Collagen types, associated-diseases and mouse models.
\r\n\tIt is a relatively simple process and a standard tool in any industry. Because of the versatility of the titration techniques, nearly all aspects of society depend on various forms of titration to analyze key chemical compounds.
\r\n\tThe aims of this book is to provide the reader with an up-to-date coverage of experimental and theoretical aspects related to titration techniques used in environmental, pharmaceutical, biomedical and food sciences.
The extracellular matrix (ECM) is the cell structural environment in tissues and organs. The ECM is a dynamic structure that it is constantly remodelled. It contributes to tissue integrity and mechanical properties. It is also essential for maintaining tissue homeostasis, morphogenesis and differentiation, which it does, through specific interactions with cells. The ECM is composed of a mixture of water and macromolecules classified into four main categories: collagens, proteoglycans, elastic proteins, and non-collagenous glycoproteins (also called adhesive glycoproteins). The nature, concentration and ratio of the different ECM components are all important factors in the regulation of the assembly of complex tissue-specific networks tuned to meet mechanical and biological requirements of tissues.
Collagens form a superfamily of 28 trimeric proteins, distinguishable from the other ECM components by their particular abundance in tissues (collagens represent up to 80-90% of total proteins in skin, tendon and bones) and their capacity to self-assemble into supramolecular organized structures (the best known being the banded fibers). The collagen superfamily is highly complex and shows a remarkable diversity in structure, tissue distribution and function (Ricard-Blum and Ruggiero, 2005).
The importance of collagens has been illustrated by the wide range of mutations in collagen genes that result in minor and severe human diseases. Various mutations (point, null or structural mutations, insertions, exon skipping, deletions) in genes encoding collagens are known to be responsible for a large spectrum of human disorders (e.g., Elhers-Danlos syndrome, epidermolysis bullosa, chondrodysplasia, osteogenesis imperfecta, Alport syndrome, Bethlem myopathy, Ulrich congenital muscular dystrophy, Fuchs\' endothelial dystrophy, Knobloch syndrome) that affect different tissues and organs, such as skin, blood vessels, cartilage, bones, kidney, muscle, cornea and retina. Considering the variety of collagen-related diseases and the complexity of collagen biology, there is a clear need to understand how mutations alter collagen synthesis, cell trafficking, cell and molecular interactions to result in tissue dysfunction. In the eighties targeted mutagenesis emerged as a new approach to help establish the structure-function relationship of collagens. Along with the emergence of protein engineering and genetically modified mice, site-directed mutagenesis has become instrumental in understanding the physiopathology of diseases, as well as in developing new and specific therapies and drugs for the treatment of human diseases. To date about 20 distinct genes encoding collagen chains have been ablated (by knock-out mutations) in mice or are involved in naturally occurring mutations. Only a few knock-in modified mice has been generated, in which a single point mutation or an exon deletion, for example, has been generated in a specific gene. This is likely due to the very large size of collagen genes. Site-directed knock-in mutations in mice have often proven to be more useful than knock-out mutations (which inactivate genes) for the analysis of the genotype-phenotype relationship, since small mutations represent the primary bases of inherited diseases.
The aim of this chapter is to describe the use of targeted mutagenesis in the understanding of the physiopathology of inherited connective tissue disorders. Specifically we are concerned with mutations in collagen genes. We will focus on the use of site-directed mutagenesis to analyze the causative effects of human-identified collagen gene mutations. Recombinant molecules were used to analyze the effects of these mutations on collagen structure, biosynthesis, posttranslational modifications and interactions with binding partners and cells. This work has considerably improved our knowledge in development and in human disorders. These results will then be compared with the limited information about the introduction of subtle targeted mutations into murine collagen genes.
The 28 members of the collagen superfamily exhibit considerable complexity and diversity in structure, assembly and function. However, collagens also share common features. (i) All members are modular proteins composed of collagenous (COL) domains flanked by non collagenous (NC) domains or linker regions. (ii) They are trimeric molecules formed by the association of three identical or different α-chains, which are characterized by repetitions of the G-x-y tripeptide (with the x and y positions often occupied by proline and hydroxyproline, respectively). (Abbreviations and single-letter codes for amino acids are given in Table 1 of the chapter by Figurski et al.) (iii) They are able to assemble into supramolecular aggregates in the extracellular space, although this property has not been proven for all recently identified collagen members. Collagens also undergo various post-translational modifications, including proteolytic processing, fibril formation, reticulation, shedding of transmembrane collagens and production of functional domains (also called matricryptins) (Ricard-Blum and Ruggiero, 2005). The mechanisms of collagen biosynthesis are far from being completely understood. Our knowledge is primarily based on the biosynthesis of fibril-forming collagens. Triple-helix formation commonly starts at the C-terminus (C-propeptide) of the α-pro-chain and proceeds toward the N-terminus (N-propeptide) in a zipper-like fashion. Prior to and simultaneously with triple-helix formation, specific prolines and lysines are chemically modified by addition of hydroxyl group. These modifications play a pivotal role in stabilization and resistance to temperature. Completed trimeric procollagens are secreted from the cells, proteolytically processed and assemble into collagen fibrils (Ricard-Blum and Ruggiero, 2005).
Based on their structure and supramolecular organization, collagens have been divided into several subfamilies (Myllyharju and Kivirikko, 2001). They are (i) the fibril-forming collagens I, II, III, V, XI, XXIV and XVII, which share the capacity to assemble into organized fibrils; (ii) the network-forming collagens IV, VIII and X and the FACIT (Fibril-Associated Collagen with Interrupted Triple-helix collagens) collagens IX, XII, XIV, XVI, XIX, XX, XI and XXII, which are known to mediate protein-protein interactions; (iii) the basement membrane multiplexin (multiple triple-helix domains and interruptions) collagens XV and XVIII; (iv) the transmembrane collagens, including the neuronal XXV collagen and types XIII, XVII, XXIII; and finally (v) other unconventional collagens, such as the anchoring fibrils collagen VII and the ubiquitous collagen VI, which assembles into characteristic beaded filaments (Table 1).
The length of the triple helical domains varies noticeably among different collagen types. Fibril-forming collagens consist of a long central COL domain with about 1000 amino acids (330 G-x-y tripeptide repeats), flanked by small terminal globular extensions (NC domains). After proteolytic processing of the N and C-terminal extensions, the mature molecules aggregate into highly ordered fibrils with a banded pattern observable by transmission electron microscopy. In other collagens, the COL domains are shorter and/or contain interruptions. The NC domains can represent the main part of the molecule, as for the FACIT collagen XII. Most, if not all, collagen types are recognized by specific cell receptors, such as the major ECM integrin receptors, collagen-specific discoidin domain receptors (DDR) and the transmembrane proteoglycan syndecans (Humphries et al., 2006; Xian et al., 2010; Leitinger et al., 2007). Through various interactions with these cell receptors, collagens can induce intracellular pathways directly or indirectly and regulate cell functions, such as migration, proliferation and differentiation. Certain collagens can also bind to growth factors and control their bioavailability by acting as reservoirs. The controlled release of growth factors by proteolytic activity or expression of a splice variant that does not contain the binding site controls morphogenesis, as described for the cartilage collagen II (Zhu et al., 1999).
A myriad of mutations has been characterized in collagen genes (Table 1). The function of the gene product and its tissue localization are criteria that lead to a number of inherited connective tissues disorders (reviewed in Bruckner-Tuderman and Bruckner, 1998; Bateman et al., 2009). Typically mutations in collagen genes are null-mutations, i.e., those resulting in the translation of an α-chain that cannot assemble into a triple helix and is consequently degraded intracellularly. Null mutations reduce the overall quantity of collagen in tissue and generally cause a human disorder. Small deletions and base substitutions can lead to synthesis of a mutated α-chain that is able to form a triple helix. The molecule is secreted, but its structure is compromised for supramolecular assembly, which normally occurs in the extracellular space. In fine collagen gene mutations result in defective matrix assembly and organization that in turn can affect cell function (Figure 1). In cases of large multimeric molecules, such as collagens, dominant-negative mutations can be more deleterious than null mutations. However, a growing body of evidence shows that the synthesis of a large quantity of abnormal collagen molecules in cells during development can induce endoplasmic reticulum stress, with consequences ranging from cell recovery to death (Tsang et al., 2010). The correlation between phenotype severity and the location of a point mutation in the gene is not clear. However, a mutation located in the coding region for the amino-terminus of the fibrillar collagen triple helix generally results in a mild phenotype, whereas a mutation in the coding region for the carboxy-terminus of the molecule is often lethal. This observation may be related to the C- to N-terminus directional propagation of the triple helix and the role of the C-propeptides in α-chain registration and triple helix nucleation. The nature of glycine substitution in the G-x-y repeats and the neighboring amino-acid sequence may have different biochemical and clinical consequences. These consequences include (i) delay of the triple-helix formation and over-glycosylation (Raghunath et al., 1994); (ii) alteration of procollagen processing (Lightfoot et al., 1994); (iii) retention of unfolded abnormal proteins intracellularly, leading to ER stress; and (iv) formation of abnormal unstable trimeric molecules, leading to disrupted fibrillogenesis.
The presence of a glycine in every third position is critical for triple-helix formation, since only glycine, the smallest amino acid, fits into the center of the triple helix. The majority of dominant-negative mutations in collagen genes are due to replacements of one of the glycines in the collagenous domains of the α-chains with a larger amino acid. Glycine substitution mutations in collagen genes underlie heritable connective tissue diseases, such as osteogenesis imperfecta (OI), chondrodysplasias, certain subtypes of Ehlers-Danlos syndrome (EDS), or Alport’s syndrome (reviewed in Bruckner-Tuderman and Bruckner, 1998; Bateman et al., 2009). Since a non-glycine amino acid does not easily fit into the interior space of the triple helix, helix formation is distorted, thereby affecting its structure and stability and impeding fibrillogenesis. Delay in triple-helix formation can result in over-modification and may affect collagen function.
Osteogenesis imperfecta (OI), also known as brittle bone disease, is caused by mutations in genes for collagen I, the most abundant collagen in organisms. OI is characterized by fragile bones that break easily and reduced bone mass. Most OI cases are believed to be associated with glycine substitution mutations in the COL1A1 or COL1A2 genes. Over 200 mutations have been reported for the COL1A1 (located on chromosome 17) and COL1A2 (located on chromosome 7) genes, which code for the collagen I pro-α1 and pro-α2 chains, respectively. This fact may explain the wide range of clinical characteristics and degrees of severity that are seen in the disease (Kuivaniemi et al., 1991, Byers and Steiner 1992, Dalgleish, 1998). Because collagen I is found in other tissues of the body, OI has non-skeletal manifestations as well. People with OI may also suffer from muscle weakness, hearing loss, fatigue, joint laxity, distensible skin, or dentinogenesis imperfecta. The fibril-forming collagen I is mostly synthesized as the [α1(I)]2α2(I) heterotrimer chain, though a minor form [α1(I)]3 is expressed in embryonic tissues. COL1A1 and COL2A1 are both susceptible to various mutations responsible for the production of quantitatively or qualitatively deficient fibrils. The clinical severity of OI relates to the extent of the conformational change in the collagen triple helix induced by the glycine substitution. These mutations result in altered fibrillogenesis. However, no general mechanism can be drawn from genotype/phenotype analyses.
