bluuebunny/crossref_metadata_2025 huggingface dataset:
212Pb/212Bi‐EDTMP — synthesis and biodistribution of a novel bone seeking alpha‐emitting radiopharmaceutical
Authors: Hassfjell Sindre P. , Hoff Per , Bruland Øyvind S. , Alstad Jorolf
Published: October 2006

At present, haematological toxicity is dose limiting in radionuclide therapy of bone metastases, and there is a need for radiopharmaceuticals with improved tumour/bone marrow dose ratios. Therefore, α‐emitters e.g. 212Bi may be more suitable than β‐emitters, because of the short range and high LET values of α‐particles. In this study, 212Bi and its mother nuclide 212Pb were produced in an isotope generator by collecting gaseous 220Rn emanating from barium (228Th) stearate. The carrier‐free 212Pb/212Bi were bound to the chelating bone‐seeking compound ethylene‐diamine‐tetra(methylene‐phosphonic acid) (EDTMP) with 90% yield. The biodistribution in Balb/c mice was investigated by injecting 100 μl of a saline PBS buffer 0.020 M in EDTMP and 10 MBq/ml in 212Pb/212Bi. Mice were killed in groups of three at 0.5, 2, 13 and 24 h post‐injection times. Both 212Pb‐EDTMP and 212Bi‐EDTMP localised strongly in the skeleton, especially in the femur, at all time points measured, with the % of injected dose per gram (%ID/g) as high as 15 for 212Pb and 13 for 212Bi. All other organs investigated showed low uptake of both radionuclides, with the exception of the kidneys, for which a ratio femur/kidney of 1.5 for 212Bi 2 h postinjection was observed. By comparison the ratio femur/blood was 20 for 212Bi 2 h postinjection. The experiment indicates a potential for 212Pb/Bi‐EDTMP in targeted radiotherapy of osteoblastic bone lesions.
Synthesis and X‐ray structure determination of N,N′‐bis‐l‐(1‐carboxy‐2‐mercaptoethyl)ethylenediamine, oxotechnetium (V), [TcO(EC)]
Authors: Pirmettis I. , Mastrostamatis S. , Papadopoulos M. , Raptopoulou C. P. , Terzis A. , Chiotellis E.
Published: October 2006

The complex of N,N′‐bis‐1‐(carboxy‐2‐mercaptoethyl)ethylene‐diamine (EC) labelled with reduced Tc‐99m was proposed as a new agent in renal tubular secretion studies. Here we present the synthesis and single‐crystal X‐ray structure determination of the N,N′‐bis‐1‐(1‐carboxy‐2‐mercaptoethyl)ethylene‐diamine oxotechnetium(V) complex, [99TcO (EC)]. Crystals are orthoro‐bic, space group P212121, with a=19.570(1), b=10.080 (1), c=6.300 (1)A, Z=4 and R=0.0294. The technetium is hexacoordinated being bonded to oxygen (1.657(3)A), the two N atoms and the two deprotonated S atoms. The sixth site, trans to the oxo‐group is occupied by one deprotonated carboxylate group. The infrared spectra is also presented.
The syntheses of radiolabelled org 5222 and its main metabolite org 30526
Authors: Vader Jan , Kaspersen Frans , Sperling Eric , Schlachter Irene , Terpstra Annie , Hilberink Peter , Wagenaars Gerard
Published: October 2006

The syntheses of trans‐5‐chloro‐2, 3, 3a‐12b‐tetrahydro‐2‐methyl‐1H‐dibenz [2, 3:6, 7] oxepino [4, 5‐c]pyrrole (Org 5222), a potential antipsychotic compound, labelled with 3H, 14C and 36C1 and trans‐5‐chloro‐2, 3, 3a, 12b‐tetrahydro‐1H‐dibenz [2, 3:6, 7]‐oxepino [4, 5‐c] pyrrole (Org 30526) labelled with 3H are described. 3H‐labelled Org 5222 of low specific activity was prepared by a base catalyzed exchange with tritiated water of an amide precursor, 3H‐labelled Org 5222 with a high specific activity by a catalytic reductive dehalogenation. 3H‐labelled Org 30526 was prepared both by demethylation of 3H‐Org 5222 and by catalytic reductive iodination of 11‐iodo‐Org 30526. 14C‐labelled Org 5222 was synthesized in 6 steps using 14C‐sarcosine as starting material. 36C1‐labelled Org 5222 was prepared by diazotation reaction in the presence of H36C1.
Labelled compounds of interest as antitumour agents. Part 41. Deuteration and tritiation of a nitroimidazole‐carborane designed for BNCT
Authors: Scobie Martin , Bew Sean P. , Threadgill Michael D.
Published: October 2006

Quenching the anion generated from a 2‐(ω‐carboranylalkyl)dithiane with 2H2O at −78°C and at 0°C introduced deuterium exclusively at C‐2 of the carborane. Extension of this model reaction to a bioreductively‐targetted carborane allowed the synthesis of 2‐[2H]‐ and 2‐[3H]‐isotopomers of a nitroimidazole‐carborane which is of interest in boron neutron capture therapy (BNCT) of cancer.
The synthesis of (−)‐4‐methyl‐8‐chloro‐trans‐1,2,3,4,4a,5,6,10b‐octahydrobenzo‐[f]‐quinolin‐3‐one‐[3‐14C] (LY300502‐14C]) via a circuitous route
Authors: Kuo Fengjiun , Wheeler William J.
Published: October 2006

The synthesis of the C‐14 labeled isotopomer of LY300502, a potent 5α‐reductase inhibitor has been accomplished in four radiochemical steps. The route involves the synthesis of LY300502‐[14C] from LY300502 via a circuitous route; the label was introduced with ethyl chloroformate‐[carbonyl‐14C].
A single‐mode microwave cavity for reducing radiolabelling reaction times, demonstrated by alkylations with [11C]alkyl halides
Authors: Stone‐Elander Sharon A. , Elander Nils , Thorell Jan‐Olov , Solås Gunnar , Svennebrink Jan
Published: October 2006

A single‐mode microwave cavity was constructed for microscale reactions with consideration taken to the space limitations typical for shielded working spaces used with positron emitting radionuclides. The effect of the microwave field in this cavity on typical reaction media is demonstrated. Reaction times for radiolabelling neuroreceptor ligands with [11C]methyl iodide and [2‐11C]isopropyl iodide were shown to be considerably reduced compared with the times reported in the literature for the corresponding thermal procedures.
Synthesis of carbon‐14 labeled Taxol® (paclitaxel)
Authors: Walker Donald G. , Swigor John E. , Kant Joydeep , Schroeder Daniel R.
Published: October 2006

