Skip to main content Accessibility help
×
Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-20T04:06:45.470Z Has data issue: false hasContentIssue false

7 - Bacterial toxins and bone remodelling

Published online by Cambridge University Press:  15 September 2009

Neil W A McGowan
Affiliation:
Department of Craniofacial Development, Dental Institute, King's College London
Dympna Harmey
Affiliation:
The Burnham Institute
Fraser P Coxon
Affiliation:
Department of Medicine and Therapeutics, University of Aberdeen
Gudrun Stenbeck
Affiliation:
Bone and Mineral Centre, University College London
Michael J Rogers
Affiliation:
Department of Medicine and Therapeutics, University of Aberdeen
Agamemnon E Grigoriadis
Affiliation:
Department of Craniofacial Development, Dental Institude, King's College London
Alistair J. Lax
Affiliation:
King's College London
Get access

Summary

Bacterial protein toxins are powerful biological poisons normally associated with impairment of cellular function and/or cellular death. The wide spectrum of physiological processes and cell types that are affected by bacterial products also includes bone tissue and bone cells. It has been known for many years that bacterial infection or exposure to certain toxins can lead to pathological bone disorders, most commonly, those associated with abnormal or excessive bone loss, such as periodontal disease (reviewed by Henderson and Nair, 2003). However, in most cases the bone-resorbing factors involved in these effects remain part of, or associated with, the bacterial surface. For example, the bone-resorbing effects of endotoxin, a component of lipopolysaccharide, are well established, although for the most part this action appears to be indirect, being dependent on the production of pro-inflammatory cytokines (IL-1, TNFα) from other cell types (Nair et al., 1996; Henderson and Nair 2003). In contrast, the effects of bacterial protein toxins on the cellular constituents of bone remain largely unknown. For simplicity, this review will focus only on bacterial toxins, in particular, those toxins that interfere with key signalling processes that have direct relevance to bone cell differentiation and function. However, a brief overview of the general biology of bone cells is necessary before discussing the mechanisms of toxin action and specific signal transduction pathways in bone.

BONE

Throughout life the vertebrate skeleton is in a constant state of turnover.

Type
Chapter
Information
Bacterial Protein Toxins
Role in the Interference with Cell Growth Regulation
, pp. 147 - 168
Publisher: Cambridge University Press
Print publication year: 2005