Collagen VII, encoded by COL7A1, is the major component of the anchoring fibrils at the dermo-epidermal junction (Burgeson, 1993). COL7A1 gene mutations cause dystrophic epidermolysis bullosa (DEB), a skin-blistering disorder (Bruckner-Tuderman, 1999). Approximately 200 mutations of COL7A1 have been characterized, leading to a very high molecular heterogeneity of collagen VII defects (Dunnill et al., 1996). Almost all cases of dominant DEB are caused by a glycine substitution in the triple helical region of collagen VII, and most of the mutations are unique to individual families. Some glycine substitutions in collagen VII interfere with biosynthesis of the protein in a dominant-negative manner, whereas others may lead to collagen VII retention within the rough endoplasmic reticulum.
Mutations in the COL5A1 and COL5A2 genes, encoding respectively the pro-α1 and pro-α2 chains of the fibril-forming collagen V, have been identified in approximately 50% of patients with a clinical diagnosis of classic Ehlers-Danlos syndrome (EDS) (Malfait et al., 2010). Collagen V contains a third chain, proα3(V); but no mutation in COL5A3 has been reported so far. Classic EDS is a heritable disorder of connective tissues characterized by skin hyperextensibility, fragile and soft skin, delayed wound healing with formation of atrophic scars, easy bruising, and generalized joint hypermobility. The majority of mutations lead to a non-functional COL5A1 allele. One mutant COL5A1 transcript showed a premature stop codon. A minority of mutations affect the structure of the central helical domain. In approximately one-third of patients, the disease is caused by a mutation leading to a non-functional COL5A1 allele, resulting in collagen V haploinsufficiency. Structural mutations in COL5A1 or COL5A2, resulting in the production of a functionally defective protein, account for a small proportion of patients.
Collagen V is a quantitatively minor fibril-forming collagen that co-polymerizes with collagen I to form heterotypic fibrils (Fichard et al., 1995). Co-polymerisation has a critical role in the nucleation and growth of fibrils in tissues. A collagen V feature is to retain in the mature molecule a major part of the α1(V) N-propeptide which projects beyond the surface of collagen fibrils. This domain was proposed to limit heterotypic fibril growth by steric hindrance and electrostatic interactions (Linsenmayer et al., 1993). Skin biopsies revealed abnormalities in fibril formation (altered diameter, contour, or shape of dermal fibrils). However, abnormalities of fibril structure affected less than 5% of fibrils (reviewed in Fichard et al., 2003). Moreover, the clinical phenotype of classical EDS supports an important role of collagen V in the biomechanical integrity of the skin, tendon and ligaments, although collagen V is only a minor component of the affected tissues. Thus, collagen V may be involved in functions other than the control of fibril growth in classical EDS. A likely hypothesis is that collagen V might be involved in the physiopathology of EDS through interactions with other fibril-associated components and/or with cell receptors. Along this line, it has been shown that mutations in the genes for the collagen V-binding partners, tenascin-X (TNXB gene) and collagen I (COL1A1 gene), resulted in EDS (Lindor and Bristow, 2005).
Although mutant gene products are thought to impair matrix structure and assembly that eventually alters tissue function, growing evidence links ER stress and the unfolded protein response (UPR) to the initiation and progression of a broad repertoire of connective tissue disorders, including those caused by collagen gene mutations. Some mutant chains cannot be incorporated into procollagen molecules, consequently causing protein degradation with important downstream effects. Misfolded or slowly folding collagens are retained within the endoplasmic reticulum (ER) and ultimately targeted for degradation by a mechanism initially called “protein suicide.” Because connective-tissue cells typically produce large quantities of collagens, the contribution of ER stress induced by misfolded collagens in disease pathogenesis has certainly been underrated. The current knowledge on the implications of unfolded protein response and ER stress in connective tissue diseases has been recently reviewed, and readers are referred to these reviews for further reading (Boot-Handford and Briggs, 2010; Tsang et al., 2010). Notably, mutations in genes encoding collagen I (COL1A1 and COL1A2) (osteogenesis imperfecta), collagen II (COL2A1) (spondyloepiphyseal dysplasia), and collagen X (COL10A1) (metaphyseal chondrodysplasia) have been shown to induce ER dilatation in patient cells. Mutations that affect the triple helix, the C-propeptide for the fibril-forming collagens, and splice donor sites, as well as single amino-acid substitutions, were shown to cause ER stress. Recently, mutations that affected the signal peptide domain of the proα1(V)-collagen chain were shown to cause classic EDS. The signal peptides are the addresses of proteins destined for secretion. The mutant procollagen V is retained within the cell, leading to a collagen V haploinsufficiency and altered collagen fibril formation. It is probable that the signal peptide mutation also causes accumulation of the mutated protein within the ER and eventually to ER stress, as described for other collagen-related disorders (Symoens et al., 2009).
Collagen types, associated-diseases and mouse models.
Schematic diagram illustrating the biological consequences of point mutations or small deletions in collagen genes on chain synthesis, protein folding and subsequent fibril assembly in the extracellular matrix.
Mutations in the three major collagen VI genes (COL6A1, COL6A2 and COL6A3) cause multiple muscle disorders, including the severe Ullrich congenital muscular dystrophy (UCMD) and the mild Bethlem myopathy, which is characterized by muscle weakness with striking joint laxity and progressive contractures. Three genetically distinct novel chains α4(VI), α5(VI), and α6(VI) have recently been identified; but very little is known about their molecular assembly and biosynthesis and their possible involvement in human diseases (Gara et al., 2011). Collagen VI biosynthesis is a complex multistep process. Monomer formation results from the heterotrimeric association of the three chains [α1(VI), α2(VI), and α3(VI)] encoded by the COL6A1, COL6A2 and COL6A3 genes. Monomers first assemble into antiparallel dimers that associate laterally to form tetramers stabilized by disulphide bonds. The tetramers associate linearly to form the unique beaded filaments, the ultimate step of collagen VI biosynthesis. Dominant and recessive autosomal mutations in COL6A1, COL6A2, and COL6A3 primarily result in dysfunctional microfibrillar collagen VI in muscle extracellular matrix. However they also affect other connective tissues, such as skin and tendons. Different mutations have been shown to have variable effects on protein assembly, secretion, and its ability to form a functioning extracellular network. As observed in other collagen-related diseases, glycine-substitution mutations in COL6A1, COL6A2, or COL6A3 that disrupt the triple-helix motif constitute a frequent pathogenic mechanism. Triple-helix distortion may exert a dominant-negative effect by reducing the ability of mutated monomers to form beaded filaments. Interestingly, mitochondrial dysfunction was implicated in the pathogenesis of a myopathic phenotype. Muscles lacking collagen VI are characterized by the presence of a dilated sarcoplasmic reticulum and dysfunctional mitochondria. This condition triggers apoptosis and leads to myofiber degeneration. Recently, it was shown that the persistence of abnormal organelles and apoptosis observed in some congenital muscular dystrophies are caused by defective activation of the autophagic machinery. Autophagy has a key role in the clearance of damaged organelles and in the turnover of cell components and is thus essential for tissue homeostasis. Recently, 56 novel mutations have been described, allowing a clinical classification and revealing the complexity of genotype-phenotype relationships (Briñas et al., 2010).
The paucity of evidence-based data regarding correlations of genotype and phenotype is in part due to the large spectrum of mutations reported for the collagen genes [e.g., about 200 mutations for the collagen I genes responsible for OI (Dalgleish, 1998); 160 mutations in the COL4A5 gene encoding collagen IV α5 chain responsible for Alport syndrome; 200 mutations in COL7A1 responsible for EDB]. Things are not as simple as one gene-numerous mutations-one phenotype. Sometimes a combination of a mutation for a connective tissue disorder and a specific collagen gene mutation will result in another disease. Some patients with UCMD show clinical characteristics typical of classical disorders of connective tissue, such as EDS. Ultrastructure of skin biopsy samples from patients with UCMD showed alterations of collagen fibril morphology in skin that resemble those described in patients with EDS (Kirschner et al., 2005). Recently, using the yeast two-hybrid approach, we showed a direct interaction between collagen V and collagen VI that may nicely explain the overlap of UCMD and classic EDS (Symoens et al., 2011). Unexpectedly an arginine-->cysteine substitution localized at position 134 of the α1(I) collagen chain resulted in classical EDS (Nuyntick et al., 2000). This finding is indicative of genetic heterogeneity in collagen-related disorders.
A powerful approach to study the biochemical consequences of mutation and the protein structure/function relationship is to engineer a specific mutation into a functional domain of the molecule. Targeted mutagenesis approaches, including the use of alanine-scanning mutagenesis techniques, have led to important insights into the effects of collagen mutations on protein structure and function. A major limitation of mutagenesis strategies to investigate collagens is the large number of collagen gene mutations to be investigated in order to have a better understanding of the molecular mechanisms of “collagenopathies.” Knowledge about the impact of collagen mutations has also been hampered by the technical difficulty of introducing targeted mutations of very large collagen genes into mice.
Production of a recombinant collagen gene represents a powerful technique to introduce a human mutation into the gene of interest by site-directed mutagenesis. It allows one to analyze the impact of the mutation on collagen assembly and secretion. Collagen biosynthesis is a complex multistep process that takes place in the intracellular and extracellular space and includes various post-translationnal modifications, such as prolyl- and lysyl-hydroxylation, glycosylation, trimerization, proteolytic processing, polymerization and cross-links. Because of recombinant technology, these large multimeric proteins have been produced in large amounts in almost all existing expression systems (Ruggiero and Koch, 2008). This technological breakthrough enabled researchers to analyze in detail the effects of collagen mutations on biosynthesis, molecular and cell interactions, processing and, in some cases, self-assembly. Researchers can also address the question of the correlation of genotype, protein structure and function.
Mutations occurring in collagen I genes are the most extensively studied mutations among all collagen types. A first set of experiments substituted glycine 859 of the proα1(I) chain with cysteine or arginine by site-directed mutagenesis to reproduce two mutations identified in OI patients. In order to study the expression of the mutant molecule in the presence or absence of the wild-type proα1(I) chain, the mutated constructs were transfected into normal fibroblasts to look for a dominant-negative effect in the presence of the wild-type gene or in fibroblasts isolated from Mov13 homozygous mice (referred to as Mov13 fibroblasts hereafter), whose cells carry a provirus that prevents transcription initiation of the natural proα1(I) gene (Schnieke et al., 1987). In agreement with observations of collagen I in OI patients, the mutated collagens were poorly secreted from the cells and exhibited reduced thermal stability and increased sensitivity to degradation. This supported the idea that the strict preservation of the G-x-y triplets is absolutely required for proper formation of the triple helix.