Reductive cleavage of the C13 side chain of Taxol® (1, paclitaxel) followed by regioselective silylation gave 7‐triethylsilylbaccatin III (4). 3‐O‐Triethylsilylation of 5 and subsequent reaction with benzoyl chloride‐C7‐14C gave azetidinone 7. Coupling of 4 and 7 followed by deprotection gave 1.26 g of Taxol®‐N3′‐14C (11) having a specific activity of 26.5 mCi/mmol and a radiochemical purity of 95%.
Labelling of benzocaina with tritium
Authors: Malik Sohail
Published: October 2006

A convenient method is described to label a local anesthetic, benzocaine, with tritium. The bromoester of para — aminobenzoic acid (PABA) was prepared from para‐nitrotoluene and was reduced with tritium. The generation of isotopic hydrogen and labelling of benzocaine was achieved in one‐step. A mixture of sodium borohydride (NaB3H4) with cobalt (II) chloride was used to generate tritium gas. 5% Pd/C was used as a catalyst. This constitutes the first report of tritium labelled benzocaine.
Synthesis of racemic [methyl‐d3]‐iabeled cis‐ and trans‐3′‐hydroxycotinine
Authors: Ravard Alain , Crooks Peter A.
Published: October 2006

A method is described for the synthesis of the racemic [methyl‐d3] forms of the nicotine metabolites cis‐3′‐hydroxycotinine and trans‐3′‐hydroxycotinine. The key intermediate was [methyl‐d3]‐N‐methylhydroxylamine, obtained from a selective hydrogenation of d3‐nitro‐methane. This intermediate was converted to [methyl‐d3]‐α‐3‐pyridyl‐N‐methylnitrone, which was condensed with methyl acrylate to give a mixture of isomeric isoxazolidines. The hydrogenolysis of this mixture afforded a 70:30 mixture of [methyl‐d3] cis‐ and trans‐3′‐hydroxycotinine, from which the pure cis‐ isomer could be isolated by recrystallization from acetone. [Methyl‐d3]‐trans‐3′‐hydroxycotinine could be prepared in high yield from the cis‐isomer via chiral inversion utilizing a Mitsunobu reaction, or by chromatographic separation from a mixture of the cis‐ and trans‐3′‐benzoyloxycotinine, followed by O‐debenzoylation in methanolic NaOH.
The synthesis of perdeuterated azone (d35‐1‐dodecylhexahydro‐2H‐azepin‐2‐one)
Authors: Groundwater Paul W. , Hadgraft Jonathan , Harrison Julian E. , Watkinson Adam C.
Published: October 2006

Perdeuteroazone (3) has been synthesised via the base‐catalysed coupling of d11‐hexahydro‐2H‐azepin‐2‐one (2) with d25‐1‐chlorododecane.
Synthesis of [2H13]heptanoic acid via the desulfurization of methyl 3‐chloro‐5,6‐dibromothieno[3,2,b]thiophene‐2‐carboxylate with nickel‐aluminium alloy in NaOD‐D2O solution
Authors: Tsuzuki Hirohisa , Mukumoto Mamoru , Tsukinoki Takehito , Mataka Shuntaro , Tashiro Masashi , Yonemitsu Tadashi , Nagano Yoshiaki
Published: October 2006

[2H13]Heptanoic acid was synthesized as its methyl ester (4) (deuterium content: 96%) with Ni‐Al alloy in 10% NaOD‐D2O via the desulfurization of methyl 3‐chloro‐5,6‐dibromothieno[3,2,b]thiophene‐2‐carboxylate (3), which was derived from 4,5‐dibromothiophene‐2‐carboaldehyde (1) in two steps.
Synthesis of [U‐14C]Arg labelled decapeptide cetrorelix, a novel lutenizing hormone — releasing hormone antagonist
Authors: Locher Mathias , Johnston James , Müller Thomas , Borbe Harald O. , Kutscher Bernhard , Engel Jürgen
Published: October 2006

The decapeptide cetrorelix is a novel luteinizing hormone — releasing hormone (LH‐RH) antagonist. For nonclinical studies concerning absorption, distribution, metabolism and excretion (ADME) in animals the [14C]‐lebelled compound is essential. Therefore, [U‐14C]Arg cetrorelix acetate salt was synthesized by Amersham International, Buckinghamshire (England) from precursor peptides provided by Degussa AG, Hanau‐Wolfgang (Germany). [U‐14C]Arg labelled cetrorelix peptide base has a specific activity of 8.13 MBq/mg (220 μCi/mg) and a molecular weight of 1442.6 g/mol at this specific activity. The chemical and radiochemical purity (92.2 % decapeptide content and 97.6%, respectively) determined by HPLC is suitable for nonclinical investigations.
The synthesis of deuterium‐labeled N‐nitrosodiethanolamine and N‐nitroso‐2‐hydroxymorpholine
Authors: Loeppky Richard N. , Xiong Heping
Published: October 2006

Two deuterated derivatives of N‐nitrosodiethanolamine and two deuterated derivatives of N‐Nitroso‐2‐hydroxymorpholine were prepared. 1,1,1′,1′‐2H4‐N‐Nitrosodiethanolamine 1a was prepared in 99% isotopic purity by simple base catalyzed H‐D exchange. 2,2,2′,2′‐2H4‐N‐Nitrosodiethanolamine 1b was synthesized in 98% isotopic purity by the LiAlD4 reduction of dimethyl iminodiethanoate, followed by acid catalyzed nitrosation. 5,5‐2H2‐N‐Nitroso‐2‐hydroxymorpholine 2a was prepared in 99% isotopic purity through a series of steps involving 1) the selective base catalyzed H‐D exchange of the CH2 adjacent to the ester function of 2,2‐diethoxyethylcarboethoxymethylnitrosamine, 2) its reduction with diisobutylaluminum hydride at −8°C followed by 3) acid catalyzed hydrolysis of the acetal and cyclization to the hemiacetal. 2‐2H‐N‐Nitroso‐2‐hydroxymorpholine 2b (98% isotopic purity) was prepared through the LiAlD4 reduction of 2‐hydroxyethylcorboethoxymethylnitrosamine at −78°C.
The preparation of high purity H235S
Authors: Collins C. D. , Bell J. N. B.
Published: October 2006

H2 35S was synthesised by the reaction of H2 with elementary [35S] sulphur at 600°C with 100 mg of pumice added as a catalyst. The resulting gas was purified by placing the preparation vessel in a pack of ice. A yield of 46% was produced.
Synthesis of an 125I analog of MK‐0591 and characterization of a 5‐lipoxygenase activating protein binding assay
Authors: Eggler J. F. , Cheng J. B. , Cooper K. , Hanak L. M. , Pillar J. S. , Zdankiewicz M.
Published: October 2006

