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University of Groningen Exploring the mechanisms underlying the phenotype of MCAD deficiency with Systems Medicine Martines, Anne-Claire

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(1)University of Groningen. Exploring the mechanisms underlying the phenotype of MCAD deficiency with Systems Medicine Martines, Anne-Claire. IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below.. Document Version Publisher's PDF, also known as Version of record. Publication date: 2019 Link to publication in University of Groningen/UMCG research database. Citation for published version (APA): Martines, A-C. (2019). Exploring the mechanisms underlying the phenotype of MCAD deficiency with Systems Medicine: from computational model to mice to man. Rijksuniversiteit Groningen.. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum.. Download date: 28-06-2021.

(2) Chapter 4 The role of hepatic fatty-acid oxidation during cold stress: remodeling of hepatic metabolism in a mouse with medium-chain acyl-CoA dehydrogenase deficiency A.M.F. Martines1,#, Wenxuan Zhang1,2,#, Albert Gerding1,3, Maaike Goris4, Marcel de Vries1, Mirjam H. Koster1,4, Laura Bongiovanni5, Alain de Bruin1,5, Rob H. Henning4, Terry G. J. Derks1,6, Rainer Bischoff2, Dirk-Jan Reijngoud1,2, Barbara M. Bakker1,* #. Authors contributed equally to this work.. 1 Laboratory. of Pediatrics, Center of Liver, Digestive and Metabolic Diseases, University of Groningen, Groningen, University Medical Center Groningen, Groningen, The Netherlands, 2 Department of Analytical Biochemistry, University of Groningen, Groningen, The Netherlands 3 Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, The Netherlands 4 Department of Anesthesiology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands 5 Department of Pathobiology, Faculty of Veterinary Medicine, Dutch Molecular Pathology Center, Utrecht University, The Netherlands 6 Section of Metabolic Diseases, Beatrix Children's Hospital, University Medical Center of Groningen, University of Groningen.. Journal publication in preparation.

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(4) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice. Abstract Š‹Ž†”‡ ™‹–Š ƒ †‡ˆ‹ ‹‡ › ‹ –Š‡ ‹–‘ Š‘†”‹ƒŽ ˆƒ––›Ǧƒ ‹† ȾǦ‘š‹†ƒ–‹‘ ȋ Ȍ ‡œ›‡ ‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ‘‡œ›‡  †‡Š›†”‘‰‡ƒ•‡ ȋȌ ƒ”‡ ƒ– ”‹• ‘ˆ Ž‹ˆ‡Ǧ–Š”‡ƒ–‡‹‰ Ž‘™ „Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž•Ǥ‘‡Ž— ‹†ƒ–‡–Š‡”‘Ž‡‘ˆŽ‹˜‡”‹–Š‡”‡•’‘•‡–‘‡‡”‰‡–‹ •–”‡••ǡ ˆƒ•–‡†Ǧ†‡ˆ‹ ‹‡–ȋǦȌƒ†™‹Ž†Ǧ–›’‡ȋȌ‹ ‡™‡”‡‡š’‘•‡†–‘ ‘Ž†ǤǦ ‹ ‡ •Š‘™‡† Ž‘™‡” „Ž‘‘† ‰Ž— ‘•‡ Ž‡˜‡Ž• ƒ† Š‡’ƒ–‹  ƒ‹‘ ƒ ‹† Ž‡˜‡Ž•ǡ ƒ† Š‹‰Š‡” Ž‹˜‡” ™‡‹‰Š–•ǡŽ‹˜‡”–”‹‰Ž› ‡”‹†‡•ƒ†„Ž‘‘†‡†‹—Ǧ Šƒ‹ƒ ›ŽǦ ƒ”‹–‹‡Ž‡˜‡Ž•Ǥ‡–ƒ‹Ž‡†Ž‹’‹†‘‹ • ƒƒŽ›•‹•”‡˜‡ƒŽ‡†ƒƒ ——Žƒ–‹‘‘ˆ–”‹‰Ž› ‡”‹†‡•’‡ ‹‡• ‘–ƒ‹‹‰‡†‹—Ǧ Šƒ‹ˆƒ––›Ǧƒ ›Ž „”ƒ Š‡•‹–Š‡Ž‹˜‡”Ǥ —””‡•—Ž–•ƒ”‡ ‘•‹•–‡–™‹–Šƒ‡›”‘Ž‡‘ˆŽ‹˜‡” ‹‰Ž— ‘•‡Š‘‡‘•–ƒ•‹•†—”‹‰ ‘Ž† •–”‡••Ǥ  Ǧ ‹ ‡ ‡šŠ‹„‹– ƒ ‘’‡•ƒ–‘”› ”‡•’‘•‡ ‹ ™Š‹ Š ƒ‹‘ ƒ ‹†• ’ƒ”–Ž› –ƒ‡ ‘˜‡”–Š‡”‘Ž‡‘ˆˆƒ––›ƒ ‹†•‹–Š‡‰‡‡”ƒ–‹‘‘ˆǡ‰Ž— ‘•‡ǡƒ†‡–‘‡„‘†‹‡•ǡ™Š‹Ž‡‡š ‡•• ‡†‹—Ǧ Šƒ‹ˆƒ––›Ǧƒ ‹†‹–‡”‡†‹ƒ–‡•ƒ”‡‡š’‘”–‡†ƒ†”‡”‘—–‡†‹–‘–”‹‰Ž› ‡”‹†‡•Ǥ  . ͳʹͻ. 4.

(5) Chapter 4. Introduction ‡’ƒ–‹  ‹–‘ Š‘†”‹ƒŽ ˆƒ––›Ǧƒ ‹† „‡–ƒǦ‘š‹†ƒ–‹‘ ȋ Ȍ ’Žƒ›• ƒ ‹’‘”–ƒ– ”‘Ž‡ ‹ ‡‡”‰› Š‘‡‘•–ƒ•‹• †—”‹‰ ˆƒ•–‹‰ ƒ† ‘†‹–‹‘• ‘ˆ Š‹‰Š ‡‡”‰› †‡ƒ†Ǥ Š‡ ’ƒ–Š™ƒ› ’”‘˜‹†‡•‡‡”‰›ˆ‘”de novo‰Ž— ‘•‡’”‘†— –‹‘ƒ†•—„•–”ƒ–‡ˆ‘”‡–‘‡„‘†›’”‘†— –‹‘ȏͳȂ ͵ȐǤ‘•‡“—‡–Ž›ǡ‹†‹˜‹†—ƒŽ•™‹–Šƒ†‡ˆ‡ –‹ ‘ˆ–‡’”‡•‡–™‹–ŠŽ‘™„Ž‘‘†‰Ž— ‘•‡ƒ† ‡–‘‡ „‘†› ‘ ‡–”ƒ–‹‘• ȋŠ›’‘‡–‘–‹  Š›’‘‰Ž› ‡‹ƒȌ ȏͳȂ͵ȐǤ Š‡ ‘•– ’”‡˜ƒŽ‡–   †‹•‘”†‡” ‹• ƒ ‘’Ž‡–‡ †‡ˆ‹ ‹‡ › ‘ˆ –Š‡ ‡œ›‡ ‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ‘ †‡Š›†”‘‰‡ƒ•‡ ȋȌǤ Š‹Ž‡ Ǧ†‡ˆ‹ ‹‡– ȋȌ Š‹Ž†”‡ ƒ› –‘Ž‡”ƒ–‡ ‘˜‡”‹‰Š– ˆƒ•–‹‰ ȏͶǡͷȐǡ ƒ ‘„‹ƒ–‹‘‘ˆ’”‘Ž‘‰‡†ˆƒ•–‹‰ƒ†ƒƒ††‹–‹‘ƒŽ–”‹‰‰‡”ǡ•— Šƒ•ȋ ‘„‹ƒ–‹‘•‘ˆȌˆ‡˜‡”ǡ †‡ ”‡ƒ•‡† ‰ƒ•–”‘‹–‡•–‹ƒŽ ‹–ƒ‡ ‘” ‹ ”‡ƒ•‡† Ž‘••‡•ǡ ‹ ”‡ƒ•‡• –Š‡ ”‹• ‘ˆ Š›’‘‡–‘–‹  Š›’‘‰Ž› ‡‹ƒƒ†•—††‡Ǧ‹ˆƒ–†‡ƒ–Šȏͳǡ͸ȂͻȐǤ ‘‹˜‡•–‹‰ƒ–‡–Š‡‡–‹‘Ž‘‰›‘ˆǡƒǦ†‡ˆ‹ ‹‡–ȋǦȌ‘—•‡‘†‡Ž Šƒ•„‡‡‰‡‡”ƒ–‡†ȏͳͲǡͳͳȐǤŠ‡Ǧ‹ ‡‡šŠ‹„‹–‡†–Š‡‡Ž‡˜ƒ–‡†„Ž‘‘†‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ ƒ”‹–‹‡• –Šƒ– ƒ”‡ ƒŽ•‘ Šƒ”ƒ –‡”‹•–‹  ‘ˆ Š—ƒ ǡ ƒ† ƒ ‡Šƒ ‡† ”ƒ–‡ ‘ˆ ‡‘ƒ–ƒŽ†‡ƒ–ŠȏͳͲȐǤ —•–ƒ• Š‹Ž†”‡ǡǦ‹ ‡™‡”‡“—‹–‡”‡•‹Ž‹‡––‘ˆƒ•–‹‰ ƒŽ‘‡ƒ†ƒ‹–ƒ‹‘”ƒŽ„Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž•†—”‹‰ʹͶŠ‘—”•‘ˆˆƒ•–‹‰ȏͳͲǡͳʹȐǤ ‘™‡˜‡”ǡ ƒ ‘„‹ƒ–‹‘ ‘ˆ ˆƒ•–‹‰ ƒ† ‘Ž† ‡š’‘•—”‡ ȋͶι Ȍ ™ƒ• Ž‡–ŠƒŽ ‘” Ž‡† –‘ ˜‡”› Ž‘™ „‘†› –‡’‡”ƒ–—”‡•‹–Š‡ǡ„—–‘–‹–Š‡™‹Ž†–›’‡ȋȌ‹ ‡ȏͳͲȐǤ‘Ž†‹–‘Ž‡”ƒ ‡‹•ƒ ‘‘ ˆ‡ƒ–—”‡ ‘ˆ   †‡ˆ‹ ‹‡– ‘—•‡ ‘†‡Ž• ƒ† ™ƒ• ƒŽ•‘ ‘„•‡”˜‡† ™Š‡ ‡‹–Š‡” ‘ˆ –Š‡ ‰‡‡• ‡ ‘†‹‰•Š‘”–ǦǡŽ‘‰Ǧǡ‘”˜‡”›ǦŽ‘‰Ǧ Šƒ‹ƒ ›ŽǦ‘†‡Š›†”‘‰‡ƒ•‡ȋǡƒ†Ȍ ™‡”‡†‡Ž‡–‡†ȋ”‡˜‹‡™‡†‹ȏͳ͵ȐȌǤ Š‡ ‹ ‡ ƒ”‡ ‡š’‘•‡† –‘ ‘Ž† •–”‡••ǡ „”‘™ ƒ†‹’‘•‡ –‹••—‡ ȋȌ —•‡• ‰Ž— ‘•‡ǡ ƒ‹‘Ǧƒ ‹†ǡƒ†ˆƒ––›Ǧƒ ‹† ƒ–ƒ„‘Ž‹•ˆ‘”‘Ǧ•Š‹˜‡”‹‰–Š‡”‘‰‡‡•‹•ȏͳͶȂͳ͸ȐǤ‘Ž†•–”‡•• ‹ ”‡ƒ•‡•–Š‡‡š’”‡••‹‘‘ˆ‰‡‡•‹˜‘Ž˜‡†‹ˆƒ––›Ǧƒ ‹†—’–ƒ‡ƒ† ‹ȏͳ͹ȐǤ

(6) –Š‹• ‘†‹–‹‘ǡ–Š‡Ž‹˜‡”’Žƒ›•ƒ‡›”‘Ž‡‹–Š‡•—’’Ž›‘ˆ•—„•–”ƒ–‡„›‹ ”‡ƒ•‡†‰Ž— ‘‡‘‰‡‡•‹• ȏͳͺȐƒ†ƒ ›ŽǦ ƒ”‹–‹‡’”‘†— –‹‘ƒ†•‡ ”‡–‹‘ȏͳͻǡʹͲȐǤ

(7) Ž‹‡™‹–Š–Š‹•ǡ–Š‡Ž‹˜‡”‡šŠ‹„‹–•ƒ Š‹‰Š‡”‡š’”‡••‹‘‘ˆ‰‡‡•‹˜‘Ž˜‡†‹ ƒ†ƒŽ‘™‡”‡š’”‡••‹‘‘ˆ‰‡‡•‹ˆƒ––›Ǧƒ ‹† •›–Š‡•‹•ȏʹͳȂʹ͵ȐǤŠ‡ ‘”ˆƒ––›Ǧƒ ‹†—’–ƒ‡‹•†‡ˆ‡ –‹˜‡ǡ–Š‡”‡Ž‹‡•‘”‡Š‡ƒ˜‹Ž› ‘‰Ž— ‘•‡ƒ†ƒ‹‘ƒ ‹†•ǤŠ‹•Ž‡ƒ†•–‘ƒŠ‹‰Š‡””‡“—‹”‡‡–ˆ‘”de novo‰Ž— ‘•‡’”‘†— –‹‘ „›–Š‡Ž‹˜‡”Ǥ ‘”†‹‰Ž›ǡˆƒ•–‹‰‹ ‘„‹ƒ–‹‘™‹–Š ‘Ž†‡š’‘•—”‡”‡•—Ž–‡†‹Ž‘™‡”„Ž‘‘† ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘•‹Ǧ ‘’ƒ”‡†–‘‹ ‡ȏʹͶȂʹ͸ȐǤ‘†ƒ–‡Š‘™‡˜‡”ǡ–Š‡”‡ ƒ”‡‘”‡’‘”–•‘–Š‡Ž‹˜‡”ˆ— –‹‘‘ˆǦ‘—•‡†—”‹‰ ‘Ž†•–”‡••ǡ‘”‘ˆƒ›‘–Š‡” ’Š›•‹‘Ž‘‰‹ ƒŽ’ƒ”ƒ‡–‡”•„—–”‡ –ƒŽ–‡’‡”ƒ–—”‡ȏͳͲǡʹ͹ȐǤŠ‡”‡ˆ‘”‡ǡ–Š‡ƒ‹‘ˆ–Š‹••–—†›™ƒ• –‘‹˜‡•–‹‰ƒ–‡Ž‹˜‡”ˆ— –‹‘‹Ǧ‹ ‡†—”‹‰ ‘„‹‡† ‘Ž†‡š’‘•—”‡ƒ†ˆƒ•–‹‰Ǥ

(8)  –Š‹• •–—†›ǡ Ǧ ‹ ‡ ™‡”‡ ˆƒ•–‡† ˆ‘” ͳͶ Š ƒ– ʹͳ ι ƒ† •—„•‡“—‡–Ž› ˆ‘” ƒ‘–Š‡”ͶǦ͸Šƒ–ͶιǤŠ‡’Š‡‘–›’‡‘ˆ›‘—‰‹ ‡ȋͺ™‡‡•Ȍ™ƒ•‘”‡ƒˆˆ‡ –‡†–Šƒ–Šƒ–‘ˆ ƒ†—Ž– ‹ ‡ ȋͳ͸ǦʹͲ ™‡‡•ȌǤ – –Š‡ ‡† ‘ˆ –Š‡ ŠƒŽŽ‡‰‡ǡ –Š‡ „Ž‘‘† ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘ ‘ˆ ›‘—‰Ǧ‹ ‡™ƒ•Ž‘™‡”–Šƒ–Šƒ–‘ˆ–Š‡‹” ‘—–‡”’ƒ”–•ǡ™Š‹Ž‡‡–‘‡Ǧ„‘†›Ž‡˜‡Ž• ™‡”‡ •‹‹Žƒ”Ǥ ‹˜‡” ƒ‹‘Ǧƒ ‹† Ž‡˜‡Ž• ™‡”‡ Ž‘™‡” ‹ Ǧ ‹ ‡ ‘’ƒ”‡† –‘ ǡ ‘•‹•–‡– ™‹–Š ƒ Š‹‰Š‡” •—„•–”ƒ–‡ —•‡ ˆ‘” ‰Ž— ‘‡‘‰‡‡•‹•Ǥ  ‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ ƒ”‹–‹‡ Ž‡˜‡Ž•ȋ‘–ƒ„Ž›ͳͲǣͳȌ™‡”‡‡Ž‡˜ƒ–‡†‹–Š‡„Ž‘‘†‘ˆ–Š‡‹ ‡ǡ„—–—ƒˆˆ‡ –‡†‹–Š‡Ž‹˜‡”Ǥ

(9) Ǧ†‡’–Š Š‡’ƒ–‹  Ž‹’‹†‘‹  ƒƒŽ›•‹• ”‡˜‡ƒŽ‡† ƒ ——Žƒ–‹‘ ‘ˆ –”‹‰Ž› ‡”‹†‡ ȋ Ȍ •’‡ ‹‡• ‘–ƒ‹‹‰ ͸ǦͳͶ ƒ ›Ž Šƒ‹•ǡ •—‰‰‡•–‹‰ –Šƒ– ‡†‹—Ǧ Šƒ‹   ‹–‡”‡†‹ƒ–‡• ™‡”‡ ”‡”‘—–‡†‹–‘ȋ’ƒ”–‹ƒŽȌ Šƒ‹‡Ž‘‰ƒ–‹‘ƒ†Ȁ‘”•–‘”ƒ‰‡ǤŠ‹•‹•–Š‡ˆ‹”•––‹‡–Šƒ––Š‡Ž‹˜‡” ‡–ƒ„‘Ž‹•‘ˆ–Š‡Ǧ‘—•‡™ƒ••–—†‹‡†—†‡”•‡˜‡”‡‡‡”‰‡–‹ •–”‡••Ǥ‡ ‘ Ž—†‡ ͳ͵Ͳ.

