Nonradioactive immunoassays incorporating an element of amplification in their detection system require the use of components that are highly purified. Flavin adenine dinucleotide-3′-phosphate (FADP) is the primary substrate used in such an amplification assay. For incorporation into a simple, single-pot assay system, the concentration of contaminating flavin adenine dinucleotide (a prosthetic group for the enzyme d-aminoacid oxidase used in the amplification cascade assay) in this primary substrate must be minimized to achieve maximum sensitivity. Production of the substrate to a high degree of purity has been achieved using apo-glucose oxidase to specifically remove contaminating flavin adenine dinucleotide from solution and hydrolysis of a cyclic intermediate as a final production protocol by ribonuclease T2 to give the product in high yield. The use of continuous ultrafiltration reactors at each stage is described and compared to a final production step utilizing immobilized ribonuclease T2. These reactors allow large volumes of material to be handled and assist in the scale-up of these processes. The suitability of each protocol is assessed for the commercial production of FADP.

In addition to hydrolysing RNA, bovine pancreatic ribonuclease splits esters of pyrimidine nucleoside 3'-phosphates, including dinucleotides. For a series of 3':5'-linked dinucleotides of general structure CpN, where N is a 5' linked nucleoside, kcat for the release of N varies enormously with the precise structure of N. Structural studies have been interpreted to indicate that the group N interacts with a subsite, B2, on the enzyme that comprises Gln69, Asn71 and Glu111. We report studies by site-directed mutagenesis that indicate that Gln69 is not involved in productive interactions with any of the dinucleotide substrates and that Asn71 is an important component of subsite B2 for all dinucleotide substrates tested. Glu111 appears to be functionally involved in catalysis for dinucleotide substrates solely when N is guanosine.

A simple to use, robust, quantitative, and extremely sensitive colorimetric assay for alkaline phosphatase (EC 3.1.3.1), designed to be used as a detection system in diagnostic assays employing antibodies or gene probes, is described. This technology is based on the novel principle of prosthetogenesis, according to which a purpose-designed substrate (a prosthetogen) for a primary analyte-linked enzyme label is hydrolyzed to produce a prosthetic group for a detector enzyme system. The prosthetogen employed here is a derivative of FAD which is phosphorylated at the 3'-position of the ribose ring (FADP), the label enzyme is alkaline phosphatase, and the detector is a D-amino-acid oxidase/horseradish peroxidase-coupled system. Essentially each turnover of every molecule of alkaline phosphatase produces a molecule of D-amino-acid oxidase for detection. Thus enormous amplification of the initial signal is achieved in short time periods because of the relatively high turnover number of alkaline phosphatase for FADP. The system can be formatted as a stable, preformed, freeze-dried preparation containing all analytical components, which is reconstituted simply by addition of buffer solution. This methodology can quantitate less than 0.1 amol of alkaline phosphatase in 30 min at 25 degrees C using microtiter plates.

An assay for alkaline phosphatase is described which is based on the hydrolysis of riboflavin phosphates (5'FMN or 4'FMN) to produce riboflavin. This is converted to 5'FMN using riboflavin kinase, and then assayed using the bacterial bioluminescent system from Vibrio harveyi or V. fischeri. The most sensitive assay is obtained using 4'FMN, which can measure less than 20 amol after a 1-hour incubation.

Alkaline phosphatase hydrolyzes riboflavin 4′-phosphate to produce riboflavin. This is converted to riboflavin 5′-phosphate, using riboflavin kinase, which reconstitutes apoglycolate oxidase to give hologlycolate oxidase. This enzyme catalyzes the oxidation of glycolate with simultaneous production of hydrogen peroxide which is detected via the formation of a colored product through the action of peroxidase. The system allows the detection of 4 amol after a 2-h incubation.