The 125I analog of MK‐0591, 1, has been prepared for use as a radioligand for developing a 5‐lipoxygenase activating protein (FLAP) binding assay. The radiosynthesis involves a two step oxidative iododestannylation‐saponification procedure. A FLAP binding assay has been developed in human neutrophil membranes. The binding of 1 to human neutrophil FLAP is rapid, reversible, of high affinity, saturable and selective for FLAP inhibitors.
Synthesis of (+)‐(3R)‐3‐(4‐fluorophenylsulfonamido)‐1,2,3,4‐tetrahydro‐9‐[5,6,7,8,12,13‐U‐14C]carbazolepropanoic acid, [14C]BAY u 3405
Authors: Radtke Martin , Boberg Michael , Pleiss Ulrich , Rosentreter Ulrich
Published: October 2006

The title compound [14C]BAY u 3405 (1) was synthesized as part of 8‐step sequence. Starting from [U‐14C]aniline hydrogensulfate the final product 1 was obtained with a specific activity of 741 MBq/mmol (20 mCi/mmol) and a radiochemical purity of > 98% in an overall yield of 6 and 10% depending on the method.
Bromination and subsequent catalytic tritiation of thienylalanine and 4‐methyltyrosine residues in the bradykinin analog RMP‐7
Authors: Akiyama Alan , Musso Gary F. , Smart Janet L. , Lang Christopher , Straub Julie Ann
Published: October 2006

A two step strategy was devised for the synthesis of 3H‐RMP‐7 for use in pharmacokinetic studies. First, RMP‐7 was brominated predominantly on the thiophene ring of thienylalanine using Br2 in AcOH. Then, reductive tritiation of the brominated RMP‐7 using 3H2 and Pd/C yielded 3H‐RMP‐7 (specific activity 1 Ci/mmol; 96% radiochemical purity). An HPLC based assay using on‐line radioactivity detection was developed for in vivo pharmacokinetic studies of 3H‐RMP‐7. Rapid clearance from the blood and metabolism of RMP‐7 to des‐Arg1‐RMP‐7 was observed in vivo in rats.
N.C.A. 11C‐labelling of benzenoid compounds in ring positions: Synthesis of nitro‐[1‐11C]benzene and [1‐11C]aniline
Authors: Steinbach J. , Mäding P. , Füchtner F. , Johannsen B.
Published: October 2006

The paper describes the first method for n.c.a 11C‐ring labelling of benzenioid compounds having a reactive group for further derivatization by use of the known principle of synchronous six‐electron cyclization of hexatriene systems into aromatics. Nitro‐[11C]methane (1) prepared from cyclotron‐produced [11C]carbon dioxide reacts in the presence of t‐BuOK with 5‐dimethylaminopenta‐2,4‐dienylidene‐dimethylammonium perchlorate (2) to form 6‐nitro‐1‐dimethylamino‐[6‐11C]hexatriene (3) follow by cyclization/ aromatization into nitro‐[1‐11C]‐benzene (4) at increased temperatures. Starting from 1, nitro‐[1‐11C]benzene of a radiochemical purity of about 92 % and a mean specific radioactivity of 1 Ci/μmol was obtained within 7 min. Related to [11C]CO2, the reproducible radiochemical yield of 4 (decay‐corrected) was 80±5 %. Reduction of 4 by heating the above reaction mixture with aqueous Na2S gave [1‐11C]aniline (5) of aradiochemical purity of about 81 %. The reproducible radiochemical yield of 5 (decay‐corrected) in relation to [11C]CO2 was 65±5 %, the synthesis time from 1 was 18 min.
Synthesis of β‐3H‐mitotane for use in a rapid assay for mitotane metabolism
Authors: Piñeiro‐Sánchez Mayra L. , Vaz Alfin D. N. , Counsell Raymond E. , Ruyan Mohamed , Schteingart David E. , Sinsheimerl Joseph E.
Published: October 2006

A 3H+‐release method has been developed for the assay of β‐hydroxylation of the adrenolytic drug mitotane. β‐3H‐mitotane was synthesized by the reduction of 1‐(2‐chlorophenyl)‐1‐(4‐chlorophenyl)‐2,2,2‐trichloroethane by an aluminium‐Hg2Cl2 couple in the presence of 3H2O. For β‐hydroxylation of mitotane, the 3H+‐release assay is more efficient and sensitive than a method utilizing 14C‐mitotane and chromatographic separation of metabolites by HPLC. The 3H+‐release assay has been used to evaluate the ability of adrenal tumors to metabolize mitotane via the β‐ hydroxylation route.
Strategies for labelling branched polypeptides with a poly(L‐lysine) backbone with radioiodines (123I, 125I, 131I) and radiometals (111In, 51Cr) for biodistribution studies and radiopharmaceutical development
Authors: Pimm Malcolm V. , Gribben Sandra J. , Mezö Gábor , Hudecz Ferenc
Published: October 2006

Methods have been developed for radiolabelling synthetic branched polypeptides, these being based on a poly(L‐lysine) backbone with short side chains of three DL‐alanine residues and one other amino acid at the end of the branches (MW ∼45‐100 kDa). Labelling has been carried out with gamma emitting radionuclides suitable for use in biodistribution studies or for gamma scintigraphy. Labelling with 125I was achieved by reaction of the polypeptides' terminal amino groups with pre‐iodinated Bolton and Hunter Reagent (N‐Succinimidyl 3‐(4‐hydroxy‐5‐[125I]iodophenyl)propionate). Alternatively, polypeptides were reacted with non‐labelled Bolton and Hunter reagent, which could subsequently be iodinated with 123I, 125I or 131I by oxidative incorporation from [123I]Nal, [125I]Nal, or [123I]Nal. For labelling with radiometals, the polypeptides' terminal amino groups were reacted with diethylenetriaminepentaacetic acid (DTPA) anhydride, and the conjugated DTPA subsequently labelled with 111In or 51Cr by chelation. An amphoteric polypeptide, having terminal glutamic acid residues on its side chains (EAK), and a polycationic polypeptide, with terminal D‐lysine (D‐KAK) were labelled in these ways. In addition EAK previously conjugated to the cis‐aconityl derivative of daunomycin (EAK‐cAD) was similarly labelled. Gel permeation chromatography on Sephacryl S‐300, which was possible with the amphoteric EAK, showed virtually identical elution profiles with 123I, 125I 131I, 111In and 51Cr labelled EAK and its cAD conjugate. Biodistribution studies in mice showed prolonged blood survival of the radionuclide of 125I, 111In or 51Cr labelled EAK and EAK‐cAD. There were, however, differences in organ levels of the radionuclides. Generally kidney, spleen and liver levels of radiometals were higher than those of radioiodine, while levels in the gastrointestinal tract were higher with radioiodine. D‐KAK labelled with any of the three radionuclides was cleared rapidly from the blood, high levels of all tracers being found in spleen, liver, kidney and lung.
Synthesis of 14C‐labelled Cefluprenam (E1077), a novel parenteral cephalosporin antibiotic
Authors: Sato Nobuaki , Kai Yasunobu , Shemilt G. I. , Chiku Shigeru
Published: October 2006