(10) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice –Šƒ–‹–‹•ƒ˜ƒŽ—ƒ„Ž‡‘†‡Ž–‘‰ƒ‹‹•‹‰Š–‹–‘‡–ƒ„‘Ž‹ ƒ†ƒ’–ƒ–‹‘•‹ƒ†’”‘˜‹†‡ ˆ—”–Š‡”‡˜‹†‡ ‡ˆ‘”–Š‡ ”— ‹ƒŽ”‘Ž‡‘ˆŽ‹˜‡” ‹’”‘–‡ –‹‘ƒ‰ƒ‹•– ‘Ž†•–”‡••Ǥ . Results Young MCAD-KO mice are more vulnerable to cold exposure than adult MCAD-KO mice. ‘‡š’‘•‡ƒ†—Ž–ȋͳ͸ǦʹͲ™‡‡•‘Ž†ȌǦ‹ ‡–‘•‡˜‡”‡‡‡”‰‡–‹ •–”‡••ǡ–Š‡›™‡”‡ˆƒ•–‡† ˆ‘”ͳͶŠ‘—”•ƒ†•—„•‡“—‡–Ž›‡š’‘•‡†–‘ƒͶ釐˜‹”‘‡–ǡ•–‹ŽŽ‹–Š‡ƒ„•‡ ‡‘ˆˆ‘‘†Ǥ‘Ž† ‡š’‘•—”‡ˆ‘”ͶǦ͸Š‘—”•‹†— ‡†ƒ•—„•–ƒ–‹ƒŽ†”‘’‹–Š‡„‘†›–‡’‡”ƒ–—”‡‘ˆͶ‘—–‘ˆͺˆƒ•–‡† ‹ ‡ǡ™Š‹Ž‡ƒŽŽ͹†‡–‡ –‡†‹ ‡ƒ‹–ƒ‹‡†ƒ•–ƒ„Ž‡–‡’‡”ƒ–—”‡ȋ ‹‰—”‡ͳȌǤ B. WT KO. 0 BF. AF. 2. 0. AF. *. 1.2 0.8 0.4 0.0. BF. AF. AFC. WT. 30. 20 20. WT KO. 0 BF. AF. 16-20 w .o. 5. AFC. BF. AF. AFC. *. 5. 16-20 w .o. 4. 3 3 0 WT KO. J. I 40. 30. Body weight (g). 40. 6. KO. H Rectal temperature ( qC). G. 10. F 1.6. 4. WT KO. 0. E Liver weight (g). Ketone bodies (mM). 25. BF. 6. Body temperature ( qC). 30. 20. AFC. D. 35. 30. 20 20 0. AFC. Rectal temperature ( qC). 20 20. 15. Blood glucose (mM). 30. C 40. i. Relative liver weight (%). i. 40. Body weight (g). Body temperature (qC). A. WT KO. 25. 20. 15 WT KO. BF. AF. 40. WT KO. 35 30. r=0.84 p<0.0001. 25. 8 w .o.. 20 20 0. AFC. 0. 10. 20. Body weight AF (g). L. i. *. 5. 0. 3 2 1. AF. AFC. *. 1.2 0.8 0.4 0.0. 0 BF. N 1.6. Liver weight (g). 10. Ketone bodies (mM). Blood glucose (mM). M 4. 15. BF. AF. AFC. WT. KO. Relative liver weight (%). K. 6. **. 5. 8 w .o.. 4. 3 3 0 WT. KO. Figure 1. Biometric results of 8- and 16-20-weeks-old mice. ‘†› –‡’‡”ƒ–—”‡ ‡ƒ•—”‡† „› ‹–”ƒ’‡”‹–‘‡ƒŽŽ›Ǧ‹’Žƒ–‡†Ž‘‰‰‡”ȋA,GȌǡ„‘†›™‡‹‰Š–ȋB,IȌǡ„Ž‘‘†‰Ž— ‘•‡ȋC,KȌƒ†‡–‘‡„‘†‹‡•ȋD,LȌ „‡ˆ‘”‡ˆƒ•–‹‰ȋ Ȍǡƒˆ–‡”ͳͶŠ”•‘ˆˆƒ•–‹‰ǡ„‡ˆ‘”‡ ‘Ž†‡š’‘•—”‡ȋ Ȍƒ†ƒˆ–‡”ˆƒ•–‹‰’Ž—• ‘Ž†‡š’‘•—”‡ ȋ Ȍǡƒ„•‘Ž—–‡Ž‹˜‡”™‡‹‰Š–ȋE,MȌƒ†”‡Žƒ–‹˜‡Ž‹˜‡”™‡‹‰Š–ȋΨ‘ˆ„‘†›™‡‹‰Š–ȌȋF,NȌ‘ˆ–Š‡ͳ͸ǦʹͲ™‡‡•Ǧ ‘Ž† ȋA-FȌ ƒ† ͺ ™‡‡•Ǧ‘Ž† ‹ ‡ ȋG,I,K-NȌ ƒ† ”‡ –ƒŽ –‡’‡”ƒ–—”‡ ȋHȌǡ ”‡ –ƒŽ –‡’‡”ƒ–—”‡ ˜‡”•—• „‘†› ™‡‹‰Š– ƒˆ–‡” ͳͶ Š”• ‘ˆ ˆƒ•–‹‰ ȋJȌ ‘ˆ –Š‡ ͺǦ ™‡‡•Ǧ‘Ž† ‹ ‡Ǥ α͸Ǧͺ ˆ‘” „‘–Š  ƒ† Ǥ Š‡ ”‡•—Ž–• ƒ”‡ ”‡’”‡•‡–‡†ƒ•—‡›„‘šƒ†™Š‹•‡”’Ž‘–•ǡ™Š‡”‡–Š‡„Žƒ Ǧˆ‹ŽŽ‡†Œƒ‰‰‡† ‹” Ž‡•‹†‹ ƒ–‡‹†‹˜‹†—ƒŽ‹ ‡ ˆƒŽŽ‹‰ ‘—–•‹†‡ –Š‡ ͹ͷΨ ’‡” ‡–‹Ž‡ ’Ž—• ͳǤͷȉ‹–‡”Ǧ“—ƒ”–‹Ž‡ ”ƒ‰‡ ȋ

(11) Ȍ ‘” ʹͷΨ ’‡” ‡–‹Ž‡ ‹—• ͳǤͷȉ

(12) Ǥ

(13) †‹ ƒ–‹˜‡ǡiǣ’δͲǤͳǡȗǣ’δͲǤͲͷǡȗȗǣ’δͲǤͲͳǡȗȗȗǣ’δͲǤͲͲͳƒ†ȗȗȗȗǣ’δͲǤͲͲͲͳ ‘’ƒ”‡†–‘ǤŽŽ„‘š’Ž‘–•ƒ† ‹†‹ ƒ–‹‘•‘ˆ•‹‰‹ˆ‹ ƒ ‡‹–Š‡”‡ƒ‹†‡”‘ˆ–Š‹•”‡’‘”–ƒ”‡”‡’”‡•‡–‡†‹–Š‡•ƒ‡™ƒ›ƒ•–Š‹•ˆ‹‰—”‡Ǥ. . ͳ͵ͳ. 4.

(14) Chapter 4 Š‡ ˜ƒ”‹ƒ–‹‘ ‹ –Š‡ Ǧ ‰”‘—’ ™ƒ• •—„•–ƒ–‹ƒŽǡ ’‘••‹„Ž› „‡ ƒ—•‡ –Š‡ •–ƒ”– ‘ˆ –Š‡ –‡’‡”ƒ–—”‡ †‡ Ž‹‡ ™ƒ• •Š‹ˆ–‡† ‹ –‹‡ „‡–™‡‡ ‹†‹˜‹†—ƒŽ• ȋ—’’Ž‡‡–ƒ”› ˆ‹‰—”‡ ͳȌǤ Š‡„‘†›™‡‹‰Š–†‡ ”‡ƒ•‡†–‘ƒ•‹‹Žƒ”‡š–‡–‹„‘–Š‰”‘—’•ǡƒ†™ƒ••‹‹Žƒ”‹Ǧ ƒ†‹ ‡ȋ ‹‰—”‡ͳȌǤ

(15) ƒ††‹–‹‘ǡ„Ž‘‘†‰Ž— ‘•‡ƒ†‡–‘‡Ǧ„‘†› ‘ ‡–”ƒ–‹‘•™‡”‡ •‹‹Žƒ”„‡–™‡‡ƒ†‹ ‡ȋ ‹‰—”‡ͳǦȌǤŠ‡ƒ„•‘Ž—–‡ƒ†”‡Žƒ–‹˜‡Ž‹˜‡”™‡‹‰Š–•™‡”‡ǡ Š‘™‡˜‡”ǡ•‹‰‹ˆ‹ ƒ–Ž›Š‹‰Š‡”‹Ǧ–Šƒ‹ ‡ƒ––Š‡‡†‘ˆ–Š‡‡š’‡”‹‡–ȋ ‹‰—”‡ ͳǡ ȌǤ ”‡˜‹‘—•Ž›ǡƒ•‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡”„‘†›–‡’‡”ƒ–—”‡Šƒ•„‡‡”‡’‘”–‡†‹ͺǦ™‡‡•‘Ž†ǡˆƒ•–‡†ǡ ‘Ž†Ǧ‡š’‘•‡†Ǧ‹ ‡”‡Žƒ–‹˜‡–‘–Š‡‹” ‘—–‡”’ƒ”–•ȏͳͲȐǤŠ‡™‡”‡’‡ƒ–‡†–Š‡ ‡š’‡”‹‡–‹ͺ™‡‡•‘Ž†‹ ‡ǡ‘”‡ƒ‹ƒŽ••Š‘™‡†ƒ†”‘’‹„‘†›–‡’‡”ƒ–—”‡ǡ‹–Š‹• ƒ•‡ƒŽ•‘‹–Š‡‰”‘—’ȋ—’’Ž‡‡–ƒ”›ˆ‹‰—”‡ͳȌǤŠ‹•‡–ƒ‹Ž‡†ǡ͵͵Ǥ͵ΨȋʹȀ͸Ȍƒ†͹ͳΨ ȋͷȀ͹Ȍ‘ˆ–Š‡ƒ†‹ ‡ǡ”‡•’‡ –‹˜‡Ž›ǡ‘ˆ™Š‹ Š–Š‡„‘†›–‡’‡”ƒ–—”‡ ‘—Ž†„‡‡ƒ•—”‡† ȋ ‹‰—”‡ ͳ ǡ Ǣ —’’Ž‡‡–ƒ”› ˆ‹‰—”‡ ͳȌǤ ‘ •‹‰‹ˆ‹ ƒ– †‹ˆˆ‡”‡ ‡„‡–™‡‡ –Š‡ ‰‡‘–›’‡• ™ƒ•‘„•‡”˜‡†ȋ ‹‰—”‡ͳ ǡ ȌǤ —”–Š‡”‘”‡ǡ–Š‡„‘†›™‡‹‰Š–†‡ ”‡ƒ•‡†‹„‘–Š‰‡‘–›’‡•–‘ƒ •‹‹Žƒ”‡š–‡–ǡƒ†ƒŽ•‘Š‡”‡™ƒ••‹‹Žƒ”Ǧƒ†‹ ‡ȋ ‹‰—”‡ͳ

(16) ȌǤ ‘””‡Žƒ–‹‘ ƒƒŽ›•‹• ”‡˜‡ƒŽ‡† –Šƒ– –Š‡ ƒ‹ƒŽ• ™‹–Š ƒ Ž‘™‡” „‘†› ™‡‹‰Š– ƒŽ•‘ Šƒ† ƒ Ž‘™‡” „‘†› –‡’‡”ƒ–—”‡ ƒˆ–‡” ‘Ž† ‡š’‘•—”‡ ȋ ‹‰—”‡ ͳ  ƒ† —’’Ž‡‡–ƒ”› ‹‰—”‡ ͳȌǤ

(17) –‡”‡•–‹‰Ž›ǡ ƒˆ–‡” ‘Ž†‡š’‘•—”‡–Š‡„Ž‘‘†Ǧ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘™ƒ••‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡”‹›‘—‰Ǧ –Šƒ‹‹ ‡ȋ ‹‰—”‡ͳȌǡƒŽ„‡‹–™‹–Š‘—–ƒ›‡ˆˆ‡ –‘–Š‡Ž‡˜‡Ž‘ˆ‡–‘‡„‘†‹‡•ȋ ‹‰—”‡ ͳȌǤ’’ƒ”‡–Ž›ǡ†‡ˆ‹ ‹‡ ›ƒˆˆ‡ –•–Š‡„Ž‘‘†‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘‘Ž›—†‡”•‡˜‡”‡ ‡‡”‰‡–‹ •–”‡••ǡ‹Ǥ‡Ǥ—’‘ƒ ‘„‹ƒ–‹‘‘ˆˆƒ•–‹‰ƒ† ‘Ž†•–”‡••‹›‘—‰ƒ‹ƒŽ•ǤŠ‹•‹• •‹‹Žƒ”–‘–Š‡•‹–—ƒ–‹‘‹’ƒ–‹‡–•ǡ™Š‘ƒŽ•‘’”‡•‡–™‹–ŠŠ›’‘‰Ž› ‡‹ƒ‘Ž›ƒ–ƒ›‘—‰ƒ‰‡ ȋ–›’‹ ƒŽŽ› ͲǦʹ ›‡ƒ”•Ȍǡ †—”‹‰ ‘˜‡”‹‰Š– ˆƒ•–‹‰ ‹ ‘„‹ƒ–‹‘ ™‹–Š ƒ‘–Š‡” •–”‡•• ˆƒ –‘”Ǥ ‡Žƒ–‹˜‡Ž‹˜‡”™‡‹‰Š–•™‡”‡•‹‰‹ˆ‹ ƒ–Ž›Š‹‰Š‡”‹›‘—‰ ‘’ƒ”‡†–‘–Š‡‹ ‡ȋ ‹‰—”‡ ͳǡȌǡƒ•‘„•‡”˜‡†ˆ‘”ƒ†—Ž–‹ ‡„—––Š‡ †‹ˆˆ‡”‡ ‡™ƒ•‡˜‡‘”‡’”‘‘— ‡†‹›‘—‰ ‹ ‡Ǥ Ž–‘‰‡–Š‡”ǡ „ƒ•‡†‘ –Š‡ ‡–ƒ„‘Ž‹  ”‡•—Ž–•ǡ –Š‡•‡ ”‡•—Ž–• •Š‘™ –Šƒ– ›‘—‰ Ǧ ‹ ‡ƒ”‡‘”‡˜—Ž‡”ƒ„Ž‡–‘ ‘Ž†‡š’‘•—”‡–Šƒƒ†—Ž–‹ ‡Ǥ . Higher plasma acyl-carnitines and NEFAs in fasted, cold-exposed MCAD-KO mice ”‡˜‹‘—•Ž›ǡ•‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡”’Žƒ•ƒƒŽƒ‹‡Ž‡˜‡Ž•Šƒ˜‡„‡‡”‡’‘”–‡†‹ˆƒ•–‡†Ǧ ‘’ƒ”‡† –‘  ‹ ‡ǡ ™Š‹Ž‡ ˜ƒŽ‹‡ǡ Ž‡— ‹‡ ƒ† ‹•‘Ž‡— ‹‡ ™‡”‡ •‹‹Žƒ” ȏʹͺȐǤ ‡ †‹† ‘– ‘„•‡”˜‡•‹‰‹ˆ‹ ƒ–†‹ˆˆ‡”‡ ‡•‹ƒ‹‘ƒ ‹† ‘ ‡–”ƒ–‹‘•„‡–™‡‡ͺ™‡‡•‘Ž† Ǧ ƒ†‹ ‡ǡ‡š ‡’–ˆ‘”ȽǦƒ‹‘„—–›”‹ ƒ ‹†ǡ™Š‹ Š™ƒ•Ž‘™‡”‹–Š‡Ǧ‰”‘—’ȋ ‹‰—”‡ ʹƒ†—’’Ž‡‡–ƒ”›ˆ‹‰—”‡ʹȌǤŽ‡˜ƒ–‡†„Ž‘‘†‡†‹—Ǧ Šƒ‹ƒ ›ŽǦ ƒ”‹–‹‡Ž‡˜‡Ž•ƒ”‡ƒ ‹’‘”–ƒ– „‹‘ƒ”‡” ˆ‘”  †‡ˆ‹ ‹‡ › ȏʹͻȂ͵͵ȐǤ