AbstractA series of dioxabicyclo[n.2.2]alkanes (n = 1,2,3 and 4) and dioxabicyclo[n.2.2]alkenes were studied under electron impact ionization. The fragmentation pathways were elucidated with the aid of accurate mass measurements, metastable scan techniojei and deuterium labelling. The preferential fragmentation of molecular ions in dioxabicyclo[n.2.2]alkanes corresponds to the loss of hydroperoxyl radical ·OOH. Hydrogens that are participating in this elimination come primarily from the syn position on the two‐membered carbon bridge. The dioxabicyclo[n.2.2]alkenes undergo the retro‐Diels–Alder process, which produces dioxygen and 1,3‐cycloalkadiene. In both saturated and unsatutated peroxides subsequent fragmentation of the hydrocarbon ring depends upon the value of n.

Chemischer InformationsdienstVolume 17, Issue 1 Preparative Organic Chemistry ChemInform Abstract: Secondary Bicyclic Peroxide Cation Radicals. S. F. NELSEN, S. F. NELSENSearch for more papers by this authorM. F. TEASLEY, M. F. TEASLEYSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author S. F. NELSEN, S. F. NELSENSearch for more papers by this authorM. F. TEASLEY, M. F. TEASLEYSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author First published: January 7, 1986 https://doi.org/10.1002/chin.198601099AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume17, Issue1January 7, 1986 RelatedInformation

The crystal structures of the title saturated bicyclic [n.2.1] peroxides confirm the previous suggestion, based on n.m.r. data and mechanistic considerations, that in each compound the bromine atom is syn to the dioxygen bridge. The C–O–O–C dihedral angles are O(1), 14(1), and 45(1)° respectively, while the corresponding O–O bond lengths are 1.498(8), 1.447(8), and 1.463(7)Å. The six-, seven-, and eight-membered carbocyclic rings adopt chair, chair, and chair-chair conformations respectively. A comparison of the [3.2.1] compound with its homologues, and with 47 other cyclic or bicyclic peroxides, suggests that its O–O bond is relatively long, and that this arises because of bond angle strain primarily at the bromine-bearing carbon atom.

Chemischer InformationsdienstVolume 17, Issue 47 Physical Organic Chemistry ChemInform Abstract: Crystal and Molecular Structures of 8-Bromo-6,7-dioxabicyclo[3.2.1]octane, 9-Bromo-7,8-dioxabicyclo[4.2.1]nonane, and 10-Bromo-8,9-dioxabicyclo[5.2.1]decane. A. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this authorH. M. DAWES, H. M. DAWESSearch for more papers by this authorM. B. HURSTHOUSE, M. B. HURSTHOUSESearch for more papers by this authorN. P. C. WALKER, N. P. C. WALKERSearch for more papers by this author A. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this authorH. M. DAWES, H. M. DAWESSearch for more papers by this authorM. B. HURSTHOUSE, M. B. HURSTHOUSESearch for more papers by this authorN. P. C. WALKER, N. P. C. WALKERSearch for more papers by this author First published: November 25, 1986 https://doi.org/10.1002/chin.198647042Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume17, Issue47November 25, 1986 RelatedInformation

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTDetection of hydrogen, hydrogen-deuterium molecule (HD), and deuterium by Raman spectroscopy: a powerful aid for the elucidation of reaction mechanismsStephen P. Best, A. J. Bloodworth, Robin J. H. Clark, and Henny J. EggelteCite this: J. Am. Chem. Soc. 1985, 107, 9, 2626–2628Publication Date (Print):May 1, 1985Publication History Published online1 May 2002Published inissue 1 May 1985https://doi.org/10.1021/ja00295a011RIGHTS & PERMISSIONSArticle Views367Altmetric-Citations7LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (386 KB) Get e-Alerts Get e-Alerts

La detection de H 2 , HD et D 2 par spectroscopie Raman en phase gazeuse a montre que la deshydrogenation photochimique et thermique de dioxabicyclo [n.2.2] alcanes (n=2,3) se produit selon un processus intramoleculaire

Chemischer InformationsdienstVolume 16, Issue 36 Preparative Organic Chemistry ChemInform Abstract: DETECTION OF HYDROGEN, HYDROGEN-DEUTERIUM MOLECULE (HD), AND DEUTERIUM BY RAMAN SPECTROSCOPY: A POWERFUL AID FOR THE ELUCIDATION OF REACTION MECHANISMS S. P. BEST, S. P. BESTSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorR. J. H. CLARK, R. J. H. CLARKSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author S. P. BEST, S. P. BESTSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorR. J. H. CLARK, R. J. H. CLARKSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author First published: September 10, 1985 https://doi.org/10.1002/chin.198536105AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume16, Issue36September 10, 1985 RelatedInformation

AbstractDurch cis‐Addition entsteht bei der Reduktion von (I) ein mit der Ringgröße zunehmender Anteil an syn‐ Addukten (II).

