{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,18]],"date-time":"2025-11-18T15:46:34Z","timestamp":1763480794250,"version":"build-2065373602"},"reference-count":64,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T00:00:00Z","timestamp":1722556800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"\u201cDeutsche Forschungsgemeinschaft\u201d","doi-asserted-by":"publisher","award":["TA291\/4-3","2021 (57559515)"],"award-info":[{"award-number":["TA291\/4-3","2021 (57559515)"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"DAAD\/BECAS Chile","award":["TA291\/4-3","2021 (57559515)"],"award-info":[{"award-number":["TA291\/4-3","2021 (57559515)"]}]},{"name":"Open-Access Publication Fund from JLU Giessen","award":["TA291\/4-3","2021 (57559515)"],"award-info":[{"award-number":["TA291\/4-3","2021 (57559515)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Besnoitia besnoiti is an obligate intracellular apicomplexan parasite and the causal agent of bovine besnoitiosis. Bovine besnoitiosis has a considerable economic impact in Africa and Asia due to reduced milk production, abortions, and bull infertility. In Europe, bovine besnoitiosis is classified as an emerging disease. Polymorphonuclear neutrophils (PMN) are one of the most abundant leukocytes in cattle blood and amongst the first immunological responders toward invading pathogens. In the case of B. besnoiti, bovine PMN produce reactive oxygen species (ROS), release neutrophil extracellular traps (NETs), and show increased autophagic activities upon exposure to tachyzoite stages. In that context, the general processes of NETosis and autophagy were previously reported as associated with AMP-activated protein kinase (AMPK) activation. Here, we study the role of AMPK in B. besnoiti tachyzoite-induced NET formation, thereby expanding the analysis to both upstream proteins, such as the calcium\/calmodulin-dependent protein kinase kinase 2 (CAMKK), and downstream signaling and effector molecules, such as the autophagy-related proteins ULK-1 and Beclin-1. Current data revealed early AMPK activation (&lt;30 min) in both B. besnoiti-exposed and AMPK activator (AICAR)-treated bovine PMN. This finding correlated with upstream responses on the level of CAMKK activation. Moreover, these reactions were accompanied by an augmented autophagic activity, as represented by enhanced expression of ULK-1 but not of Beclin-1. Referring to neutrophil effector functions, AICAR treatments induced both AMPK phosphorylation and NET formation, without affecting cell viability. In B. besnoiti tachyzoite-exposed PMN, AICAR treatments failed to affect oxidative responses, but led to enhanced NET formation, thereby indicating that AMPK and autophagic activation synergize with B. besnoiti-driven NETosis.<\/jats:p>","DOI":"10.3390\/ijms25158442","type":"journal-article","created":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T13:14:42Z","timestamp":1722604482000},"page":"8442","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The CAMKK\/AMPK Pathway Contributes to Besnoitia besnoiti-Induced NETosis in Bovine Polymorphonuclear Neutrophils"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9995-4757","authenticated-orcid":false,"given":"Iv\u00e1n","family":"Conejeros","sequence":"first","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1360-2209","authenticated-orcid":false,"given":"Zahady D.","family":"Vel\u00e1squez","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7268-3590","authenticated-orcid":false,"given":"Lisbeth","family":"Rojas-Bar\u00f3n","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8041-8097","authenticated-orcid":false,"given":"Gabriel","family":"Espinosa","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4550-8492","authenticated-orcid":false,"given":"Carlos","family":"Hermosilla","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8649-1020","authenticated-orcid":false,"given":"Anja","family":"Taubert","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Justus Liebig University of Giessen, 35392 Giessen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ramakrishnan, C., Krishnan, A., Francisco, S., Schmid, M.W., Russo, G., Leit\u00e3o, A., Hemphill, A., Soldati-Favre, D., and Hehl, A.B. (2022). Dissection of Besnoitia besnoiti Intermediate Host Life Cycle Stages: From