(18) †‡‡†ǡ –Š‡ ͸Ǧ –‘ ͳͲǦƒ ›ŽǦ ƒ”‹–‹‡ ‘ ‡–”ƒ–‹‘•™‡”‡Š‹‰Š‡”‹Ǧ ‘’ƒ”‡†–‘‹ ‡ǡ—†‡”ƒŽŽ ‘†‹–‹‘•ƒ†‹ „‘–Šƒ‰‡‰”‘—’•ȋ ‹‰—”‡ʹǦƒ†—’’Ž‡‡–ƒ”›ˆ‹‰—”‡ʹǦȌǤ

(19) ƒ‰”‡‡‡–™‹–Š’”‡˜‹‘—• ˆ‹†‹‰•‹Ǧ‹ ‡ȏͳͲǡʹͺǡ͵ͶȐǡ’ƒ”–‹ —Žƒ”Ž›ͳͲǣͳ™ƒ•‡Ž‡˜ƒ–‡†ǤŠ‹•‹•‹ ‘–”ƒ•––‘ Š—ƒ’ƒ–‹‡–•ǡ‹™Š‹ ŠͺǦƒ ›ŽǦ ƒ”‹–‹‡ƒ ——Žƒ–‡•‘•–ȏʹͻȂ͵͵ȐǤŠ‡†‹ˆˆ‡”‡ ‡ „‡–™‡‡Ǧƒ†™ƒ•‘•–’”‘‘— ‡†ƒˆ–‡”ˆƒ•–‹‰ǡ‹””‡•’‡ –‹˜‡‘ˆ ‘Ž†‡š’‘•—”‡ ȋ ‹‰—”‡ ʹǦǡ —’’Ž‡‡–ƒ”› ˆ‹‰—”‡ ʹǦ ƒ† —’’Ž‡‡–ƒ”› –ƒ„Ž‡ ͳȌǤ ‹ƒŽŽ›ǡ ’Žƒ•ƒ  Ž‡˜‡Ž•ƒˆ–‡” ‘Ž†‡š’‘•—”‡™‡”‡•‹‰‹ˆ‹ ƒ–Ž›Š‹‰Š‡”‹–Š‡›‘—‰Ǧ‹ ‡ȋ ‹‰—”‡ ʹȌǤ—”‹‰ ‘Ž†•–”‡••‡Šƒ ‡†–”‹‰Ž› ‡”‹†‡Ž‹’‘Ž›•‹•„›ƒ†‹’‘•‡–‹••—‡”‡•—Ž–•‹‹ ”‡ƒ•‡† ”‡Ž‡ƒ•‡‘ˆ •ȏ͵ͷȐǤ. ͳ͵ʹ.

(20) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice. [Blood amino acid] ( PM). A 1500. 1000. 500. Al an Ar ine As gin pa in ra e gi Cy ne G sti lu ne ta G ma lu ta te m G ine ly Hi cin s e Is tidi ol ne eu c Le ine uc in e M Lys Ph eth ine en ion yl in al e an in Pr e ol in Se e Th ri re ne Tr on yp in to e p Ty han ro si n Va e lin e. 0. Blood C n-acyl-carnitine ( PM). B. before fasting. C. 0.3. 0.3. ***. ***. 0.3. ****. 0.2. **. **. i. ** *. 0.1. 0.1. 0.0. 0.0 C6. C8. C10:1 C10. C12:1 C12. C6. after cold exposure. 0.4. ***. * 0.2. D. 14-hr fasted. 0.4. 0.4. 4. C8. C10:1 C10. C12:1 C12. 0.2. *** **. *. 0.1 0.0 C6. C8. C10:1 C10. C12:1 C12. E Serum NEFA (PM). 1500. **. 1000. 500. 0 16-20 w.o.. 8 w.o.. Figure 2. Blood metabolite concentrations.Žƒ•ƒƒ‹‘ƒ ‹† ‘ ‡–”ƒ–‹‘•ƒˆ–‡” ‘Ž†‡š’‘•—”‡ȋAȌǡ „Ž‘‘† ƒ ›ŽǦ ƒ”‹–‹‡ ‘ ‡–”ƒ–‹‘• ȋB-DȌ ‘ˆ –Š‡ ͺǦ™‡‡•Ǧ‘Ž† ‹ ‡ǡ ƒ† •‡”—   Ž‡˜‡Ž• ƒˆ–‡” ‘Ž† ‡š’‘•—”‡ȋEȌǤ.  Š‡†‹ˆˆ‡”‡ ‡‹’Žƒ•ƒ Ž‡˜‡Ž•ƒ› –Š‡”‡ˆ‘”‡‡‹–Š‡”„‡†—‡–‘–Š‡Š‹‰Š‡””‡Ž‡ƒ•‡‘ˆ  •„›ƒ†‹’‘•‡–‹••—‡ǡ‘”–‘–Š‡†‡ˆ‡ –‹˜‡ ’ƒ–Š™ƒ›‹–Š‡Ž‹˜‡”ǡ‘”ƒ ‘„‹ƒ–‹‘‘ˆ „‘–ŠǤ . Altered amino-acid and lipid handling in the liver of cold-exposed MCAD-KO mice —„•‡“—‡–Ž›ǡ ™‡ ˆ‘ —•‡† ‘ ‡–ƒ„‘Ž‹  ƒ†ƒ’–ƒ–‹‘• ‹ –Š‡ Ž‹˜‡” ‘ˆ ˆƒ•–‡†ǡ ‘Ž†Ǧ‡š’‘•‡† ƒ‹ƒŽ•Ǥ ‡’ƒ–‹ ƒ‹‘ƒ ‹†•ƒ›•‡”˜‡ƒ•ƒŽ–‡”ƒ–‹˜‡•—„•–”ƒ–‡•ƒ†ˆ—‡Ž•ˆ‘”‡–‘‰‡‡•‹• ƒ†‰Ž— ‘‡‘‰‡‡•‹•‹–Š‡Ǧ‹ ‡Ǥ

(21) Ž‹‡™‹–Š–Š‹•ǡ™‡‘„•‡”˜‡†•‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡” ƒ‹‘Ǧƒ ‹† ‘ ‡–”ƒ–‹‘• ‹ Ž‹˜‡”• ‘ˆ ‘Ž†Ǧ‡š’‘•‡† ˆƒ•–‡† Ǧ ‹ ‡ ȋ ‹‰—”‡ ͵ǡ —’’Ž‡‡–ƒ”› ˆ‹‰—”‡ ͵ ƒ† —’’Ž‡‡–ƒ”› –ƒ„Ž‡ ͳȌǤ Š‡ †‹ˆˆ‡”‡ ‡ ™ƒ• ƒ‰ƒ‹ ‘”‡ ’”‘‘— ‡† ‹ –Š‡ ›‘—‰ ‹ ‡Ǥ

(22)  –Š‡•‡ ‹ ‡ǡ –Š‡ Ž‡˜‡Ž• ™‡”‡ •‹‰‹ˆ‹ ƒ–Ž› Ž‘™‡” ˆ‘” ‰Ž—–ƒ‹‡ǡ ‰Ž› ‹‡ǡ Ž‡— ‹‡ǡ Ž›•‹‡ǡ •‡”‹‡ǡ –Š”‡‘‹‡ǡ –›”‘•‹‡ ƒ† Ǧ ›•–ƒ–Š‹‘‹‡ ƒ† ‹†‹ ƒ–‹˜‡ȋiǡ’δͲǤͳȌˆ‘”ƒŽƒ‹‡ǡ‰Ž—–ƒ‹ ƒ ‹†ǡȽǦƒ‹‘„—–›”‹ ƒ ‹†ƒ†‘”‹–Š‹‡Ǥ

(23) –Š‡ƒ†—Ž– ‹ ‡ǡ‘Ž›‰Ž› ‹‡Ž‡˜‡Ž•™‡”‡•‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡”ǡƒŽ–Š‘—‰Š–Š‡•ƒ‡–”‡†™ƒ•‘„•‡”˜‡† ˆ‘”‘–Š‡”ƒ‹‘ƒ ‹†•ȋ—’’Ž‡‡–ƒ”›ˆ‹‰—”‡͵ǡȌǤ‹˜‡”ƒ ›ŽǦ ƒ”‹–‹‡ ‘ ‡–”ƒ–‹‘•™‡”‡ •‹‹Žƒ” „‡–™‡‡  ƒ†  ‹ ‡ ȋ ‹‰—”‡ ͵ǡ ƒ† —’’Ž‡‡–ƒ”› ƒ„Ž‡ ͳȌǤ ”‡˜‹‘—•Ž›ǡ •‘‡™Šƒ–Š‹‰Š‡” ‘ ‡–”ƒ–‹‘•‘ˆ‡†‹—Ǧ Šƒ‹Ž‹˜‡”ƒ ›ŽǦ ƒ”‹–‹‡•Šƒ˜‡„‡‡”‡’‘”–‡†‹ ˆƒ•–‡†Ǧ‹ ‡ǡƒŽ„‡‹––Š‡†‹ˆˆ‡”‡ ‡™ƒ•Ž‡••’”‘‘— ‡†–Šƒ‹„Ž‘‘†ȏ͵ͶȐǤ. ͳ͵͵.

(24) Chapter 4. 8 6. i. **. **. *. ***. ** i. 4. **. 2. *. i. 0. Al an A ine As rgin i As par ne pa agi rti ne c a Cy cid s G tei G lut ne lu am ta m ine ic ac G id ly Hi cin s e Is tidi ol ne eu c Le ine uc in e M Lys Ph eth ine en ion yl in al e an in Pr e ol in S e Th eri re ne on Ty ine ro si n Va e lin e. [Liver amino acid] (nmol/g). A. C 1.0. 100. TG (P mol/g liver). Liver C n-acyl-carnitine (nmol/g). B 0.8 0.6 0.4 0.2 0.0. *. **. 80 60 40 20 0. C6. C8. C10:1 C10. C12:1 C12. 16-20 w.o.. 8 w.o..  Figure 3. Hepatic metabolite profiles.‹˜‡”ƒ‹‘Ǧƒ ‹†ȋAȌƒ†͸Ǧͳʹƒ ›ŽǦ ƒ”‹–‹‡ ‘ ‡–”ƒ–‹‘•ˆ‘” ͺǦ™‡‡•‘Ž†‹ ‡ȋBȌǡƒ†Ž‹˜‡”–”‹‰Ž› ‡”‹†‡ȋ Ȍ ‘ ‡–”ƒ–‹‘‘ˆͳ͸ǦʹͲǦƒ†ͺǦ™‡‡•Ǧ‘Ž†‹ ‡ȋCȌǤ.  ’’ƒ”‡–Ž›ǡ ‘Ž†Ǧ‡š’‘•—”‡–”‹‰‰‡”•ƒƒ†ƒ’–ƒ–‹‘‘ˆŽ‹’‹†‡–ƒ„‘Ž‹•‹–Š‡Ǧ‹ ‡ǡ Ž‡ƒ†‹‰ –‘ ƒ ›ŽǦ ƒ”‹–‹‡ Š‘‡‘•–ƒ•‹•Ǥ ‹ƒŽŽ›ǡ Ž‹˜‡” –”‹‰Ž› ‡”‹†‡ ȋ Ȍ ‘ ‡–”ƒ–‹‘• ™‡”‡ Š‹‰Š‡”‹–Š‡Ǧ–Šƒ‹–Š‡‹ ‡ǡ‹„‘–Šƒ‰‡‰”‘—’•ȋ ‹‰—”‡͵ȌǡƒŽ•‘•—‰‰‡•–‹‰ ƒŽ–‡”‡†Ž‹’‹†Šƒ†Ž‹‰‹–Š‡Ž‹˜‡”Ǥ . Rerouting of medium-chain acyl-CoAs into chain-elongation and triglyceride synthesis Š‡Š‹‰Š‡”Ž‹˜‡”  ‘ ‡–”ƒ–‹‘•‹–Š‡Ž‹˜‡”ƒ† ‘’ƒ”ƒ„Ž‡Ž‹˜‡”ƒ ›ŽǦ ƒ”‹–‹‡’”‘ˆ‹Ž‡• •—‰‰‡•–ƒƒŽ–‡”‡†Ž‹’‹†‡–ƒ„‘Ž‹•‹–Š‡Ǧ‹ ‡ǤŠ‡”‡ˆ‘”‡ǡ™‡ƒƒŽ›œ‡†–Š‡Ž‹’‹†• ‡š–”ƒ –‡† ˆ”‘ –Š‡ Ž‹˜‡” „› —–ƒ”‰‡–‡† Ž‹’‹†‘‹ •Ǥ ˜‡” ͸ͲͲ •‹‰Ž‡ Ž‹’‹† •’‡ ‹‡• ‘—Ž† „‡ ƒ‘–ƒ–‡†ǡ ‘˜‡”‹‰ ƒŽŽ ƒŒ‘” Ž‹’‹† Žƒ••‡• ȋ ‹‰—”‡ Ͷ ƒ† —’’Ž‡‡–ƒ”› ƒ„Ž‡ ʹȌǤ ”‹ ‹’ƒŽ‘’‘‡–ƒŽ›•‹•ȋȌ•Š‘™‡†ƒ Ž‡ƒ”•‡’ƒ”ƒ–‹‘„‡–™‡‡ƒ†Ǧ ‹ ‡ „ƒ•‡† ‘ –Š‡‹” Š‡’ƒ–‹  Ž‹’‹† ’”‘ˆ‹Ž‡• ȋ ‹‰—”‡ ͶȌǤ Š‡ Ž‹’‹†• –Šƒ– „”‘—‰Š– ƒ„‘—– –Š‡ •‡’ƒ”ƒ–‹‘ ‘ˆ –Š‡ ‰”‘—’• ™‡”‡ ƒŽ‘•– ‡š Ž—•‹˜‡Ž›  •Ǥ Š‡ ‰”‘—’ ‘ˆ Ž‹’‹†• ™‹–Š ‘•– ’”‘‘— ‡††‹ˆˆ‡”‡ ‡•„‡–™‡‡ƒ†ȋ’δͲǤͲͷƒ†ˆ‘Ž† Šƒ‰‡εʹǢ ‹‰—”‡Ͷ—’’‡” ”‹‰Š–“—ƒ†”ƒ–Ȍ ‘•‹•–‡†‘ˆ͵ͻ •ǡͳ†‹‰Ž› ‡”‹†‡ǡƒ†ͳ•’Š‹‰‘Ž‹’‹†ȋ—’’Ž‡‡–ƒ”›–ƒ„Ž‡ ʹȌǤŠ‡ •™‹–Š–Š‡Š‹‰Š‡•–ˆ‘Ž†Ǧ Šƒ‰‡•ǡ‡˜‡—’–‘ʹͶǦˆ‘Ž†ǡŠƒ†ƒ•—‘ˆ ƒ”„‘ƒ–‘• ‹–Š‡ƒ ›Ž Šƒ‹•‹–Š‡”ƒ‰‡ͶͲǦͶͺȋ ‹‰—”‡ͶǦ ȌǤŠ‹••—‰‰‡•–•–Šƒ–‘•–‘ˆ–Š‡ ‘–ƒ‹‡† ƒ–Ž‡ƒ•–‘‡ƒ ›Ž Šƒ‹•Š‘”–‡”–Šƒͳ͸Ǥ—„•‡“—‡–Ž›ǡ•–”— –—”ƒŽƒƒŽ›•‹•”‡˜‡ƒŽ‡†–Š‡ƒ ›ŽǦ Šƒ‹ ‘’‘•‹–‹‘‘ˆ͵ͳ‘—–‘ˆ–Š‡ͶʹŽ‹’‹†•™Š‹ Š™‡”‡•‹‰‹ˆ‹ ƒ–Ž›‡Ž‡˜ƒ–‡†‹Ǧ ‹ ‡ ȋ—’’‡” ”‹‰Š– “—ƒ†”ƒ–ǡ ‹‰—”‡ ͶǤ Š‡ sn-’‘•‹–‹‘ ‘ˆ –Š‡ ƒ ›Ž Šƒ‹• ‘—Ž† ‘– „‡ †‡–‡”‹‡†Ǥ˜‡”ͳ͵Ͳ ‹•‘‡”•ƒ†ͳ ™‡”‡ƒ‘–ƒ–‡†ȋ—’’Ž‡‡–ƒ”›–ƒ„Ž‡͵ȌǤͻͷΨ ‘ˆ–Š‡‹†‡–‹ˆ‹‡† ‹•‘‡”• ‘–ƒ‹‡†ƒ–Ž‡ƒ•–‘‡ˆƒ––›Ǧƒ ›Ž Šƒ‹™‹–Šƒ ƒ”„‘—„‡”‹ –Š‡”ƒ‰‡͸–‘ͳͶǤʹ͸Ψ‘ˆ–Š‡•‡ ‘–ƒ‹‡†ƒ•‡ ‘†ͳͶˆƒ––›ƒ ›ŽǦ Šƒ‹ȋ ‹‰—”‡Ͷ

(25) ȌǤŠ‡ ƒŒ‘”‹–›‘ˆ • ‘–ƒ‹‡†ƒ–Ž‡ƒ•–‘‡ Šƒ‹ˆƒ––›ƒ ›ŽǦ”‡•‹†—‡tͳ͸ǤŠ‡†‡ˆ‹ ‹‡ ›‘ˆ–Š‡ ‡œ›‡‘•–Ž›ƒˆˆ‡ –•–Š‡‘š‹†ƒ–‹‘‘ˆˆƒ––›ƒ ‹†•‘ˆ Šƒ‹Ž‡‰–Š͸ǦͳͲǤ. ͳ͵Ͷ.

