Journal of Food and Nutrition Research
ISSN (Print): 2333-1119 ISSN (Online): 2333-1240 Website: https://www.sciepub.com/journal/jfnr Editor-in-chief: Prabhat Kumar Mandal
Open Access
Journal Browser
Go
Journal of Food and Nutrition Research. 2018, 6(7), 419-432
DOI: 10.12691/jfnr-6-7-1
Open AccessArticle

Dietary Modifications of Nitrogen Intake Decreases Inflammation and Promotes Rejuvenation of Spleen in Aged Mice

Claudia Romano1, Giovanni Corsetti1, , Evasio Pasini2, Vincenzo Flati3 and Francesco S Dioguardi4

1Department of Clinical & Experimental Sciences, Division of Human Anatomy and Physiopathology, University of Brescia, Brescia, Italy

2“S. Maugeri Foundation”, IRCCS, Cardiology Division, Medical Centre of Lumezzane, Brescia, Italy

3Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, L’Aquila, Italy

4Department of Clinical Sciences and Community Health, University of Milan, Milan, Italy

Pub. Date: July 14, 2018

Cite this paper:
Claudia Romano, Giovanni Corsetti, Evasio Pasini, Vincenzo Flati and Francesco S Dioguardi. Dietary Modifications of Nitrogen Intake Decreases Inflammation and Promotes Rejuvenation of Spleen in Aged Mice. Journal of Food and Nutrition Research. 2018; 6(7):419-432. doi: 10.12691/jfnr-6-7-1

Abstract

The spleen is a lymphoid organ with multiple functions including blood filtration and immune activity. Aging changes the spleen’s anatomy for both immune and stromal cells and can lead to immunesenescence, contributing to the increased rates of mortality and morbidity commonly observed in the elderly. Much evidence indicates that the combination of food quantity and quality can influence chronic inflammatory states in the spleen. Quantitative amino-acids (AA) adequacy is pivotal to maintain cell integrity in mammals. Aged mice feed with balanced essential-AA (EAA) formulation improved mitochondrial biogenesis and morphological and molecular changes in many organs, as well as increased lifespan. Here, we evaluated the inflammatory state of the spleen in aged male mice (fifteen months old) chronically fed for twelve months with a particular EAA-rich diet compared to a standard laboratory diet. This study found that chronic consumption of an EAA-rich diet decreased inflammation and modulated reticular and mitochondrial chaperones, mitochondrial function and cells survival, maintaining the correct architecture of the spleen. These changes could also be beneficial for immune system integrity, providing a possible theoretical-speculative basis for the role of EAA improving the quality of life of the elderly by probably slowing immune-senescence.

Keywords:
Essential Amino-acids spleen inflammation chaperones Klotho aging

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

Figure of 9

References:

[1]  Mebius, R.E., and Kraal., G., “Structure and function of the spleen”, Nature review. Immunology, 5 (8). 606-616. Aug 2005.
 
[2]  Brendolan, A., Rosado, M.M., Carsetti, R., Selleri, L., and Dear, T.N., “Development and function of the mammalian spleen”, BioEssays 29 (2). 166-177. February 2007.
 
[3]  Martin, F., and Kearney, J.F., “Marginal-zone B cells”, Nature Review. Immunology, 2 (5). 323-335. Review. May 2002.
 
[4]  Balogh, P., Horvath, G., and Szakal, A.K., “Immunoarchitecture of distinct reticular fibroblastic domains in the white pulp of mouse spleen”, Journal of histochemistry & cytochemistry. 52 (10). 1287-1298. October 2004.
 
[5]  Junt, T., Scandella, E., and Ludewig, B. Form follows function: lymphoid tissue microarchitecture in antimicrobial immune defence. Nature review. Immunology, 8 (10). 764-775. Oct 2008.
 
[6]  Aw, D., Hilliard, L., Nishikawa, Y., Cadman, E.T., Lawrence, R.A. and Palmer, D.B., “Disorganization of the splenic microanatomy in ageing mice”, Immunology, 148 (1). 92-101. May 2016.
 
