tissues, whereas 5′-proviral LTRs displayed two- to fivefold higher promoter activities. The relative content of promoter-active LTRs in gene-rich regions was significantly higher than that in gene-poor loci. This content was maximal in those regions where LTRs “overlapped” readthrough transcripts. Although many promoter-active LTRs were mapped near known genes, no clear-cut correlation was observed between transcriptional activities of genes and neighboring LTRs. Our data also suggest a selective suppression of transcription for LTRs located in gene introns.
Viral and Cellular Determinants of the Hepatitis C Virus Envelope-Heparan SulfateInteraction
Authors: Barth Heidi , Schnober Eva K. , Zhang Fuming , Linhardt Robert J. , Depla Erik , Boson Bertrand , Cosset Francois-Loic , Patel Arvind H. , Blum Hubert E. , Baumert Thomas F.
Published: October 2006

Cellular
binding and entry of hepatitis C virus (HCV) are the first steps of
viral infection and represent a major target for antiviral antibodies
and novel therapeutic strategies. We have recently demonstrated that
heparan sulfate (HS) plays a key role in the binding of HCV envelope
glycoprotein E2 to target cells (Barth et al., J. Biol. Chem.
278:
41003-41012, 2003). In this study, we characterized
the HCV-HS interaction and analyzed its inhibition by antiviral host
immune responses. Using recombinant envelope glycoproteins, virus-like
particles, and HCV pseudoparticles as model systems for the early steps
of viral infection, we mapped viral and cellular determinants of HCV-HS
interaction. HCV-HS binding required a specific HS structure that
included N-sulfo groups and a minimum of 10 to 14 saccharide subunits.
HCV envelope binding to HS was mediated by four viral epitopes
overlapping the E2 hypervariable region 1 and E2-CD81 binding domains.
In functional studies using HCV pseudoparticles, we demonstrate that
HCV binding and entry are specifically inhibited by highly sulfated HS.
Finally, HCV-HS binding was markedly inhibited by antiviral antibodies
derived from HCV-infected individuals. In conclusion, our results
demonstrate that binding of the viral envelope to a specific HS
configuration represents an important step for the initiation of viral
infection and is a target of antiviral host immune responses in vivo.
Mapping of viral and cellular determinants of HCV-HS interaction sets
the stage for the development of novel HS-based antiviral strategies
targeting viral attachment and
entry.
Loss of VHL Confers Hypoxia-Inducible Factor (HIF)-Dependent Resistance to Vesicular Stomatitis Virus: Role of HIF in Antiviral Response
Authors: Hwang Irene I. L. , Watson Ian R. , Der Sandy D. , Ohh Michael
Published: October 2006

Hypoxia-inducible factor (HIF) is a central regulator of cellular responses to hypoxia, and under normal oxygen tension the catalytic α subunit of HIF is targeted for ubiquitin-mediated destruction via the VHL-containing E3 ubiquitin ligase complex. Principally known for its association with oncogenesis, HIF has been documented to have a role in the antibacterial response. Interferons, cytokines with antiviral functions, have been shown to upregulate the expression of HIF-1α, but the significance of HIF in the antiviral response has not been established. Here, using renal carcinoma cells devoid of VHL or reconstituted with functional wild-type VHL or VHL mutants with various abilities to negatively regulate HIF as an ideal model system of HIF activity, we show that elevated HIF activity confers dramatically enhanced resistance to vesicular stomatitis virus (VSV)-mediated cytotoxicity. Inhibition of HIF activity using a small-molecule inhibitor, chetomin, enhanced cellular sensitivity to VSV, while treatment with hypoxia mimetic CoCl2promoted resistance. Similarly, targeting HIF-2α by RNA interference also enhanced susceptibility to VSV. Expression profiling studies show that upon VSV infection, the induction of genes with known antiviral activity, such as that encoding beta interferon (IFN-β), is significantly enhanced by HIF. These results reveal a previously unrecognized role of HIF in the antiviral response by promoting the expression of the IFN-β gene and other genes with antiviral activity upon viral infection.
In Vitro Synthesis of Minus-Strand RNA by an Isolated
Cereal Yellow Dwarf Virus
RNA-Dependent RNA Polymerase Requires VPg and a Stem-Loop Structure at the 3′ End of the Virus RNA
Authors: Osman Toba A. M. , Coutts Robert H. A. , Buck Kenneth W.
Published: October 2006

Cereal yellow dwarf virus
(CYDV) RNA has a 5′-terminal genome-linked protein (VPg). We have expressed the VPg region of the CYDV genome in bacteria and used the purified protein (bVPg) to raise an antiserum which was able to detect free VPg in extracts of CYDV-infected oat plants. A template-dependent RNA-dependent RNA polymerase (RdRp) has been produced from a CYDV membrane-bound RNA polymerase by treatment with BAL 31 nuclease. The RdRp was template specific, being able to utilize templates from CYDV plus- and minus-strand RNAs but not those of three unrelated viruses,
Red clover necrotic mosaic virus
,
Cucumber mosaic virus
, and
Tobacco mosaic virus
. RNA synthesis catalyzed by the RdRp required a 3′-terminal GU sequence and the presence of bVPg. Additionally, synthesis of minus-strand RNA on a plus-strand RNA template required the presence of a putative stem-loop structure near the 3′ terminus of CYDV RNA. The base-paired stem, a single-nucleotide (A) bulge in the stem, and the sequence of a tetraloop were all required for the template activity. Evidence was produced showing that minus-strand synthesis in vitro was initiated by priming by bVPg at the 3′ end of the template. The data are consistent with a model in which the RdRp binds to the stem-loop structure which positions the active site to recognize the 3′-terminal GU sequence for initiation of RNA synthesis by the addition of an A residue to VPg.
Integrity of Membrane Lipid Rafts Is Necessary for the Ordered Assembly and Release of Infectious Newcastle Disease Virus Particles
Authors: Laliberte Jason P. , McGinnes Lori W. , Peeples Mark E. , Morrison Trudy G.
Published: October 2006