Cefluprenam (E1077) is a new parenteral cephalosporin with a well‐balanced antibacterial spectrum and potent antibacterial activity. It was synthesized labelled in the side chain at the 3‐position of the cephem with carbon‐14, starting from potassium [14C]cyanide, according to the method illustrated in Schemes 1,2. [14C]Cefluprenam having a specific activity of 3.9 MBq/mg, was obtained in 28.6 % overall radiochemical yield, with a radiochemical purity of more than 97.3 %.
New [99mTc]‐cytectrene amine compounds as specific brain imaging agents
Authors: Kuntschke Dirk , Wenzel Martin , Schulze Paul
Published: October 2006

Lipophilic tertiary amines attached to cyclopentadienyl technetium‐99m tricarbonyl (cytectrene) have been prepared with high radiochemical yield and purity. Biodistribution studies in mice showed that [99mTc]‐cytectrenes, containing in their structure an N‐methylpiperidine, were accumulated in the brain up to 2.8 % of injected dose with high brain‐to‐blood ratios at 15 min p.i.1. They therefore indicate some potential as brain imaging agents.

It has to be pointed out that the N‐methylpiperidine ester showed similar biological behaviour as the keto derivative. This indicates that the conversion to polar metabolite(s) via hydrolysis of the ester group ‐ as described for [99mTc]‐ECD ‐ is not essential for brain retention.
Synthesis of deuteriated benzaldehydes
Authors: Ishimoto Keiko , Tsukinoki Takehito , Tsuge Otohiko , Tashiro Masashi
Published: October 2006

A new method for the preparation of d1‐benzaldehydes from α‐keto acids in 2 steps is reported.
Synthesis of N‐[4‐[2‐(2,4‐dimethylphenoxy)phenyl]‐2‐thiazolyl]‐hexahydro‐2‐pyrimidinimine, [14C]BAY w 6341
Authors: Pleiss Ulrich , Conrad Michael , Baasner Bernd
Published: October 2006

The title compound [14C]BAY w 6341 was synthesized as part of a 4‐step sequence. Starting from diethyl [2‐14C]malonate the final product (8) was obtained with a specific activity of 1.2 GBq/mmol (32.9 mCi/mmol) and a radiochemical purity of > 99 % in an overall yield of 19 %.
Synthesis of tritium labelled 2‐vinyl dihydrosphingosine‐1‐phosphate
Authors: Boumendjel Ahcene , Miller Stephen P. F.
Published: October 2006

The synthesis of [1,1,3‐3H]‐2‐vinyl dihydrosphingosine‐1‐phosphate is described. The key step in this synthesis is the reduction of methyl 2‐trifluoro acetamido‐2‐vinyl nonadecanoate‐3‐one by means of 3H‐calcium diborohydride. This compound was synthesized to determine whether 2‐vinyl dihydrosphingosine‐1‐phosphate inhibits the PLP‐dependent sphingosine‐1‐phosphate lyase irreversibly via covalent modification.
Synthesis of [u‐13C,15N]‐cysteine hydrochloride: An important tool for heteronuclear, multi‐dimensional NMR studies of proteins
Authors: Panigot Michael J. , Fesik Stephen W. , Curley Robert W.
Published: October 2006

Protein structure determination by modern NMR techniques is greatly facilitated using 15N‐ and 13C‐labeled proteins. Labeling of proteins that are overexpressed in mammalian cells is a difficult task that requires a growth medium consisting of algal hydrolysates supplemented with labeled amino acids. Although most of the amino acids can be obtained to prepare an isotopically labeled growth medium for mammalian cells,[U‐13C,15N]‐cysteine is not available, hampering the backbone and cysteine side‐chain assignments and structure determination in the vicinity of the cysteine residues. A synthesis of D,L‐[U‐13C,15N]‐cysteine hydrochloride in good overall yield is described which makes use of readily available 15N‐ and 13C‐labeled starting materials and will facilitate heteronuclear multidimensional NMR studies of proteins that are overexpressed in mammalian cells.
Synthesis of paclitaxel‐C3′‐14C
Authors: Walker Donald G. , Standridge Robert T. , Swigor John E.
Published: October 2006

Reductive cleavage of the C13 side chain of paclitaxel (3) followed by regioselective silylation gave 7‐triethylsilylbaccatin III (6). Successive reaction of L‐threonine methyl ester hydrochloride (7) with tertbutoxydiphenylchlorosilane, benzaldehyde‐C7‐14C and acetoxyacetyl chloride/ triethylamine gave a 92:8 ratio (NMR) of azetidinones 10:11 in 57% yield from 7. Removal of the chiral auxiliary led to 16, which after 3‐O‐triethylsilylation and N‐benzoylation provided (3R,4S)‐cis‐1‐benzoyl‐3‐O‐(triethylsilyl)‐4‐phenylazetidin‐2‐one‐C4‐14C (18). Coupling of 18 and 6 followed by deprotection gave 1.12 g of paclitaxel‐C3′‐14C (2) having a specific activity of 16.4 mCi/mmol and a radiochemical purity of 96%.
Synthesis of [14C2] SDZ FOX 988, a hypoglycemic agent
Authors: Sunay Ustun B. , Talbot Kenrick , Prasad Kapa , Lee George , Jones Lawrence
Published: October 2006

4‐[2,2‐Dimethyl‐1‐oxopropyl‐1‐[14C]‐4‐[2,2‐dimethyl‐1‐oxopropyl‐ 1‐[14C]‐phenyl(methoxyimino)‐benzoic acid, methyl ester, [14C2] SDZ FOX 988, doubly labelled in the two keto carbons, was prepared from 4‐bromotoluene in four steps. The final condensation featured a novel method for preparation of N‐oxyimidic acid derivatives.
A new practical method for the preparation of [3H6]‐leukotrienes C4 and D4
Authors: Belley Michel L. , Gareau Yves , Zamboni Robert J. , Ahern David G. , Hong Yang
Published: October 2006