The integrity of the C-propeptide is pivotal for the trimerization of all fibril-forming collagens. The C-propeptides of the proα1(I) and proα2(I) chains contain an Asn-Ile-Thr sequence. That sequence fits a consensus sequence for the addition of N-linked oligosaccharides. To analyze the role of this post-translational modification, the asparagine residue of the proα1(I) chain was changed to glycine by site-directed mutagenesis. The expression of the corresponding molecule was analyzed in transfected normal and Mov13 fibroblasts (Lamandé and Bateman, 1995). The mutation did not impair heterotrimeric assembly and secretion of hybrid procollagen I into the extracelllular space. Only a slight effect on C-proteinase cleavage efficiency was observed with the unglycosylated molecule. To circumvent the difficulty of producing a large repertoire of full-length mutated collagens I in order to undertake a genotype/phenotype analysis, a recombinant trimeric mini-collagen I was recently expressed in an Escherichia coli system. Recombinant mini-collagens can be obtained by fusing the sequence encoding a fragment of the proα1(I) chain triple-helix to the sequence encoding the C-terminal domain (called “foldon”) of the bacteriophage T4 fibritin, which is capable of trimerization (Xu et al., 2008). Two mutations (G901S and G913S), corresponding to mild and severe types of OI, respectively, were introduced into the recombinant mini-collagen I. Biophysical measurements and protease cleavage analysis revealed that the G913S mutant chain resulted in the formation of an unstable collagen I triple helix by disrupting salt bridges important for maintaining the chains in a triple-helix conformation (Yang et al., 1997; Xu et al., 2008). A very recent study utilized a recombinant bacterial collagen to develop a mutagenesis scheme in which a glycine residue within the triple-helix sequence is substituted with arginine or serine. The purpose was to analyze the positional effect of glycine mutations on triple-helix formation and stability (Cheng et al., 2011). Interestingly, all glycine mutations provoked a significant delay in the triple-helix formation. However, a more severe defect was observed when the mutation was located near the trimerization domain of the triple-helix where folding is initiated.
COL7A1 mutations cause dystrophic epidermolysis bullosa (DEB), a skin blistering disorder. Woodley and collaborators (2008) have used site-directed mutagenesis to elucidate the effect of human mutations on the function of collagen VII, which is the major component of the epidermal anchoring fibrils. To undertake a comprehensive analysis of the impact of human mutations in the formation, folding and stability of collagen VII and, particularly relevant to the DEB phenotype, its effect on cell attachment and migration, four distinct substitutions occurring in collagen VII (G2049E, R2063W, G2569R, and G2575R) were introduced using COL7A1 cDNA. The authors demonstrated that the G2049E and R2063W mutants caused local destabilization of the triple helix and reduced the capability of collagen VII to elicit cell adhesion and migration. The G2569R and G2575R mutants interfered with triple-helix formation and stability. Alterations of protein stability and/or cell attachment to collagen VII mutants help explain the fragility of the dermal-epidermal junction observed in DEB patients. Naturally occurring COL7A1 mutations were investigated in a separate study (Hammami-Hauasli et al., 1998). As commonly described for glycine-substitution mutants of collagens, the authors showed that three glycine substitutions located in the same triple-helix portion affected folding, stability and secretion of procollagen VII in a dominant-negative manner. However, the glycine substitution G1519D located in another segment of the triple helix had no effect on procollagen VII secretion or its ability to anchor fibril assembly. These data showed that the biological impact of glycine substitutions can depend on their position within the triple helix, as shown for collagen I (Cheng et al., 2011).
Human collagen IV mutations, thought to affect the biosynthesis of this basement membrane collagen, were extensively investigated. These mutations were known to cause Alport syndrome, a severe renal disease leading eventually to kidney failure. Collagen IV chains, α1(IV)-α6(IV), are encoded by 6 genes, COL4A1-COL4A6, respectively. Although mutations have also been identified in COL4A3 and COL4A4, about 30% of known missense mutations occur in the COL4A5 gene, which encodes the human α5(IV) chain. Most of them are glycine substitutions. One glycine-substitution mutation in COL4A5 could prevent correct α-chain folding or/and the association with other α-chains to form a stable triple helix. To address this question, the authors took advantage of the bacterial system. A DNA encoding a 22-kDa recombinant domain of the α5(IV) triple helix in its wild-type form or harboring the G1015V or G1030S mutations was expressed in E. coli (Wang et al., 2004). The recombinant wild-type and mutant proteins were purified and assayed for changes in triple-helix assembly and stability by circular dichroism. The two different glycine-substitution mutants displayed different defects in the secondary structures of their protein products that matched with the severity of the patient phenotypes. However, the use of a bacterial system to analyze the effects of specific human mutations on mini-collagen assembly and stability presents several disadvantages. Because collagens are large multimeric proteins, full-length molecules cannot be produced in a bacterial host. Most importantly, the bacterial system is limited. Not all post-translational modifications needed for the triple-helix formation and stability, such as hydroxylation, glycosylation, and disulfide-bond formation, are present in bacteria. A few years later, the bacterial limitations were bypassed by the development of the production of full-length recombinant collagen molecules in mammalian cells (Fichard et al., 1997; Ruggiero and Koch, 2008). No less than eighteen human mutations (11 substitutions and 7 deletions) were introduced into the sequence encoding the trimerization NC1 domain of the α5(IV) chain gene. The constructs were transfected into cells together with constructs containing the wild-type sequences of α3(IV) and α4(IV) chains to analyze the impact of the mutations in the NC1 domain on the formation of the α3α4α5 collagen IV heterotrimer. Twelve out of 15 mutant chains did loose their capacity to assemble into heterotrimeric molecules. The three remaining mutants formed heterotrimers, but the mutations prevented their secretion into the extracellular space (Kobayashi et al., 2008). The authors nicely demonstrated, using site-directed mutagenesis, that amino acid substitutions in the α5(IV) NC1 trimerization domain are specifically responsible for impairment of collagen IV heterotrimer assembly. This defect may be a main molecular mechanism for the pathogenesis of Alport syndrome. Interestingly, an interactome (a map of known and predicted molecular interactions, as well as phenotypic and structural landmarks) of collagen IV was recently constructed to identify functional and disease-associated domains and genotype-phenotype relationships (Parkin et al., 2011). Construction of such interactomes will greatly improve our capacity to integrate all data from different site-directed mutagenesis experiments. This advance will greatly help our understanding of the molecular mechanisms underlying “collagenopathies”; and, consequently, it may lead to the development of specific treatments.
Collagens undergo a great variety of proteolytic modifications. The fate and functions of the released fragments derived from collagens are still under intensive investigation, but the consequences of mutations in the coding regions for the cleavage sites on collagen structure, self-assembly and function have not been investigated in detail. A large repertoire of proteinases is responsible for these processing interactions. Included among such enzymes are the ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) and the BMP-1/tolloid families of metalloproteinases and more recently the furin-like proprotein convertases (Ricard-Blum and Ruggiero, 2005). To investigate collagen processing, fastidious extraction and purification steps were often necessary to obtain limited amounts of unprocessed proteins and enzymes with full activity in order to undertake in vitro enzymatic assays. To circumvent this problem, we recently described a new cell system allowing a rapid and straightforward analysis of processing interactions. Our system relies on the use of site-directed mutagenesis. This strategy was particularly instrumental in analyzing the complex procollagen V processing during maturation. We showed it to be unique among the fibril-forming collagens (Bonod-Bidaud et al., 2007). Collagen V is a minor fibrillar collagen that can be distinguished from the others by its capacity to control fibrillogenesis (Fichard et al., 1995). In addition this molecule undergoes a particular form of processing; and it is involved in fundamental processes, such as development and human connective tissues disorders. The proα1(V) N-terminus can be processed by the procollagen proteinases ADAMTS-2 and BMP-1 (Colige et al., 2005; Bonod-Bidaud et al., 2007), whereas the C-propeptide can be cleaved by furin and BMP-1 (Kessler et al., 2001). The proα1(V) C-propeptide furin cleavage site, which occurs immediately downstream of the recognition sequence RTRR, was double-mutated to alanine residues (R1584A/R1585A) to abolish furin cleavage. All constructs were introduced into cells, along with a BMP-1-expressing construct; and the cleavage products were directly analyzed in conditioned medium of the transiently transfected cells. We were able to show that BMP-1 is capable of processing the α1(V) C-propeptide in absence of furin activity (Bonod-Bidaud et al., 2007). In the same way, the determinant for α1(V) N-propeptide processing by BMP-1 activity was identified by introducing in the coding region for the cleavage site (S254/Q255-D256) three single mutations (S254A, Q255A and D256A), two double mutations (S254A/Q255N and Q255A/D256A) and one triple mutation (S254A/Q255A/D256A). The data highlighted the unexpected importance of the aspartic acid in the P2’ position of the BMP-1 cleavage site (Bonod-Bidaud et al., 2007). Processing, proteolytic release of functional domains and shedding of collagens are involved in fundamental processes. It is likely that substitutions located in the proteolytic cleavage sites may represent a molecular cause of connective tissues disorders. A reported mutation in the α1(V) N-propeptide in one patient with classic EDS resulted in a protein product missing the sequence of exon 5 that encompasses the BMP-1 cleavage site. The abnormal-sized N-propeptide present in the mutated collagen V caused dramatic alterations in fibril structure (Takahara et al., 2002).
In vitro studies are useful and necessary approaches to understand the mechanisms of collagen biosynthesis and to establish structure-function relationship. However, they do not always reflect the normal and pathological in vivo situations. Genetically modified mice appear to be a powerful technique to better understand the physiopathology of connective tissue disorders. Several different genetically modified mice have been created during the last 10 years (reviewed in Aszódi et al., 2006). This clearly opened doors to better understand collagen function in developing tissues and provide reliable mouse models for inherited collagen diseases. Along this line, a targeted disruption of Col4a3 gene led to renal failure and eventually to the death of mice at 3-4 months of age (Cosgrove et al., 1996; Miner and Sanes, 1996). This result is consistent with defects described for Alport disease.
In most cases, the gene of interest was disrupted and knock-out mice were preferably generated. Few transgenic mice harbouring point mutations or small deletion in collagen genes have been generated (Table 1). Naturally occurring mutations in mice disrupting collagen genes have also been identified and characterized. The oim mice present a spontaneously acquired deletion in the Col1a2 gene that leads to an accumulation of [α(I)]3 collagen homotrimer in the extracellular matrix. These mice develop a phenotype similar to moderate OI in humans, providing a good model for this collagen disorder (Chipman et al., 1993). It was shown that homozygous Mov13 embryos harboring an inactivated proα1(I) chain (due to the insertion of the Moloney murine leukaemia virus into the first exon of the Col1a1 gene) died in utero around day 12 because of vascular failure (Löhler et al., 1984). However, in 1999 Forlino et al. developed the first knock-in mouse model for human OI by introducing a G349C mutation into the Col1a1 gene. Along this line, a knock-in mouse model for OI, harboring a point mutation (G610C) in Col1a2 was recently created (Daley et al., 2010). These mice had reduced body mass and bone strength and exhibited bone fracture susceptibility consistent with the clinical features of human OI. Thus, the G610C knock-in mouse represents a novel model for the study of OI pathogenesis and also for testing potential therapies for OI.
