e2abf1bf5c564496ddf171f54d4960cc pmc_34944191.pdf 8e49051b68d10202330adb790eac914fef6b7ebd pmc_34944191.pdf d98c62f5acd8e6946a96b849da149cf015c4d06665a1eb8432ca804d83fccc0c pmc_34944191.pdf Title: The Potential of Metalloproteinase-9 Administration to Accelerate Mammary Involution and Boost the Immune System at Dry-Off Subject: The dry period is decisive for the milking performance of dairy cows. The promptness of mammary gland involution at dry-off affects not only the productivity in the next lactation, but also the risk of new intra-mammary infections since it is closely related with the activity of the immune system. Matrix metalloproteinase-9 (MMP-9) is an enzyme present in the mammary gland and has an active role during involution by disrupting the extracellular matrix, mediating cell survival and the recruitment of immune cells. The objective of this study was to determine the potential of exogenous administration of a soluble and recombinant version of a truncated MMP-9 (rtMMP-9) to accelerate mammary involution and boost the immune system at dry-off, avoiding the use of antibiotics. Twelve Holstein cows were dried abruptly, and two quarters of each cow received an intra-mammary infusion of either soluble rtMMP-9 or a positive control based on immunostimulant inclusion bodies (IBs). The contralateral quarters were infused with saline solution as negative control. Samples of mammary secretion were collected during the week following dry-off to determine SCC, metalloproteinase activity, bovine serum albumin, lactoferrin, sodium, and potassium concentrations. The soluble form of rtMMP-9 increased endogenous metalloproteinase activity in the mammary gland compared with saline quarters but did not accelerate either the immune response or involution in comparison with control quarters. The results demonstrated that the strategy to increase the mammary gland immunocompetence by recombinant infusion of rtMMP-9 was unsuccessful. Keywords: dry period; mammary gland involution; MMP-9; immunity Author: Sílvia Parés, Olivia Cano-Garrido, Alex Bach, Neus Ferrer-Miralles, Antonio Villaverde, Elena Garcia-Fruitós and Anna Arís Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Dec 14 07:49:26 2021 CET ModDate: Tue Dec 14 07:55:37 2021 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1369271 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HKQHZQ+VnURWPalladioL Type 1 Custom yes yes no 10 0 NPJJQB+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 QQDLWK+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 KYBVYZ+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 SZMRXJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 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PANGPF+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 259 0 PANHDE+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 265 0 PANHDF+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 271 0 PANHDG+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 274 0 PANHDH+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 277 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-05-27 02:53:11 CEST RepresentationInformation: pmc_34944191.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:06:32 CEST Size: 1369271 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 437 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Bacterial Strains, Plasmids, Recombinant Proteins and Growth Conditions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Purification and Quantification of rtMMP-9 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Electron Microscopy (EM) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Analysis of Metalloproteinase Activity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Intra-Mammary Infusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Mammary Secretion Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: rMMP-9 Production, Activity Determination and Mammary Gland Infusion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Induced Metalloproteinase Activity in the Mammary Gland Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Mammary Immune Response and Involution Markers Monitorization after Intramammary Administration of rtMMP-9 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Info: Title: The Potential of Metalloproteinase-9 Administration to Accelerate Mammary Involution and Boost the Immune System at Dry-Off Author: Sílvia Parés, Olivia Cano-Garrido, Alex Bach, Neus Ferrer-Miralles, Antonio Villaverde, Elena Garcia-Fruitós and Anna Arís Subject: The dry period is decisive for the milking performance of dairy cows. The promptness of mammary gland involution at dry-off affects not only the productivity in the next lactation, but also the risk of new intra-mammary infections since it is closely related with the activity of the immune system. Matrix metalloproteinase-9 (MMP-9) is an enzyme present in the mammary gland and has an active role during involution by disrupting the extracellular matrix, mediating cell survival and the recruitment of immune cells. The objective of this study was to determine the potential of exogenous administration of a soluble and recombinant version of a truncated MMP-9 (rtMMP-9) to accelerate mammary involution and boost the immune system at dry-off, avoiding the use of antibiotics. Twelve Holstein cows were dried abruptly, and two quarters of each cow received an intra-mammary infusion of either soluble rtMMP-9 or a positive control based on immunostimulant inclusion bodies (IBs). The contralateral quarters were infused with saline solution as negative control. Samples of mammary secretion were collected during the week following dry-off to determine SCC, metalloproteinase activity, bovine serum albumin, lactoferrin, sodium, and potassium concentrations. The soluble form of rtMMP-9 increased endogenous metalloproteinase activity in the mammary gland compared with saline quarters but did not accelerate either the immune response or involution in comparison with control quarters. The results demonstrated that the strategy to increase the mammary gland immunocompetence by recombinant infusion of rtMMP-9 was unsuccessful. 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