A new method for the synthesis of highly deuterated or tritiated leukotrienes was developed. The higher reactivity of a terminal alkene compared to a 1,2‐disubstituted one permitted the selective deuteration or tritiation of the diyne 14,15,19,19,20,20‐hexadehydro LTC4 triester 1. After hydrolysis, LTC4 was obtained in 36% overall yield. An average of seven deuterium or tritium atoms was incorporated and the specific activity of the tritiated LTC4 was greater than 180 Ci/mmol. 1 was obtained from the addition of glutathione to 14,15,19,19,20,20‐hexadehydro LTA4 ethyl ester which was the product of a Wittig reaction between (3,8‐nonanediyn‐1‐yl)triphenylphosphonium iodide and 5(S), 6(S), 7(E), 9(E) ethyl 5,6‐epoxy‐11‐oxo‐7,9‐undecadienoate.
Synthesis of tritiated 1α,25‐dihydroxy‐22‐oxavitamin D3
Authors: Watanabe Hiroyoshi , Akiyama Masashi , Kawanishi Takehiko , Kubodera Noboru
Published: October 2006

Synthesis of two tritiated 1α,25‐dihydroxy‐22‐oxavitamin D3 (OCT), [26‐3H3]OCT (3) and [2β‐3H]OCT (4), is described. [26‐3H3]OCT (3) was prepared by tritiation at the side chain with tritiated methylmagnesium iodide and [2β‐3H]OCT (4) was labeled at the A‐ring by tritiation with sodium borotritiide.
Synthesis of 1,1′‐carbonyldimidazole‐1‐14C and its use in preparing a methoxy(polyethylene)glycol semicarbazide linker
Authors: Walker Donald G. , Leister William H. , Weaner Larry E.
Published: October 2006

Reaction of N‐(trimethylsilyl)imidazole (2) with phosgene‐14C in toluene afforded 92 w/w % pure (1H NMR) 1,1′‐carbonyldimidazole‐1‐14C (3) in near quantitative yield. Activation of MPEG‐amine 4 by reaction with 3 in dichloromethane afforded intermediate 5 in situ. Further reaction with hydrazine in toluene at 50°C and reprecipitation from 2‐propanol afforded crude 1 (81%). A two‐step chromatographic purification of this material followed by reprecipitation from 2‐propanol gave MPEG‐semicarbazide linker1. This material had a chemical purity estimated at 98%, a radiochemical purity of 95%, a specific activity of 7.7 μCi/mg (37.6 mCi/mmol) and a number‐average molecular weight of 4885 (1H NMR).
A one‐step synthesis of [3‐2H]oxazepam
Authors: Yang Shen K. , Bao Ziping , Shou Magang
Published: October 2006

The proton at 3‐position of oxazepam (7‐chloro‐1,3‐dihydro‐3‐hydroxy‐5‐phenyl‐2H‐1,4‐benzodiazepin‐2‐one) undergoes a deuterium exchange in deuterated alkaline methanol. A base‐catalyzed keto‐enol tautomerism is proposed to be responsible for the observed deuterium exchange. This simple method is a considerable improvement of the multi‐step synthetic procedure reported in the literature.
Synthesis of a tritium labeled rat enterostatin, Val‐[3,4‐3H‐Pro]‐Gly‐[3,4‐3H‐Pro]‐Arg‐Oh
Authors: Kozlowski Edward S. , Dischino Douglas D. , Johnson Graham
Published: October 2006

A tritium labeled rat Enterostatin, Val‐[3,4‐3H‐Pro]‐Gly‐[3,4‐3H‐Pro]‐Arg‐OH, was prepared by the catalytic tritiation of the didehydroproline analog of rat Enterostatin, Val‐[3,4‐dehydroPro]‐Gly‐[3,4‐dehydroPro]‐Arg‐OH, over Pd/C in methanol. The product had a specific activity of 59.2 Ci/mmol and a radiochemical purity of greater than 99%.
Pesticides labelled with 14C I. Synthesis of [6‐14C]hexazinone
Authors: Koltai Ernǒ , Rutkai György , Kling Ferenc
Published: October 2006

Hexazinone ‐ a s‐triazine type contact herbicide ‐ was labelled with 14C for the investigation of metabolic pathways. A convenient synthesis was elaborated for labelling of the carbon atom in the 6 position of the triazine ring. The molar activity of Hexazinone was 797.0 MBq/mmole (21.54 mCi/mmole).
Synthesis of deuterium‐labeled acrivastine and an acrivastine metabolite
Authors: Thompson J. B. , Blake K. W. , McNutt R. W.
Published: October 2006

The antihistamine acrivastine and a metabolite dihydroacrivastine were prepared as their [2H7]‐congeners from [2H8]‐tolualdehyde for use as internal standards in gas chromatography/mass spectroscopy of biological samples.
Preparation of [2′,3′,5′,6′‐2H4]pteroylglutamic acid
Authors: Dueker Stephen R. , Jones A. Daniel , Smith Gary M. , Clifford Andrew J.
Published: October 2006

Folic acid plays a key role in nucleic acid biosynthesis, essential for normal cell proliferation and function. Localized folate deficiencies may be related to changes in cytology associated with cancer development; analogs of folic acid, such as methotrexate, are potent chemotherapeutic agents and are widely used either alone or in combination therapy for many types of cancer. In this report we describe the synthesis of a tetra‐deuterated folic acid from perdeuterated toluene. The primary intermediate, N‐(4‐amino [2,3,5,6‐2H4]benzoyl)‐L‐glutamic acid diethyl ester was coupled to N(2′)‐acetyl‐6‐formylpterin to create [2′,3′,5′,6′‐2H4]folic acid. A similar scheme can be used for the preparation of [1′,2′,3′,4′,5′,6′‐13C6]folic acid from [13C6] ring labeled toluene.
Radiolysis of radioimmunoconjugates. Reduction in antigen‐binding ability by α‐particle radiation
Authors: Larsen Roy H. , Bruland Øyvind S.
Published: October 2006

The potential of delivering a high and localized radiation dose using α‐particle‐emitting radioimmunoconjugates (RIC's) is investigated as treatment for some types of cancer. High specific activity of the RIC may for some uses be desirable, allowing enrichment of the radionuclide on membrane antigens on target cells. Radiation exposure during the radiolabelling procedures may, however, reduce the antigen binding ability of RIC's of high specific activity. The influence of α‐particle dose on the cell binding fraction was therefore investigated for two monoclonal antibodies, i.e., TP‐1 F(ab')2 and TP‐3 IgG. Samples of 125I‐labelled MoAb were added different amounts of [211At]astatide to give accumulated doses ranging from 50 to 50,000 Gy. After the 211At had decayed 15 half‐lives, the cell binding fractions were measured. The results show that [211At] astatide activities up to 25 kBq/μl, corresponding to an accumulated α‐particle radiation dose of approximately 1,000 Gy, could be tolerated without significantly reducing the cell binding fraction, whereas higher doses gradually reduced the immunoreactivity. Although there may be differences in dose sensitivity of different radioimmunoconjugate due to differences in radionuclide conjugation methods, size of the antibody and structure of the antigen binding portion, the results presented here indicate that it is possible to produce radioimmunoconjugates of high specific activity with currently used methods of preparation. The potential problem of radiolysis should, however, be taken into consideration when planning procedures for preparation and use of α‐particle‐emitting radioimmunoconjugates.
Synthesis of (2E,6E)‐[10‐3H]farnesol and (2E6E)‐[10‐3H]farnesal for insect dehydrogenase studies
Authors: Sen Stephanie E. , Garvin Gail M.
Published: October 2006