Another example concerns collagen V deficiency/dysfunction, which is responsible for Ehlers-Danlos syndrome (EDS). In the absence of the Col5a1 gene, the mice died at the onset of organogenesis at approximately embryonic day 10 (Wenstrup et al., 2004). Interestingly, a targeted deletion in the Col5a2 gene, encoding the proα2(V) chain, recapitulated many of the clinical, biomechanical, morphologic, and biochemical features of the classical EDS. The deletion removes the sequence encoding the N-telopeptide (pN), a 20-residue region that confers flexibility to the N-terminal part of the molecule (Andrikopoulos et al., 1995). A detailed study of the skin at the morphological, histological, ultrastructural and biochemical levels indicated that the Col5a2 deletion impairs assembly and/or secretion of the [α1(V)]2α2(V) heterotrimer. Consequently, the [α1(V)]3 homotrimer, and not the [α1(V)]2α2(V) heterotrimer, is the predominant species deposited into the matrix, which in turn severely impaired extracellular matrix organization (Chanut-Delalande et al., 2004). These data underscored the importance of the collagen V [α1(V)]2α2(V) heterotrimer in dermal fibrillogenesis and can explain defects observed in the dermis of EDS patients.
Site-directed mutagenesis has been extensively used in collagen engineering and has shed light on collagen structure, expression, folding, secretion, interactions and self-assembly in the extracellular space. It also opened the way for the analysis of specific functional domains. It allowed the study of the wide variety of collagen types, including those expressed in trace amounts in tissues but nevertheless display pivotal functions. While it is true that site-directed mutagenesis has yielded important information on the functional consequences of a range of collagen mutations responsible for human diseases, only few studies have approached the consequences of collagen gene mutations on cell adaptation to ER stress. Collagen gene mutations affect protein synthesis, folding and secretion imbalance, which eventually induces ER stress. In vitro studies have been done on transfected cells, in which expression and trafficking of mutant collagen can be easily manipulated and analysed at the cellular level. The effects of gene manipulation can be studied in vivo using mice. The effect of collagen gene mutations on induction of an ER stress response could be straightforwardly addressed in the near future. It may be a key factor in pathogenesis (Boot-Handford and Briggs, 2010).
Mouse models are particularly useful for analysing the biological significance of collagens in pathological situations. Knock-out mice often lead to embryonic lethality, which hampers in-depth analysis of the phenotype. A few knock-in mice have been created with subtle mutations or small deletions that reproduce human mutations. The major reason for the paucity of knock-in mice is certainly that collagen genes are very large. Thus, they are difficult to manipulate. The introduction of a small deletion or a single point mutation in murine collagen genes still represents a considerable challenge. Nevertheless, the few examples of knock-in mouse lines tend to prove that mouse models can bring new information about in vivo consequences of collagen dysfunction that cannot be predicted by in vitro approaches. Knock-in mice are also indispensable models for assessing the effects of subtle mutations on tissue function, development, and aging. They are also valuable for developing specific gene therapy approaches to combat collagen-related disorders. The combination of site-directed mutagenesis in transfected cells and knock-in approaches in mice to address the impact of specific mutations will enable us to identify mechanisms underlying the vast repertoire of collagen-related diseases. The implications may lead to the development of a specific therapy.
The study of architectural history is and always has been one of the fundamentals in each architect’s education; up to the 1920’s circa also the design classes were embedded by the replica of the Greek temple orders and the imitations taken from ancient reference books such as Vitruvius, Palladio, Viollet-le-duc and many other. This changed after the 1920’s in favor of a more multidisciplinary and technical orientated training, promoted by “Ornament and Crime” (A. Loos, 1908), “Form follows function” (R. Sullivan, 1900 ca.) and other design maxims. It caused a proportionate impoverishment of building history knowledge and of traditional design methods. One said the architect-artist’s mind had to be liberated from all historic ballast, and should be able to create within his free individual creativity. This is a discussable principle with potentially quite negative consequences as architecture is not only a question of artistic creativity or aesthetical harmony, nor a pure functional or technical discipline: “Architectura … nascitur et fabrica et ratiocinatione …” (“Architecture is born by craftsmanship and balanced rationality”, Vitruvius, I°sec. b.C.) in ([1], I,1). Architecture (with capital) needs both approaches and apart from the Vitruvian “utilitas, venustas et firmitas [1]”, Architecture always had an existential and universal dimension dealing with bringing sense and structure in the surrounding space, including physical communication with meanings and messages’ to the observer [2].
The often poor knowledge on historic design criteria nowadays, inevitably leads in many cases to a considerable loss of ‘sense’ and a different type of ‘meaning and message’ in contemporary projects. Many heritage buildings get their conservation status because of tangible cultural and historic characteristics, and in many cases it is completed with a large intangible content expressed through symbolisms and allegories. Unfortunately, very often this symbolism and allegories get lost today as man is not familiar any more with the ancient allegorical languages. Also the other way around, modern design rarely uses those so called ‘old-fashioned’ allegorical indications in such speechless but most effective communication between designer and observer. Medieval buildings are particularly representative for the presence of this mostly forgotten intangible communication content, expressed through the symbolism of form, number, proportion, material or color. Based on the analysis of some representative medieval buildings, this chapter illustrates and tries to detect such design indicators to inspire the contemporary designer, not suggesting a flat imitation but a personal modern interpretation and use of the very same ancient design indicators. The two mayor instruments to all kind of allegorical allusions in medieval design are the geometry of the architectural form and the arithmetic’s within the different quantities and dimensions.
This book aims contributing in sustainable construction. The easy re-use or reconversion without great structural change or loss of architectural identity is part of all sustainability and certainly one of the most crucial assignments today. Recent experiences on the reconversion of existing fabric or the recuperation of ancient abandoned structures, mostly for evident economic reasons, have proved abundantly that reconversion or recuperation is much easier and less invasive with ancient well-modulated traditional buildings as it is the case with some contemporary building or probably shall be with one of the super eye-catching designed ones, created by great archistars as e.g. the Bilbao Guggenheim Museum or the Baku Heydar Aliyer Center. Certainly, those superlatives are strong signs of digital design and technical knowledge, but their quality remains one-sided, limited to never seen forms and materials. They do not show great flexibility nor long lasting esthetic pleasure; any probable later intervention, as proof of sustainable (re-)use, risks to damage considerably their actual identity. Society needs avant-garde, but this has to be applied with cure and caution. Contemporary design should reconsider the historic canons, take profit of the three thousand years’ experience, evaluate and integrate the old principles for harmony and sustainable use in the modern design algorithms to guarantee qualitative architecture and long lasting construction.
Unlike what many people thinks, architectural design is not so much a question of spontaneous creativity but much more of theoretical and technical knowledge. “Ars sine scientia nihil est” according the well know exclamation of the French master builder Jean Mignot consulting the Milan Cathedral builders in 1400 ca. The theoretical “scientia” at Mignot’s time was little and approximate, the artistic drive on the contrary was all the stronger. The success of so many ancient buildings, in particular the audacious finely jointed gothic structures were the result of practical experience during about 3000 years of building since the first Mesopotamian temples. Those ancient buildings were always expression not only of specific needs but also of the then living spiritual concepts about society, religion and esthetics. As in many other disciplines, also architecture and design owe a lot to ancient Greek philosophers from the early 10th century BC (and the Egyptians before them) as founders of West-European culture. Within the larger context of the Mediterranean Basin they developed a world view, not precisely as told in Genesis, but quite similar, i.e. created by a Supreme Divinity who organized and structured the initial chaos using calculated and measured geometric forms. This cosmos of a well ordered celestial and terrestrial creation by the Divine Geometer was the example that man had to follow in structuring his own small local chaos of space. All architectural project implies structure of space, and for that reason, all architectural design must be based on calculation, arithmetic and geometry. This idea was further developed, especially by the Christian scholastics (ca. 9th–13th century) and became an existential obligation for all architectural projects. This explains the permanent presence of numbers and geometries in architectural design for more than 3000 years. In this prospective, one could consider Plato, Aristotle, Pythagoras and Euclid (and the unnamed Egyptian and Mesopotamian priests) as the founders of European design principles.
To study the cosmos’s structure, Greek philosophers developed arithmetic number systems and geometric procedures to explain the phenomena of life and nature [3] p. 7. Numbers are abstract concepts related with the quantity of things, but in relation with real sets or groups they become a tangible reality which, in ancient metaphysic thinking, got very often an intangible connotation or symbolic value, variable in history. In architecture, this number symbolisms got related with the physical quantity of distinct built elements or with the measured quantity of length, width, height or volume. Also the procedure to establish a single number, i.e. the type of calculus by simple arithmetic using the four basic operations (addition, detraction, multiplication and dividing) or more complicate ones (square or cubic root), and the position of each single number as part of a sequence (arithmetic, harmonic or geometric progressions) got specific symbolic meaning and became associated with human or natural phenomena or events. In particular the ‘harmonic progression’, i.e. where each number of a simple sequence stays in ‘harmonious’ proportion to the previous and the following number, were popular and looked for1. The numbers, visible in real quantities (e.g. number of piers, of bays, of rooms, of corners, of stair-steps, ecc.) were a fundamental part of each building design; the same numbers served as the metaphoric indicator par excellence for expressing intangible values or messages such as power, devotion, glory, utility, science, beauty, harmony and other.
This is not the place to enter in detail about the number systems and the numeral calculus, nor about the wide range of symbolic values in Greek and/or medieval numbering. This chapter only stresses their presence and application since ancestral times, and their fundamental role in the genesis of all pre-industrial building projects. Understanding the ancient metaphors, hidden behind the physical quantities and dimensions in the building, is not so easy as the correct lecture and interpretation of the dimensions presumes the often missing knowledge about the metric unit (yard, foot, cubit?), about the eventual modulus (fixed group or set of units) and about the measuring and building conventions at the time and the place of the design. On top of this uncertainty, the modern observer is seldom familiar enough with the ancient design canons and number or figure symbolisms. The Pythagoreans (IVth-IIIth century BC) knew many types of numbers: real or rational ones, integers, fractions, even and uneven ones, primes, perfect numbers2, as well as irrational and complex ones (roots, unlimited ratio’s such as
Number ‘one’ is seen as the most important number, being the origin of everything, not only in arithmetic calculation but also in the natural world and the cosmos (also the justification for monotheism; although many cults worshipped a Divine Threesome in one Union, i.e. the Holy Trinity in Christianity). Number ‘two’, first and only even prime, represents dualism, the base of philosophy and all science; number ‘three’ means the female and number ‘four’ the male element in the 3–4-5 triangle. Number ‘four’ also refers to all groups of four elements in nature: the basic elements of everything (earth, water, fire, air); four cardinal directions, four seasons and, in Christian context, e.g. the four evangelists). The sum of these first four initial numbers 1 + 2 + 3 + 4 gives the number 10 (the sequence called “tetractys”), creating the sacral number ‘ten’, representing the universal order. Because of this special property, ‘ten’ got a special ‘mystic’ value and the Pythagoreans cultivated a particular preference for decades and pentades in arithmetic calculus and their homonymous polygons in geometry. The tetractys sequence generated the concept of calculated harmony in a eight-divided music-scale (from second to octave, the double of four tone intervals4), and the ancient eight-divided foot unit as well as the modern decimal measuring system. Also Vitruvius, explaining and defending the use of anthropomorphic dimensions, presented the number ‘ten’ as a sacral and most ‘beautiful’ number ([1], III,275)5 . The theory and philosophy on the use and allegorical value of numbers in ancient times is large and filled with unexpected results, but their decisive role in pre-industrial design and sometime also in post-industrial projects, is evident.