(26) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice. B. A. C 10. -log (p-value). 5 0 -5 -10 -10. Total=624. 0. F. E. Liver TG (n.s.i. ˜ 106 ). TG(48:8). 5. 10. 100. ***. ****. 1.8. G ****. -5. 0. TG(42:2). 4. 5. DG(28:2) 1.5. ****. 1.2. 60. 2. H. 12. 80 1.0. 4. log2(fold change). TG(42:4). TG(44:2). 1.5. n.s. TG Other. 0 -5. PC1 (35%). D. 6. WT KO. PC2 (25.5%). TG (240) PC (124) DG (80) PE (49) SP (48) PS (18) CE (18) MG (13) PG (12) PI (11) CL (11). **** 1.0. 6 40. 0.5. 0.6. 0.5. 20 0.0 KO. I Number of isomers. 0.0. 0 WT. 0.0. 0 WT KO. WT KO. WT KO. WT KO. TG composition C6 C8 C9 C10 C12 C14 C15 C16 >C16. 100. 50. 0 c. b. a.  Figure 4. Hepatic lipidomics profile of 8-weeks old mice. ‹‡ Šƒ”–‘ˆ‹†‡–‹ˆ‹‡†Ž‹’‹† Žƒ••‡•ȋAȌǡ ’Ž‘–‘ˆ‹†‡–‹ˆ‹‡†Ž‹’‹†•’‡ ‹‡•ˆ‘”Ž‹˜‡”ǡ™‹–Šƒ†‹ ‡†‡’‹ –‡†‹”‡†ƒ†„Žƒ ǡ”‡•’‡ –‹˜‡Ž›ȋBȌǡ ‘Ž ƒ‘’Ž‘–‘ˆ–Š‡‹†‡–‹ˆ‹‡†Ž‹’‹†•’‡ ‹‡•ˆ‘”˜•ȋCȌǤ‹’‹†•ˆ‘”™Š‹ Š–Š‡–Š”‡•Š‘Ž†‘ˆ’δͲǤͲͷ™‡”‡ ‘–”‡ƒ Š‡†ƒ”‡†‡’‹ –‡†‹„Žƒ Ǥ ‘”–Š‡‘–Š‡”Ž‹’‹†•ǡ •ƒ”‡†‡’‹ –‡†‹”‡†ƒ†–Š‡”‡ƒ‹‹‰Ž‹’‹†•‹ „Ž—‡—‡›’Ž‘–•‘ˆ–Š‡–‘’ͷŽ‹’‹†•ȋ™‹–Š–Š‡Žƒ”‰‡•–ƒ„—†ƒ ‡†‹ˆˆ‡”‡ ‡„‡–™‡‡ƒ†ȋD-HȌǡƒ† –Š‡ƒ ›ŽǦ Šƒ‹ ‘’‘•‹–‹‘‘ˆ†‹ˆˆ‡”‡–‹ƒŽŽ›‡š’”‡••‡† ‹•‘‡”•ȋIȌǤ‹’‹†•‹’ƒ‡ŽI™‡”‡ƒ‘–ƒ–‡†ƒ• ȋǣȌǡ™Š‡”‡‹†‹ ƒ–‡•Ž‹’‹† Žƒ••ǡ–Š‡–‘–ƒŽ ƒ”„‘—„‡”••—‡†‘˜‡”ƒŽŽƒ ›Ž Šƒ‹•ǡƒ†–Š‡ —„‡”‘ˆ†‘—„Ž‡„‘†•ȏ͵͸ȐǤŠ‡ƒ ›Ž Šƒ‹•‹‡ƒ Š‹•‘‡”™‡”‡‘”†‡”‡†ˆ”‘ƒ–‘ ǡˆ”‘ •Š‘”–‡•––‘ Ž‘‰‡•– Šƒ‹Ž‡‰–ŠǤ•ǣ‘–•‹‰‹ˆ‹ ƒ–ǡȗȗȗ’δͲǤͲͲͳǡȗȗȗȗ’δͲǤͲͲͲͳǤαͺ’‡”‰”‘—’ǤǤ•Ǥ‹Ǥ‘”ƒŽ‹œ‡†•‹‰ƒŽ ‹–‡•‹–›Ǥ ‡‡ ‹‰—”‡ ͳ ˆ‘” ˆ—”–Š‡” ‡š’Žƒƒ–‹‘ ‘ˆ ’Ž‘–•Ǥ ‡‡ —’’Ž‡‡–ƒ”› ƒ„Ž‡ ʹ ˆ‘” ƒ††‹–‹‘ƒŽ Ž‹’‹†‘‹ ”‡•—Ž–•Ǥ.  Š‡Ž‹’‹†‘‹ •”‡•—Ž–••—‰‰‡•––Šƒ– •‘‡‘ˆ–Š‡ƒ„‡””ƒ–‡†‹—Ǧ Šƒ‹ˆƒ––›ƒ ›ŽǦ”‡•‹†—‡• ™‡”‡‹ ‘”’‘”ƒ–‡†‹–‘ •ǡ‡‹–Š‡”†‹”‡ –Ž›‘”‹†‹”‡ –Ž›˜‹ƒ’ƒ”–‹ƒŽ Šƒ‹‡Ž‘‰ƒ–‹‘ǡ–Š‡”‡„› ‘–”‹„—–‹‰–‘–Š‡’”‡˜‡–‹‘‘ˆ–Š‡‹”ƒ ——Žƒ–‹‘‹–Š‡ ‡ŽŽ•Ǥ . Zooming into an MCAD-KO mouse with a severe liver phenotype ‹ƒŽŽ›ǡ ™‡ •–—†‹‡† –Š‡  ‘—•‡ ™‹–Š –Š‡ ‘•– ‡š–”‡‡ ’Š‡‘–›’‡ ‹ ‘”‡ †‡’–Šǡ ‘•‹†‡”‹‰–Šƒ–‹–ƒ›„‡ƒ‘†‡Žˆ‘”ƒŠ—ƒ’ƒ–‹‡– Ž‘•‡–‘ƒ‡–ƒ„‘Ž‹  ”‹•‹•Ǥ‡ ˆ‘ —•‡† ‘ •‡˜‡”‡ Š‡’ƒ–‘•–‡ƒ–‘•‹• ȋ‘„•‡”˜‡† ‹  ’ƒ–‹‡–• ™Š‘ †‹‡† ‘ˆ ƒ ‡–ƒ„‘Ž‹  ”‹•‹• ȏ͵͹ȐȌ ƒ† ‘ –Š‡ ‡ˆˆ‡ – ‘ˆ ‘Ž† ‡š’‘•—”‡ ‘ „Ž‘‘† ‰Ž— ‘•‡Ǥ ”‘ “—ƒŽ‹–ƒ–‹˜‡ ƒ† “—ƒ–‹–ƒ–‹˜‡Š‡’ƒ–‘Š‹•–‘Ž‘‰‹ ƒŽƒƒŽ›•‡•ƒ†„Ž‘‘†‰Ž— ‘•‡ Šƒ‰‡†—”‹‰ ‘Ž†‡š’‘•—”‡ǡ™‡ ‹†‡–‹ˆ‹‡†ƒ‘—•‡ȋͳͲȌ™‹–Š„‘–Š•‡˜‡”‡•–‡ƒ–‘Š‡’ƒ–‹–‹•ȋi.e.‹ˆŽƒƒ–‹‘‘ˆ–Š‡Ž‹˜‡”. ͳ͵ͷ.

(27) Chapter 4 †—‡–‘ˆƒ–ƒ ——Žƒ–‹‘Ȍȋ ‹‰—”‡ͷƒ†—’’Ž‡‡–ƒ”›–ƒ„Ž‡ͶȌƒ† ‘Ž†Ǧ‹†— ‡††‡ ”‡ƒ•‡ ‘ˆ –Š‡ „Ž‘‘† ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘Ǥ Š‹• ’Š‡‘–›’‡ ™ƒ• •’‡ ‹ˆ‹  ˆ‘” ͳͲǡ •‹ ‡ ‡‹–Š‡” ƒ Š‡’ƒ–‘’ƒ–Š‘Ž‘‰‹ ƒŽ ’Š‡‘–›’‡ ȋ—’’Ž‡‡–ƒ”› ˆ‹‰—”‡ ͶȌ ‘” ƒ ”‡†— –‹‘ ‘ˆ –Š‡ „Ž‘‘† ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘†—”‹‰ ‘Ž†‡š’‘•—”‡ȋ—’’Ž‡‡–ƒ”›ˆ‹‰—”‡ͶȌ™ƒ•–›’‹ ƒŽ‘ˆ–Š‡ Ǧ‰”‘—’ǤͳͲ™ƒ•–Š‡•ƒŽŽ‡•–‘—•‡„‡ˆ‘”‡ ‘Ž†‡š’‘•—”‡ȋͳͺǤͶ‰Ȍƒ†Šƒ†–Š‡ Ž‘™‡•– „‘†› –‡’‡”ƒ–—”‡ ƒ– –‡”‹ƒ–‹‘ ȋʹͳǤ͵ ιȌǤ ™‘Ǧ†‹‡•‹‘ƒŽ Ž—•–‡”‹‰ ‘ˆ •‡Ž‡ –‡† •‡–•‘ˆ‘ǦŽ‹’‹†‘‹ ‡–ƒ„‘Ž‹–‡•ȋ ˆǤƒ–‡”‹ƒŽ•ƒ†‡–Š‘†•Ȍ•Š‘™‡†–Šƒ–’ƒ”–‹ —Žƒ”Ž›–Š‡ ‘ ‡–”ƒ–‹‘•‘ˆŠ‡’ƒ–‹ ƒ‹‘ƒ ‹†•ƒ†Š‡’ƒ–‹ ˆ”‡‡ƒ†–‘–ƒŽ ƒ”‹–‹‡™‡”‡–Š‡Ž‘™‡•–‹ ͳͲ ‘’ƒ”‡† –‘ –Š‡ ‘–Š‡” Ǧ ‹ ‡ ȋ ‹‰—”‡ ͷȌǤ  ŽŽ –Š‡ ƒ‹‘ ƒ ‹†• ‹ –Š‡ ‘””‡•’‘†‹‰ Ž—•–‡”ǡ ‡š ‡’– ˆ‘” Ž‡— ‹‡ǡ ƒ”‡ ‰Ž— ‘‡‘‰‡‹ Ǥ Š‹• •—‰‰‡•–• ƒ •Š‘”–ƒ‰‡ ‘ˆ •—„•–”ƒ–‡ˆ‘”‰Ž— ‘‡‘‰‡‡•‹•ƒ†‹•‹ƒ‰”‡‡‡–™‹–Š–Š‡Ž‘™‡”„Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž‹–Š‹• ‘—•‡Ǥ

(28) ƒ††‹–‹‘ǡ–Š‡‘—•‡Šƒ•–Š‡Ž‘™‡•– ‘ ‡–”ƒ–‹‘•‘ˆ‡–Š‹‘‹‡ǡŠ‹•–‹†‹‡ƒ† Ǧ ›•–ƒ–Š‹‘‡ǡƒŽŽ‘ˆ™Š‹ Š ‘–”‹„—–‡–‘–Š‡ƒ–‹‘š‹†ƒ– ƒ’ƒ ‹–›Ǥ ‹ƒŽŽ›ǡ–Š‹•‘—•‡Šƒ†–Š‡ Ž‘™‡•–ƒ•’ƒ”–ƒ–‡ƒ†•‡”‹‡Ž‡˜‡Ž•‘ˆƒŽŽǦ‹ ‡ǤŠ‡•‡ƒ‹‘ƒ ‹†•’”‘˜‹†‡„—‹Ž†‹‰ „Ž‘ •ˆ‘”’—”‹‡•ǡ™Š‹ Šƒ”‡‹–—”„—‹Ž†‹‰„Ž‘ •ˆ‘”„‘–Š‘ ƒ†ΪǤ‘–Š‘  ƒ† Ϊ ƒ”‡ ‹’‘”–ƒ– ‘ˆƒ –‘”• ˆ‘”   ƒ† ƒ”‡ ƒŽ•‘ ‹˜‘Ž˜‡† ‹ ƒ–‹‘š‹†ƒ– ƒ† †‡–‘š‹ˆ‹ ƒ–‹‘‡ Šƒ‹••Ǥ . Discussion

(29)  –Š‹• •–—†›ǡ ™‡ ‹˜‡•–‹‰ƒ–‡† Ž‹˜‡” ‡–ƒ„‘Ž‹• ‹   †‡ˆ‹ ‹‡– Ǧ ‹ ‡ †—”‹‰ˆƒ•–‹‰ƒ† ‘Ž† ŠƒŽŽ‡‰‡ǡƒ•ƒ‘†‡Žˆ‘”•‡˜‡”‡‡‡”‰‡–‹ •–”‡••Ǥ–‹Ž‘™ǡǦ ‹ ‡™‡”‡ˆ‘—†–‘Šƒ˜‡ƒ”‡Žƒ–‹˜‡Ž›‹Ž†’Š‡‘–›’‡ƒ†–‘ƒ‹–ƒ‹‘”ƒŽ„Ž‘‘†‰Ž— ‘•‡ Ž‡˜‡Ž•†—”‹‰ˆƒ•–‹‰Ǥ”‡˜‹‘—•‡“ƒ†’”‘–‡‘‹ ƒƒŽ›•‹•‘ˆŽ‹˜‡”–‹••—‡‘ˆˆƒ•–‡†Ǧ Ž‹˜‡”–‹••—‡‹ ‡•Š‘™‡†‘Ž›‹‘”ƒ†ƒ’–ƒ–‹‘•ȏƒ”–‹‡•‡–ƒŽǤScientific Reportsǡ—†‡” ”‡˜‹•‹‘Ȑǡ •—‰‰‡•–‹‰ –Šƒ– –Š‡ ‘”ƒŽ ƒ’ƒ ‹–› ‘ˆ •Š‘”–Ǧ ƒ† Ž‘‰Ǧ Šƒ‹ ‹•‘‡œ›‡• ȋǡ ǡȌ™ƒ••—ˆˆ‹ ‹‡––‘‡‡––Š‡Š‡’ƒ–‹ ‡‡”‰›†‡ƒ††—”‹‰ˆƒ•–‹‰ǤŠ‡‘”‡ •‡˜‡”‡ ‡‡”‰‡–‹  •–”‡•• ‹’‘•‡† ‹ –Š‹• •–—†› Ȃ ƒ ‘„‹ƒ–‹‘ ‘ˆ ˆƒ•–‹‰ ƒ† ‘Ž† •–”‡•• Ȃ ”‡˜‡ƒŽ‡†ˆ‘”–Š‡ˆ‹”•––‹‡•‹‰‹ˆ‹ ƒ–Ž›Ž‘™‡”„Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž•‹Ǧ ‘’ƒ”‡†–‘ ‹ ‡Ǥ

(30)  ‘–”ƒ•––‘ƒ‡ƒ”Ž‹‡”•–—†›ȏͳͲȐǡ™‡†‹†‘–Ž‡ƒ˜‡–Š‡‹ ‡‹–Š‡ ‘Ž†—–‹Ž–Š‡› †‹‡†ǡ•‹ ‡™‡™‡”‡‹–‡”‡•–‡†‹–Š‡‹”‡–ƒ„‘Ž‹ ”‡•’‘•‡„‡ˆ‘”‡–Š‡‡‡”‰‡–‹ •–”‡••™‘—Ž† „‡Ž‡–ŠƒŽǤ’ƒ”–ˆ”‘–Š‡Ž‘™‡”„Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž•ǡ ‘’ƒ”‡†–‘–Š‡‹ ‡ǡ–Š‡Ǧ ‹ ‡•Š‘™‡†Ž‘™‡”ƒ‹‘Ǧƒ ‹†Ž‡˜‡Ž•‹–Š‡Ž‹˜‡”„—–‘–‹„Ž‘‘†ǡŠ‹‰Š‡”„Ž‘‘† Ž‡˜‡Ž•ǡ ƒ†Š‹‰Š‡”•–‘”ƒ‰‡‘ˆ • ‘–ƒ‹‹‰͸ǦͳͶƒ ›Ž Šƒ‹•‹–Š‡Ž‹˜‡”Ǥ

