{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T21:10:23Z","timestamp":1775769023606,"version":"3.50.1"},"reference-count":36,"publisher":"American Veterinary Medical Association (AVMA)","issue":"7","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["ajvr"],"published-print":{"date-parts":[[2004,7,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:bold>Objectives<\/jats:bold>\n                    \u2014To compare stability, antigenicity, and\naggregation characteristics of\n                    <jats:italic>Moraxella bovis<\/jats:italic>\n                    cytolysins among isolates from geographically diverse\nareas.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Study Population<\/jats:bold>\n                    \u20148 isolates of\n                    <jats:italic>M bovis<\/jats:italic>\n                    .\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Procedure<\/jats:bold>\n                    \u2014Filter-sterilized broth culture supernatants\nof\n                    <jats:italic>M bovis<\/jats:italic>\n                    were concentrated, diafiltered, and\nchromatographed. The endotoxin and cytolysin activities\nin samples were measured. Chromatographed\ncytolysins of\n                    <jats:italic>M bovis<\/jats:italic>\n                    were examined by immunoblotting.\nHemolytic and leukotoxic activities were measured\nfrom samples collected at each step of purification\nand before and after storage. Hemolysis was measured\ndirectly by use of washed bovine erythrocyte\ntargets. Leukotoxicity was measured by use of a\n                    <jats:sup>51<\/jats:sup>\n                    Cr\nrelease assay.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Results<\/jats:bold>\n                    \u2014Cytolysin was retained by a filter with 100-kd nominal molecular weight limit. Hemolytic activity,\nleukotoxic activity, and endotoxin were eluted together\nin void volume of a gel-filtration column (molecular\nmass exclusion limit = 4 \u00d7 10\n                    <jats:sup>7<\/jats:sup>\n                    d). Gel-column chromatographed\ndiafiltered retentate had the greatest\nspecific cytolytic activity and the highest endotoxinto-\nprotein ratio. Frozen diafiltered retentate(\u201380\u00b0C, 4\nmonths) was cytolytic after thawing. Immunoblots of\ngel-column chromatographed cytolysin contained 4\nproteins with molecular masses between 90 and 68\nkd. Fractions with high lytic activities also had additional\nprotein bands with molecular masses of 98 and\n63 kd. Immunoblots of gel-column chromatographed\ndiafiltered retentate revealed proteins with molecular\nmasses between 90 and 68 kd.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Conclusions and Clinical Relevance<\/jats:bold>\n                    \u2014Diafiltered\n                    <jats:italic>M bovis<\/jats:italic>\n                    cytolysin is aggregated with endotoxin.\nAntigenicity and cytolytic activities in diafiltered retentate\nare conserved among\n                    <jats:italic>M bovis<\/jats:italic>\n                    isolates.\nDiafiltration could be useful for bulk semipurification\nof\n                    <jats:italic>M bovis<\/jats:italic>\n                    cytolysin. Cytolysin-enriched vaccines of\n                    <jats:italic>M bovis<\/jats:italic>\n                    could be contaminated by endotoxin. (\n                    <jats:italic>Am J Vet Res<\/jats:italic>\n                    2004;65:977\u2013983)\n                  <\/jats:p>","DOI":"10.2460\/ajvr.2004.65.977","type":"journal-article","created":{"date-parts":[[2005,12,22]],"date-time":"2005-12-22T05:39:17Z","timestamp":1135229957000},"page":"977-983","source":"Crossref","is-referenced-by-count":7,"title":["Stability, antigenicity, and aggregation of Moraxella bovis cytolysin after purification and storage"],"prefix":"10.2460","volume":"65","author":[{"given":"Lisle W.","family":"George","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John A.","family":"Angelos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William W.","family":"Ruehl","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1272","reference":[{"key":"p_1_413","doi-asserted-by":"publisher","DOI":"10.1016\/S0378-1135(02)00410-8"},{"key":"p_2_414","doi-asserted-by":"publisher","DOI":"10.2460\/ajvr.2001.62.1222"},{"key":"p_3_415","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1135(94)00084-A"},{"key":"p_4_416","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1135(94)90074-4"},{"key":"p_5_417","first-page":"1848","volume":"45","author":"Ostle AG","year":"1984","journal-title":"Am J Vet Res"},{"key":"p_6_418","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1128\/IAI.59.3.1148-1152.1991","volume":"59","author":"Clinkenbeard KD","year":"1991","journal-title":"Infect Immun"},{"key":"p_7_419","doi-asserted-by":"crossref","first-page":"191","DOI":"10.2460\/ajvr.1990.51.02.191","volume":"51","author":"Hoien-Dalen PS","year":"1990","journal-title":"Am J Vet Res"},{"key":"p_8_420","first-page":"18","volume":"50","author":"Kagonyera GM","year":"1990","journal-title":"Am J Vet Res"},{"key":"p_9_421","first-page":"883","volume":"38","author":"Sandhu TS","year":"1977","journal-title":"Am J Vet Res"},{"key":"p_10_422","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1292\/jvms1939.41.363","volume":"41","author":"Nakazawa M","year":"1979","journal-title":"Jap J Vet Sci"},{"key":"p_11_423","first-page":"10","volume":"50","author":"Kagonyera GM","year":"1989","journal-title":"Am J Vet Res"},{"key":"p_12_424","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1097(94)90334-4"},{"key":"p_13_425","first-page":"1011","volume":"46","author":"Ostle AG","year":"1985","journal-title":"Am J Vet Res"},{"key":"p_14_426","first-page":"393","author":"Weyant RS","year":"1996","journal-title":"Baltimore: The Williams & Wilkins Co"},{"key":"p_15_427","first-page":"629","volume":"80","author":"Bovre K","year":"1972","journal-title":"Acta Pathol Microbiol Scand"},{"key":"p_16_428","first-page":"79","author":"Anonymous","year":"2004","journal-title":"NJ: Amersham Biosciences"},{"key":"p_17_429","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"p_18_430","doi-asserted-by":"crossref","first-page":"4350","DOI":"10.1073\/pnas.76.9.4350","volume":"76","author":"Towbin H","year":"1979","journal-title":"Proc Natl Acad Sci U S A"},{"key":"p_19_431","unstructured":"Harlow E, Lane D. 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