The most curious invention from ancient Greece, without any doubt, regards the concept of ‘figured numbers’. This means that the number (except number ‘one’) should not be seen as a single independed entity, but as a set or distribution, or as a part within a progression, and can be represented in space (linear, superficial or volumetric). The abstract number indicates the ratio between a certain quantity and the unit or dimension of that quantity on which it is relying (in this case on two-dimensional figures or surfaces or three-dimensional volumes). The philosophical background of the concept is more complex and relies on Plato’s theory on the proportions of volumes in the dialog Thééthète and presumes the alchemic mixture of arithmetic and geometry [4], p. 45. The concept of ‘figured numbers’ is particularly useful and explanatory in case of irrational numbers such as root
The combination of architecture and number philosophy has nothing to do with “numerology”, being a predominant esoteric discipline of fortune-telling and kabalistic or astrologic reading of phenomena about man and nature. It does not apply the scientific and rational ‘theory of numbers’ as intended by Greek philosophers, although even they did not use always the most objective logic, as e.g. by naming male and female numbers, inherited from Egypt. Part of this “numerology” is the practice of the old Hebraic ‘gematric’-modus (i.e. giving a numerical value to each letter of the alphabet, making it possible to convert letter-words into a mathematical value), used sometimes in the design of mayor buildings but forbidden in church design by the ecclesiastic authorities.
Finally, the rather primitive measuring instruments and the long lasting construction programs, forced ancient building practice to use preferably integer quotes and simple fractions (half, quarter, third), to facilitate tracing and execution on the building site. This explains the preference for integer numbers in the design of plan and elevation of a building. One also has to consider metric rounding after theoretic calculation and the difference between theory and practice to facilitate execution. Such condition on top of the normal building tolerances, on top of the physical degradation and deformation of historic buildings, ask for benevolent interpretation margins.
The geometry is indispensable in structuring any chaotic space. Similar with the procedures used by the Divine Geometer, also man had to create order and harmony by using appropriate geometric figures and proportions. Euclid of Alexandria (ca. 325–265 b.C.) wrote the first systematic manual on this matter, and from that period, a large gamma of regular and irregular geometric forms was developed. The numeric quantity and form of the angles and sides of the figures, in combination with other geometric properties as size, symmetry, congruency, similarity or opposition, they got special symbolic meaning in their architectural application. The most evident figures used in architectural design are the different types of lines (strait, bowed, dotted, alternated), the regular bi-dimensional figures (square, circle, triangle, polygons), and their tri-dimensional derivate. Plato’s description in Timaeus on the symbolic content of the regular polyhedra found many applications: the tetrahedron (fire), cube (earth), octahedron (air), dodecahedron (heaven with 12 constellations), and icosahedron (water) [3].
The wohltemperierte amalgamation of geometry and numbering were the necessary conditions for all harmonious architecture; they were the real determinants and driving forces in the design process, and the real generators of all architectural styles. Everything must be calculated, measured and proportionate, as the Holy Bible’s verse “Omnia in mensura et numero et pondere disposuisti” (Thou hast ordered all things in measure, number and weight – Book of Wisdom 11:21). This also explains the frequent presence of specific proportions such as the ‘Golden Mean’ or ‘Divina Proportione
As said, the most frequent geometries were the circle, the square and the triangle, as this were the most easy figures to draw up with simple instruments as wooden rod and cord, compass and plumb, but also because of their specific semantics generated since ancestral times. Before Columbus (ca. 1492), the image people had about the structure of the cosmos was that of a flat and square earth (with Jerusalem in the center) and a celestial half globe. It seems logic that the square and the circle, representing the earth and the heaven, were the first geometric figures used in architectural creations. Plato’s vision on the origin of the cosmos and the ‘elementary triangle’ as the fundament of all matter, together with his exaltation of mathematics and geometry at the expense of artistic creativity, contributed considerably in the use of different kind of triangles and the five polyhedra. Christian philosophers extended the ancient semantics with biblical or religious connotations as e.g. the circle became the representation of human society with God in the center and the people staying on the circle line, equidistant from the center and meaning that everyone is equally considered and protected by God. The square represented the walled Terrestrial Paradise or the walled terrestrial and celestial city of Jerusalem.
Apart from the circle, the triangle (equilateral, isosceles, rectangular, proportioned) and the four-angle polygon (square, rectangle, parallelogram, trapezium and rhomb) are the most frequent figures in architectural design, because of their large semantic spectrum and the easy designing technique. For that reason, they are the most popular geometries in architecture. The design ‘ad quadratum’ i.e. using different squarely formats connected, turned around, divided or superposed, was very popular in all kind of utility-building, ‘ad circulum’ was frequently used in centralized buildings (e.g. sepulchral monuments, baptisteries); ‘ad triangulum’ was most appropriate for the design of the building elevation and applied in many church buildings. Also in the panoptic of triangle-types, all had his specific symbolic meaning related with their arithmetic and geometric properties, e.g. the equilateral or ‘perfect’ triangle (symbol for the divine Trinity: three gods equipotential, united in one figure), the rectangular Pythagorean triangle with figured numbers on each side and, with female base and male height; the isosceles triangle symbolizes Christ: divine and human at the same time. Also triangles with specific ratios were used, as e.g. the ‘Egyptian’ triangle (as it signs the profile of the Cheops pyramid) is a isosceles triangle with the height equal to 0,625 (= 5/8) of the base (the most beautiful triangle according to Plutarch). Similar semantic discours got connected with all mentioned polygons (pentagon, hexagon, octagon and other).
As numbers and geometries are the mayor determinants in free architectural design (i.e. without conditions from the commissioner, from materials, or from topography), the definition of the right number and the proper geometry will determine the quality and the legibility of the final product. The correct selection and the proper combination of both determinants within the context of a given assignment, signs the art and the discipline of good design. Design is a research activity, similar at all other scientific research, and this chapter has not to enter in research methodologies or procedures, but intends to look after those tangible criteria used by earlier generations. Moreover, every designer, working on a specific commission, sitting in front of an empty piece of paper (or a white computer display) knows very well the process of trial and error, characteristic for all design processes.
This was not different in ancient times; one has to go back to the Roman legionary-architect Marcus Vitruvius Pollio (ca.81–15 b.C.) to read about procedures and criteria in architectural design. According Vitruvius, the decision on ‘what number to choose?’ is given in the proportions of the human body. The numbers of a good design should respect the metric relations between different parts of the body, to be multiplied according the necessity of the project. The use of anthropomorphic proportions and the human body as guiding principle in architectural design was an ancestral tradition adopted from Mesopotamia and Egypt and further used in all West-European cultures, up to the Modulor of Le Corbusier dd.1930 ca. (Figure 1a-e).
The proportions of the human body as reference for harmonic design. (a) Ancient Egypt (ca.2000 b.C.), (b) Vitruvius (1st century B.C.; illustration from the Italian edition by F. de Franceschi & J. Criegher in Venice dd. 1567), (c) Villard de Honnecourt (ca. 1235), (d) Leonardo da Vinci, the ‘Vitruvian man’ (ca.1490), (e) Le Corbusier, the ‘Modulor’ (ca. 1935).
The same Vitruvius gives indications about the geometry in the architectural project, not directly by speaking about geometric figures, but by explaining the disposition and distribution of each individual quantity. He puts ‘ordinatio et quantitas’ (in Greek: taxis and posotys) as the first of five conditions, what confirms what was said before on the importance of the ‘number’. The second design determinant, the geometry, is included in the ‘ordinatio’ and ‘dispositio’ (in Greek: diathesin) meaning the appropriate attention on the three design aspects and image-interpretation, i.e. iconographic, orthographic and scenographic criteria. Furthermore, Vitruvius explains the need for ‘eurythmia’ (general visual harmony), ‘symmetria et analoghia’ (harmony and similarity between elements by using a common ‘modulus’ for each part of the building, and ‘convenientia et distributio (greek: oikonomia)’ i.e. equilibrated administration of the available resources and space and, during the execution of the project, a proper division of the costs by calculation ([1], I,1,2). This topics are all well-known fundamentals of architectural theory, and consequently also the fundamentals in architectural design.
As Vitruvius expresses general and universal principles for correct and good architecture, it’s evident to find clear applications of his maxims in the design of all medieval buildings, even when there is no written testimony about the spread of his “Ten books of Architecture” in medieval times. The first evident reference to his manual is the drawing of Leonardo’s ‘Vitruvian Man’ (dd. 1490 ca.) as the new standard for artistic proportion and design. This means that the input of Vitruvius’s maxims never disappeared, not even after quasi 1500 years.
The one and only authentic written source on medieval design are the 65 pages the Picardian masterbuilder Villard de Honnecourt drew during his travels in Flanders and the Nord of France in the beginning of XIII° century. This sketchbook reveals some of the design techniques of his time, with particular attention for the proportion of building elements and sculptural decoration. Villard does not mention many numbers but shows in his drawings all geometric auxiliary lines and frames to guarantee the correct “euritmia” and “eumetria” in the project. He introduces the aid of a geometric pattern in the design of figures, building plans and elevations, and uses quasi exclusively square or triangular grids. His ideal plan for the “glize desquarie” (squared church) for the Cistercian convent is of particular interest as it represents the model of many West-European medieval churches. Basically, the plan adopts the three naves Latin basilica type with enlarged choir section, flat-ended apse and transept. It is easy to recognize the Vitruvian proportions of a double square module in the central nave axes (longitudinal sequence 6:2:4; transept 4:2:4) and a transversal section in the sequence 1:2:1, with the addition of co-modulated single square lateral naves in both directions. Villard also adds three alternatives for richer gothic choir and apse projects ([6], Taf.28) (Figure 2a-c).
Sketches from VILLARD DE HONNECOURT, before 1235 [6]; (a) linear modulated plan of the ‘squared church’ and ground floor of a large choir with deambulation and radiating chapels; (b) design of a men’s head on a squared modulated grid, (c) group of figures, using geometric patterns.
In the metric analysis of the St. Francis Church in Assisi [7], we also found the presence of a double grid design i.e. the superposition of a first principal square grid defining the sequence of open spaces for practical use, and a secondary in-between- grid for structural elements (walls and columns) (largely preceding the SAR-design method 7 presented as much innovative in the 1960’s).