(31) –Š‹••‡ –‹‘™‡™‹ŽŽ ‹–‡”’”‡– ‘—” ˆ‹†‹‰•‹ –Š‡ Ž‹‰Š– ‘ˆ ™Šƒ– ‹•‘™ ƒ„‘—– –Š‡ ”‘Ž‡ ‘ˆ –Š‡ Ž‹˜‡” †—”‹‰ ‘Ž† •–”‡••ǤŠ‡™‡™‹ŽŽ†‹• —••Š‘™–Š‡‘„•‡”˜‡††‹ˆˆ‡”‡ ‡•‹ƒ‹‘ƒ ‹†ƒ†Ž‹’‹†‡–ƒ„‘Ž‹• „‡–™‡‡ –Š‡  ƒ†  ‹ ‡ ‘„˜‹ƒ–‡ –Š‡ ‡–ƒ„‘Ž‹  ‘•‡“—‡ ‡• ‘ˆ  †‡ˆ‹ ‹‡ ›Ǥ ‹ƒŽŽ›ǡ™‡™‹ŽŽ†‹• —••–Š‡‹’ƒ –‘ˆ‘—”ˆ‹†‹‰•ˆ‘”’ƒ–‹‡–’ƒ–Š‘’Š›•‹‘Ž‘‰›Ǥ ‘•–•–—†‹‡•‘–Š‡”‘Ž‡‘ˆ †—”‹‰ ‘Ž†•–”‡••ˆ‘ —•‘ǡ–Š‡’”‹ƒ”›–‹••—‡ ”‡•’‘•‹„Ž‡ˆ‘”‘Ǧ•Š‹˜‡”‹‰–Š‡”‘‰‡‡•‹•ȏͳͶȂͳ͸ǡʹͲȐǤŠ‡”‡‹•‡˜‹†‡ ‡ǡŠ‘™‡˜‡”ǡˆ‘”ƒ †‹”‡ –”‘Ž‡‘ˆ—• Ž‡ȏ͵ͺȐƒ†Ž‹˜‡”ȏ͵ͻȐ‹‘Ǧ•Š‹˜‡”‹‰–Š‡”‘‰‡‡•‹•ƒ•™‡ŽŽǤŠ‡”‘Ž‡‘ˆ –Š‡Ž‹˜‡”–‘•—’’Ž›—–”‹‡–•ˆ‘”ˆ— –‹‘Šƒ•Šƒ”†Ž›„‡‡•–—†‹‡†ǡ„—–”‡ ‡–Ž›‰ƒ‹‡† ”‡‡™‡†‹–‡”‡•–ȏͳͻǡʹͲȐǤ‹ ‘š‡–ƒŽǤȏͳͻǡʹͲȐ”‡˜‡ƒŽ‡†Š‘™ ‘Ž†‡š’‘•—”‡‡Šƒ ‡•Ž‹’‘Ž›•‹• ‹™Š‹–‡ƒ†‹’‘•‡–‹••—‡ƒ†•—’’Ž‹‡•–Š‡Ž‹˜‡”™‹–Š •Ǥ. ͳ͵͸.

(32) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice. A. H&E. ORO. KO typical KO severe (KO10). 4. B. Ń Severe (KO10). . Figure 5. KO individual with extreme phenotype. ‹•–‘Ž‘‰‹ ƒŽ ”‡’”‡•‡–ƒ–‹‘ ‘ˆ ƒ ”‡’”‡•‡–ƒ–‹˜‡  ‘—•‡Ž‹˜‡”ƒ†‘ˆ–Š‡Ž‹˜‡”‘ˆ–Š‡‘—•‡™‹–Š–Š‡‘•–‡š–”‡‡’Š‡‘–›’‡ȋͳͲȌȋAȌƒ†Š‡ƒ–ƒ’‘ˆ ƒ•‡Ž‡ –‡†•‡–‘ˆ‡–ƒ„‘Ž‹–‡•‹ͺǦ™‡‡•‘Ž†Ǧ‹ ‡ȋBȌǤ‹˜‡”•™‡”‡•–ƒ‹‡†„› ƒ‡‘–‘š›Ž‹ƒ† ‘•‹ȋŽ‡ˆ–Ȍƒ†‹Ž‡†ȋ”‹‰Š–ȌǤ. . ͳ͵͹.

(33) Chapter 4

(34)  –—”ǡ –Š‡  • ƒ –‹˜ƒ–‡ Ž‹˜‡”   ƒ† ‡Šƒ ‡ •‡ ”‡–‹‘ ‘ˆ ƒ ›ŽǦ ƒ”‹–‹‡• ‹–‘ –Š‡ „Ž‘‘†•–”‡ƒǤŠ‡•‡ƒ ›ŽǦ ƒ”‹–‹‡•’”‘˜‡†–‘„‡ ”‹–‹ ƒŽ•—„•–”ƒ–‡•ˆ‘”–Š‡”‘‰‡‡•‹•„› ȏͳͻǡʹͲȐǤ

(35) ‘—”‹ ‡ǡMcad†‡ˆ‹ ‹‡ ›Ž‡†–‘Š‹‰Š‡”Ž‡˜‡Ž•‘ˆƒ ›ŽǦ ƒ”‹–‹‡•‹–Š‡„Ž‘‘†Ǥ

(36) –‹• Ž‹‡Ž›–Šƒ––Š‡›•‡”˜‡ƒ••—„•–”ƒ–‡•ˆ‘”ǡ‡˜‡–Š‘—‰Š–Š‡›ƒ›„‡‘Ž›’ƒ”–Ž›‘š‹†‹œ‡††—‡ –‘–Š‡Mcad†‡ˆ‹ ‹‡ ›ǤŠ‹Ž‡™‡Šƒ˜‡‘‹ˆ‘”ƒ–‹‘ƒ„‘—–‡†‹—Ǧ Šƒ‹  ƒ’ƒ ‹–›‹ ‘ˆ–Š‡Mcad-†‡ˆ‹ ‹‡–‹ ‡ǡ‹–Šƒ•„‡‡”‡’‘”–‡†–Šƒ– ‰‡‡•ǡ‹ Ž—†‹‰Acadvl, Acadl and Acadm‰‡‡•ǡƒ”‡—’”‡‰—Žƒ–‡†‹ƒ†Ž‹˜‡”•‘ˆǦƒ†Ǧ‹ ‡–Šƒ–Šƒ˜‡ „‡‡ˆƒ•–‡†ƒ† ‘Ž†Ǧ‡š’‘•‡†ȏʹʹȐǤ Š‡–‹‡ ‘—”•‡•‘ˆ–Š‡„‘†›–‡’‡”ƒ–—”‡•Ȃ™‹–Š•‘‡‹ ‡•Š‘™‹‰‹‹ƒŽƒ† •‘‡•—„•–ƒ–‹ƒŽ–‡’‡”ƒ–—”‡†”‘’•‹„‘–Š‰”‘—’•ȋ—’’Ž‡‡–ƒ”›ˆ‹‰—”‡•ͳǦȌȂ•—‰‰‡•– –Šƒ–™‡ƒ”‡ƒ„Ž‡–‘„”‹‰‘Ž›ƒ•—„•‡–‘ˆǦ‹ ‡–‘–Š‡„”‹‘ˆƒ‡–ƒ„‘Ž‹  ‘ŽŽƒ’•‡Ǥ Š‹•™‘—Ž†‡š’Žƒ‹™Š›™‡‘„•‡”˜‡ƒ˜ƒ•–Š‡–‡”‘‰‡‡‹–›‘ˆ„‘†›–‡’‡”ƒ–—”‡•ƒ––Š‡‡†‘ˆ –Š‡ ‘Ž† ’‡”‹‘† ȋ ‹‰—”‡• ͳ ƒ† ȌǤ

(37)  –Š‹• ‘†‹–‹‘ǡ ™‡ ‘„•‡”˜‡† ƒ”‡† †‹ˆˆ‡”‡ ‡• ‹ ƒ‹‘Ǧƒ ‹†ƒ†Ž‹’‹†‡–ƒ„‘Ž‹•„‡–™‡‡–Š‡ƒ†‹ ‡ǤŽ‹‹–ƒ–‹‘‘ˆ–Š‹••–—†›‹• –Šƒ–™‡†‹†‘–‡ƒ•—”‡‡–ƒ„‘Ž‹ ˆŽ—š‡•„›‹•‘–‘’‡Žƒ„‡ŽŽ‹‰Ǥ ‘™‡˜‡”ǡ–Š‡ ‘’”‡Š‡•‹˜‡ ƒƒŽ›•‹• ‘ˆ „Ž‘‘† ƒ† Ž‹˜‡” ‡–ƒ„‘Ž‹–‡• ƒŽ”‡ƒ†› ›‹‡Ž†• ‹–‡”‡•–‹‰ ‹•‹‰Š–• ‹ ‡–ƒ„‘Ž‹  ƒ†ƒ’–ƒ–‹‘•ǡƒ•†‹• —••‡†„‡Ž‘™Ǥ Š‡–Š‡‘š‹†ƒ–‹‘‘ˆˆƒ––›ƒ ‹†•‹•†‡ˆ‹ ‹‡–ǡƒ‹‘ƒ ‹†•ƒ›•‡”˜‡ƒ•ƒƒŽ–‡”ƒ–‹˜‡ •—„•–”ƒ–‡ˆ‘”‘š‹†ƒ–‹‘ƒ†’”‘†— –‹‘Ǥ’‡ ‹ˆ‹ ‡–‘‰‡‹ ƒ‹‘ƒ ‹†•ȋ‘–ƒ„Ž›Ž‡— ‹‡ ƒ†Ž›•‹‡Ȍ ƒƒŽ•‘•‡”˜‡–‘”‡’Žƒ ‡ˆƒ––›ƒ ‹†•ƒ•ƒ•—„•–”ƒ–‡ˆ‘”‡–‘‡Ǧ„‘†›’”‘†— –‹‘ †—”‹‰ˆƒ•–‹‰Ǥ‘•‹•–‡–™‹–Š•— Š‹ ”‡ƒ•‡†—–‹Ž‹œƒ–‹‘ǡ‘•–ƒ‹‘ƒ ‹† ‘ ‡–”ƒ–‹‘•ǡ ‹ Ž—†‹‰Ž‡— ‹‡ƒ†Ž›•‹‡ǡ™‡”‡•—„•–ƒ–‹ƒŽŽ›Ž‘™‡”‹ǦŽ‹˜‡”• ‘’ƒ”‡†–‘ ‹ ‡ǡ’ƒ”–‹ —Žƒ”Ž›‹›‘—‰‹ ‡ȋ ‹‰—”‡͵ƒ†—’’Ž‡‡–ƒ”›ˆ‹‰—”‡͵ȌǤŽ‘‘†‡–‘‡Ǧ „‘†› Ž‡˜‡Ž• ™‡”‡ ‘– ƒŽ–‡”‡† ‹ Ǧ ‹ ‡ǡ •—‰‰‡•–‹‰ –Šƒ– ƒ‹‘ ƒ ‹†• ‡ˆˆ‹ ‹‡–Ž› ”‡’Žƒ ‡† ˆƒ––› ƒ ‹†• ƒ• •—„•–”ƒ–‡• ˆ‘” ‡–‘‰‡‡•‹•Ǥ Š‡ ˆƒ – –Šƒ– ’Žƒ•ƒ ƒ‹‘ ƒ ‹†• ™‡”‡ ‘”ƒŽ‹–Š‡Ǧ‹ ‡ǡ•—‰‰‡•–•–Šƒ––Š‡Š‡’ƒ–‹ ƒ‹‘Ǧƒ ‹††‡ˆ‹ ‹–‘”‹‰‹ƒ–‡•‹–Š‡ Ž‹˜‡”ƒ†ƒ›„‡ ƒ—•‡†„›ƒ‹ ”‡ƒ•‡††‡ƒ†Ǥ Š‡ Š‹‰Š‡” ’Žƒ•ƒ   ‘ ‡–”ƒ–‹‘• ‹ Ǧ ‘’ƒ”‡† –‘  ‹ ‡ ‹• ‘’ƒ–‹„Ž‡ ™‹–Š ‘Ž†Ǧ‹†— ‡† Ž‹’‘Ž›•‹• ȏͳͻǡʹͲȐ ‹ ‘„‹ƒ–‹‘ ™‹–Š ƒ ”‡†— ‡† ˆƒ––›Ǧƒ ‹† —–‹Ž‹œƒ–‹‘ †—‡ –‘ –Š‡  †‡ˆ‹ ‹‡ ›Ǥ  †‡ˆ‹ ‹‡ › ‘ˆ‡”•ƒ ”‹• ‘ˆ ƒ ——Žƒ–‹‘ ‘ˆ ‡†‹—Ǧ Šƒ‹ƒ ›ŽǦ‘‡•–‡”•ƒ††‡’Ž‡–‹‘‘ˆ‘Ǥ

(38) –Šƒ•„‡‡Š›’‘–Š‡•‹œ‡†–Šƒ––Š‹•ƒ› ƒ—•‡ –Š‡ ‡–ƒ„‘Ž‹  ‘ŽŽƒ’•‡ ‘ˆ  ’ƒ–‹‡–• ȏͶͲȂͶʹȐǤ

(39) –‡”‡•–‹‰Ž›ǡ ƒŽŽ Ž‹’‹†‘‹  ƒŽ–‡”ƒ–‹‘•–Šƒ–™‡‘„•‡”˜‡‹Ǧ‹ ‡ǡƒ›„‡‹–‡”’”‡–‡†ƒ•ƒ’”‘–‡ –‹‘ƒ‰ƒ‹•–•— Š ‘†‡’Ž‡–‹‘Ǥ ‹”•–ǡ™‡‘„•‡”˜‡†–Šƒ–Ž‹˜‡”ƒ ›ŽǦ ƒ”‹–‹‡•™‡”‡‘”ƒŽ‹Ǧ‹ ‡Ǥ  ›ŽǦ ƒ”‹–‹‡•ƒ”‡‘ˆ–‡˜‹‡™‡†ƒ•ƒ”‡ˆŽ‡ –‹‘‘ˆ–Š‡‹” ‘‰ƒ–‡ƒ ›ŽǦ‘‡•–‡”•ǤŠ‹•™‘—Ž† •—‰‰‡•– –Šƒ– Ǧ ‹ ‡ ƒ‹–ƒ‹ ƒ ›ŽǦ‘ Š‘‡‘•–ƒ•‹• —†‡” ‘Ž† •–”‡••Ǥ ‡ ‘†ǡ ™‡ ‘„•‡”˜‡†‹ ”‡ƒ•‡†ƒ ›Ž ƒ”‹–‹‡Ž‡˜‡Ž•‹–Š‡„Ž‘‘†‘ˆǦ‹ ‡ȋ ‹‰—”‡ʹǦȌǤŠ‹•‹• ‘’ƒ–‹„Ž‡ ™‹–Š ‡Šƒ ‡† ‘Ž†Ǧ‹†— ‡† ƒ ›Ž ƒ”‹–‹‡ •‡ ”‡–‹‘ ˆ”‘ –Š‡ Ž‹˜‡” ȏͳͻǡʹͲȐ ƒ† ™‘—Ž† ‘–”‹„—–‡–‘ƒŽŽ‡˜‹ƒ–‹‰Š‡’ƒ–‹ ƒ ›ŽǦ ƒ”‹–‹‡ƒ†ƒ ›ŽǦ‘ƒ ——Žƒ–‹‘Ǥ

(40)  ‘–”ƒ•–ǡ Ǧ‹ ‡–Šƒ–™‡”‡‘Ž›ˆƒ•–‡†ǡ„—–‘–‡š’‘•‡†–‘ ‘Ž†ǡ†‹†‘–ƒ‹–ƒ‹Š‡’ƒ–‹ ƒ ›ŽǦ ƒ”‹–‹‡ Š‘‡‘•–ƒ•‹• ȏͶ͵Ȑǡ •—‰‰‡•–‹‰ –Šƒ– ƒ ›ŽǦ ƒ”‹–‹‡ •‡ ”‡–‹‘ †—”‹‰ ‘Ž†Ǧ‡š’‘•—”‡ ƒ††•–‘–Š‡Š‘‡‘•–ƒ•‹•Ǥ‡š–ǡ™‡‘„•‡”˜‡†Š‹‰Š‡” Ž‡˜‡Ž•ǡ™Š‹ Š™ƒ•‘•–’”‘‘— ‡† ˆ‘” • ‘–ƒ‹‹‰͸ǦͳͶƒ ›Ž Šƒ‹•ȋ ‹‰—”‡ͶǦ ȌǤ

(41)  ‘”’‘”ƒ–‹‘‘ˆ‡š ‡••ƒ ›Ž‘‹‡–‹‡•‹  • ™‘—Ž† ƒŽ•‘ ƒ†† –‘ –Š‡ ’”‡˜‡–‹‘ ‘ˆ ƒ ›ŽǦ‘ ƒ ——Žƒ–‹‘ ƒ† •—„•‡“—‡– ‘ †‡’Ž‡–‹‘Ǥ͸ǦͳͲƒ ›ŽǦ‘‡•–‡”•ƒ”‡–Š‡’”‡ˆ‡””‡†•—„•–”ƒ–‡•‘ˆ–Š‡‡œ›‡ƒ†ƒ”‡ ͳ͵ͺ.

