{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:44:29Z","timestamp":1753879469462,"version":"3.41.2"},"reference-count":17,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T00:00:00Z","timestamp":1734652800000},"content-version":"vor","delay-in-days":19,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Reprod Domestic Animals"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Urine samples were systematically collected from inseminated Murrah buffaloes (<jats:styled-content style=\"fixed-case\"><jats:italic>Bubalus bubalis<\/jats:italic><\/jats:styled-content>) on days 0, 7, 14, 21 and 28 (with day 0 representing the day of artificial insemination). Following confirmation of pregnancy via trans rectal palpation 45\u2009days of insemination, the animals were categorised into pregnant and non\u2010pregnant groups (<jats:italic>n<\/jats:italic>\u2009=\u200910 each). The urine samples on 0, 7, 14, 21 and 28\u2009days of pregnant and one sample from non\u2010pregnant preferably collected on 28th day was used for SDS\u2010PAGE after diafiltration. We focused on urine samples obtained on 28th day of insemination as it could contain high fetuin\u2010A levels associated with pregnancy. Each of the two urine samples from both the pregnant and non\u2010pregnant groups underwent purification through wheat germ agglutinin (WGA) affinity separation. The resulting fraction\/elute was subsequently employed for the determination and validation of fetuin\u2010A through 1D SDS\u2010PAGE, western blotting and MALDI\u2010TOF analysis. The samples were used for measuring the concentration of fetuin\u2010A by ELISA. Fetuin\u2010A was significantly higher (1020.50\u2009\u00b1\u200919.75\u2009mg\/L) in pregnant Murrah buffalo urine than their non\u2010pregnant counterparts (86.5\u2009\u00b1\u20096.33\u2009mg\/L), with a <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05. On SDS\u2010PAGE analysis, the WGA fractions revealed distinct protein bands, notably between ~54 to 70\u2009kDa and slightly above ~91\u2009kDa, in pregnant Murrah buffaloes compared with their non\u2010pregnant counterparts. Following confirmation of fetuin\u2010A at ~58\u2009kDa through western blotting, the corresponding protein band was excised from the SDS gel and subjected to mass spectrometry for MALDI\u2010TOF analysis. Peptide mass fingerprinting, coupled with protein BLAST, confirmed the upregulated protein as Alpha2\u2010HS glycoprotein, also known as fetuin\u2010A. These findings are crucial for elucidating the presence of fetuin\u2010A in urine samples from pregnant Murrah buffaloes, offering valuable insights into their physiological status.<\/jats:p>","DOI":"10.1111\/rda.70001","type":"journal-article","created":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T10:09:50Z","timestamp":1734689390000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Upregulation of Urinary Alpha\u20102\u2010<scp>HS<\/scp> Glycoprotein During Pregnancy of Murrah Buffaloes"],"prefix":"10.1111","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8889-0471","authenticated-orcid":false,"given":"T.","family":"Prasada Rao","sequence":"first","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V.","family":"Paramesh","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S.","family":"Pranavi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V.","family":"Manasa","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T. V. Sai","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K.","family":"Padmaja","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biochemistry College of Veterinary Science, S.V. Veterinary University  Tirupati Andhra Pradesh India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,12,20]]},"reference":[{"doi-asserted-by":"publisher","key":"e_1_2_10_2_1","DOI":"10.1016\/j.jprot.2015.05.026"},{"doi-asserted-by":"publisher","key":"e_1_2_10_3_1","DOI":"10.1080\/14767050802326255"},{"doi-asserted-by":"publisher","key":"e_1_2_10_4_1","DOI":"10.1042\/bj20030676"},{"doi-asserted-by":"publisher","key":"e_1_2_10_5_1","DOI":"10.1038\/256430a0"},{"doi-asserted-by":"publisher","key":"e_1_2_10_6_1","DOI":"10.1007\/BF02473242"},{"doi-asserted-by":"publisher","key":"e_1_2_10_7_1","DOI":"10.1111\/j.1600-0897.1997.tb00244.x"},{"doi-asserted-by":"publisher","key":"e_1_2_10_8_1","DOI":"10.1530\/eje.0.1470243"},{"doi-asserted-by":"publisher","key":"e_1_2_10_9_1","DOI":"10.1111\/j.1462-5822.2007.00901.x"},{"doi-asserted-by":"publisher","key":"e_1_2_10_10_1","DOI":"10.2337\/diacare.29.02.06.dc05-1484"},{"doi-asserted-by":"publisher","key":"e_1_2_10_11_1","DOI":"10.1016\/S0021-9258(17)30046-7"},{"doi-asserted-by":"publisher","key":"e_1_2_10_12_1","DOI":"10.1038\/154575a0"},{"doi-asserted-by":"publisher","key":"e_1_2_10_13_1","DOI":"10.1111\/1348-0421.12468"},{"key":"e_1_2_10_14_1","first-page":"1","article-title":"Identification of Potential Protein Biomarkers for Early Detection of Pregnancy in Cow Urine Using 2D DIGE and Label Free Quantification","volume":"13","author":"Preeti R.","year":"2016","journal-title":"Clinical Proteomics"},{"key":"e_1_2_10_15_1","first-page":"1","article-title":"Methods of Pregnancy Diagnosis in Domestic Animals: The Current Status","volume":"1","author":"Purohit G.","year":"2010","journal-title":"Webmedcentral"},{"issue":"2","key":"e_1_2_10_16_1","first-page":"83","article-title":"One\u2010Step Affinity Purification of Fetuin From Fetal Bovine Serum","volume":"35","author":"Simone C.","year":"2010","journal-title":"Biotechnology and Applied Biochemistry"},{"doi-asserted-by":"publisher","key":"e_1_2_10_17_1","DOI":"10.1681\/ASN.2008060572"},{"doi-asserted-by":"publisher","key":"e_1_2_10_18_1","DOI":"10.1186\/1471-2164-14-777"}],"container-title":["Reproduction in Domestic Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/rda.70001","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T11:02:15Z","timestamp":1735210935000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/rda.70001"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12]]},"references-count":17,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2024,12]]}},"alternative-id":["10.1111\/rda.70001"],"URL":"https:\/\/doi.org\/10.1111\/rda.70001","archive":["Portico"],"relation":{},"ISSN":["0936-6768","1439-0531"],"issn-type":[{"type":"print","value":"0936-6768"},{"type":"electronic","value":"1439-0531"}],"subject":[],"published":{"date-parts":[[2024,12]]},"assertion":[{"value":"2024-06-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-11-17","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-12-20","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70001"}}