[7]  Anderson, G., and Jenkinson, E.J., “Lymphostromal interactions in thymic development and function”, Nature Review. Immunology, 1 (1). 31-40. 1:31-40 October 2001.
 
[8]  Su, D.M., Aw, D., and Palmer, D.B., “Immunosenescence: a product of the environment?” Current Opinion in Immunology, 25 (4). 498-503. August 2013.
 
[9]  Aw, D., Silva, A.B., and Palmer, D.B., “Immunosenescence: emerging challenges for an ageing population”, Immunology, 120 (4). 435-446. Apr 2007.
 
[10]  Haynes, L., and Swain, S.L., “Why aging T cells fail: implications for vaccination”, Immunity, 24 (6). 663-666. June 2006.
 
[11]  Pawelec, G., Akbar, A., Caruso., C., Solana, R., Grubeck-Loebenstein, B., and Wikby, A., “Human immunosenescence: is it infectious?”, Immunological Review 205. 257-268. Jun 2005.
 
[12]  Chen, W.H., Kozlovsky, B.F., Effros, R.B., Grubeck-Loebenstein, B., Edelman, R., and Sztein, M.B., “Vaccination in the elderly: an immunological perspective”, Trends in Immunology, 30 (7). 351-319. Jul 2009
 
[13]  Gardner, I.D., “The effect of aging on susceptibility to infection”, Review of Infectious Diseases; 2 (5). 801-810. Sep-Oct 1980.
 
[14]  McElhaney, J.E., Zhou, X., Talbot, H.K., Soethout, E., Bleackley, R.C., Granville, D.J., and Pawelec, G., “The unmet need in the elderly: how immunosenescence, CMV infection, co-morbidities and frailty are a challenge for the development of more effective influenza vaccines”, Vaccine, 30 (12). 2060-2067. March 2012.
 
[15]  Neustadt, J., “Western diet and inflammation”, Integrative Medicine, 5. 14-18. Aug-Sept 2006.
 
[16]  Galland, L., “Diet and inflammation”, Nutrition in Clinical Practice, 25 (6). 634-640. Dec 2010.
 
[17]  Pasini, E., Corsetti, G., Aquilani, R., Romano, C., Picca, A., Calvani, R., and Dioguardi, F.S., “Protein-amino acid metabolism disarrangements: the hidden enemy of chronic age-related conditions” Nutrients, 10 (4). Pii: E391. March 2018.
 
[18]  Hou, Y., Yin, Y., and Wu, G., “Dietary essentiality of “nutritionally non-essential amino acids” for animals and humans”, Experimental Biology and Medicine (Maywood), 240 (8). 997-1007. August 2015.
 
[19]  Corsetti, G., Pasini, E., D’Antona, G., Nisoli, E., Flati, V., Assanelli, D., Dioguardi, F.S., and Bianchi R., “Morphometric changes induced by amino acid supplementation in skeletal and cardiac muscles of old mice”, The American journal of cardiology, 101 (11A). E26-E34. June 2008.
 
[20]  Flati, V., Pasini, E., D’Antona, G., Speca, S., Toniato, E., and Martinotti, S., “Intracellular mechanisms of metabolism regulation: the role of signaling via the mammalian target of rapamycin pathway and other routes”, The American journal of cardiology, 101 (11A). 16E-21E. June 2008.
 
[21]  D’Antona, G., Ragni, M., Cardile, A., Tedesco, L., Dossena, M., Bruttini, F., Caliaro, F., Corsetti, G., Bottinelli, R., Carruba, M.O., Valerio, A., and Nisoli E., “Branched-chain amino acid supplementation promotes survival and supports cardiac and skeletal muscle mitochondrial biogenesis in middle-aged mice”, Cell metabolism, 12 (4). 362-372. October 2010.
 
[22]  Stacchiotti A, Corsetti G, Lavazza A, Rezzani R. “Microscopic features of mitochondria rejuvenation by amino acids”, in: Antonio Méndez-Vilas editor, Current microscopy contributions to advances in science and technology. Formatex Research Center, Microscopy Series nº 5, vol. 2. 286-294. 2012.
 