(42) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice ƒ’’ƒ”‡–Ž›ƒŽ•‘”‡”‘—–‡†‹–‘ •›–Š‡•‹•ǤŠ‡‹ ”‡ƒ•‡†ͳʹǦͳͶƒ ›Ž ‘–‡–‘ˆ–Š‡ ’‘‘Ž ƒ„‡‡š’Žƒ‹‡†‡‹–Š‡”„›†‹”‡ –”‡”‘—–‹‰‘ˆ–Š‡•‡‹–‡”‡†‹ƒ–‡•ˆ”‘ ‘š‹†ƒ–‹‘‘”„› ’ƒ”–‹ƒŽ Šƒ‹‡Ž‘‰ƒ–‹‘‘ˆ͸ǦͳͲ‹–‡”‡†‹ƒ–‡•’”‹‘”–‘–Š‡‹”‹ ‘”’‘”ƒ–‹‘‹–‘ •Ǥ ‹ƒŽŽ›ǡ ™‡ ™‘†‡”‡† ™Šƒ– ‹• –Š‡ ”‡Ž‡˜ƒ ‡ ‘ˆ –Š‹• •–—†› ˆ‘” –Š‡ —†‡”•–ƒ†‹‰ ‘ˆ Š—ƒǤŠ‡ˆƒ ––Šƒ–‘Ž››‘—‰Ǧ‹ ‡‡šŠ‹„‹–‡†Ž‘™‡”„Ž‘‘†‰Ž— ‘•‡Ž‡˜‡Ž• –Šƒ–Š‡‹”™‹Ž†Ǧ–›’‡ ‘—–‡”’ƒ”–•†—”‹‰•‡˜‡”‡‡‡”‰‡–‹ •–”‡••ǡ•—‰‰‡•–•–Šƒ––Š‡›ƒ”‡ƒ‰‘‘† ‘†‡Žˆ‘”Š—ƒ†‹•‡ƒ•‡ǤŽ•‘‹Š—ƒǡ—–”‡ƒ–‡† Š‹Ž†”‡ƒ”‡–›’‹ ƒŽŽ›ƒ–”‹•‘ˆ Š›’‘‰Ž› ‡‹ƒ †—”‹‰ –Š‡ ˆ‹”•– ›‡ƒ”• ‘ˆ Ž‹ˆ‡ ™Š‡ ˆƒ•–‡† ‹ ‘„‹ƒ–‹‘ ™‹–Š ƒ ƒ††‹–‹‘ƒŽ –”‹‰‰‡”ǡ „—– Ž‡•• ˜—Ž‡”ƒ„Ž‡ ™Š‡ –Š‡› ‰”‘™ ‘Ž†‡” ȏͶͶȐǤ

(43)  Š—ƒ  Š›’‘‡–‘–‹  Š›’‘‰Ž› ‡‹ƒ‹•‘•–‘ˆ–‡”‡’‘”–‡†–‘‘ —”—’‘ƒ ‘„‹ƒ–‹‘‘ˆˆƒ•–‹‰ƒ†‹–‡” —””‡– ‹ŽŽ‡••ȏͳǡ͸ȂͻȐǤŠ‹•‹•ǡŠ‘™‡˜‡”ǡ†‹ˆˆ‹ —Ž––‘‹‹ ‹–Š‡Žƒ„ǤŠ‡”‡ƒ”‡ƒŽ•‘”‡’‘”–•‘ˆ ‘Ž† •‡•‹–‹˜‹–› ‘ˆ  ’ƒ–‹‡–• ȏͺǡ͵ͲǡͶͷȐǡ ƒ‹‰ ‘Ž† •–”‡•• ƒ ”‡Ž‡˜ƒ– ƒ† ‡š’‡”‹‡–ƒŽŽ› ƒ ‡••‹„Ž‡ Žƒ„‘”ƒ–‘”› ‹–‡”˜‡–‹‘Ǥ ›’‘–Š‡•‹œ‹‰ –Šƒ– –Š‡ ‘—•‡ ™‹–Š –Š‡ ‘•– •‡˜‡”‡ ’Š‡‘–›’‡ ‘—Ž†›‹‡Ž† ‹•‹‰Š–•‹–‘ –Š‡ ’”‘ ‡••‡• †—”‹‰ ‘” Ž‘•‡ –‘ ƒ ‡–ƒ„‘Ž‹  ”‹•‹•ǡ ™‡ œ‘‘‡†‹‘Ǧ‘—•‡ͳͲȋͳͲȌǤŠ‹•‘—•‡•Š‘™‡†„‘–ŠŠ‡’ƒ–‘•–‡ƒ–‘•‹•ƒ†ƒ †‡ ”‡ƒ•‡ ‘ˆ „Ž‘‘† ‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘ †—”‹‰ ‘Ž† ‡š’‘•—”‡Ǥ Š‹• ‹• ‹ Ž‹‡ ™‹–Š Š‡’ƒ–‘•–‡ƒ–‘•‹•‘„•‡”˜‡†‹’ƒ–‹‡–•™Š‘†‹‡†ˆ”‘‡–ƒ„‘Ž‹  ”‹•‹•ȋ„‹‘ Š‡‹ ƒŽŽ› Šƒ”ƒ –‡”‹œ‡†„›Š›’‘‡–‘–‹ Š›’‘‰Ž› ‡‹ƒȌǤͳͲ•Š‘™‡†Ž‘™‡” Ž‡˜‡Ž•‘ˆ–Š‡ƒ‹‘ƒ ‹†• –Šƒ– ‘–”‹„—–‡–‘–Š‡ƒ–‹‘š‹†ƒ– ƒ’ƒ ‹–›Ǥ ‘”†‹‰Ž›ǡŽ‘™‡”ƒ–‹‘š‹†ƒ– ƒ’ƒ ‹–›Šƒ•„‡‡ ”‡’‘”–‡†‹’ƒ–‹‡–•ȏͶ͸ȐǤŠ‡”‡ˆ‘”‡ǡ™Š‡–Š‡„Ž‘‘†‰Ž— ‘•‡ ‘ ‡–”ƒ–‹‘‹ ‘Ž†Ǧ ‡š’‘•‡† Ǧ ‹ ‡ †”‘’•ǡ ƒ• ‘„•‡”˜‡† ‹ ͳͲǡ •— Š ‹ ‡ ƒ› „‡ ƒ†‡“—ƒ–‡ ƒ‹ƒŽ ‘†‡Ž•ˆ‘”–Š‡‘•‡–‘ˆ–Š‡‡–ƒ„‘Ž‹  ”‹•‹•‹Š—ƒ†‹•‡ƒ•‡Ǥ ‹ƒŽŽ›ǡͳͲŠƒ†–Š‡Ž‘™‡•– „‘†› –‡’‡”ƒ–—”‡ ƒ– –‡”‹ƒ–‹‘Ǥ ‹ ‡ –‡’‡”ƒ–—”‡ ƒ „‡ ˆ‘ŽŽ‘™‡† ‘–‹—‘—•Ž› „› –Š‡ ‹’Žƒ–‡†Ž‘‰‰‡”•ǡ‹–•‡‡•ˆ‡ƒ•‹„Ž‡–‘—•‡‹–ƒ•ƒ”‡ƒ†‘—––‘•–—†›‹ ‡‹ƒ ‘–”‘ŽŽ‡†™ƒ› †—”‹‰‘”‡•‡˜‡”‡‡–ƒ„‘Ž‹  ”‹•‹•Ǥ ‡ ‘ Ž—†‡–Šƒ–›‘—‰ǡˆƒ•–‡†ǡƒ† ‘Ž†‡š’‘•‡†Ǧ‹ ‡ƒ”‡ƒ”‡Ž‡˜ƒ–‘†‡Žˆ‘”–Š‡ ‘•‡–‘ˆ‡–ƒ„‘Ž‹  ”‹•‹•‹’ƒ–‹‡–•Ǥ‡‘„•‡”˜‡†ƒ”‡†‡–ƒ„‘Ž‹ ƒŽ–‡”ƒ–‹‘•‹ Ž‹˜‡” ‡–ƒ„‘Ž‹• ‘ˆ ‘Ž†Ǧ‡š’‘•‡† ˆƒ•–‡† Ǧ ‹ ‡Ǥ Š‡ ”‡•—Ž–• ‘””‘„‘”ƒ–‡ ”‡ ‡– ˆ‹†‹‰• ‘ˆ –Š‡ ‹’‘”–ƒ ‡ ‘ˆ Š‡’ƒ–‹  ˆƒ––›Ǧƒ ‹† ‡–ƒ„‘Ž‹• †—”‹‰ ‘Ž† •–”‡•• ȏͳͻǡʹͲȐǤ Ž–‡”ƒ–‹‘•‹ƒ‹‘ƒ ‹†ƒ†Ž‹’‹†‡–ƒ„‘Ž‹•ƒ”‡‹Ž‹‡™‹–Šƒ ‘’‡•ƒ–‘”›”‡•’‘•‡‹ ™Š‹ Šƒ‹‘ƒ ‹†•’ƒ”–Ž›–ƒ‡‘˜‡”–Š‡”‘Ž‡‘ˆˆƒ––›ƒ ‹†•ƒ••—„•–”ƒ–‡•ˆ‘”ƒ†Ȁ‘”‡–‘‡ „‘†›•›–Š‡•‹•ǡƒ† •›–Š‡•‹••‡”˜‡•ƒ•ƒ•‹ˆ‘”‡š ‡••ƒ ›Ž‘‹‡–‹‡•ǡ–Š‡”‡„›’”‡˜‡–‹‰ †ƒ‰‡”‘—•ƒ ›ŽǦ‘ƒ ——Žƒ–‹‘ȏͶͲȂͶʹȐǤ. Materials and Methods Animals, Experimental design and Tissue sampling ƒŽ‡Ǧƒ†™‹Ž†Ǧ–›’‡ȋȌ‹ ‡‘ƒͷ͹Ȁ͸ „ƒ ‰”‘—†ȋ„ƒ  ”‘••‡†ˆ‘”ͳͲ‰‡‡”ƒ–‹‘• ȏͶ͹ȐȌ™‡”‡Š‘—•‡†‹ƒ–‡’‡”ƒ–—”‡ǦȋʹͳιȌƒ†Ž‹‰Š–Ǧ ‘–”‘ŽŽ‡†ˆƒ ‹Ž‹–›Ǥ‹ ‡™‡”‡ͳ͸ǦʹͲ™‡‡•‘ˆ ƒ‰‡ȋ›‘—‰ƒ†—Ž–‹ ‡Ȍ‘”ͺ™‡‡•‘ˆƒ‰‡ȋ›‘—‰‹ ‡Ȍƒ––Š‡•–ƒ”–‘ˆ–Š‡‡š’‡”‹‡–ǤŽŽ‡š’‡”‹‡–• ™‡”‡ƒ’’”‘˜‡†„›–Š‡–Š‹ •‘‹––‡‡ˆ‘”‹ƒŽš’‡”‹‡–•‘ˆ–Š‡‹˜‡”•‹–›‘ˆ ”‘‹‰‡Ǥ”‹‘” –‘–Š‡•–ƒ”–‘ˆ–Š‡‡š’‡”‹‡–ǡ‹ ‡™‡”‡ˆ‡†ƒ Š‘™ǦŽ‹‡•‡‹Ǧ•›–Š‡–‹ †‹‡–ȋͳʹͶͷͲǡ‡•‡ƒ” Š‹‡– ‡”˜‹ ‡•ǡ‹Œ‹Œ——”•–‡†‡ǡŠ‡‡–Š‡”Žƒ†•Ȍˆ‘”ͶǦ͸™‡‡•ǡƒ†Šƒ†ˆ”‡‡ƒ ‡••–‘†”‹‹‰™ƒ–‡”Ǥ —„•‡“—‡–Ž›ǡ–Š‡‹ ‡™‡”‡ˆƒ•–‡†ˆ‘”ͳͶŠ”•‘˜‡”‹‰Š–ƒ†–Š‡”‡ƒˆ–‡”’Žƒ ‡†‹ƒͷ釐˜‹”‘‡– ˆ‘”ͶǦ͸Š”•ǤŠ‡‹ ‡™‡”‡–‡”‹ƒ–‡†„› ƒ”†‹ƒ ’— –—”‡—†‡”‹•‘ˆŽ—”ƒ‡ƒ‡•–Š‡•‹ƒǤ™‘™‡‡• „‡ˆ‘”‡ ˆƒ•–‹‰ǡ ‹‹ƒ–—”‹œ‡† ‹’‹ŽŽ ‡’‡”ƒ–—”‡

(44) ’Žƒ– Ž‘‰‰‡”• ȋƒ–ƒ  ‹‡ ‡•

(45) –‡”ƒ–‹‘ƒŽǡ. ͳ͵ͻ. 4.

(46) Chapter 4 ‹‡•‘–ƒǡ Ȍ ™‡”‡ ‹’Žƒ–‡† ‹–”ƒ’‡”‹–‘‡ƒŽŽ› —†‡” ‹•‘ˆŽ—”ƒ‡ ƒ† ƒƒŽ‰‡•‹ƒ ™‹–Š ˆŽ—‹š‹Ȁ‡‰Ž—‹‡ ȋʹ ‰Ȁ‰ȌǤ – –Š‡ ‡† ‘ˆ –Š‡ ‘Ž† ‡š’‘•—”‡ ƒ† „‡ˆ‘”‡ –‡”‹ƒ–‹‘ǡ –Š‡ „‘†› –‡’‡”ƒ–—”‡™ƒ•ƒŽ•‘‡ƒ•—”‡†™‹–Šƒ”‡ –ƒŽ’”‘„‡ȋŠ›•‹–‡’

(47) •–”—‡–•ǡŽ‹ˆ–‘ǡ ǡȌǤ—”‹‰ –Š‡ ‡š’‡”‹‡–ǡ „Ž‘‘† ‰Ž— ‘•‡ ƒ† ‡–‘‡ „‘†› ‘ ‡–”ƒ–‹‘• ™‡”‡ ‡ƒ•—”‡† „› ƒ ‡‘— Š Ž–”ƒƒ•›̺‘‹–‘”ȋ‹ˆ‡ ƒ‡‡Ž—šǡ‡‡”•‡ǡ‡Ž‰‹—Ȍƒ†’–‹— ‡‡†‘‹–‘”ȋ„‘––‹ƒ„‡–‡• ƒ”‡–†Ȍǡ”‡•’‡ –‹˜‡Ž›Ǥˆ–‡”–‡”‹ƒ–‹‘–Š‡Ž‹˜‡”™ƒ•”‡‘˜‡†“—‹ Ž›ǡ™‡‹‰Š‡†ǡƒ†ƒ’‹‡ ‡‘ˆ –Š‡Ž‹˜‡”ˆ”‡‡œ‡Ǧ Žƒ’‡†Ǣƒ•‡ ‘†’‹‡ ‡™ƒ•‹‡”•‡†‹ͳͲΨ„—ˆˆ‡”‡†ˆ‘”ƒŽ‹ƒ†ƒ–Š‹”†’‹‡ ‡™ƒ• ˆ‹šƒ–‡†‹‹••—‡Ǧ‡̺ƒ†ˆ”‘œ‡ǤŽ‘‘†•’‘–•™‡”‡–ƒ‡‹‡†‹ƒ–‡Ž›„‡ˆ‘”‡ˆƒ•–‹‰ǡƒ–ͳͶŠ‘—”• ƒˆ–‡”–Š‡•–ƒ”–‘ˆˆƒ•–‹‰ȋ‹Ǥ‡Ǥ„‡ˆ‘”‡‡š’‘•‹‰–Š‡‹ ‡–‘ ‘Ž†Ȍƒ†ƒ––‡”‹ƒ–‹‘ȋƒˆ–‡” ‘Ž†‡š’‘•—”‡ȌǤ . Metabolite analysis ‘ †‡–‡”‹‡ Š‡’ƒ–‹  –”‹‰Ž› ‡”‹†‡ Ž‡˜‡Ž•ǡ ˆ”‡‡œ‡Ǧ Žƒ’‡† Ž‹˜‡” ™ƒ• ”—•Š‡† ‹ Ž‹“—‹† ‹–”‘‰‡Ǥ —„•‡“—‡–Ž›ʹͲΨȋ™Ȁ˜ȌŽ‹˜‡”Š‘‘‰‡ƒ–‡•™‡”‡’”‡’ƒ”‡†‹ȋ’ ͹ǤͶȌǤ‘–ƒŽŽ‹˜‡”Ž‹’‹†•™‡”‡ ‡š–”ƒ –‡†ƒ ‘”†‹‰–‘ȏͶͺȐǡ”‡†‹••‘Ž˜‡†‹ʹΨ”‹–‘ǦͳͲͲƒ†ƒƒŽ›•‡†™‹–Šƒ ‘‡” ‹ƒŽŽ›ƒ˜ƒ‹Žƒ„Ž‡ ‹– ˆ‘” –”‹‰Ž› ‡”‹†‡ ȋ‘ Š‡ ‹ƒ‰‘•–‹ •ǡ ƒŠ‡‹ǡ ‡”ƒ›Ȍ ƒ ‘”†‹‰ –‘ –Š‡ ƒ—ˆƒ –—”‡”ǯ• ‹•–”— –‹‘•Ǥ ‹‘Ǧƒ ‹† Ž‡˜‡Ž• ™‡”‡ ‡ƒ•—”‡† ƒ ‘”†‹‰ –‘‘‘”‡ et al. ȏͶͻȐǤ ›ŽǦ ƒ”‹–‹‡ Ž‡˜‡Ž• ™‡”‡ ‡ƒ•—”‡† ƒ ‘”†‹‰ –‘ ‡”• et al. ȏͷͲȐǤ   ‘ ‡–”ƒ–‹‘• ™‡”‡ †‡–‡”‹‡† ™‹–Š ƒ ‘‡” ‹ƒŽŽ›ƒ˜ƒ‹Žƒ„Ž‡‹ƒ•›• ‹–ȋ‹ƒ•›•‹ƒ‰‘•–‹ ›•–‡• „ ǡ ‡”ƒ›ȌǤ . Liver lipid extraction ‹˜‡”–‹••—‡™ƒ•Š‘‘‰‡‹œ‡†‹ͲǤͻΨƒŽ–‘’”‡’ƒ”‡ƒʹͲΨȋ™Ȁ˜ȌŠ‘‘‰‡ƒ–‡Ǥ‡—•‡†ƒ‰”‘—’‘ˆ Ž‹’‹†• ȋ’—” Šƒ•‡† ˆ”‘ ˜ƒ–‹ ‘Žƒ” Ž‹’‹†•ǡ Žƒ„ƒ•–‡”ǡ Ȍ ƒ• ‹–‡”ƒŽ •–ƒ†ƒ”†• ˆ‘” ‘””‡ –‹‰ –Š‡ ˜ƒ”‹ƒ–‹‘‹Ž‹’‹†‡š–”ƒ –‹‘ƒ†ƒƒŽ›–‹ ƒŽƒƒŽ›•‹•Ǥ