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ackermann, M R, Adams, D A, Gerken, L L, Beckman, M J, and Rimler, R B (1993). Purified Pasteurella multocida protein toxin reduces acid phosphatase-positive osteoclasts in the ventral nasal concha of gnotobiotic pigs. Calcif. Tissue Int., 52, 455–459CrossRefGoogle ScholarPubMed
Ackermann, M R, Rimler, R B, and Thurston, J R (1991). Experimental model of atrophic rhinitis in gnotobiotic pigs. Infect. Immun., 59, 3626–3629Google ScholarPubMed
Anderson, D M, Maraskovsky, E, Billingsley, W L, Dougall, W C, Tometsko, M E, Roux, E R, Teepe, M C, DuBose, R F, Cosman, D, and Galibert, L (1997). A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature, 390, 175–179CrossRefGoogle ScholarPubMed
Athanasou, N A (1996). Cellular biology of bone-resorbing cells. J. Bone. Joint. Surg. Am., 78, 1096–1112CrossRefGoogle ScholarPubMed
Aubin, J E (1998). Advances in the osteoblast lineage. Biochem. Cell Biol., 76, 899–910CrossRefGoogle ScholarPubMed
Bishop, A L and Hall, A (2000). Rho GTPases and their effector proteins. Biochem. J., 348, 241–255CrossRefGoogle ScholarPubMed
Blair, H C, Teitelbaum, S L, Ghiselli, R, and Gluck, S (1989). Osteoclastic bone resorption by a polarized vacuolar proton pump. Science, 245, 855–857CrossRefGoogle ScholarPubMed
Boquet, P (2000). The cytotoxic necrotizing factor 1 (CNF1). From uropathogenic Escherichia Coli. Adv. Exp. Med. Biol., 485, 45–51CrossRefGoogle Scholar
Boyce, B F, Yoneda, T, Lowe, C, Soriano, P, and Mundy, G R (1992). Requirement of pp 60c-src expression for osteoclasts to form ruffled borders and resorb bone in mice. J. Clin. Invest, 90, 1622–1627CrossRefGoogle Scholar
Chellaiah, M A, Soga, N, Swanson, S, McAllister, S, Alvarez, U, Wang, D, Dowdy, S F, and Hruska, K A (2000). Rho-A is critical for osteoclast podosome organization, motility, and bone resorption. J. Biol. Chem., 275, 11993–12002CrossRefGoogle ScholarPubMed
Coxon, F P and Rogers, M J (2003). The Role of Prenylated Small GTP-Binding Proteins in the Regulation of Osteoclast Function. Calcif. Tissue Int., 72, 80–84CrossRefGoogle ScholarPubMed
Darnay, B G, Haridas, V, Ni, J, Moore, P A, and Aggarwal, B B (1998). Characterization of the intracellular domain of receptor activator of NF-κB (RANK). Interaction with tumor necrosis factor receptor-associated factors and activation of NF-κB and c-Jun N-terminal kinase. J. Biol. Chem., 273, 20551–20555CrossRefGoogle ScholarPubMed
Darnay, B G, Ni, J, Moore, P A, and Aggarwal, B B (1999). Activation of NF-κB by RANK requires tumor necrosis factor receptor-associated factor (TRAF) 6 and NF-κB-inducing kinase. Identification of a novel TRAF6 interaction motif. J Biol. Chem., 274, 7724–7731CrossRefGoogle ScholarPubMed
Ducy, P, Schinke, T, and Karsenty, G (2000). The osteoblast: a sophisticated fibroblast under central surveillance. Science, 289, 1501–1504CrossRefGoogle ScholarPubMed
Duong, L T, Lakkakorpi, P, Nakamura, I, and Rodan, G A (2000). Integrins and signaling in osteoclast function. Matrix Biol., 19, 97–105CrossRefGoogle ScholarPubMed
Etienne-Manneville, S and Hall, A (2002). Rho GTPases in cell biology. Nature, 420, 629–635CrossRefGoogle ScholarPubMed
Felix, R, Fleisch, H, and Frandsen, P L (1992). Effect of Pasteurella multocida toxin on bone resorption in vitro. Infect. Immun., 60, 4984–4988Google Scholar