[23]  Corsetti, G., Stacchiotti, A., Tedesco, L., D’Antona, G., Pasini, E., Dioguardi, F.S., Nisoli, E., Rezzani R., “Essential amino acid supplementation decreases liver damage induced by chronic ethanol consumption in rats”, International journal of immunopathology and pharmacology, 24 (3). 611-619. Jul-Sep 2011.
 
[24]  Tedesco, L., Corsetti, G., Ruocco, C., Ragni, M., Rossi, F., Carruba, M.O., Valerio, A., and Nisoli, E., “A specific amino acid formula prevents alcoholic liver disease in rodents”, American journal of physiology. Gastrointestinal and liver physiology, 314 (5). G566-G582. May 2018.
 
[25]  Corsetti, G., D’Antona, G., Ruocco, C., Stacchiotti, A., Romano, C., Tedesco, L., Dioguardi, F., Rezzani, R., and Nisoli, E., “Dietary supplementation with essential amino acids boots the beneficial effects of rosuvastatin on mouse kidney”, Amino acids, 46 (9). 2189-2203. September 2014.
 
[26]  Corsetti, G., Romano, C., Pasini, E., Marzetti, E., Calvani, R., Picca, A., Flati, V., and Dioguardi, F.S., “Diet enrichment with a specific essential free amino acid mixture improves healing of undressed wounds in aged rats”, Experimental gerontology, 96. 138-145. October 2017.
 
[27]  Bonfili, L., Cecarini, V., Cuccioloni, M., Angeletti, M., Flati, V., Corsetti, G., Pasini, E., Dioguardi, F.S., and Eleuteri, A.M., “Essential amino acid mixtures drive cancer cells to apoptosis through proteasome inhibition and autophagy activation”, The FEBS journal, 284 (11). 1726-1737. June 2017.
 
[28]  Corsetti, G., Pasini, E., Romano, C., Calvani, R., Picca, A., Marzetti, E., Flati, V., and Dioguardi. F.S. “Body weight loss and tissue wasting in late middle-aged mice on slightly imbalanced essential/non-essential amino acids diet”, Frontiers in Medicine, 5. 1-11. May 2018.
 
[29]  Hall, P., Davies, W., Stamp, K., Clamp, I., and Bigley, A. - “Comparison of computerized image analysis with traditional semiquantitative scoring of Perls’ Prussian Blue stained hepatic iron deposition”. Toxicology and Pathology, 41 (7). 992-1000. February 2013.
 
[30]  Prats-Puig, A., Gispert-Saüch, M., Díaz-Roldán, F., Carreras-Badosa, G., Osiniri, I., Planella-Colomer, M., et al. “Neutrophil-to-lymphocyte ratio: an inflammation marker related to cardiovascular risk in children”, Thromb Haemost, 114(4). 727-34. July 2015.
 
[31]  Ouellet, G., Malhotra, R., Penne, E.L., Usvya, L., Levin, N.W., Kotanko, P. “Neutrophil-lymphocyte ratio as a novel predictor of survival in chronic hemodialysis patients”, Clinical Nephrology, 85(4). 191-198. April 2016.
 
[32]  Kuro-o, M., Matsumura, Y., Aizawa, H., Kawaguchi, H., Suga, T., Utsugi, T., Ohyama, Y., Kurabayashi, M., Kaname, T., Kume, E., Iwasaki, H., Iida, A., Shiraki-Iida, T., Nishikawa, S., Nagai, R., Nabeshima, Y.I.. “Mutation of the mouse klotho gene leads to a syndrome resembling ageing”, Nature, 390. 45-51. November 1997.
 
[33]  Nakashima, Y., Mima, T., Yashiro, M., Sonou, T., Ohya, M., Masumoto, A., Yamanaka, S., Koreeda, D., Tatsuta, K., Hanba, Y., Moribata, M., Negi, S., Shigematsu, T. “Expression and localization of fibroblast growth factor (FGF) 23 and Klotho in the spleen: its physiological and functional implications”, Growth Factors. 34 (5-6). 196-202. December 2016.
 