(48) –‡”ƒŽ•–ƒ†ƒ”†• ‘–ƒ‹Ͷ‘Ž‘ˆ‡ƒ Š‘ˆ–Š‡ ˆ‘ŽŽ‘™‹‰ Ž‹’‹†•  ȋͳͶǣͲȌǡ  ȋͳͶǣͲȌǡȋͳ͹ǣͳȌǡ  ȋͳ͹ǣͲȀͳ͹ǣͲȌǡ ȋʹ͵ǣͲȀʹ͵ǣͲȌǡ  ȋͳͶǣͲȀͳͶǣͲȌǡ ȋͳ͹ǣͲȀͳ͹ǣͲȌǡ  ȋͳ͹ǣͲȀͳ͹ǣͲȌǡ   ȋͳ͹ǣͲȀͳ͹ǣͲȌǡ  ȋͳͶǣͲȌͶǡ   ȋͳͻǣʹȀͳͻǣʹȀͳͻǣʹȌǡ  ȋͳ͹ǣͲȌǡ  ȋͳͶǣͲȌƒ†ͺ‘Ž‘ˆ ȋͳͶǣͲȌǤ

(49) –‡”ƒŽ•–ƒ†ƒ”†•™‡”‡†‹••‘Ž˜‡†‹ʹͲͲɊ Ž͵ǣ‡ ȋͳǣͳǡ˜Ȁ˜Ȍ ƒ† ™‡”‡ ƒ††‡† –‘ ‡ƒ Š ’’‡†‘”ˆ –—„‡ ƒ† ’”‡Ǧ†”‹‡† ‹ ˜ƒ —— ‡–”‹ˆ—‰‡ ƒ– Ͷͷι „‡ˆ‘”‡ Ž‹’‹† ‡š–”ƒ –‹‘Ǥ‹˜‡”Š‘‘‰‡ƒ–‡ȋͷͲɊ ‘””‡•’‘†‹‰–‘γͳͲ‰Ȍ™ƒ•ƒ††‡†–‘–Š‡–—„‡• ‘–ƒ‹‹‰–Š‡ ’”‡Ǧ†”‹‡†Ž‹’‹†•–ƒ†ƒ”†•Ǥ‹’‹†‡š–”ƒ –‹‘™ƒ•’‡”ˆ‘”‡†ƒ ‘”†‹‰–‘ȏͷͳȐ™‹–Š•Ž‹‰Š–‘†‹ˆ‹ ƒ–‹‘Ǥ

(50)  „”‹‡ˆǡ ͵ͲͲ Ɋ ‡  ™ƒ• ‹š‡† ™‹–Š ͷͲ Ɋ Ž‹˜‡” Š‘‘‰‡ƒ–‡ ƒ† „ƒ–ŠǦ•‘‹ ƒ–‡† ˆ‘” ͷ ‹Ǥ —„•‡“—‡–Ž›ǡͳͲͲͲ Ɋ™ƒ•ƒ††‡†–‘–Š‡‹š–—”‡ƒ†‹ —„ƒ–‡†‘ƒ•Šƒ‡”ƒ–ͻͲͲ”’ˆ‘” ʹͲ‹ƒ–”‘‘–‡’‡”ƒ–—”‡Ǥƒ’Ž‡•™‡”‡ ‡–”‹ˆ—‰‡†ƒ–͵ͲͲͲ‰ˆ‘”ͳͲ‹–‘‹†— ‡ ‘’Ž‡–‡’Šƒ•‡ •‡’ƒ”ƒ–‹‘ǤŠ‡—’’‡”’Šƒ•‡ȋͻͲͲɊȌ™ƒ•–”ƒ•ˆ‡””‡†–‘ƒ‡™–—„‡ǤŠ‡Ž‘™‡”’Šƒ•‡™ƒ•”‡Ǧ‡š–”ƒ –‡† ™‹–Š͸ͲͲɊȀ‡ Ȁ ʹȋͳͲǣ͵ǣʹǤͷǡ˜Ȁ˜Ȁ˜ȌǤŠ‡ ‘„‹‡†ͳͶͲͲɊ—’’‡”’Šƒ•‡™ƒ•†”‹‡†‹ƒ ˜ƒ —— ‡–”‹ˆ—‰‡ƒ–ͶͷιǤŠ‡Ž‹’‹†’‡ŽŽ‡–™ƒ•†‹••‘Ž˜‡†‹ Ž͵Ȁ‡ ȋͳǣͳǡ˜Ȁ˜Ȍ‹‡†‹ƒ–‡Ž›’”‹‘” –‘Ǧ‡ƒ•—”‡‡–Ǥ . LC-MS measurement ‹’‹†‘‹ ƒƒŽ›•‹•™ƒ•’‡”ˆ‘”‡†‘Ž–‹ƒ–‡͵ͲͲͲŠ‹‰Š’‡”ˆ‘”ƒ ‡Ž‹“—‹† Š”‘ƒ–‘‰”ƒ’Š›ȋ Ȍ ‘—’Ž‡† ™‹–Š Ǧšƒ –‹˜‡ ”„‹–”ƒ’ ‹‰Š ‡•‘Ž—–‹‘ ƒ•• •’‡ –”‘‡–”› ȋȌ ȋŠ‡”‘  ‹‡–‹ˆ‹ ǡ ƒŽ–ŠƒǡȌǤ•‡’ƒ”ƒ–‹‘™ƒ•’‡”ˆ‘”‡†ƒ•†‡• ”‹„‡†’”‡˜‹‘—•Ž›ȏͷʹȐǤ†‡–‡ –‹‘™ƒ•†‘‡‹ ’‘•‹–‹˜‡‡Ž‡ –”‘•’”ƒ›‹‘‹œƒ–‹‘‘†‡™‹–Š†ƒ–ƒ†‡’‡†‡–Ȁƒ “—‹•‹–‹‘ƒ ”‘••–Š‡’”‡ —”•‘” ‹‘”ƒ‰‡ȀœͷͲΫͳ͹ͷͲǤ‡• ƒ™ƒ•’‡”ˆ‘”‡†ˆ‘ŽŽ‘™‡††‹”‡ –Ž›„›ͳͲȀ• ƒ•”‡•—Ž–‹‰ ‹ƒ†—–› › Ž‡‘ˆ̱ͳǤʹ•Ǥ‡•‘Ž—–‹‘ˆ‘”ˆ—ŽŽ• ƒƒ†ˆ”ƒ‰‡–•’‡ –”ƒƒ “—‹•‹–‹‘™‡”‡•‡–ƒ–͹ͲǡͲͲͲ ƒ†ͳ͹ǡͷͲͲ  ǡ”‡•’‡ –‹˜‡Ž›ǤŠ‡•‘—” ‡’ƒ”ƒ‡–‡”•™‡”‡•‡–ƒ•ˆ‘ŽŽ‘™•ǣ•‘—” ‡–‡’‡”ƒ–—”‡ͳͷͲ ιǡ•’”ƒ›˜‘Ž–ƒ‰‡ǣ͵Ǥͷǡ ƒ’‹ŽŽƒ”›–‡’‡”ƒ–—”‡ǣ͵ͲͲιǤ . ͳͶͲ.

(51) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice Lipidomic data analysis Š‡”‘  ƒŽ‹„—”̺ •‘ˆ–™ƒ”‡ ȋ˜‡”•‹‘ ͵ǤʹǤ͸͵ ȋŠ‡”‘  ‹‡–‹ˆ‹ ǡ ƒŽ–Šƒǡ Ȍ ™ƒ• —•‡† ˆ‘” †ƒ–ƒ ƒ “—‹•‹–‹‘Ǥ ”‘‰‡‡•‹• 

(52) ̺ •‘ˆ–™ƒ”‡ ȋƒ–‡”• ‘”’‘”ƒ–‹‘ǡ ‹Žˆ‘”†ǡ Ȍ ™ƒ• —•‡† ˆ‘” †ƒ–ƒ ’”‡’”‘ ‡••‹‰ ‹ Ž—†‹‰ ”‡–‡–‹‘ –‹‡ ƒŽ‹‰‡–ǡ ’‡ƒ ’‹ ‹‰ ƒ† ƒ‘–ƒ–‹‘ ‘ˆ –Š‡ Ǧ †ƒ–ƒǤ ‡ƒ’‹ ‹‰™ƒ•’‡”ˆ‘”‡†™‹–Šƒƒ„•‘Ž—–‡‹‘‹–‡•‹–›ˆ‹Ž–‡”‘ˆʹͲͲǡͲͲͲ ‘—–•Ǥ‹’‹†ƒ‘–ƒ–‹‘ ™ƒ• ’‡”ˆ‘”‡† „› •‡ƒ” Š‹‰ ’—„Ž‹•Š‡† Ž‹’‹† †ƒ–ƒ„ƒ•‡• ȋ —ƒ ‡–ƒ„‘Ž‘‡ ƒ–ƒ„ƒ•‡ǡ ‹’‹†ƒ’•ǡ ‹’‹†„Žƒ•–Ȍ„ƒ•‡†‘ƒ••ƒ —”ƒ ›ȋδͷ’’Ȍǡ‹•‘–‘’‹ •‹‹Žƒ”‹–‹‡•ǡƒ††— ––›’‡ǡȀ•’‡ –”ƒƒ† ‡Ž—–‹‘„‡Šƒ˜‹‘”‹ ‘’ƒ”‹•‘™‹–Š‘™Ž‹’‹†•‘ˆƒŠ‘‡Ǧƒ†‡†ƒ–ƒ„ƒ•‡ǤƒŽ›•‹•‘ˆ–Š‡ˆƒ––›Ǧƒ ›Ž ‘’‘•‹–‹‘‘ˆ ™ƒ•’‡”ˆ‘”‡†—•‹‰‹’‹†Š—–‡”ʹ•‘ˆ–™ƒ”‡ȏͷ͵Ȑ™‹–Šƒ’”‡†‡ˆ‹‡†–ƒ„Ž‡‘ˆ’‘••‹„Ž‡ ˆ”ƒ‰‡–• ȋ—’’Ž‡‡–ƒ”› –ƒ„Ž‡ ͷȌǤ ‹’‹† ‹–‡•‹–› ™ƒ• ‘”ƒŽ‹œ‡† –‘ –Š‡ ‹–‡•‹–› ‘ˆ –Š‡ ‘””‡•’‘†‹‰Ž‹’‹†•–ƒ†ƒ”†•‹–Š‡•ƒ‡ Žƒ••Ǥ

(53)  ƒ•‡‘ˆ–Š‡ƒ„•‡ ‡‘ˆŽ‹’‹†•–ƒ†ƒ”†•ǡ–Š‡‹–‡•‹–› ™ƒ•‘”ƒŽ‹œ‡†–‘–Š‡ƒ˜‡”ƒ‰‡‘ˆƒŽŽ•–ƒ†ƒ”†•Ǥ”‹ ‹’Ž‡ ‘’‘‡–ƒƒŽ›•‹•ȋȌ™ƒ•’‡”ˆ‘”‡† —•‹‰‡–ƒ„‘ƒŽ›•–ȏͷͶȐǤ . Histology ‘”ƒŽ‹Ǧˆ‹š‡†Ž‹˜‡”•™‡”‡’”‘ ‡••‡†ˆ‘”’ƒ”ƒˆˆ‹•‡ –‹‘‹‰ǡƒ†•–ƒ‹‡†™‹–ŠŠ‡ƒ–‘š›Ž‹ƒ†‡‘•‹Ǥ ”‘œ‡ǡ‹••—‡Ǧ‡̺ˆ‹š‡†Ž‹˜‡”•™‡”‡’”‘ ‡••‡†—•‹‰•–ƒ†ƒ”†‡–Š‘†•ƒ†•‡ –‹‘•™‡”‡•–ƒ‹‡† ™‹–Š ‹ŽǦ‡†ǦǤ Š‡ •Ž‹†‡• ™Š‡”‡  • ‘”‡† ˆ‘” ‘ƒŽ ‘Š‘Ž‹  ˆƒ––› Ž‹˜‡” †‹•‡ƒ•‡ ȋ Ȍ ƒ ‘”†‹‰ –‘ Ž‡‹‡”‡–ƒŽǤȏͷͷȐǤ . Statistical analysis Š‡•‹‰‹ˆ‹ ƒ ‡‘ˆ†‹ˆˆ‡”‡ ‡•ǡ‡š ‡’–ˆ‘”Ž‹’‹†‘‹ †ƒ–ƒǡ™‡”‡ƒ••‡••‡†—•‹‰’ƒ”ƒ‡–”‹ ȋ–—†‡–ǯ•–Ǧ –‡•–Ȍ‘”‘’ƒ”ƒ‡–”‹ –‡•–•ȋƒŠ‹–‡›–‡•–Ȍ†‡’‡†‹‰‘–Š‡‹”†‹•–”‹„—–‹‘Ǥ ‘” Šƒ‰‡•‹ –‹‡ǡ”‡’‡ƒ–‡†Ǧ‡ƒ•—”‡•™ƒ•—•‡†Ǥ–ƒ”‰‡–‡†Ž‹’‹†‘‹ †ƒ–ƒ™‡”‡ˆ‹”•–Ž‘‰Ǧ–”ƒ•ˆ‘”‡†ƒ† •—„•‡“—‡–Ž›†‹ˆˆ‡”‡ ‡•™‡”‡ƒ••‡••‡†™‹–Š–—†‡–ǯ•–Ǧ–‡•–ƒ†ƒ†Œ—•–‡†ˆ‘”—Ž–‹’Ž‡ ‘’ƒ”‹•‘Ǥ ’˜ƒŽ—‡‘ˆδͲǤͲͷ™ƒ• ‘•‹†‡”‡†•–ƒ–‹•–‹ ƒŽŽ›•‹‰‹ˆ‹ ƒ–Ǥ–ƒ–‹•–‹ ƒŽƒƒŽ›•‡•™‡”‡’‡”ˆ‘”‡†—•‹‰ ȋ˜ʹʹȌƒ†–Š‡†ƒ–ƒ™ƒ•˜‹•—ƒŽ‹œ‡†—•‹‰ ”ƒ’Šƒ†”‹••‘ˆ–™ƒ”‡ȋ ”ƒ’Šƒ†‘ˆ–™ƒ”‡

(54)  Ǥǡ˜‡”•‹‘ ͷǤͲͲǡʹͲͲ͹Ȍƒ†–Š‡‡–ƒ„‘ƒŽ›•–™‡„•‡”˜‡”ȏͷͶȐǤ . Author contributions  ǡǡ ǡ ǡ ƒ††‡•‹‰‡†–Š‡‡š’‡”‹‡–•Ǥ ǡ ƒ† ’‡”ˆ‘”‡†–Š‡ƒ‹ƒŽ ‡š’‡”‹‡–•Ǥ ǡǡ ǡ ǡǡǡǡǡ ’”‡’ƒ”‡†ƒ†ƒƒŽ›œ‡†–Š‡•ƒ’Ž‡•Ǥ ǡ ƒ†  ƒƒŽ›œ‡† –Š‡ †ƒ–ƒǤ  ǡ ǡ ǡ ǡ ǡ  ǡ  ǡ ǡ  ǡ  ™‡”‡ ‹˜‘Ž˜‡† ‹ ‹–‡”’”‡–ƒ–‹‘ ‘ˆ ”‡•—Ž–•Ǥ  ǡ ǡ  ǡ ƒ†  ™”‘–‡ –Š‡ ƒ—• ”‹’–Ǥ ŽŽ ƒ—–Š‘”• ‡†‹–‡† –Š‡ ƒ—• ”‹’–Ǥ . Acknowledgements Š‹•™‘”™ƒ••—’’‘”–‡†„›–Š‡‹˜‡”•‹–›‡†‹ ƒŽ‡–‡” ”‘‹‰‡ǤŠ‡ˆ—†‡”•Šƒ†‘”‘Ž‡‹•–—†› †‡•‹‰ǡ†ƒ–ƒ ‘ŽŽ‡ –‹‘ƒ†ƒƒŽ›•‹•ǡ†‡ ‹•‹‘–‘’—„Ž‹•Šǡ‘”’”‡’ƒ”ƒ–‹‘‘ˆ–Š‡ƒ—• ”‹’–ǤŠ‡ƒ—–Š‘”• Ž‹‡ –‘ –Šƒ ‹ ‡– Ǥ Ž‘• ˆ‘” ƒ”‡ˆ—Ž ƒ† ‹†‡’‡†‡– Š‡ ‹‰ ‘ˆ –Š‡ •–ƒ–‹•–‹ ƒŽ ƒƒŽ›•‹• ƒ† ƒ”‹‡‡‹–ˆ‘”ƒ••‹•–‹‰™‹–Š–‡”‹ƒ–‹‘•Ǥ . Declaration of Interests Š‡ƒ—–Š‘”•†‡ Žƒ”‡‘ ‘’‡–‹‰‹–‡”‡•–•Ǥ . References ͳǤ. ‹ƒŽ†‘ ǡ ƒ–‡” ǡ ‡‡––  Ǥ ƒ––›  ‹† ‘š‹†ƒ–‹‘ ‹•‘”†‡”•Ǥ — ‡˜ Š›•‹‘ŽǤ ʹͲͲʹǢ͸ͶȋͳȌǣͶ͹͹ȂͷͲʹǤ. ͳͶͳ. 4.