Flatau, G, Lemichez, E, Gauthier, M, Chardin, P, Paris, S, Fiorentini, C, and Boquet, P (1997). Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine. Nature, 387, 729–733CrossRefGoogle Scholar
Fujikawa, Y, Sabokbar, A, Neale, S D, Itonaga, I, Torisu, T, and Athanasou, N A (2001). The effect of macrophage-colony stimulating factor and other humoral factors (interleukin-1, -3, -6, and -11, tumor necrosis factor-alpha, and granulocyte macrophage-colony stimulating factor) on human osteoclast formation from circulating cells. Bone, 28, 261–267CrossRefGoogle ScholarPubMed
Grigoriadis, A E, Wang, Z Q, Cecchini, M G, Hofstetter, W, Felix, R, Fleisch, H A, and Wagner, E F (1994). c-Fos: A key regulator of osteoclast-macrophage lineage determination and bone remodeling. Science, 266, 443–448CrossRefGoogle ScholarPubMed
Gwaltney, S M, Galvin, R J, Register, K B, Rimler, R B, and Ackermann, M R (1997). Effects of Pasteurella multocida toxin on porcine bone marrow cell differentiation into osteoclasts and osteoblasts. Vet. Pathol., 34, 421–430CrossRefGoogle ScholarPubMed
Harmey, D, Stenbeck, G, Nobes, C D, Lax, A J, and Grigoriadis, A E (2004). Regulation of osteoblast differentiation by Pasteurella multocida toxin (PMT): A role for Rho GTPase in bone formation. J. Bone Miner. Res., 19, 661–670CrossRefGoogle ScholarPubMed
Helfrich M H and Horton M A (1999). Integrins and adhesion molecules. In Dynamics of Bone and Cartilage Metabolism, ed. M J Siebel, S P Robins, and J P Bileziken, pp. 111–125, Academic Press, San Diego
Henderson, B and Nair, S P (2003). Hard labour: Bacterial infection of the skeleton. Trends Microbiol., 11, 570–577CrossRefGoogle ScholarPubMed
Hofbauer, L C and Heufelder, A E (1998). Osteoprotegerin and its cognate ligand: A new paradigm of osteoclastogenesis. Eur. J. Endocrinol., 139, 152–154CrossRefGoogle ScholarPubMed
Horiguchi, Y, Inoue, N, Masuda, M, Kashimoto, T, Katahira, J, Sugimoto, N, and Matsuda, M (1997). Bordetella bronchiseptica dermonecrotizing toxin induces reorganization of actin stress fibers through deamidation of Gln-63 of the GTP-binding protein Rho. Proc. Natl. Acad. Sci. USA, 94, 11623–11626CrossRefGoogle ScholarPubMed
Horiguchi, Y, Okada, T, Sugimoto, N, Morikawa, Y, Katahira, J, and Matsuda, M (1995). Effects of Bordetella bronchiseptica dermonecrotizing toxin on bone formation in calvaria of neonatal rats. FEMS Immunol. Med. Mic., 12, 29–32CrossRefGoogle Scholar
Hotokezaka, H, Sakai, E, Kanaoka, K, Saito, K, Matsuo, K, Kitaura, H, Yoshida, N, and Nakayama, K (2002). U0126 and PD98059, specific inhibitors of MEK, accelerate differentiation of RAW264.7 cells into osteoclast-like cells. J. Biol. Chem., 277, 47366–47372CrossRefGoogle ScholarPubMed
Hsu, H, Lacey, D L, Dunstan, C R, Solovyev, I, Colombero, A, Timms, E, Tan, H L, Elliott, G, Kelley, M J, Sarosi, I, Wang, L, Xia, X Z, Elliott, R, Chiu, L, Black, T, Scully, S, Capparelli, C, Morony, S, Shimamoto, G, Bass, M B, and Boyle, W J (1999). Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc. Natl. Acad. Sci. USA, 96, 3540–3545CrossRefGoogle ScholarPubMed
Iotsova, V, Caamano, J, Loy, J, Yang, Y, Lewin, A, and Bravo, R (1997). Osteopetrosis in mice lacking NF-κB1 and NF-κB2. Nat. Med., 3, 1285–1289CrossRefGoogle ScholarPubMed