As far as we could check, nor Vitruvius nor Villard use the word modulus in their texts, but it is obvious that the ‘modulus’ must be the key in any design project. This is what Vitruvius intends by combining “ordinatio et quantitas”, and what he makes explicit in his definition on “symmetria … est ex ipsius operis membris conveniens consensus, ex partibusque separatis ad universae figurae speciem ratae partis responses, …” (symmetry means bringing convenient consensus between all parts of the project, in the separate elements as well as by applying universal forms and figures….) ([6], I, 20–21). He continues by insisting on the use of the dimensions and the proportions within the perfect human body (mutual ratio’s between head, chest, arm, palm, foot, finger ecc…) as reference for all design. Villard does not say it by so many words, but presupposes the criterion of mathematical harmonious proportions in the totality of the project as well as in the mutual relation between separated components. He shows it through his graphic analysis and fragmentation of the design steps in his figures (without giving numeric quotes). Also for Villard, symmetry and analogy seem the main criteria.
The choice of the right modulus is the first indicator for the appropriate “ratiocinatione” (= the result of a rational decision-making process) by a most polyvalent-educated architect. In consult with his commissioner, he has to decide about the architectural typology of the building, determined by geometry and size, considering the symbolic capacity of both criteria. In medieval church building, the width of the choir indicated the module for the whole building; the numeric length of the modulus was most symbolic and took normally a number from the Holy Bible or some event related with the purpose of the project. In civil building, the length of the modulus could be of any kind related with the commissioner or the function of the building, as can be seen in the examples below.
Our ‘youngest’ example dates from ca. 1250 a.D. (Castel del Monte, Andria), but evidently, the modulated design practice did not stop after that period; quite on the contrary! The renaissance architects rediscovered and re-interpreted Vitruvius; and in one way or another, up to the end of the XIXth century all architectural design, working with traditional materials and traditional structures, took profit of the old master’s procedures, and ‘translated’ them in their own contemporary language. But the analysis of the design praxis in the Modern Times is a topic for another study.
Long before the Pythagorean philosophers, the design of this monumental tomb expresses a stupefacient simplicity and coherency in geometry and dimension, fully compatible with the Egyptian vision on the society and the cosmos, witnessing the exceptional culture and knowledge of some 5.000 years ago (Figure 3a-c).
Pyramid of pharaoh Cheops at Giza a. general view [8], photo: Giza-legacy.Ch; b + c: Geometries (drawings: Pinterest).
The symbolic geometry is obvious: the combination of a square ground floor, symbolizing the flat plane of the earth, and the upwards rising triangular flanks directed versus the celestial globe, with the mummy of the pharaoh and his wife, waiting for rebirth, at the center of the monument. The sides of the square are closely aligned to the four cardinal points. According to some researchers, inside passages and corridors are orientated versus astrologic constellations at the time of building. The King’s and Queen’s tomb chambers are located in the geometric gravity center of the construction, on emplacement and distances of interior corridors related to the golden mean proportion
The physical environment of the pyramid, the access way, the sphinx statue and the position of other monumental tombs of pharaoh related people or animals show a well-considered geometric and measured design.
The pyramid’s external dimensions witness an equally exceptional design with a selection of most allegorical and harmonious numbering, combining length of sides, diameter, vertical and sloping heights, including also particular prime numbers and the irrational ratio’s
groundfloor: square (440 x 440) Egyptian Royal Cubits (Erc) of ca.0,524 m = 230,56 m x 230,56 m;
height (original): 280 Erc = 146,70 m;
base angle ca. 51,575°, top angle: ca. 76,85°, length slope 220 Erc/cos 51,575° = 354Erc = 185,48 m
diameter groundfloor ca. 622,25 Erc = 326 m
ratio (height: side) = 280/440 Erc = 0,636 = ~ 5: 8 (= ~ 0,618 = golden mean) giving origin at the so called ‘egyptian triangle’, according Plutarch, the most beautiful triangle as it is derived from the equilateral triangle, see Figure 3b)
the ratio (height: ½ side) or the ratio between both cathedes of half of the isosceles profile signs = Erc (280/220) = 1,272727…=
ratio (perimeter: height) = 1760/280 Erc = 6,2857 equates to 2π to an accuracy of better than 0.05 percent (corresponding to the approximation of π as 22:7).
other surprising arithmetic ratios and golden- mean- related dimensions are signed in Figure 3c.
The integer dimensions (440 and 280 Erc) as well as the irrational ratio’s 5:8 and 2
This ancient Roman temple, dedicated to all God’s, is a most interesting monument for many reasons, a.o. for his unusual design showing a multilayered intangible content(Figure 4a-d). The building was founded as a rectangular temple about 27 b.C. by Marco Vipsanio Agrippa (ca. 63–12 b.C.), rebuilt two times, in his present form by the Emperor Hadrian in ca.125 a.C.. The monument shows two distinct parts: the mayor part or ‘Rotunda’ with cupola, and a squared classic temple, today serving as entrance area. The overall image shows two buildings, with two separated spatial identities, two formats and two functions.
Roma, pantheon: (a) aerial view (photo Pinterest); (b) ground floor of rotunda and so-called portico (Wikimedia & author); (c) transversal section (Pinterest); (d) longitudinal section (Getty images); (e) general view rotunda inside (Wikimedia).
The Rotunda represents the allegoric bricked envelop of an impressive regular globe. The horizontal diametrical plane of the globe divides the interior in two equally high volumes: the upper part shows a half-sphere dome, structured with cassettes and at the top an open oculus, the only entrance of natural light; the lower half is a cylindrical volume, for his part divided, according the golden mean ratio, in a lower section including a sequence of niches, apses and columns, and a upper dome-tambour section elaborated with squared cornice patterns. The cylindrical walls of the lower half of the interior are richly decorated by different kind of materials and forms. The Rotunda is the evident metaphor for the cosmos with the dome as the celestial half-round and the cylindrical lower space representing the terrestrial world, everything dominated by the central oculus representing the Supreme Divinity, generating life and dynamism through the zenithal light entering from the oculus .
The second part, the so-called portico, minor but nevertheless substantial part of the design, seems conform with one of the traditional Greek-temple-inspired models described in Vitruvius manual ([1], I, III, 284). However, the design does not follow this models too much; on the contrary, it is much closer with the most ancient tripartite Etruscan-Italian temple as described below. This deviant design from the conventional temple-pattern, seems an explicit demonstration of the own native Etruscan and Italian temple-building origins, different from those imported by the first Greek colonists in the 3th and 2th century b.C. It is as if Agrippa Vipsanio, commissioner of the first temple and son in law of the first roman emperor Antony-Augustus (with divine status), or one of his successors involved in the reconstruction of the temple during the 1st and 2d century a.C., liked to stress the native identity of the italic people, denying the fact of roman sacral architecture being indebted to the Greek (although this clearly appears from the first temples e.g. in Sicily and Paestum, or from the great basilica’s built on the Fori Romani). The squared portico part has a net floor area of circa half that of the Rotunda, which is much too large for being only ‘portico’ (or ‘pronaos’ as it is wrongly called in some literature). The approximate ground floor surface ratio 1:2, and the change from the rectangular versus the circular format, rather seems a conscious combination of old and new, an indication on the start of a new era for Rome, reminding the remote origins of the first Sannite (origin of the temple’s founder Agrippa Vip-sanio?), Tuscan or other settlers in Central Italy, and Rome’s passage as the capital of a new Mediterranean and European empire. The creation of the square antichambre-like temple before entering the incomparable grandeur of the Rotunda reinforces the expressivity of this last one. Both united entities are a rare example of architectural design as a political statement, materializing history and social order.
The geometry of the double monument
The circular Rotunda: as easily deductible from (Figure 4a-e), the geometry of the Rotunda is a most regular compilation of circles, squares and even a equilateral triangle (transversal section) in bi-dimensional and tridimensional edition, all of them having their specific semantic content. The rhythmic alternation of the tripartite ‘negative’ savings and ‘positive’ porches on the lower cylindrical ring, and the polychrome with the changing incidence of natural light from the oculus at the top, creates a great sense of wealth and dynamism. The frequency of the number eight in the design, by quantity (e.g. 8 mayor interior savings and 8 minor exterior ones in the perimeter wall) and by dimension (e.g. the overall inside diameter of 146 roman feet – see below), as well as the ancestral semantic of this number (being the first cubic number as 8 = 2 3) do believe that eight (and his composing factors 2–3-4) is the numeric modulus for the design of the complete building (to be completed with the number 5 as we’ll see further). The modulus should be found also in the net width of the opening of niches and apses, but the necessary metric information to prove such assumption, was not available. Apart from the 8 mayor niches/apses, the interior parietal composition includes 8 jutting out porches; both artifacts create a imaginary cylindrical space-filling web of 8 + 8 = 16 isosceles spiked volumes, pointing to the central vertical axis connecting nadir and zenith of the as imaginary interior sphere.
The squared entrance-temple is designed according the archetype of the three-cellae open Tuscan temple surrounded by columns and divided by two intermediate rows of three columns. The net inside width of the three cellae-areas is traced according the golden mean ratio. The two lateral cells have a small apse at the end for some simulacrum, the wider middle area covers the common central longitudinal axis with the double entrance door of the Rotunda, and is orientated versus the head apse of this Rotunda with the statue or the seat of the emperor. The entrance-temple is surrounded by eight columns at the front and three columns at each side with a short in- between piece of wall connecting with the Rotunda. The temple front looks similar with the Vitruvian models but other research is needed to identify all metric differences. After conversion of the Pantheon ensemble in a Christian church in 7th century, specially this entrance part and the in-between connecting structure suffered various amputations of materials and additions with demolition afterwards of crowning towers, but the present image should approach the authentic one.
The arithmetic at the double monument
The choice of the numbers, i.e. the dimensions and the quantities of the decorative elements (niches, apses, columns, porches, cornices, marble paneling), as well as their spatial distribution are additional indicators for the intangible messages. This chapter cannot enter into the detailed design aspects of this elements, but one can safely conclude that the Pantheon ensemble is probably the very first example of a fully integrated Gesamtkunstwerk based on the ancestral symbolic geometry and numbering; this last by using the very basic integer quantities 1–2–3-4-5-(1,618=
Three decisive choices at the start of the design process
The very first step concerns the choice on the architectural typology and form, based on the functional requirements and the symbolic content, mentioned before. This resulted in the option to create an innovative cosmic sphere imitation in a format which was never done before; to be connected with a reminder of the presumed architectural origins of Rome, i.e. the Tuscan temple. The innovation is proved by the technical audacity and capacity to build a dome structure signing the incredible span of ca. 43,40 m or 146 rf, which is the largest span ever in building from antiquity up to mid XIX° century! The 146rf dimension is certainly not arbitrary; the radius of 73rf = 1 + 8 + (8x8) might refer to the number 8 = 2x2x2 as the probable numeric modulus for the entire composition. Unfortunately, we do not know what symbolic content Agrippa or Hadrian connected with the number 8 modulus.