[34]  Knutson, M., Wessling-Resnick, M. “Iron metabolism in the reticuloendothelial system”, Critical Reviews in Biochemistry and Molecular Biology, 38. 61-88. September 2003.
 
[35]  Barlas, N., Aydogan, M. “Histopathologic effects of maternal 4-tert-octylphenol exposure on liver, kidney and spleen of rats at adulthood”, Archives of Toxicology, 83 (4). 341-349. April 2009.
 
[36]  Iancu, T.C. “Ferritin and hemosiderin in pathological tissues”, Electron Microscopy Review. 5 (2). 209-229. July 1992.
 
[37]  Dobryszycka, W. “Biological functions of haptoglobin-new pieces to an old puzzle”, Eur J Clin Chem Clin Biochem 35 (9). 647-54. September 1997.
 
[38]  Vanuytsel, T., Vermeire, S., Cleynen, I. “The role of Haptoglobin and its related protein, Zonulin, in inflammatory bowel disease”, Tissue Barriers. 1 (5). e27321. December 2013.
 
[39]  Galicia G, Maes W, Verbinnen B, Kasran, A., Bullens, D., Arredouani, M., Ceuppens J.L. “Haptoglobin deficiency facilitates the development of autoimmune inflammation”, European Journal of Immunology. 39. 3404-3412. December 2009.
 
[40]  Van Vlierberghe, H., Langlois, M., Delanghe, J. “Haptoglobin polymorphisms and iron homeostasis in health and in disease”, Clinica Chimica Acta. 345 (1-2). 35-42. July 2004.
 
[41]  Miller R.A. “Aging and immune function”, International Review of Cytology. 124. 187-214. 1991.
 
[42]  Paulus W.J. “Cytokines and heart failure”, Heart Failure Monitor. 1 (2). 50-56. 2000.
 
[43]  Dinarello C.A. “Biologic basis for interleukin-1 in disease”, Blood, 87 (6). 2095-2147. March 1996.
 
[44]  Dinarello C.A. “Anti-inflammatory agents: present and future”, Cell.140 (6). 935-950. March 2010.
 
[45]  Zhou, D., Chrest, F.J., Adler, W., Munster, A., Winchurch, R.A. “Increased production of TGF-beta and Il-6 by aged spleen cells”, Immunology Letters 36 (1). 7-11. April 1993.
 
[46]  Couper N.K., Blount D.G., Riley E.M. “IL-10: The Master Regulator of Immunity to Infection”, Journal of Immunology. 180 (9). 5771-5777. May 2008.
 
[47]  Redpath, S.A., Fonseca, N.M., Perona-Wright, G. “Protection and pathology during parasite infection: IL-10 strikes the balance”, Parasite Immunology. 36. 233-252. June 2014.
 
[48]  Moore, K.W., de Waal,M.R., Coffman, R.L., O’Garra, A. “Interleukin-10 and the interleukin-10 receptor”, Annual Review of Immunology. 19. 683-765. 2001.
 
[49]  Banchereau, J., Pascual, V., O’Garra, A. “From IL-2 to IL-37: the expanding spectrum ofanti-inflammatory cytokines”. Nature Immunology. 13(10). 925-931. October 2012.
 
[50]  Taga, K., Mostowski, H., Tosato, G. “Human interleukin-10 can directly inhibit T-cell growth”, Blood. 81 (11). 2964-71. June 1993.
 
[51]  Spencer, N.F.L., Norton, S.D., Harrison, L.L., Li, G.Z., Daynes, R.A. “Dysregulation of IL-10 production with aging: Possible linkage to the age-associated decline in DHEA and its sulfated derivative”, Experimental Gerontology. 31 (3). 393-408. May-June 1996.
 
[52]  Laroux F.S., Pavlick K.P., Hines I.N., Kawachi S, Harada H., Bharwani S., Hoffman J.M., Grisham M.B. “Role of nitric oxide in inflammation”, Acta Physiologica Scandinava. 173 (1). 113-118. September 2001.
 
[53]  Ni, M., Lee, A.S. “ER chaperones in mammalian development and human diseases”, FEBS Letters. 581 (19). 3641-3651. July 2007.
 