[10‐3H]Farnesol ((2E,6E)‐3,7,11‐trimethyl‐2,6,10‐dodecatrien‐1‐ol) and [10‐3H]farnesal ((2E,6E)‐3,7,11‐trimethyl‐2,6,10‐dodecatrien‐1‐al) were synthesized by sequential reduction and oxidation of the corresponding tert‐butyldiphenylsilyl protected 11,12,13‐trisnoraldehyde, followed by Wittig homologation using isopropyltriphenylphosphorane. Direct reduction of the C‐12 allylic bromide or allylic chloride proved to be an nonviable method, resulting in either multiple decomposition products or significant double bond transposition. Oxidation of the [10‐3H]labelled trisnoralcohol with pyridinium chlorochromate resulted in essentially complete retention of the radiolabel as was established for the corresponding deuterated material. [3H]labelled farnesol and farnesal were obtained in 78% and 74% yields, respectively, from the trisnoraldehyde. These materials were used as radiotracers for examining the enzymatic activity of insect farnesol and farnesal dehydrogenase, key enzymes in the biosynthesis of juvenile hormone.
The synthesis of [U‐14C phenyl] LS 840606, an agricultural fungicide
Authors: Madegard G. , Mestre P. , Raimond P. , Noël J‐P.
Published: October 2006

2,2′4′‐Trichloro‐[ring U‐14C]acetophenone 3 was the key intermediate of this synthesis patterned after the industrial route. An unexpected poor yield was observed during the preparation of 3 by the Friedel‐Crafts reaction of chloroacetyl chloride with 1,3‐dichloro‐[U‐14C]benzene 10, possibly the result of an isotope effect although this poor yield might be explained by other factors.

Two routes were checked for the preparation of 1,3‐dichloro‐[U‐14C]benzene 10. The action of CCL4 with 1,3‐dinitro‐[U‐14C]benzene at 280°C was entailed with explosions. A safer route started from [U‐14C]aniline via 2,4‐dichloro‐[ring U‐14C]acetanilide.

Friedel‐Crafts reaction of 10 with acetyl chloride gave rise in 52% yield to 2′,4′‐dichloro‐[ring U‐14C]acetophenone 16 which was brominated to 2‐bromo‐2′,4′‐dichloro‐[ring U‐14C]acetophenone 17;17 was condensed with 2,2‐(ethylenedioxy)ethylmagnesium bromide to compound 18; 18 was condensed with 1,2,4‐triazole to 5 then successively treated with HCl:water:dioxane and 2,2,2‐trifluoroethanol/HCl. Separation of the two diastereomers by medium pressure liquid chromatography. 7% overall radioactive yield from [U‐14C]aniline. Radiochemical purity 99%.
The synthesis and purification of 2, 6‐di‐tert‐buytl‐p‐cresol‐−14C6
Authors: Shipp B. D. , Data J. B. , Christian J. E.
Published: October 2006

2, 6‐Di‐tert‐butyl‐p‐cresol‐14C6 was synthesised by reacoting p‐cresol‐14C6 with isobutylene using concetentrated sulfurica acid as catalyst.

Purification of the Labelled butylated p‐cresol involved very careful recrystalliation of this product from absolute alcohol.

Proof of radiochemical purity was determined using thin layer chromatography and autoradiography. Internal liquid acintillation counting techniques were used to determine the apeacific activity of the compound.
Die synthese von mit 14C‐isotop markierten aryl‐isothiocyanten
Authors: Zòlyomi G. , Toldy L. , Bánfi D.
Published: October 2006

Die thermische Zersetung von Aryl‐thioharmstoffe, zwecks Darstellung der mit 14C‐Isotop markierten Aryl‐isothiocyanate wurde untersucht. Aus 2, 6‐Dichlorphenyl‐, bzw. 2, 6‐Dimethyl‐phenyl‐isothiocyanate ausgehend wurden substituierte, mit radioaktivem Kohlenstoff markierte Thiazolidin‐Derivate synthetisiert.
Preparation of labeled aflatoxin B1 from acetate‐1, 2‐14C
Authors: Mabee M. S. , Chipley J. R. , Applegate K. L.
Published: October 2006

Aflatoxin B1‐14C was produced by resting cell cultures of Aspergillus flavus (NRRL 3145) from a sodium acetate ‐ 1, 2‐14C precursor. A specific activity of 455 uCi/uM was determined for the purified aflatoxin B1‐14C.
Synthese d'allylamines deuteriees en position specifique
Authors: le Coustumer F. , Lautié M. F. , Dizabo P.
Published: October 2006

Les allylamines CH2 = CH ‐ CD2NH2, = CD ‐ CH2NH2, CH2 = CD ‐ CD2NH2, ont été préparées par réduction par AlLiH4 (ou D4), des adduits anthracéniques des acrylonitiles CH2 = CH ‐ CN et CH2 = CD ‐ CN. Pour les allylamines CD2 = CH ‐ CH2NH2 et CD2 = CH ‐ CD2NH2, deux méthodes sont décrites: réduction soit de l'adduit anthracè;ne‐acrylonitrile CD2 = CH‐CN, soit de l'aduit anthracène‐acrylamide CD2 = CH ‐ CONH2.