Chemischer InformationsdienstVolume 16, Issue 10 Physical Organic Chemistry ChemInform Abstract: PHOTOELECTRON SPECTRA OF DIOXABICYCLO(N.2.1)ALKANES R. GLEITER, R. GLEITERSearch for more papers by this authorW. DOBLER, W. DOBLERSearch for more papers by this authorM. ECKERT-MAKSIC, M. ECKERT-MAKSICSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this authorD. CREMER, D. CREMERSearch for more papers by this author R. GLEITER, R. GLEITERSearch for more papers by this authorW. DOBLER, W. DOBLERSearch for more papers by this authorM. ECKERT-MAKSIC, M. ECKERT-MAKSICSearch for more papers by this authorA. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this authorD. CREMER, D. CREMERSearch for more papers by this author First published: March 12, 1985 https://doi.org/10.1002/chin.198510036Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References R. GLEITER, W. DOBLER, M. ECKERT-MAKSIC, A. J. BLOODWORTH, H. J. EGGELTE, D. CREMER, PHOTOELECTRON SPECTRA OF DIOXABICYCLO(N.2.1)ALKANES, J. Org. Chem., 1984, 49, 3716. DOI: 10.1021/jo00194a009; 10.1021/jo00194a009 CASWeb of Science®Google Scholar Volume16, Issue10March 12, 1985 ReferencesRelatedInformation

The 2H n.m.r. spectra of the mixtures of endoperoxides obtained by photo-oxygenation of pyrolysed 1,4-diacetoxy-1,4-dideuteriocyclo-hexane, -heptane, and -octane demonstrate that 1.5-migrations of hydrogen and deuterium in the corresponding dideuteriated cycloalka-1,3-dienes proceed to equilibrium in less than 5 s at 480 °C.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTSecondary bicyclic peroxide cation radicalsStephen F. Nelsen, Mark F. Teasley, A. John Bloodworth, and Henny J. EggelteCite this: J. Org. Chem. 1985, 50, 18, 3299–3302Publication Date (Print):September 1, 1985Publication History Published online1 May 2002Published inissue 1 September 1985https://pubs.acs.org/doi/10.1021/jo00218a011https://doi.org/10.1021/jo00218a011research-articleACS PublicationsRequest reuse permissionsArticle Views77Altmetric-Citations10LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail Other access optionsGet e-AlertscloseSupporting Info (1)»Supporting Information Supporting Information Get e-Alerts

Chemischer InformationsdienstVolume 16, Issue 43 Preparative Organic Chemistry ChemInform Abstract: USE OF DEUTERATION, ENDOPEROXIDATION, AND DEUTERIUM NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY TO DEMONSTRATE THERMAL (1,5) SIGMATROPIC REARRANGEMENTS IN CYCLOALKA-1,3-DIENES A. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author A. J. BLOODWORTH, A. J. BLOODWORTHSearch for more papers by this authorH. J. EGGELTE, H. J. EGGELTESearch for more papers by this author First published: October 29, 1985 https://doi.org/10.1002/chin.198543118Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume16, Issue43October 29, 1985 RelatedInformation

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTPhotoelectron spectra of dioxabicyclo[n.2.1]alkanesRolf Gleiter, Walter Dobler, Mirjana Eckert-Maksic, A. John Bloodworth, Henny J. Eggelte, and Dieter CremerCite this: J. Org. Chem. 1984, 49, 20, 3716–3720Publication Date (Print):October 1, 1984Publication History Published online1 May 2002Published inissue 1 October 1984https://pubs.acs.org/doi/10.1021/jo00194a009https://doi.org/10.1021/jo00194a009research-articleACS PublicationsRequest reuse permissionsArticle Views34Altmetric-Citations7LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail Other access optionsGet e-Alertsclose Get e-Alerts