[54]  Wang, M., Wey, S., Zhang, Y., Ye, R., Lee, A.S. “Role of the unfolded protein response regulator GRP78/BiP in development, cancer, and neurological disorders”, Antioxidants & Redox Signaling. 11 (9). 2307-2316. September 2009.
 
[55]  Hendershot, L.M, “The ER function BiP is a master regulator of ER function”, Mount Sinai Journal of Medicine, 71 (5): 289-297. October 2004.
 
[56]  Matsumoto, A., Hanawalt, P.C, “Histone H3 and heat shock protein GRP78 are selectively cross-linked to DNA by photoactivated gilvocarcin V in human fibroblasts”, Cancer Research, 60 (14). 3921-3926. July 2000.
 
[57]  Sun, F.C., Wei, S., Li, C.W., Chang, Y.S., Chao, C.C., Lai, Y.K, “Localization of GRP78 to mitochondria under the unfolded protein response”, Biochemical Journal, 396 (1). 31-39. May 2006.
 
[58]  Ni, M., Zhou, H., Wey, S., Baumeister, P., Lee, A.S, “Regulation of PERK signaling and leukemic cell survival by a novel cytosolic isoform of the UPR regulator GRP78/BiP”, PLoS One, 4 (8). e6868. August 2009.
 
[59]  Quinones, Q., de Ridder, G., Pizzo, S, “GRP78: a chaperone with diverse roles beyond the endoplasmic reticulum”, Histology and Histopathology, 23 (11). 1409-1416. November 2008.
 
[60]  Ni, M., Zhang, Y., Lee, A, “Beyond the endoplasmic reticulum: atypical GRP78 in cell viability, signalling and therapeutic targeting”, Biochemical Journal, 434 (2).181-188. March 2011.
 
[61]  Zhang, Y., Liu, R., Ni, M., Gill, P., Lee, A.S, “Cell Surface Relocalization of the Endoplasmic Reticulum Chaperone and Unfolded Protein Response Regulator GRP78/BiP”, Journal of Biological Chemistry, 285 (20). 15065-15075. May 2010.
 
[62]  Zughayer, S., Stauffer, B., Mc Carty, N, “Inflammation and ER stress downregulate BDH2 expression and dysregulate intracellular iron in macrophages”, Journal of Immunology Research, 2014. ID140728. December 2014.
 
[63]  Wadhwa, R., Taira, K., Kaul, S.C, “An Hsp70 family chaperone, mortalin / mthsp70 / PBP74 / Grp75: what, when, and where?”, Cell Stress & Chaperones, 7 (3): 309-316. July 2002.
 
[64]  Burbulla, L.F., Krüger, R, “Mortalin Biology: Life, Stress and Death”, In book: Mortalin Biology: Life, Stress and Death, March 2012.
 
[65]  Burbulla, L.F., Fitzgerald, J.C., Stegen, K., Westermeier, J., Thost, A.K., Kato, H., Mokranjac, D., Sauerwald, J., Martins, L.M., Woitalla, D., Rapaport, D., Riess, O., Proikas-Cezanne, T., Rasse, T.M., Krüger, R, “Mitochondrial proteolytic stress induced by loss of mortalin function is rescued by Parkin and PINK1”, Cell Death & Disease, 5 (4). e1180. April 2014.
 
[66]  Ornatsky, O.I., Connor, M.K., Hood, D.A, “Expression of stress proteins and mitochondrial chaperonins in chronically stimulated skeletal muscle”, Biochemical Journal, 311 (Pt 1). 119-123. October 1995.
 
[67]  Takahashi, M., Chesley, A., Freyssenet, D., Hood, D.A, “Contractile activity-induced adaptations in the mitochondrial protein import system”, American Journal of Physiology, 274 (5 Pt 1). C1380-1387. May 1998.
 
[68]  Kaul, S.C., Matsui, M., Takano, S., Sugihara, T., Mitsui, Y., Wadhwa, R, “Expression Analysis of Mortalin, a Unique Member of the Hsp70 Family of Proteins, in Rat Tissues”, Experimental Cell Research, 232(1). 56 -63. April 1997.
 