Les allylamines récupérées par décomposition de leurs adduits anthracéniques ont été purifiées par chromatographie en phase vapeur et leur pureté contrôlée par R.M.N.
The chemical and enzymatic syntheses of specifically deuterated dinucleoside monophosphates
Authors: Kondo N. S. , Leung A. , Danyluk S. S.
Published: October 2006

Proton magnetic resonance (pmr) studies of nucleic acid structure are currently hampered by extensive overlapping of signals, particularly in the ribose region. An approach with considerable promise involves the preparation of oligomers with specifically deuterated nucleotidyl units at known positions. In this publication, we report the synthesis of five dinucleoside monophosphates in which the 3′‐nucleotidyl unit is deuterated. These compounds were prepared by standard chemical procedures and an enzymatic approach using ribonuclease T1. The deuterated dimers give pmr spectra which are much simpler than their protio analogs, thus permitting unambiguous peak assigments.
Syntheses with stable isotopes: Synthesis of deuterium and 13C labeled bile acids
Authors: Hachey D. L. , Szczepanik P. A. , Berngruber O. W. , Klein P. D.
Published: October 2006

A series of 5β‐cholanic acids labeled with deuterium in the A ring were prepared by exchange labeling of the corresponding ketone by column chromatography on deuterated alumina. Factors affecting yield and labeling efficiency are discussed. 5β‐Cholanic acids labeled with 13C in the carboxyl position were prepared by treatment of the corresponding 23‐chloro‐24‐norcholane with sodium cyanide‐13C followed by alkaline hydrolysis of the nitrile. The intermediates in the synthesis were characterized by high resolution NMR spectroscopy. Mass spectra are also reported for the 13C labeled products.
Methode de synthese et D'etude par spectrometrie infrarouge des composes de coordination de molecules marquees application aux complexes de l'acetone et du trifluorure de bore
Authors: Cornut Jean Claude , Fouassier Monique , Forel Marie‐Thérèse
Published: October 2006

La synthése et l'étude spectroscopique des composés d'addition de molécules marquées avec des isotopes lourds présentent de nombreuses difficultés en raison des faibles quantités de produit qui doivent être utilisées. Une méthode de piégeages à basse température est proposée, elle est illustrée par l'étude des complexes de l'acétone 18 et du triifluorure de bore 10B.
Determination of purity of some tritiated steroids
Authors: Merill Edward J. , Vernice Gerald G.
Published: October 2006

When radiolabeled steroids at high specific activities are required for biological studies, the particular requirements of purity must be determined. The analyses to which the compound is subjected is based on these requirements. Analytical methods are complicated because of the radiochemical instability and the lack of aufficient sample. Methods have been developed which will allow IR and UV spectra to be determined with subsequent recovery of the sample. A prime requisite for this is specially purified solvents. The acceptability of certain solvents for this use can be determined by concentrating an aliquot of the solvent and then examining the residue by TLC. Spraying the developed chromatogram with a 5% solution of phosphomolybdic acid in ethanol, followed by heating will reveal the presence of impurities in the solvents. These impurities appear as spots and can readily be misinterpreted as contaminants in the sample if unpurified solvents are used to manipulate a pure steroid. Problems associated with the determination of radiochemical purity via GLC and TLC will be discussed. The synthesis of tritiated ethynyl estraniol its cyclopentul ether and the cyclopentyl ether of norethinarone acetate will illustrate the above problems.
Molecular Cloning and Characterization of NAD+‐Dependent Xylitol Dehydrogenase from Candida tropicalis ATCC 20913
Authors: Ko Byoung Sam , Jung Heung Chae , Kim Jung Hoe
Published: October 2006

Induction of xylitol dehydrogenase of Candida tropicalis ATCC 20913 by various carbon sources was investigated. The enzyme activity was induced when the yeast was grown on l‐arabinose and d‐xylose. A novel gene encoding the enzyme was cloned and characterized. The 1,095‐bp coding sequence of the gene encodes a polypeptide of 364 amino acids, with a molecular mass of 39.4 kDa. Sequence analysis of the putative protein showed it to be a member of the zinc‐containing alcohol dehydrogenase family and to have homology to xylitol dehydrogenase genes from other yeasts and fungi. The recombinant xylitol dehydrogenase expressed in Escherichia coli oxidized polyols such as xylitol and d‐sorbitol and reduced ketoses such as d‐xylulose and d‐fructose. It required exclusively NAD or NADH as a cofactor.
Abiotic Stress Generates ROS That Signal Expression of Anionic Glutamate Dehydrogenases to Form Glutamate for Proline Synthesis in Tobacco and Grapevine
Authors: Skopelitis Damianos S. , Paranychianakis Nikolaos V. , Paschalidis Konstantinos A. , Pliakonis Eleni D. , Delis Ioannis D. , Yakoumakis Dimitris I. , Kouvarakis Antonios , Papadakis Anastasia K. , Stephanou Euripides G. , Roubelakis-Angelakis Kalliopi A.
Published: October 2006

Glutamate dehydrogenase (GDH) may be a stress-responsive enzyme, as GDH exhibits considerable thermal stability, and de novo synthesis of the α-GDH subunit is induced by exogenous ammonium and senescence. NaCl treatment induces reactive oxygen species (ROS), intracellular ammonia, expression of tobacco (Nicotiana tabacum cv Xanthi) gdh-NAD;A1 encoding the α-subunit of GDH, increase in immunoreactive α-polypeptide, assembly of the anionic isoenzymes, and in vitro GDH aminating activity in tissues from hypergeous plant organs. In vivo aminating GDH activity was confirmed by gas chromatorgraphy–mass spectrometry monitoring of 15N-Glu, 15N-Gln, and 15N-Pro in the presence of methionine sulfoximine and amino oxyacetic acid, inhibitors of Gln synthetase and transaminases, respectively. Along with upregulation of α-GDH by NaCl, isocitrate dehydrogenase genes, which provide 2-oxoglutarate, are also induced. Treatment with menadione also elicits a severalfold increase in ROS and immunoreactive α-polypeptide and GDH activity. This suggests that ROS participate in the signaling pathway for GDH expression and protease activation, which contribute to intracellular hyperammonia. Ammonium ions also mimic the effects of salinity in induction of gdh-NAD;A1 expression. These results, confirmed in tobacco and grape (Vitis vinifera cv Sultanina) tissues, support the hypothesis that the salinity-generated ROS signal induces α-GDH subunit expression, and the anionic iso-GDHs assimilate ammonia, acting as antistress enzymes in ammonia detoxification and production of Glu for Pro synthesis.
Identification of a novel mutation in hereditary vitamin D resistant rickets causing exon skipping
Authors: Hawa N. S. , Cockerill F. J. , Vadher S. , Hewison M. , Rut A. R. , Pike J. W. , O'Riordan J. L. H. , Farrow S. M.
Published: October 2006

OBJECTIVE Hereditary vitamin D resistant rickets (HVDRR) is an autosomal recessive disorder resulting in target organ resistance to the actions of 1,25‐dihydroxyvitamin D3 (1,25(OH)2D3). In many cases, this disorder has been shown to be due to mutations in the gene encoding vitamin D receptors (VDR). In a patient with characteristic features of this disorder, we investigated the functional defect and sequenced the coding region of the gene for mutations.