Morphology to Gene Expression. PLoS Pathog., 18.","DOI":"10.1371\/journal.ppat.1010955"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.1017\/S0031182014000262","article-title":"A Review on Bovine Besnoitiosis: A Disease with Economic Impact in Herd Health Management, Caused by Besnoitia besnoiti (Franco and Borges)","volume":"141","author":"Cortes","year":"2014","journal-title":"Parasitology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/j.pt.2013.06.002","article-title":"A Century of Bovine Besnoitiosis: An Unknown Disease Re-Emerging in Europe","volume":"29","author":"Frey","year":"2013","journal-title":"Trends Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.vetpar.2010.08.013","article-title":"Bovine Besnoitiosis: Epidemiological and Clinical Aspects","volume":"174","author":"Jacquiet","year":"2010","journal-title":"Vet. Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.vetpar.2011.01.027","article-title":"Exploring the Life Cycle of Besnoitia besnoiti\u2014Experimental Infection of Putative Definitive and Intermediate Host Species","volume":"178","author":"Basso","year":"2011","journal-title":"Vet. Parasitol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Langenmayer, M.C., Scharr, J.C., Sauter-Louis, C., Schares, G., and Gollnick, N.S. (2015). Natural Besnoitia besnoiti Infections in Cattle: Hematological Alterations and Changes in Serum Chemistry and Enzyme Activities. BMC Vet. Res., 11.","DOI":"10.1186\/s12917-015-0326-8"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1377","DOI":"10.1016\/j.immuni.2021.06.006","article-title":"The Neutrophil","volume":"54","author":"Burn","year":"2021","journal-title":"Immunity"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1038\/nri1785","article-title":"Neutrophils and Immunity: Challenges and Opportunities","volume":"6","author":"Nathan","year":"2006","journal-title":"Nat. Rev. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1126\/science.1092385","article-title":"Neutrophil Extracellular Traps Kill Bacteria","volume":"303","author":"Brinkmann","year":"2004","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1083\/jcb.201203170","article-title":"Neutrophil Extracellular Traps: Is Immunity the Second Function of Chromatin?","volume":"198","author":"Brinkmann","year":"2012","journal-title":"J. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1083\/jcb.200606027","article-title":"Novel Cell Death Program Leads to Neutrophil Extracellular Traps","volume":"176","author":"Fuchs","year":"2007","journal-title":"J. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7326","DOI":"10.1073\/pnas.1909546117","article-title":"NETosis Proceeds by Cytoskeleton and Endomembrane Disassembly and PAD4-Mediated Chromatin Decondensation and Nuclear Envelope Rupture","volume":"117","author":"Thiam","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3767","DOI":"10.1038\/s41467-018-06263-5","article-title":"Chromatin Swelling Drives Neutrophil Extracellular Trap Release","volume":"9","author":"Neubert","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Conejeros, I., L\u00f3pez-Osorio, S., Zhou, E., Vel\u00e1squez, Z.D., Del R\u00edo, M.C., Burgos, R.A., Alarc\u00f3n, P., Chaparro-Guti\u00e9rrez, J.J., Hermosilla, C., and Taubert, A. (2022). Glycolysis, Monocarboxylate Transport, and Purinergic Signaling Are Key Events in Eimeria bovis-Induced NETosis. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.842482"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2797","DOI":"10.1007\/s00436-014-3939-0","article-title":"The Apicomplexan Parasite Eimeria arloingi Induces Caprine Neutrophil Extracellular Traps","volume":"113","author":"Silva","year":"2014","journal-title":"Parasitol. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1128\/IAI.05730-11","article-title":"Toxoplasma gondii Triggers Release of Human and Mouse Neutrophil Extracellular Traps","volume":"80","author":"Lin","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Vel\u00e1squez, Z.D., Peixoto, R., G\u00e4rtner, U., Hermosilla, C., Taubert, A., and Conejeros, I. (2023). Dynamics of Cell Cycle Proteins Involved in Toxoplasma gondii-Induced Bovine NET Formation. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1125667"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.dci.2015.05.007","article-title":"NADPH Oxidase, MPO, NE, ERK1\/2, P38 MAPK and Ca2+ Influx Are Essential for Cryptosporidium parvum-Induced NET Formation","volume":"52","author":"Lendner","year":"2015","journal-title":"Dev. Comp. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.ijppaw.2017.09.002","article-title":"Bottlenose Dolphins (Tursiops Truncatus) Do Also Cast Neutrophil Extracellular Traps against the Apicomplexan Parasite Neospora caninum","volume":"6","author":"Silva","year":"2017","journal-title":"Int. J. Parasitol. Parasites Wildl."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Grob, D., Conejeros, I., Vel\u00e1squez, Z.D., Preu\u00dfer, C., G\u00e4rtner, U., Alarc\u00f3n, P., Burgos, R.A., Hermosilla, C., and Taubert, A. (2020). Trypanosoma brucei brucei Induces Polymorphonuclear Neutrophil Activation and Neutrophil Extracellular Traps Release. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.559561"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Conejeros, I., Vel\u00e1squez, Z.D., Grob, D., Zhou, E., Salecker, H., Hermosilla, C., and Taubert, A. (2019). Histone H2A and Bovine Neutrophil Extracellular Traps Induce Damage of Besnoitia besnoiti-Infected Host Endothelial Cells but Fail to Affect Total Parasite Proliferation. Biology, 8.","DOI":"10.3390\/biology8040078"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Mu\u00f1oz Caro, T., Hermosilla, C., Silva, L.M.R., Cortes, H., and Taubert, A. (2014). Neutrophil Extracellular Traps as Innate Immune Reaction against the Emerging Apicomplexan Parasite Besnoitia besnoiti. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0091415"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhou, E., Conejeros, I., Vel\u00e1squez, Z.D., Mu\u00f1oz-Caro, T., G\u00e4rtner, U., Hermosilla, C., and Taubert, A. (2019). Simultaneous and Positively Correlated NET Formation and Autophagy in Besnoitia besnoiti Tachyzoite-Exposed Bovine Polymorphonuclear Neutrophils. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.01131"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Espinosa, G., Conejeros, I., Rojas-Bar\u00f3n, L., Hermosilla, C.R., and Taubert, A. (2023). Besnoitia besnoiti-Induced Neutrophil Clustering and Neutrophil Extracellular Trap Formation Depend on P2X1 Purinergic Receptor Signaling. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1244068"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1017\/S0031182019001707","article-title":"Besnoitia besnoiti Bradyzoite Stages Induce Suicidal- and Rapid Vital-NETosis","volume":"147","author":"Zhou","year":"2020","journal-title":"Parasitology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1007\/s00436-019-06543-z","article-title":"Metabolic Requirements of Besnoitia besnoiti Tachyzoite-Triggered NETosis","volume":"119","author":"Zhou","year":"2020","journal-title":"Parasitol. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/cr.2010.150","article-title":"Neutrophil Extracellular Trap Cell Death Requires Both Autophagy and Superoxide Generation","volume":"21","author":"Remijsen","year":"2011","journal-title":"Cell Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1164\/rccm.201603-0596OC","article-title":"Autophagy Primes Neutrophils for Neutrophil Extracellular Trap Formation during Sepsis","volume":"196","author":"Park","year":"2017","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Skendros, P., Mitroulis, I., and Ritis, K. (2018). Autophagy in Neutrophils: From Granulopoiesis to Neutrophil Extracellular Traps. Front. Cell Dev. Biol., 6.","DOI":"10.3389\/fcell.2018.00109"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.cell.2012.03.017","article-title":"mTOR Signaling in Growth Control and Disease","volume":"149","author":"Laplante","year":"2012","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e224","DOI":"10.1038\/emm.2016.16","article-title":"AMPK Activators: Mechanisms of Action and Physiological Activities","volume":"48","author":"Kim","year":"2016","journal-title":"Exp. Mol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"L497","DOI":"10.1152\/ajplung.90210.2008","article-title":"Activation of AMPK Attenuates Neutrophil Proinflammatory Activity and Decreases the Severity of Acute Lung Injury","volume":"295","author":"Zhao","year":"2008","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5179","DOI":"10.1210\/en.2003-0982","article-title":"Minireview: The AMP-Activated Protein Kinase Cascade: The Key Sensor of Cellular Energy Status","volume":"144","author":"Hardie","year":"2003","journal-title":"Endocrinology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"387","DOI":"10.2119\/molmed.2013.00065","article-title":"Activation of AMPK Enhances Neutrophil Chemotaxis and Bacterial Killing","volume":"19","author":"Park","year":"2013","journal-title":"Mol. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4358","DOI":"10.1096\/fj.11-190587","article-title":"AMP-Activated Protein Kinase Enhances the Phagocytic Ability of Macrophages and Neutrophils","volume":"25","author":"Bae","year":"2011","journal-title":"FASEB J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.febslet.2004.07.077","article-title":"Stimulators of AMP-Activated Protein Kinase Inhibit the Respiratory Burst in Human Neutrophils","volume":"573","author":"Alba","year":"2004","journal-title":"FEBS Lett."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Brinkmann, V., Goosmann, C., K\u00fchn, L.I., and Zychlinsky, A. (2013). Automatic Quantification of in Vitro NET Formation. Front. Immunol., 3.","DOI":"10.3389\/fimmu.2012.00413"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Neumann, A., Brogden, G., and von K\u00f6ckritz-Blickwede, M. (2020). Extracellular Traps: An Ancient Weapon of Multiple Kingdoms. biology, 9.","DOI":"10.3390\/biology9020034"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Worku, M., Rehrah, D., Ismail, H.D., Asiamah, E., and Adjei-Fremah, S. (2021). A Review of the Neutrophil Extracellular Traps (NETs) from Cow, Sheep and Goat Models. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22158046"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"de Buhr, N., Bonilla, M.C., Jimenez-Soto, M., von K\u00f6ckritz-Blickwede, M., and Dolz, G. (2018). Extracellular Trap Formation in Response to Trypanosoma cruzi Infection in Granulocytes Isolated From Dogs and Common Opossums, Natural Reservoir Hosts. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00966"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Sousa-Rocha, D., Thomaz-Tobias, M., Diniz, L.F.A., Souza, P.S.S., Pinge-Filho, P., and Toledo, K.A. (2015). Trypanosoma cruzi and Its Soluble Antigens Induce NET Release by Stimulating Toll-Like Receptors. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0139569"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1111\/pim.12332","article-title":"Entamoeba histolytica Induces Human Neutrophils to Form NETs","volume":"38","year":"2016","journal-title":"Parasite Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"\u00c1vila, E.E., Salaiza, N., Pulido, J., Rodr\u00edguez, M.C., D\u00edaz-God\u00ednez, C., Laclette, J.P., Becker, I., and Carrero, J.C. (2016). Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0158979"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Abdallah, D.S.A., and Denkers, E.Y. (2012). Neutrophils Cast Extracellular Traps in Response to Protozoan Parasites. Front. Immunol., 3.","DOI":"10.3389\/fimmu.2012.00382"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4189","DOI":"10.1007\/s00436-014-4094-3","article-title":"Besnoitia besnoiti Tachyzoites Induce Monocyte Extracellular Trap Formation","volume":"113","author":"Silva","year":"2014","journal-title":"Parasitol. Res."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Ong, C.W.M., Elkington, P.T., Brilha, S., Ugarte-Gil, C., Tome-Esteban, M.T., Tezera, L.B., Pabisiak, P.J., Moores, R.C., Sathyamoorthy, T., and Patel, V. (2015). Neutrophil-Derived MMP-8 Drives AMPK-Dependent Matrix Destruction in Human Pulmonary Tuberculosis. PLOS Pathog., 11.","DOI":"10.1371\/journal.ppat.1004917"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Carretta, M.D., Barr\u00eda, Y., Borquez, K., Urra, B., Rivera, A., Alarc\u00f3n, P., Hidalgo, M.A., and Burgos, R.A. (2020). \u03b2-Hydroxybutyrate and Hydroxycarboxylic Acid Receptor 2 Agonists Activate the AKT, ERK and AMPK Pathways, Which Are Involved in Bovine Neutrophil Chemotaxis. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-69500-2"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Wang, X., Zhang, Y., Li, Y., Tang, M., Deng, Q., Mao, J., and Du, L. (2021). Estrogen Regulates Glucose Metabolism in Cattle Neutrophils Through Autophagy. Front. Vet. Sci., 8.","DOI":"10.3389\/fvets.2021.773514"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1038\/ncb2152","article-title":"AMPK and mTOR Regulate Autophagy through Direct Phosphorylation of Ulk1","volume":"13","author":"Kim","year":"2011","journal-title":"Nat. Cell Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1161\/CIRCRESAHA.116.309937","article-title":"AMP-Activated Protein Kinase A2 in Neutrophils Regulates Vascular Repair via Hypoxia-Inducible Factor-1\u03b1 and a Network of Proteins Affecting Metabolism and Apoptosis","volume":"120","author":"Zippel","year":"2017","journal-title":"Circ. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6693921","DOI":"10.1155\/2021\/6693921","article-title":"Effects of Fluorine on Neutrophil Extracellular Trap Formation through Regulating AMPK\/P38 Signaling Pathway","volume":"2021","author":"Song","year":"2021","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"4625","DOI":"10.1046\/j.1432-1033.2002.03162.x","article-title":"Role of Ca2+\/Calmodulin Regulated Signaling Pathways in Chemoattractant Induced Neutrophil Effector Functions","volume":"269","author":"Verploegen","year":"2002","journal-title":"Eur. J. Biochem."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Zhang, B., Ma, X., Huang, B., Jiang, Q., Loor, J.J., Lv, X., Zhang, W., Li, M., Wen, J., and Yin, Y. (2022). Transcriptomics of Circulating Neutrophils in Dairy Cows with Subclinical Hypocalcemia. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.959831"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.cell.2007.12.018","article-title":"Autophagy in the Pathogenesis of Disease","volume":"132","author":"Levine","year":"2008","journal-title":"Cell"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1038\/nature09782","article-title":"Autophagy in Immunity and Inflammation","volume":"469","author":"Levine","year":"2011","journal-title":"Nature"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1038\/icb.2012.82","article-title":"Autophagy and Inflammatory Diseases","volume":"91","author":"Jones","year":"2013","journal-title":"Immunol. Cell Biol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1126\/science.1196371","article-title":"Phosphorylation of ULK1 (hATG1) by AMP-Activated Protein Kinase Connects Energy Sensing to Mitophagy","volume":"331","author":"Egan","year":"2011","journal-title":"Science"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.semcdb.2019.04.016","article-title":"LC3-Associated Phagocytosis\u2014The Highway to Hell for Phagocytosed Microbes","volume":"101","author":"Herb","year":"2020","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1002\/eji.200940025","article-title":"Regulation of the Autophagic Machinery in Human Neutrophils","volume":"40","author":"Mitroulis","year":"2010","journal-title":"Eur. J. Immunol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1007\/s10495-013-0893-8","article-title":"Adiponectin Inhibits Neutrophil Apoptosis via Activation of AMP Kinase, PKB and ERK 1\/2 MAP Kinase","volume":"18","author":"Rossi","year":"2013","journal-title":"Apoptosis"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/0165-2427(91)90115-S","article-title":"Comparison of the Response of Bovine and Human Neutrophils to Various Stimuli","volume":"28","author":"Brown","year":"1991","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1007\/s00441-018-2789-y","article-title":"Bovine Neutrophils in Health and Disease","volume":"371","author":"Bassel","year":"2018","journal-title":"Cell Tissue Res."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An Open-Source Platform for Biological-Image Analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1109\/TSMC.1979.4310076","article-title":"A Threshold Selection Method from Gray-Level Histograms","volume":"9","author":"Otsu","year":"1979","journal-title":"IEEE Trans. Syst. Man Cybern."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8442\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:28:44Z","timestamp":1760110124000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8442"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,2]]},"references-count":64,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["ijms25158442"],"URL":"https:\/\/doi.org\/10.3390\/ijms25158442","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2024,8,2]]}}}