[69]  Flachbartová, Z., Kovacech, B, “Mortalin, a multipotent chaperone regulating cellular processes ranging from viral infection to neurodegeneration”, Acta Virologica, 57 (1). 3-15. 2013.
 
[70]  Yang, L., Liu, X., Hao, J., Yang, Y., Zhao, M., Zuo, J., Liu, W, “Glucose-regulated protein 75 suppresses apoptosis induced by glucose deprivation in PC12 cells through inhibition of Bax conformational change”, Acta Biochimica et Biophysica Sinica (Shanghai), 40 (4). 339-348. April 2008.
 
[71]  Lv, L.J., Li, J., Qiao, H.B., Nie, B.J., Lu, P., Xue, F., Zhang, Z.M. Overexpression of GRP75 inhibits inflammation in a rat model of intracerebral haemorrhage. Molecular Medicine Reports, 15 (3). 1368-1372. March 2017.
 
[72]  Liu, Y., Liu, W., Song, X.D., Zuo, J, “Effect of GRP75/mthsp70/PBP74/mortalin overexpression on intracellular ATP level, mitochondrial membrane potential and ROS accumulation following glucose deprivation in PC12 cells”, Molecular and Cellular Biochemistry, 268 (1-2). 45-51. January 2005.
 
[73]  Someya, S., Yu, W., Hallows, W.C., Xu, J., Vann, J.M., Leeuwenburgh, C., Tanokura, M., Denu, J.M., Prolla, T.A, “Sirt3 mediates reduction of oxidative damage and prevention of age-related hearing loss under caloric restriction”. Cell, 143 (5). 802-812. November 2010.
 
[74]  Qiu, X., Brown, K., Hirschey, M.D., Verdin, E., Chen, D, “Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation”. Cell Metabolism, 12 (6). 662-667. December 2010.
 
[75]  Tao, R., Coleman, M.C., Pennington, J.D., Ozden, O., Park, S.H., Jiang, H., Kim, H.S., Flynn, C.R., Hill, S., Hayes McDonald, W., Olivier, A.K., Spitz, D.R., Gius, D, “Sirt3-mediated deacetylation of evolutionarily conserved lysine 122 regulates Mn-SOD activity in response to stress”, Molecular Cell, 40 (6). 893-904. December 2010.
 
[76]  Kwon, Y., Kim, J., Lee, C-Y., Kim, H, “Expression of SIRT1 and SIRT3 varies according to age in mice”, Anatomy & Cell Biology, 48 (1). 54-61. March 2015.
 
[77]  Weindruch, R.H., Cheung, M.C., Verity, M.A., Walford, L.R, “Modification of mitochondrial respiration by aging and dietary restriction”, Mechanisms of Ageing and Development, 12 (4): 375-392. April 1980.
 
[78]  Nisoli, E., Cozzi V., Carruba M.O, “Amino Acids and Mitochondrial Biogenesis”, American Journal of Cardiology, 101(11A). 22E-25E. June 2008.
 
[79]  Saito, Y., Nakamura, T., Ohyama, Y., Suzuki, T., Iida, A., Shiraki-Iida, T., Kuro-o, M., Nabeshima, Y., Kurabayashi, M., Nagai, R, “In vivo klotho gene delivery protects against endothelial dysfunction in multiple risk factor syndrome”, Biochemical and Biophysical Research Communication, 276(2). 767-72. September 2000.
 
[80]  Ikushima, M., Rakugi, H., Ishikawa, K., Maekawa, Y., Yamamoto, K., Ohta, J., Chihara, Y., Kida, I., Ogihara, T, “Anti-apoptotic and anti-senescence effects of Klotho on vascular endothelial cells”, Biochemical and Biophysical Research Communication, 339 (3). 827-832. January 2006.
 
[81]  Lorenzi, O., Veyrat-Durebex, C., Wollheim, C.B., Villemin, P., Rohner-Jeanrenaud, F., Zanchi, A., Vischer, U.M, “Evidence against a direct role of klotho in insulin resistance”, Pflugers Arch: European Journal of Physiology, 459 (3). 465-473. February 2010.
 