(55) Chapter 4 ʹǤ. ͵Ǥ. ͶǤ ͷǤ ͸Ǥ ͹Ǥ ͺǤ ͻǤ ͳͲǤ. ͳͳǤ. ͳʹǤ. ͳ͵Ǥ ͳͶǤ ͳͷǤ. ͳ͸Ǥ. ͳ͹Ǥ. ͳͺǤ ͳͻǤ. ͳͶʹ. ‘—–‡ ǡ ‹‘Žƒ–‡ ǡ ‡–—”ƒ  Ǥǡ ƒ†‡”•  Ǥ Š‡ ‹‘ Š‡‹•–”› ƒ† Š›•‹‘Ž‘‰› ‘ˆ ‹–‘ Š‘†”‹ƒŽ ƒ––›  ‹† ȾǦš‹†ƒ–‹‘ ƒ†

(56) –• ‡‡–‹  ‹•‘”†‡”•Ǥ — ‡˜ Š›•‹‘ŽǤ ʹͲͳ͸Ǣ͹ͺȋͳȌǣʹ͵ȂͶͶǤ ‘––‡”—• ǡŽ‡‡‡” ǡò•–

(57) ǡ ‡”†‹ƒ†—••‡ǡ

(58) ŒŽ•–ǡ‹Œ„—”‰ ǡƒ†‡”• ǡ‹••‡”. ǡ ‘—–‘‘’‡” Ǥ‹•‘”†‡”•‘ˆ‹–‘ Š‘†”‹ƒŽŽ‘‰Ǧ Šƒ‹ˆƒ––›ƒ ‹†‘š‹†ƒ–‹‘ƒ†–Š‡ ƒ”‹–‹‡ •Š—––Ž‡ǤʹͲͳͺǢ ‡”• ǡƒ’”‘•‡ ǡƒ‡ ǡƒ‡” ‹Ž•–ǡ’ƒǡ‹– Ǥ ƒˆ‡ƒ†—•ƒˆ‡ †—”ƒ–‹‘‘ˆˆƒ•–‹‰ˆ‘” Š‹Ž†”‡™‹–Š†‡ˆ‹ ‹‡ ›Ǥ—” ‡†‹ƒ–”ǤʹͲͲ͹Ǣͳ͸͸ȋͳȌǣͷȂͳͳǤ ƒŽ–‡” Ǥ ‘Ž‡”ƒ ‡ –‘ ˆƒ•–ǣ ƒ–‹‘ƒŽ ƒ† ’”ƒ –‹ ƒŽ ‡˜ƒŽ—ƒ–‹‘ ‹ Š‹Ž†”‡ ™‹–Š Š›’‘‡–‘ƒ‡‹ƒǤ 

(59) Š‡”‹–‡–ƒ„‹•ǤʹͲͲͻǢ͵ʹȋʹȌǣʹͳͶȂ͹Ǥ ‡‡––  Ǥ ƒ–Š‘’Š›•‹‘Ž‘‰› ‘ˆ ˆƒ––› ƒ ‹† ‘š‹†ƒ–‹‘ †‹•‘”†‡”•Ǥ 

(60) Š‡”‹– ‡–ƒ„ ‹•Ǥ ʹͲͳͲǢ͵͵ȋͷȌǣͷ͵͵Ȃ͹Ǥ ‹•Š™ƒƒ–Š Ǥ ƒ––› ƒ ‹† „‡–ƒǦ‘š‹†ƒ–‹‘ †‹•‘”†‡”•ǣ  „”‹‡ˆ ”‡˜‹‡™Ǥ  ‡—”‘• ‹Ǥ ʹͲͳ͸Ǣʹ͵ȋͳȌǣͷͳȂͷǤ  Šƒ–œƒǤǡ•‡ƒ—‡”ǤŠ‡ Ž‹‹ ƒŽƒ‹ˆ‡•–ƒ–‹‘‘ˆ†‡ˆ‹ ‹‡ ›ǣŠƒŽŽ‡‰‡•–‘™ƒ”†• ƒ†—Ž–Š‘‘†‹–Š‡• ”‡‡‡†’‘’—Žƒ–‹‘Ǥ 

(61) Š‡”‹–‡–ƒ„‹•ǤʹͲͳͲǢ͵͵ȋͷȌǣͷͳ͵ȂʹͲǤ ‘Ž‡”Ǧ–‘—™Ǥ†‡ˆ‹ ‹‡ ›‘„‡ǡ‘”‘––‘„‡ƒ–”‹•ǤʹͲͳͶǤ ‘Ž™ƒ‹ ǡ ƒǡ‹ƒǡŠƒ”‡” ǡ‘ Ž‡› ǡ‹ƒŽ†‘ǡƒ–‡”ǡ Š‘‡„ǡ‘‘†Ǥ ‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ‘ †‡Š›†”‘‰‡ƒ•‡ †‡ˆ‹ ‹‡ › ‹ ‰‡‡Ǧ–ƒ”‰‡–‡† ‹ ‡Ǥ ‘ ‡‡–Ǥ ʹͲͲͷǢͳȋʹȌǣͲʹͲͷȂͳʹǤ ˜ƒ—‡ǡ‘Ž‡”Ǧ‘—™ǡ ‡”†‹‰ǡŽ‡‡‡”ǡ‘Ž–‡”• ǡ˜ƒ‹Œ– ǡƒ”–‹‡•Ǧ ǡ ‹‡œ‡Ǧ‘‹‰ǡ ‡‹‡”ǡ‡”‡–‹‡” ǡ ”‘‡ǡ‡‹Œ‰‘—†Ǧ ǡ‡”• ǡƒ‡”Ǥ ‹˜‹‰‘–Š‡‡†‰‡ǣ•—„•–”ƒ–‡ ‘’‡–‹–‹‘‡š’Žƒ‹•Ž‘••‘ˆ”‘„—•–‡••‹‹–‘ Š‘†”‹ƒŽˆƒ––›Ǧ ƒ ‹†‘š‹†ƒ–‹‘†‹•‘”†‡”•Ǥ‹‘ŽǤʹͲͳ͸ǢͳͶȋͳȌǣͳͲ͹Ǥ ‡””‡ƒ ǡ‡”• ǡƒ‹Œ ǡŽ‘•ǡ ‡”†‹‰ǡ ƒ˜‹‰ƒǡ‹‡–‰‡ ǡǫǫŽŽ‡”ǡ‹–. ǡ —‹’‡”• ǡ ‡‹Œ‰‘—†  Ǥ ‹•–—”„‡† Š‡’ƒ–‹  ƒ”„‘Š›†”ƒ–‡ ƒƒ‰‡‡– †—”‹‰ Š‹‰Š ‡–ƒ„‘Ž‹  †‡ƒ† ‹ ‡†‹—Ǧ Šƒ‹ ƒ ›ŽǦ‘ †‡Š›†”‘‰‡ƒ•‡ ȋȌǦ†‡ˆ‹ ‹‡– ‹ ‡Ǥ ‡’ƒ–‘Ž‘‰›ǤʹͲͲͺǢͶ͹ȋ͸ȌǣͳͺͻͶȂͻͲͶǤ ’‹‡‡”‘‡––‡”ǡ‘‘†Ǥ‹–‘ Š‘†”‹ƒŽˆƒ––›ƒ ‹†‘š‹†ƒ–‹‘†‹•‘”†‡”•ǣƒ–Š‘’Š›•‹‘Ž‘‰‹ ƒŽ •–—†‹‡•‹‘—•‡‘†‡Ž•Ǥ 

(62) Š‡”‹–‡–ƒ„‹•ǤʹͲͳͲǢ͵͵ȋͷȌǣͷ͵ͻȂͶ͸Ǥ Š‹„ƒ–ƒ ǡ±”—••‡ ǡƒŽŽ‡”ƒ†ƒǡ—‘™‹‡ ‹ Ǥ‘Ž†‡š’‘•—”‡”‡˜‡”•‡•‹Š‹„‹–‘”›‡ˆˆ‡ –•‘ˆ ˆƒ•–‹‰‘’‡”‹’Š‡”ƒŽ‰Ž— ‘•‡—’–ƒ‡‹”ƒ–•Ǥ Š›•‹‘ŽǤͳͻͺͻǢʹͷ͹ȋͳ–ʹȌǣͻ͸ȂͳͲͳǤ ג‡œǦ‘”‹ƒ‘ ǡ ‡”ž†‡œǦג‡œ ǡƒ’‡Žǡ‹ŽŽƒ””‘›ƒ ǡ

(63) ‰Ž‡•‹ƒ•ǡŽ‡ƒ›Ǥ‹‘ ƒ ‹†ƒ†‰Ž— ‘•‡—’–ƒ‡„›”ƒ–„”‘™ƒ†‹’‘•‡–‹••—‡Ǥˆˆ‡ –‘ˆ ‘Ž†Ǧ‡š’‘•—”‡ƒ†ƒ Ž‹ƒ–‹‘Ǥ ‹‘ Š‡ ǤͳͻͺͺǢʹͷʹȋ͵ȌǣͺͶ͵ȂͻǤ ‡‹” ǡƒƒ‰‡ǡ›‘ŽǡŠ‘†‡• ǡ›Ž‡ ǡ Ž‡– Š‡”ǡ”ƒ˜‡ ǡƒ‡Ž‹ ǡ”ƒ‡ ǡ. ”‡‰‘”‹ƒ†‡•ǡ•Š–‘ǡ‡‹”ǡ˜ƒ‡‡ ǡƒ”’‡ ǡƒŽ‡”ǡ–‹•‘ Ǥ—„•–ƒ–‹ƒŽ ‡–ƒ„‘Ž‹  –‹˜‹–›‘ˆ —ƒ”‘™†‹’‘•‡‹••—‡†—”‹‰ƒ”‘†‹–‹‘•ƒ†‘Ž†Ǧ

(64) †— ‡† ‹’‘Ž›•‹•‘ˆ‘ ƒŽ”‹‰Ž› ‡”‹†‡•Ǥ‡ŽŽ‡–ƒ„ǤʹͲͳͺǢʹ͹ȋ͸Ȍǣͳ͵ͶͺǦͳ͵ͷͷǤ‡ͶǤ ƒ”–‡Ž–ǡ”—•ǡ‡‹‡”ǡ ‘Š‡„‡”‰ ǡ

(65) ––”‹ Š ǡ‡Ž†• Š—•ǡƒ—Ž ǡ”‘•†‘”ˆ

(66) ǡ ‡ŽŽ‡” ǡ ƒ—”‹• Š ǡ › ŠòŽŽ‡” ǡ ‘”†–• ǡ ‹‹‰‡” ǡ ”—‡‰‡Žƒ ǡ ”‡—† ǡ ‹‡Ž•‡ǡ‡”‡Žǡ ‡‡”‡ Ǥ”‘™ƒ†‹’‘•‡–‹••—‡ƒ –‹˜‹–› ‘–”‘Ž•–”‹‰Ž› ‡”‹†‡ Ž‡ƒ”ƒ ‡Ǥƒ– ‡†ǤʹͲͳͳǢͳ͹ȋʹȌǣʹͲͲȂ͸Ǥ Š‹‘–ƒǡƒƒƒǡ—‰ƒ‘Ǥˆˆ‡ –‘ˆ‘”‡’‹‡’Š”‹‡‘‰Ž— ‘‡‘‰‡‡•‹•‹’‡”ˆ—•‡†Ž‹˜‡”• ‘ˆ ‘Ž†Ǧ‡š’‘•‡†”ƒ–•Ǥ Š›•‹‘Ž‡–ƒ„ǤʹͲͳ͹ǢʹͶͻȋ͵ȌǣʹͺͳȂ͸Ǥ ‹ ‘š ǡ ‡‘‰Š‡‰ƒ ǡ ƒ• Š‡ ǡ ‡•ƒ”† ǡ ƒ•“—ƒŽ‹ ǡ ‹ƒ‘ ǡ ‡‡ ǡ ‹ƒ‰ ǡ — 

(67) ǡ ‡”•Šƒ™ǡ‘ƒ–‘ ǡ’–‡ǡ‘‰‘ǡ—––‡” ǡ Š”‡‹„‡”ǡ‡ Š‡”ǡ‘š ǡ‹ŽŽƒ—‡˜ƒ Ǥ. Ž‘„ƒŽƒŽ›•‹•‘ˆŽƒ•ƒ‹’‹†•

(68) †‡–‹ˆ‹‡•‹˜‡”Ǧ‡”‹˜‡† ›Ž ƒ”‹–‹‡•ƒ•ƒ —‡Ž‘—” ‡ˆ‘” ”‘™ ƒ–Š‡”‘‰‡‡•‹•Ǥ‡ŽŽ‡–ƒ„ǤʹͲͳ͹Ǣʹ͸ȋ͵ȌǣͷͲͻǦͷʹʹǤ‡͸Ǥ.

(69) Effect of fasting and cold stress on hepatic metabolism in MCAD-KO mice ʹͲǤ ʹͳǤ. ʹʹǤ. ʹ͵Ǥ. ʹͶǤ. ʹͷǤ. ʹ͸Ǥ ʹ͹Ǥ. ʹͺǤ. ʹͻǤ ͵ͲǤ. ͵ͳǤ ͵ʹǤ ͵͵Ǥ ͵ͶǤ. ͵ͷǤ ͵͸Ǥ. ͵͹Ǥ. „—”ƒ†ǤŠ‡‹˜‡”ƒ•ƒ —„‹Š‡”‘‰‡‡•‹•Ǥ‡ŽŽ‡–ƒ„ǤʹͲͳ͹Ǣʹ͸ȋ͵ȌǣͶͷͶȂͷǤ. ”‡ˆŠ‘”•–ǡ˜ƒ†‡‡—‡Ž ǡ‹Œǡ–‡‡„‡”‰‡ ǡ‘‘”–ƒ ǡ‡‡—™‡„—”‰Šǡ‹••‡” ǡ ‡”•–‡ǡ ”‹‡•‡ƒ ǡŠ‡‡ǡ‹••‡” Ǥ—Ž–‹’Ž‡‡ˆˆ‡ –•‘ˆ ‘Ž†‡š’‘•—”‡‘Ž‹˜‡”• ‘ˆƒŽ‡‹ ‡Ǥ †‘ ”‹‘ŽǤʹͲͳͺ—‰Ǣʹ͵ͺȋʹȌǣͻͳȂͳͲ͸Ǥ. ‘‡–œƒǡ‹ƒǡ‘‘†Ǥ‹ˆˆ‡”‡–‹ƒŽ‹†— –‹‘‘ˆ‰‡‡•‹Ž‹˜‡”ƒ†„”‘™ƒ†‹’‘•‡–‹••—‡ ”‡‰—Žƒ–‡†„›’‡”‘š‹•‘‡’”‘Ž‹ˆ‡”ƒ–‘”Ǧƒ –‹˜ƒ–‡†”‡ ‡’–‘”ǦȽ†—”‹‰ˆƒ•–‹‰ƒ† ‘Ž†‡š’‘•—”‡‹ ƒ ›ŽǦ‘†‡Š›†”‘‰‡ƒ•‡Ǧ†‡ˆ‹ ‹‡–‹ ‡Ǥ‘Ž ‡‡–‡–ƒ„ǤʹͲͲͷǢͺͶȋͳȌǣ͵ͻȂͶ͹Ǥ Š‘”‡ǡƒ”ƒ‹–”‹ǡ‡’ǡ’‡ƒƒ ǡ ”ƒŠƒǡ‘ƒšǤ‘Ž†Ǧ

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