Jutras, I, and Martineau-Doize, B (1996). Stimulation of osteoclast-like cell formation by Pasteurella multocida toxin from hemopoietic progenitor cells in mouse bone marrow cultures. Can. J. Vet. Res., 60, 34–39Google ScholarPubMed
Karsenty, G, and Wagner, E F (2002). Reaching a genetic and molecular understanding of skeletal development. Dev. Cell, 2, 389–406CrossRefGoogle ScholarPubMed
Kimman, T G, Lowik, C W, , Wee-Pals L J, Thesingh, C W, Defize, P, Kamp, E M, and Bijvoet, O L (1987). Stimulation of bone resorption by inflamed nasal mucosa, dermonecrotic toxin-containing conditioned medium from Pasteurella multocida, and purified dermonecrotic toxin from P. multocida. Infect. Immun., 55, 2110–2116Google ScholarPubMed
Lacerda, H M, Pullinger, G D, Lax, A J, and Rozengurt, E (1997). Cytotoxic necrotizing factor 1 from Escherichia coli and dermonecrotic toxin from Bordetella bronchiseptica induce p21(rho)-dependent tyrosine phosphorylation of focal adhesion kinase and paxillin in Swiss 3T3 cells. J. Biol. Chem., 272, 9587–9596CrossRefGoogle ScholarPubMed
Lacey, D L, Timms, E, Tan, H L, Kelley, M J, Dunstan, C R, Burgess, T, Elliott, R, Colombero, A, Elliott, G, Scully, S, Hsu, H, Sullivan, J, Hawkins, N, Davy, E, Capparelli, C, Eli, A, Qian, Y X, Kaufman, S, Sarosi, I, Shalhoub, V, Senaldi, G, Guo, J, Delaney, J, and Boyle, W J (1998). Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell, 93, 165–176CrossRefGoogle ScholarPubMed
Lax, A J, and Chanter, N (1990). Cloning of the toxin gene from Pasteurella multocida and its role in atrophic rhinitis. J. Gen. Microbiol., 136, 81–87CrossRefGoogle ScholarPubMed
Lee, S E, Woo, K M, Kim, S Y, Kim, H M, Kwack, K, Lee, Z H, and Kim, H H (2002). The phosphatidylinositol 3-kinase, p38, and extracellular signal-regulated kinase pathways are involved in osteoclast differentiation. Bone, 30, 71–77CrossRefGoogle ScholarPubMed
Lerm, M, Schmidt, G, and Aktories, K (2000). Bacterial protein in toxins targetting rho GTPases. FEMS Microbiol. Lett., 188, 1–6CrossRefGoogle Scholar
Lerm, M, Schmidt, G, Goehring, U M, Schirmer, J, and Aktories, K (1999). Identification of the region of rho involved in substrate recognition by Escherichia coli cytotoxic necrotizing factor 1 (CNF1). J. Biol. Chem., 274, 28999–29004CrossRefGoogle Scholar
Lomaga, M A, Yeh, W C, Sarosi, I, Duncan, G S, Furlonger, C, Ho, A, Morony, S, Capparelli, C, Van, G, Kaufman, S, , Heiden A, Itie, A, Wakeham, A, Khoo, W, Sasaki, T, Cao, Z D, Penninger, J M, Paige, C J, Lacey, D L, Dunstan, C R, Boyle, W J, Goeddel, D V, and Mak, T W (1999). TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling. Gene Dev., 13, 1015–1024CrossRefGoogle ScholarPubMed
Lowe, C, Yoneda, T, Boyce, B F, Chen, H, Mundy, G R, and Soriano, P (1993). Osteopetrosis in Src-deficient mice is due to an autonomous defect of osteoclasts. Proc. Natl. Acad. Sci. USA, 90, 4485–4489CrossRefGoogle Scholar
Mackay, D J and Hall, A (1998). Rho GTPases. J. Biol. Chem., 273, 20685–20688CrossRefGoogle ScholarPubMed
Manolagas, S C (2000). Birth and death of bone cells: Basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr. Rev., 21, 115–137Google ScholarPubMed
Martineau-Doize, B, Caya, I, Gagne, S, Jutras, I, and Dumas, G (1993). Effects of Pasteurella multocida toxin on the osteoclast population of the rat. J. Comp. Pathol., 108, 81–91CrossRefGoogle ScholarPubMed