DESIGN Skin fibroblasts from patient and control were used to measure binding of 1,25(OH)2D3 and functional responses to the hormone. These cells were also used to prepare RNA from which cDNA was prepared and sequenced. Furthermore, genomic DNA was prepared from the fibroblasts and the intronlexon boundarles sequenced.

PATIENT A child with classic features of HVDRR with alopecia diagnosed as having rickets due to resistance to 1,25(OH)2D3.

MEASUREMENTS Nuclear association of 1,25(OH)2D3 was determined in patient and control cells and the functional response to 1,25(OH)2D3 was assessed by measurement of 25‐hydroxyvitamln D‐24‐hydroxylase(24‐hydroxylase) activity. VDR cDNA and genomic DNA prepared from patient and control cells were sequenced.

RESULTS Cells from the patient with HVDRR had undetectable amounts of VDR compared to control cells and did not show induction of 24‐hydroxylase activity following treatment with 1,25(OH), D3. Sequencing of the VDR coding region after RT‐PCR of RNA revealed an absence of exon 4 in patient RNA which was not due to a deletion in genomic DNA but was caused by exon skipping during RNA processing. In addition, the deletion of exon 4 sequences from RNA leads to a frameshift in translation resulting in a premature stop codon. Amplification of genomic DNA around the intron/exon boundary of exon 4 revealed a point mutation in the 5’donor splice site of intron 4.

CONCLUSION In this study, we have identified a novel mutation in the gene for vitamin D receptors in a patient with the Characteristic phenotype of hereditary vitamin D resistant rickets. The mutation at the + 5 position in intron 4 is most likely to cause skipping of exon 4 in this patient.
Changes in free fatty acid composition during storage of whole milk powder
Authors: PÁEZ R , PENSEL N , SABBAG N , TAVERNA M , CUATRÍN A , ZALAZAR C
Published: October 2006

Changes in fat matter due to the microbial thermostable lipases are among the most significant deteriorations influencing the shelf life of whole milk powder. As a consequence, free fatty acids are produced, being the short‐chain acids mainly responsible for the rancidity flavour. The aim of this work was to evaluate changes in different fatty acid concentrations during the storage of whole milk powder produced in two different seasons. Samples of whole milk powder produced in winter and summer and packed under inert atmosphere were used in the study. The samples were stored at 21°C and 40°C. Samples stored at 21°C were evaluated every 3 months, whereas samples stored at 40°C were evaluated monthly by free fatty acids profile and sensory analysis. Data were processed by principal component analysis and anova. Results showed that changes in the free fatty acids of milk powders were correlated with the season of manufacture. Temperature and storage time had little influence on that profile.
Signal Transduction by Human Herpesvirus 8 Viral Interleukin-6 (vIL-6) IsModulated by the Nonsignaling gp80 Subunit of the IL-6 Receptor Complex and Is Distinct from Signaling Induced by Human IL-6
Authors: Hu Fang , Nicholas John
Published: October 2006

Human
herpesvirus 8 (HHV-8) viral interleukin-6 (vIL-6) mediates signaling
through the gp130 signal transducer but unlike human IL-6 (hIL-6) does
not require the nonsignaling gp80 α subunit of the IL-6
receptor complex. By utilizing a gp80-refractory vIL-6 variant,
vIL-6(R189L), we found that signal transduction, as measured by STAT1
and STAT3 activation and gp130 tyrosine phosphorylation in
gp80
+
/gp130
+
HEK293T cells, was
modulated by gp80. Furthermore, the signaling and BAF-130 cell
growth-promoting activities of vIL-6 and hIL-6 could be distinguished,
and exogenous addition of soluble gp80 enhanced cell growth supported
by vIL-6. Our findings demonstrate that gp80 can modulate vIL-6
activity and that vIL-6 and hIL-6 signaling are not directly
equivalent.
Kaposi's Sarcoma-Associated Herpesvirus LANA-1 Interacts with the Short Variant of BRD4 and Releases Cells from a BRD4- and BRD2/RING3-Induced G
1
Cell CycleArrest
Authors: Ottinger Matthias , Christalla Thomas , Nathan Kavita , Brinkmann Melanie M. , Viejo-Borbolla Abel , Schulz Thomas F.
Published: October 2006

The Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated
nuclear antigen 1 (LANA-1) is required for the replication of episomal
viral genomes. Regions in its N-terminal and C-terminal domains mediate
the interaction with host cell chromatin. Several cellular nuclear
proteins, e.g., BRD2/RING3, histones H2A and H2B, MeCP2, DEK, and
HP1α, have been suggested to mediate this interaction. In this
work, we identify the double-bromodomain proteins BRD4 and BRD3/ORFX as
additional LANA-1 interaction partners. The carboxy-terminal region of
the short variant of BRD4 (BRD4
S
) containing the highly
conserved extraterminal domain directly interacts with an element in
the LANA-1 carboxy-terminal domain. We show that ectopically expressed
BRD4
S
and BRD2/RING3 delay progression into the S phase of
the cell cycle in epithelial and B-cell lines and increase cyclin E
promoter activity. LANA-1 partly releases epithelial and B cells from a
BRD4
S
- and BRD2/RING3-induced G
1
cell cycle
arrest and also promotes S-phase entry in the presence of
BRD4
S
and BRD2/RING3. This is accompanied by a reduction in
BRD4
S
-mediated cyclin E promoter activity. Our data are in
keeping with the notion that the direct interaction of KSHV LANA-1 with
BRD4 and other BRD proteins could play a role in the G
1
/S
phase-promoting functions of KSHV LANA-1. Further, our data support a
model in which the LANA-1 C terminus contributes to a functional
attachment to acetylated histones H3 and H4 via BRD4 and BRD2, in
addition to the recently demonstrated direct interaction (A.
J. Barbera, J. V. Chodaparambil, B. Kelley-Clarke, V. Joukov,
J. C. Walter, K. Luger, and K. M. Kaye, Science
311:856-861, 2006) of the LANA-1 N terminus with histones H2A
and
H2B.
Pseudorabies Virus EP0 Protein Counteracts an Interferon-Induced Antiviral State in a Species-Specific Manner
Authors: Brukman Alla , Enquist L. W.
Published: October 2006

Pseudorabies virus (PRV), an alphaherpesvirus related to herpes simplex virus type 1 and varicella-zoster virus, infects a broad host range of mammals. A striking characteristic of PRV infection is the different symptoms and outcomes of infection in natural and nonnatural hosts. Adult pigs, the natural hosts of PRV, survive infection with only mild respiratory symptoms, while nonnatural hosts, including rodents and cattle, invariably die after exhibiting neurological symptoms. Here, we show that the PRV EP0 protein is necessary to overcome an interferon-mediated antiviral response in primary cells from the natural host of PRV but is not necessary in nonnatural-host cells.