It’s interesting to notice also the open oculus diameter of ca. 8,35 m = ~ 28rf which signs ca. 28:146 = ~ 1:5 ratio to the central diametric plane of the sphere, and a reference to the semantic of number five, the golden mean ratio’s (including
The second decision concerns the likewise exceptional thickness of the external wall of ca. 5,90 m = ~ 20,00 rf (=2x2x5). Considering the presence of the deep niche savings, the minimum thickness of the external bricked shell is reduced at ca. 2,36 m ~ 8,00 rf (including the todays disappeared external marble wall cladding), which seems comparable with other antique buildings. This quote is not the result of any structural calculation (although technical experience must have been involved), but of an exclusive geometric property. Indeed, the external diameter signs ca. 55,20 m or 186,50 rf,, and his ratio to the inside 146rf diameter equals 186/146 = 1274 = ~ 1,2727 =
The third decision regards the dimensions of the entrance-temple, signing a double squared ground floor of ca. (31x17,5)m = ~(104x59)rf or a normal length to width ratio of approximate 2:1. Archeological excavations at the end of 19th century have proved the presence of more steps and a normal stylobate space to get on the columned entrance area; this means that the squaring could have been slightly different from today. More important however is the geometric connection between the Rotunda and this open temple structure. After searching and calculating possibilities, the author found out that the net frontal width BC (free passage between the side walls) of the portico (ca. 31,0 m) is given by the base of the ‘golden’ or ‘sublime’ triangle ABC inscribed in the Rotunda, with the top A in the center of the head-apse and the base along the inner line of the double (formerly bronze) entrance door of the Rotunda. (Figure 4b). The ‘golden triangle’ is found in the spikes of regular pentagons and decagons, and is a isosceles triangle such that the ratio of the hypotenuse to base is equal to the golden mean. The calculated hypotenuse signs 48,45 m (diameter from apse to entrance door)
The next design step draws a double square, sided ½ 31 m, at the left and the right of the central longitudinal axis to create the overall entrance-temple area DEFG. Further on, each of both composing squares get divided according golden mean ratio with the width of the lateral bay (with end-apse) as the ratios mayor and half of the central area as the ratios minor. The joining of both minors results in the central area along the common longitudinal axis of the ensemble. By this procedure, the entrance temple and the Rotunda get physical (through geometry and numbering) and spiritual (through various semantic) most intimately connected.
We notice, once again, the application of the golden mean ratio. The frequency of this ratio in so many design procedures indicates his particularly powerful allegoric meaning as indicator of cosmic harmony in life and society and of rebirth and infinity of man.
This concise analysis discovered several unexpected qualities of the Pantheon ensemble. They are not simple architectural ‘curiosities’ but existential part of the building’s identity. The multilayered image of this 2000 year’s old ensemble shows the resilience of simple but conscious design and his timeless capacity for tangible and intangible communication.
The chapel of the imperial palace of Charlemagne is another example of the impact of the geometry and the arithmetic in the dialog between the building and his observer. The commissioner is the first West- European emperor Charlemagne (742-814 a.D.) after the fall of Rome. For the chapel of his palace, he chooses the model of what he might have seen on his conquests (e.g. S. Vitale at Ravenna dated ca. 530 a.D.) and what linked him with his illustrious predecessors. He adopts, for the first time applied on this scale in the northern-of-the-Alps countries, similar innovative design which includes a lot of Christian and imperial symbolisms and allegories. We refer to the architectural history books for all details – e.g. see [11]. Our limited notes mark the most evident design characteristics employed as tangible instruments in the communication of intangible contents .
The geometry (unit of 1 cubit = 0,4281 m [12])
The plan: although the external image of the Carolingian building looks almost circular, the fundamental plan concept is of a squared design, i.e. a central octagon, which is the result of two superposed identic squares of which one is rotated over 45°. The central octagonal area is surrounded by a ring of eight squared chapels connected by eight triangular interspaces, generating the hexadecagonal external envelop. The central area is connected with the ring area through eight arched passages, covering quasi the full length of the octagon’s side (Figure 5b and e).
(a) Aachen, our Lady’s cathedral, longitudinal section with the carolingian chapel at the west half and the gothic choir (14th -15th century) at the east half of the ensemble. The tower above the west entrance and the raised dome above the octoganal Centre date from 18th & 19th century (drawing Kunstdenkmäler Rheinland, 1916). (b) Aachen, our Lady’s cathedral, ground floor of the end-8th century Kaiserkapelle (drawing from [12]). (c) squared proportions of different sections of the building and indication (star) of the emperor’s chair at the center of the lower circle (d): Arithmetic modulation and ratio’s in the octagon; (e): Mutual proportions in the squared ring-chapels / drawings from [12].
The vertical section shows two concentric volumes; the central octagonal higher one, open up to the top, and the surrounding hexadecagonal ring of two levels: the ground floor spanned by cross ribbed vaults and a upper gallery similar to the Romanesque matroneum, looking into the central octagonal space through a double superposed tripartite arched opening, divided by two columns. With the outside windows at the top of the octagon, the central area gives the impression of a four-level structure, where the physical evidence of the surrounding ring-volume counts only two floors (Figure 5c).
The center of the chapel forms a regular octagonal prism; the overall circumscribing volume marks a regular virtual cube, with the sides equal to the hexadecagonal’s diameter, inside of ca. 30,82 m = 72 cubits, and outside of 32,96 m = 77 cubits. The prism’s inside height from floor to the top also signs 72 cubits. The outside top-cornice of the hexadecagonal ring signs the precise horizontal middle plane (height 36 cubits) of the chapel, separating virtually the ground floor with the upper gallery of the imaginary ‘terrestrial world’ from the single globe on the top, inscribed in the dome and tambour volume, the imaginary space with the presence of the one God. Some parallel can be made with the domed upper half (with oculus) of the Roman Pantheon, apart from the octagonal perimeter versus the roman circular one. The sequence of spaces in the lower surrounding ring volume applies a most regular and symmetric geometry with congruent cubic volumes alternated by triangular spikes on both levels of the ring space (Figure 5c).
The amalgamation of the three most fundamental geometries: the square (part of the octagon), the circle (inside and outside virtual volumes, plus the staircase towers flanking the entrance), and the triangle (the eight spikes inside the hexadecagonal ring-space) generates an exquisite allegoric ensemble, open to all kind of profane and religious interpretations. In addition to the predominant centralizing plan-concept, the entrance extension with two massive towers in the west generates a additional east–west longitudinal axis (different from the Ravenna example10) with the emperor’s marble coronation armchair at the center of the upper squared floor section, looking east towards the central golden altar. Although the setting of the armchair might have been different at Charles’s time, it signs the beginning of a later developed Romanesque tradition to build a special imperial room on top of the west-entrance of more double oriented churches, as one can still admire in some large Meuse and Rhine churches (e.g. St. Servaes at Maastricht, Netherlands)11 .
The hexadecagonal chapel was a integrated part of a squared parceled design covering the complete imperial palace site, including an open forecourt to the chapel, two small and one larger basilica’s for the emperor’s activities as well as several residential buildings. On top of the sophisticated chapel geometry and the squared environmental design, there should have been involved also astronomic considerations related with the orientation of the single buildings and, concerning the chapel, with the incidence of natural light, the original emplacement of the altar, the location of the emperor’s chair, and other particularities, but this seems not enough studied yet.
The dimensions and quantities in the Imperial Chapel reveal the intended communication by specific dimensions. It starts with the choice for the octagonal plan, derived from the square, and including the semantics of the number four, eight, sixteen, and the multiplication factor two, appearing frequently in the building. The single entrance with the two flanking towers maybe symbolizes the double nature of his commissioner as worldly and religious sovereign, or also refers to the two columns crowning the entrance of King Solomon’s temple (1Kings,7,15–22) indicating imperial dynamism and power. The overall design might have been guided by Alcuinus of York (732–804), chief philosopher and theologian at Charlemagne’s court, as all mayor dimensions, after conversion into the then local cubitus (= ~ yard of ca.41,81 cm), result symbolic for biblical quotes. In this perspective, the design modulation does not appear that much a physical condition for related quotes, but a mental and metaphysical set of numbers taken from the Holy Bible and the ancestral arithmetic symbolisms passed down from the Pythagorean philosophers. Nevertheless, there is great logic and uniformity in the geometric figures as well as in the choice of integer and simple numbering for the design of length, width and height12 .
This logic is found in the harmonic multiples of small (metric) quantities, and simple mathematical sequences (e.g. the arched openings connecting the octagon and the hexadecagon sign 10–12-14 cub. (Figure 5e). As said, there is preference for prime numbers and biblical numbers, many of them borrowed from the Apocalypse and the there description of the celestial Jerusalem (Apo.21, 9). This Jerusalem is a squared city (the octagon signs the superposition of the terrestrial and the celestial Jerusalem), and was equally long, large and high, with a wall of 144 (=12x12) cub. Also the virtual cube of the chapel volume is equally high and large (inside 72 cub., outside 77 cub.13); the longitudinal side of the squares in the ring-volume signs 12 cub. and the transversal opening signs 11 cub.. The ratio 12:11 has his own semantic with 12 referring to the 12 apostles of Christ or to his presumed 33 year’s life time on earth (to read as 12 = 2x(3 + 3), meaning the double nature of Christ (divine and human) living during 33 years; and the 11-quote indicates men’s imperfection while Christ’s perfection is signed by 12; to be seen also as 11=1/3 x 33 of Christ’s lifetime and the metaphysical passage (12=4x3) from a squared central space into the triangular inserted spaces of the hexadecagonal ring.
The passages from the octagonal area into the surrounding area sign an opening of10 cub.(with 10 being the ‘perfect’ quantity). In most medieval churches, the architectural modulus is indicated by the passage opening between apse and choir; this brings us to conclude that also in this case, the metric modulus has been ten cubits (= ~ 4,18 m), eventually in combination with a secondary 12 cub. Quote as a recall at the left and right of the passage opening (Figure 5e). The octagon’s diameter of 33 ½ cub. is another reference to Christ’s life time on earth (Figure 5d); the diagonal of the hexadecagonal and the altimetry design, signing the same quote of cub. 72 = 2x3x12 = 3(2x3x4); the external diameters of the octagon (38 ½ cub.) and the hexadecagonal (77 cub.) sign the evident proportion 1:2. Also the number 77 should not be fortuitous because, according the calculations by S. Augustine of Hippo (354–430), (based on the Old Testament history data), the number of generations from Adam to Christ counts 77! [12]. The lecture of this building’s dimensions seems a florilège of biblical quotes, a very typical phenomenon in medieval church design .
The design of this French gothic key monument followed the then current design criteria: a ground floor ad quadratum according the église carrée of Villard (Figure 2a), and an elevation ad triangulum. Although the available metric documentation [13, 14, 15, 16, 17]; [18], p. 84, is not uniform, the evidence of a coherent modulated design is undeniable, including the inevitable smaller or bigger differences and deviations, caused by ca. 100 years construction activity by different master-building teams (Figure 6a). Similar to what we’ll see in Assisi, also in Chartres there is the very first question about what might have been the metric standard applied on the building site because of the confusion in the historic terminology and the uncertainty on the real length unit in practice at that time: or the French “coudée” or cubit of 0,5236 m, or the ancient “pied du Roy” of 0,3236 m or maybe still another local standard .