Matsumoto, M, Sudo, T, Saito, T, Osada, H, and Tsujimoto, M (2000). Involvement of p38 mitogen-activated protein kinase signaling pathway in osteoclastogenesis mediated by receptor activator of NF-κB ligand (RANKL). J. Biol. Chem., 275, 31155–31161CrossRefGoogle Scholar
Miyazaki, T, Katagiri, H, Kanegae, Y, Takayanagi, H, Sawada, Y, Yamamoto, A, Pando, M P, Asano, T, Verma, I M, Oda, H, Nakamura, K, and Tanaka, S (2000). Reciprocal role of ERK and NF-κB pathways in survival and activation of osteoclasts. J. Cell Biol., 148, 333–342CrossRefGoogle ScholarPubMed
Mullan, P B and Lax, A J (1996). Pasteurella multocida toxin is a mitogen for bone cells in primary culture. Infect. Immun., 64, 959–965Google ScholarPubMed
Mullan, P B and Lax, A J (1998). Pasteurella multocida toxin stimulates bone resorption by osteoclasts via interaction with osteoblasts. Calcified. Tissue Int., 63, 340–345CrossRefGoogle ScholarPubMed
Mundy G R (1998). Bone remodelling. In Primer on the metabolic bone diseases and disorders of mineral metabolism, 4th ed., ed. M J Favus, pp. 30–39, Lippincott Williams & Wilkins, Philadelphia
Nair, S P, Meghji, S, Wilson, M, Reddi, K, White, P, and Henderson, B (1996). Bacterially induced bone destruction: Mechanisms and misconceptions. Infect. Immun., 64, 2371–2380Google ScholarPubMed
Nesbitt, S A and Horton, M A (1997). Trafficking of matrix collagens through bone-resorbing osteoclasts. Science, 276, 266–269CrossRefGoogle ScholarPubMed
Nobes, C D and Hall, A (1995). Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell, 81, 53–62CrossRefGoogle ScholarPubMed
Raisz, L G (1999). Physiology and pathophysiology of bone remodeling. Clin. Chem., 45, 1353–1358Google ScholarPubMed
Rodan, G A and Martin, T J (1981). Role of osteoblasts in hormonal control of bone resorption–A hypothesis. Calcif. Tissue Int., 33, 349–351CrossRefGoogle Scholar
Rozengurt, E, Higgins, T, Chanter, N, Lax, A J, and Staddon, J M (1990). Pasteurella multocida toxin: Potent mitogen for cultured fibroblasts. Proc. Natl. Acad. Sci. USA, 87, 123–127CrossRefGoogle ScholarPubMed
Salo, J, Lehenkari, P, Mulari, M, Metsikko, K, and Vaananen, H K (1997). Removal of osteoclast bone resorption products by transcytosis. Science, 276, 270–273CrossRefGoogle ScholarPubMed
Sarma, U and Flanagan, A M (1996). Macrophage colony-stimulating factor induces substantial osteoclast generation and bone resorption in human bone marrow cultures. Blood, 88, 2531–2540Google ScholarPubMed
Schmidt, G, Goehring, U M, Schirmer, J, Lerm, M, and Aktories, K (1999). Identification of the C-terminal part of Bordetella dermonecrotic toxin as a transglutaminase for rho GTPases. J. Biol. Chem., 274, 31875–31881CrossRefGoogle ScholarPubMed
Schmidt, G, Sehr, P, Wilm, M, Selzer, J, Mann, M, and Aktories, K (1997). Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1. Nature, 387, 725–729CrossRefGoogle ScholarPubMed
Schmidt, G, Selzer, J, Lerm, M, and Aktories, K (1998). The Rho-deamidating cytotoxic necrotizing factor 1 from Escherichia coli possesses transglutaminase activity. Cysteine 866 and histidine 881 are essential for enzyme activity. J. Biol. Chem., 273, 13669–13674CrossRefGoogle ScholarPubMed
Simonet, W S, Lacey, D L, Dunstan, C R, Kelley, M, Chang, M S, Luthy, R, Nguyen, H Q, Wooden, S, Bennett, L, Boone, T, Shimamoto, G, DeRose, M, Elliott, R, Colombero, A, Tan, H L, Trail, G, Sullivan, J, Davy, E, Bucay, N, Renshaw-Gegg, L, Hughes, T M, Hill, D, Pattison, W, Campbell, P, and Boyle, W J (1997). Osteoprotegerin: A novel secreted protein involved in the regulation of bone density. Cell, 89, 309–319CrossRefGoogle ScholarPubMed