Membrane lipid raft domains are thought to be sites of assembly for many enveloped viruses. The roles of both classical lipid rafts and lipid rafts associated with the membrane cytoskeleton in the assembly of Newcastle disease virus (NDV) were investigated. The lipid raft-associated proteins caveolin-1, flotillin-2, and actin were incorporated into virions, while the non-lipid raft-associated transferrin receptor was excluded. Kinetic analyses of the distribution of viral proteins in lipid rafts, as defined by detergent-resistant membranes (DRMs), in non-lipid raft membranes, and in virions showed an accumulation of HN, F, and NP viral proteins in lipid rafts early after synthesis. Subsequently, these proteins exited the DRMs and were recovered quantitatively in purified virions, while levels of these proteins in detergent-soluble cell fractions remained relatively constant. Cholesterol depletion of infected cells drastically altered the association of viral proteins with DRMs and resulted in an enhanced release of virus particles with reduced infectivity. Decreased infectivity was not due to effects on subsequent virus entry, since the extraction of cholesterol from intact virus did not significantly reduce infectivity. Particles released from cholesterol-depleted cells had very heterogeneous densities and altered ratios of NP and glycoproteins, demonstrating structural abnormalities which potentially contributed to their lowered infectivity. Taken together, these results indicate that lipid rafts, including cytoskeleton-associated lipid rafts, are sites of NDV assembly and that these domains are important for ordered assembly and release of infectious Newcastle disease virus particles.
Generation and Analysis of Infectious Virus-Like Particles of Uukuniemi Virus (
Bunyaviridae
): a Useful System for Studying Bunyaviral Packaging and Budding
Authors: Överby Anna K. , Popov Vsevolod , Neve Etienne P. A. , Pettersson Ralf F.
Published: October 2006

In the present report we describe an infectious virus-like particle (VLP) system for the Uukuniemi (UUK) virus, a member of the
Bunyaviridae
family. It utilizes our recently developed reverse genetic system based on the RNA polymerase I minigenome system for UUK virus used to study replication, encapsidation, and transcription by monitoring reporter gene expression. Here, we have added the glycoprotein precursor expression plasmid together with the minigenome, nucleoprotein, and polymerase to generate VLPs, which incorporate the minigenome and are released into the supernatant. The particles are able to infect new cells, and reporter gene expression can be monitored if the
trans
-acting viral proteins (RNA polymerase and nucleoprotein) are also expressed in these cells. No minigenome transfer occurred in the absence of glycoproteins, demonstrating that the glycoproteins are absolutely required for the generation of infectious particles. Moreover, expression of glycoproteins alone was sufficient to produce and release VLPs. We show that the ribonucleoproteins (RNPs) are incorporated into VLPs but are not required for the generation of particles. Morphological analysis of the particles by electron microscopy revealed that VLPs, either with or without minigenomes, display a surface morphology indistinguishable from that of the authentic UUK virus and that they bud into Golgi vesicles in the same way as UUK virus does. This infectious VLP system will be very useful for studying the bunyaviral structural components required for budding and packaging of RNPs and receptor binding and may also be useful for the development of new vaccines for the human pathogens from this family.
Immune suppression prevents renal damage and dysfunction and reduces arterial pressure in salt-sensitive hypertension
Authors: Tian N. , Gu J.-W. , Jordan S. , Rose R. A. , Hughson M. D. , Manning R. D.
Published: October 2006

The goal of this study was to test the hypothesis that renal infiltration of immune cells in Dahl S rats on increased dietary sodium intake contributes to the progression of renal damage, decreases in renal hemodynamics, and development of hypertension. We specifically studied whether anti-immune therapy, using mycophenolate mofetil (MMF), could help prevent increases in renal NF-κB activation, renal infiltration of monocytes/macrophages, renal damage, decreases in glomerular filtration rate (GFR) and renal plasma flow, and increases in arterial pressure. Seventy-four 7-to 8-wk-old Dahl S, Rapp strain rats were maintained on an 8% Na, 8% Na + MMF (20 mg·kg−1·day−1), 0.3% Na, or 0.3% Na + MMF diet for 5 wk. Arterial and venous catheters were implanted at day 21. By day 35, renal NF-κB in 8% Na rats was 47% higher than in 0.3% Na rats and renal NF-κB was 41% lower in 8% Na + MMF rats compared with the 8% Na group. MMF treatment significantly decreased renal monocyte/macrophage infiltration and renal damage and increased GFR and renal plasma flow. In high-NA Dahl S rats mean arterial pressure increased to 182 ± 5 mmHg, and MMF reduced this arterial pressure to 124 ± 3 mmHg. In summary, in Dahl S rats on high sodium intake, treatment with MMF decreases renal NF-κB and renal monocyte/macrophage infiltration and improves renal function, lessens renal injury, and decreases arterial pressure. This suggests that renal infiltration of immune cells is associated with increased arterial pressure and renal damage and decreasing GFR and renal plasma flow in Dahl salt-sensitive hypertension.