[82]  Corsetti, G., Pasini, E., Scarabelli, T.M., Romano, C., Agrawal, P.R., Chen-Scarabelli, C., Knight, R., Saravolatz, L., Narula, J., Ferrari-Vivaldi, M., Flati, V., Assanelli, D., Dioguardi, F.S, “Decreased expression of klotho in cardiac atria biopsy samples from patients at higher risk of atherosclerotic cardiovascular disease” Journal of Geriatric Cardiology, 13(8): 701-711. August 2016.
 
[83]  Yamamoto, M., Clark, J.D., Pastor, J.V., Gurnani, P., Nandi, A., Kurosu, H., Miyoshi, M., Ogawa, Y., Castrillon, D.H., Rosenblatt, K.P., Kuro-o, M, “Regulation of oxidative stress by the anti-aging hormone klotho”, Journal of Biological Chemistry, 280 (45). 38029-38034. November 2005.
 
[84]  Song, S., Gao, P., Xiao, H., Xu, Y., Yi Si, L, “Klotho suppresses cardiomyocyte apoptosis in mice with stress-induced cardiac injury via down regulation of endoplasmic reticulum stress”, PLoS One, 8 (12). e82968. December 2013.
 
[85]  Kuro-o, M, “Klotho”, Pflugers Arch: European Journal of Physiology, 459 (2). 333-343. January 2010.
 
[86]  Garcia De Vinuesa, C., Gulbranson‐Judge, A., Khan, M., O'Leary, P., Cascalho, M., Wabl, M., Klaus, G.G.B., Owen, M.J., MacLennan, I.C.M, “Dendritic cells associated with plasmablast survival”, European Journal of Immunology, 29. 3712-3721. October 1999.
 
[87]  Arnold, R., Calvo, W., Heymer, B., Schmeiser, T., Heimpel, H., Kubanek, B, “Extramedullary haematopoiesis after bone marrow transplantation”, European Journal of Haematology, 34. 9-12. January 1985.
 
[88]  Scognamiglio, R., Negut, C., Piccolotto, R., Dioguardi, F.S., Tiengo, A., Avogaro, A, “Effects of oral amino acid supplementation on myocardial function in patients with type 2 diabetes mellitus”, American Heart Journal, 147(6). 1106-1112. June 2004.
 
[89]  Scherbakov, N., Ebner, N., Sandek, A., Meisel, A., Haeusler, K.G., von Haehling, S., Anker, S.D., Dirnagl, U., Joebges, M., Doehner, W, “Influence of essential amino acids on muscle mass and muscle strength in patients with cerebral stroke during early rehabilitation: protocol and rationale of a randomized clinical trial (AMINO-Stroke Study)”, BMC Neurology 16. 10. January 2016.
 
[90]  Aquilani, R., Zuccarelli, G.C., Dioguardi, F.S., Baiardi, P., Frustaglia, A., Rutili, C., Comi, E., Catani, M., Iadarola, P., Viglio, S., Barbieri, A., D'Agostino, L., Verri, M., Pasini, E., Boschi, F, “Effects of oral amino acid supplementation on long-term-care-acquired infections in elderly patients”, Archives of Gerontology and Geriatrics, 52 (3). e123-128. May-June 2011.
 
[91]  Fafournoux, P., Bruhat, A., Jousse, C, “Amino acid regulation of gene expression”, Biochemical Journal, 351 (Pt 1). 1-12. October 2000.
 
[92]  Chandra, R.K, “Nutrition and the immune system from birth to old age”, European Journal of Clinical Nutrition, 56 (Suppl 3). S73-S76. August 2002.
 
[93]  Roth, E, “Non-nutritive effects of glutamine”, Journal of Nutrition, 138 (10). 2025S-2031S. October 2008.
 
[94]  Corsetti, G., Romano, C., Stacchiotti, A., Pasini, E., Dioguardi, F.S., “Endoplasmic Reticulum Stress and Apoptosis Triggered by Sub-Chronic Lead Exposure in Mice Spleen: a Histopathological Study”, Biological Trace Element Research, 178 (1). 86-97. July 2017.