Soriano, P, Montgomery, C, Geske, R, and Bradley, A (1991). Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell, 64, 693–702CrossRefGoogle ScholarPubMed
Sterner-Kock, A, Lanske, B, Uberschar, S, and Atkinson, M J (1995). Effects of the Pasteurella multocida toxin on osteoblastic cells in vitro. Vet. Pathol., 32, 274–279CrossRefGoogle ScholarPubMed
Suda, T, Nakamura, I, Jimi, E, and Takahashi, N (1997). Regulation of osteoclast function. J. Bone Miner. Res., 12, 869–879CrossRefGoogle ScholarPubMed
Suda, T, Takahashi, N, and Martin, T J (1992). Modulation of osteoclast differentiation. Endocr. Rev., 13, 66–80Google ScholarPubMed
Suda, T, Takahashi, N, Udagawa, N, Jimi, E, Gillespie, M T, and Martin, T J (1999). Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr. Rev., 20, 345–357CrossRefGoogle ScholarPubMed
Takahashi, N, Akatsu, T, Udagawa, N, Sasaki, T, Yamaguchi, A, Moseley, J M, Martin, T J, and Suda, T (1988). Osteoblastic cells are involved in osteoclast formation. Endocrinology, 123, 2600–2602CrossRefGoogle ScholarPubMed
Takai, Y, Sasaki, T, and Matozaki, T (2001). Small GTP-binding proteins. Physiol. Rev., 81, 153–208CrossRefGoogle ScholarPubMed
Teitelbaum, S L (2000). Bone resorption by osteoclasts. Science, 289, 1504–1508CrossRefGoogle ScholarPubMed
Vaananen, H K and Horton, M (1995). The osteoclast clear zone is a specialized cell-extracellular matrix adhesion structure. J. Cell Sci., 108, 2729–2732Google ScholarPubMed
Ward, P N, Miles, A J, Sumner, I G, Thomas, L H, and Lax, A J (1998). Activity of the mitogenic Pasteurella multocida toxin requires an essential C-terminal residue. Infect. Immun., 66, 5636–5642Google ScholarPubMed
Wei, S, Teitelbaum, S L, Wang, M W, and Ross, F P (2001). Receptor activator of nuclear factor-kappa b ligand activates nuclear factor-κB in osteoclast precursors. Endocrinology, 142, 1290–1295CrossRefGoogle Scholar
Yasuda, H, Shima, N, Nakagawa, N, Yamaguchi, K, Kinosaki, M, Mochizuki, S, Tomoyasu, A, Yano, K, Goto, M, Murakami, A, Tsuda, E, Morinaga, T, Higashio, K, Udagawa, N, Takahashi, N and Suda, T (1998). Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc. Natl. Acad. Sci. USA, 95, 3597–3602CrossRefGoogle ScholarPubMed
Zhang, D, Udagawa, N, Nakamura, I, Murakami, H, Saito, S, Yamasaki, K, Shibasaki, Y, Morii, N, Narumiya, S, and Takahashi, N (1995). The small GTP-binding protein, rho p21, is involved in bone resorption by regulating cytoskeletal organization in osteoclasts. J. Cell Sci., 108, 2285–2292Google ScholarPubMed
Zhao, H, Laitala-Leinonen, T, Parikka, V and Vaananen, H K (2001). Downregulation of small GTPase Rab7 impairs osteoclast polarization and bone resorption. J. Biol. Chem., 276, 39295–39302CrossRefGoogle ScholarPubMed
Zywietz, A, Gohla, A, Schmelz, M, Schultz, G, and Offermanns, S (2001). Pleiotropic effects of Pasteurella multocida toxin are mediated by Gq-dependent and -independent mechanisms: Involvement of Gq but not G11. J. Biol. Chem., 276, 3840–3845CrossRefGoogle Scholar

Save book to Kindle

To save this book to your Kindle, first ensure [email protected] is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×