Chartres, Our Lady’s Cathedral. (a): Plan [13]. (b) Transversal section [16] and author. (c) Ratio’s transversal section (drawing from [16]). (d) Hypothetical altimetry design according musical canon [Wikicommons + A. Rubino + author].
As said, the metric modulus of the medieval church was given by the net passage opening between the apse and the choir or the choir and the nave, in this case signing 14,78 m (and not the 16,40 m distance between the center-to-center columns axis’s, as kept on in most publications)! Our reference publications propose different ‘fabricated’ measure units, quite close to the “pied du Roy”, as e.g. 0,369 m in [14] or 0,333 m in [18], partly inspired by the Christian semantics associated with number three and his multiples. However, none of those are coherent nor with any historic evidence. By reasoning back to front, comparing with other medieval churches, and inspired by the case of the St. Francis Church in Assisi, it seems probable and plausible that the Chartres’s bishop in charge, out of respect to the Roman Pope, absolute head of the Christian church, and maybe also out of piety tradition, imposed to practice the ancient roman foot standard of 0,296 m, applied in Rome and in many Christian church buildings. This same standard was called in France also “pied de Cluny” for the evident reason that the Cluny convent and the Cistercian monks, since late 10th century, were the most active church builders in Europe. Converting on this base, we obtain a most sense full and most Christian symbolic modulus of 14,78 m: 0,296 = 50rf (roman foot). The acceptability of this “pied de Rome” unit is confirmed in the converting of some representative dimensions of the building (always considering the only free passable spaces).
However, for want of a recent complete measured drawing, we only note following data (Figure 6a):
Architectural modulus: 14,78 m → 50rf = 2x5x5
Transversal width of choir-bay (estimated) 6,25 m →21rf = 3x7
The overall design grid in the choir and the nave signs a sequence of oblong rectangles and squares with slightly variable dimensions, apparently depending on the section of the columns and alternative ratio’s in the tracing of a virtual geometric grid. The grid indicates the net squared areas of the quadrangles, separated by the construction strips, corresponding with the width of the vault-arches. The quadrangles can include different semantics such as ‘golden rectangles’ (
According our scaled drawings, the nave and choir rectangles sign theoretically (50x21)rf, the aisles (25x21)rf and the crossing (50x42)rf. All quadrangles are semantically considered equivalent with regular squares.
Global inside length (from the entrance porte royal to the closing wall of the apse): 130,20 m → = 440 rf or 400rf of church passage and 40 rf of entrance zone between the towers;
Global inside width of nave and two aisles: 32,40 m → = 109,46 rf or 100rf of passable church area and 9,46 rf for two rows of pillars construction zone;
Global height inside the choir (from first outside stair- threshold of the entrance, i.e. the public space area, to the underside of the vault-keystone: 36,93 m → =125 rf = (5x5x5)rf (Figure 6b)
Global height from crypt-floor to nave vault: 39,46 m → 133,33 rf (to read as separated ciphers, with double reference to the 33 years Christ lived on earth)
As shown in (Figure 6b) there is no doubt about the elevation design ad triangulum with the modulus BC equal the half-side of the equilateral triangle with top A at quote 25,58 m (= 29,50x1/2
The top E signs the intersection of the two vault curves ‘at the fifth point’, at the height from outside threshold of 37 m in a ratio versus the width of the nave (= modulus) of 14,78 m:37 m = 1: 2,5. The modulus signs half of the length of the diagonal AB (=side equilateral triangle).
The external transversal section of the nave(including the crypt areas) describes a quasi-squared plane (height 40 m x large 44 m) = 10:11, which signs the already noticed semantics. The capitals of the triforium corridor (quote 19,73 m) sign the geometric middle of the full inside elevation; the overall transversal section on the nave forms a 5:2 rectangle (Figure 6b and c).
In this limited list, we see more references to number five and his multiples, including the number 100 and 400 as reference to the exclusive hecatompedon quantity (i.e. 100 = 10x10 what means a more than perfect number) and a supplementary indicator for the 50rf as present modulus.
To conclude, we have to mention the interesting hypothesis on the relation between architectural geometry canons and the music harmony canons, which seems to find confirmation in the elevation of Chartres’s nave (Figure 6d). Just as good architecture should bring order in a chaotic space, also harmonic music is as a cosmos imposed upon chaos [19], 251ff. Considering the side-length of the equilateral triangle (= double modulus) as an octave interval, one connects some significant altimetry quotes or virtual lengths from the opposite floor-bottom, with the tierce (lower capitals of central piers), the quart (keystone vault aisles), the quint (passageway triforium), sext or sept (lower threshold nave windows), first octave (impost vault-arch nave), second octave (~ keystone vault-arch nave). This needs further study before being confirmed.
The S. Francis Basilica at Assisi was built to worship the sepulcher of Francis of Assisi (1182–1226), founder of the mendicant Order of the Minorite Fathers. The work started March 1228, but after two years, mid 1230, the project mission was extended with a second assignment i.e. becoming the representative mother church of the new religious Order. Integrating this new function in the same, still under construction, sepulchral and pilgrimage church, did not seem possible and for want of space on the same site, it was decided to build a fully separated second church on top of the first one, keeping the same external wall’s perimeter and a similar inside spatial distribution. Such audacious project got realized after reinforcing the already built exterior walls and after inserting a new type of cross-rib vaulting for the lower church. This phased realization resulted in the actually superposed double-church, characterized not only by two different functions (a Lower devotional Church for S. Francis’s tomb and a Upper Church for the Fathers Convent’s services), but also by a double architectural and artistic identity: a first late-Romanesque Umbrian Lower Church and a second early-Gothic European Upper Church.
The design of the Lower Church (excluding later extensions) adopted the traditional middle sized Umbrian single-nave and single level church model. The geometry and the arithmetic’s on dimensions and quantities of the Lower Church integrated the ancient Pythagorean traditions, modifying them according Christian semantics with tangible imitation of some iconic Christian churches of that time i.e. the S. Peters Basilica of Rome and, as it was built in full crusaders period, also the S. Sepulcher of Jerusalem, and an allegoric record of the since long demolished biblical Temple of King Solomon on Mount Moriah in Jerusalem. This multifaceted mission was realized in a multilayered design, building a quite old-fashioned Romanesque Lower Church and an innovative Upper Church, this last one in the new transalpine (French) gothic style and structure, the very first application of this new architecture in Italy. It’s interesting to notice the remarkable coordination in the design of dimensions, forms and structures within two architecturally and structurally so different buildings. Under this prospective, the S. Francis double church is a interesting example of design resilience and flexibility avant la lettre [7, 20, 21].
The geometry
Notwithstanding the two-phased origin of the S. Francis Basilica14, the geometry and the arithmetic follow the ancestral symbolisms of figures and numbers, independent from the clearly different stylistic character of each church and the references in orientation, distribution and architectural forms to the three mentioned iconic Christian churches .The plan-geometry of the S. Francis Basilica is a evident ad quadratum design, persisting in the early XIII° century four bays Romanesque Lower church, continued in the mid XIII° century Upper Church and the XIV°- XVII° side chapels. The modulus is given by the width (11,84 m = 40 roman feet of 0,296 m) of the passage in between the apse and the crossing. As a one-nave church, the initial three nave-bays sign a perfect (40x40)rf square, separated from each other by a narrow strip for the transversal arches. The crossing signs (43x43)rf in order to realize an inside area with hecatompedon overall length from the XIII°century entrance door to the end of the apse. The bell-tower adopts the same modulation, applied however on the outside perimeter. The design of the later additions and modifications (the superposed second church, the east entrance-transept of the lower church and seven of the twelve lateral chapels) followed similar squared design. The lateral chapels however applied a reduced modulus of 23 or 24 roman feet (Figure 7b). The geometry of the two transept sections (added in a second campaign) are two ‘golden rectangles’, applying the same 40 rf modulus in longitudinal direction and defining the transversal width according the golden mean proportion (40 rf x 0,618 = 24,72 rf = ~ 7,32 m). The overall analysis of the modulation permitted the author to discover some revisions and changes in the design, realized in the course of building, and to identify the probable chronology of each section of this medieval project [7].
Double St. Francis basilica, Assisi. (a) General view from south-east [Wikidata]. (b) Squared ground floors lower basilica (left) and upper basilica (right); the black delineation indicates the mid-thirteenth initial three-bays church, the gray volumes the later additions [22] & author. Francis basilica, Assisi. (c) longitudinal section over both churches, extended in XIX° century with crypt. (d) transversal section over the transept (drawings [22] and author). S. Francis basilica, Assisi. (e) Lower church, view on the main altar, versus west [Wikimedia]. (f) Transversal section over main altar lower church and XIX° century crypt with location of sarcophagus in the nadir of reduced diophantic sphere (FOART measuring & author). (g, h) Geometric design of the east façade upper church (drawing and photo: Author).
The vertical section of the Lower Church signs a surprising ad circulum design, i.e. each bay includes a regular sphere with a diameter equal to the 40 rf modulus. The nadir of each sphere does not coincide with the pavement level, what should be the normal design, but with the quote of the Saint’s sarcophagus, below the pavement. The sarcophagus is located at the base of a virtual sphere, inscribed in the crossing of the nave and the transept. The same sarcophagus marks also the starting point for an imaginary vertical axis, passing the middle of the mayor altar, rising to the zenith, as to indicate that the remains of S. Francis, a unique relic treasure with exceptional thaumaturgy capacities, is the best go-between to resolve men’s problems. To stress this capacity, the difference between the extrados and the intrados perimeter of the circular transversal ribs delimitating the crossing, have been calculated according to a 1/3 reduced width of the diophantic proportion (similar to the Pantheon’s enveloping wall) indicating the double irrational and golden mean reference of
As the initial design provided with two identic churches, both ground floors are similar apart from the nave’s interior width, which is little larger in the Upper Church as the exterior wall thickness of the Upper Church is only half of those of the Lower Church. However, the opening between apse and crossing also in the Upper Church keeps strict on the architectural modulus of 40 rf, notwithstanding the different geometry of the apse (circular in the Lower Church and decagonal in the Upper Church). The stylistic differences in ornamental design (gothic versus romanesque) is also very evident as this emerges e.g. in the massive trilobate wall-piers of the Lower Church in contrast with the five-lobate clustered piers in the Upper Church. We also noticed several metric design irregularities, e.g. the vertical axis’ of the Lower and Upper Church wall-piers are not well centralized, nor the length of the bays and squared plan-grid of the Upper Church are very regular. This are obvious indications for the separate design of both churches, spread over more years and more chief-master builders; maybe also connected with the medieval concept on metric tolerance’s and the not so perfect measuring instruments. However, the visual impact of this metric irregularities is negligible as they get disguised by the full polychromic decoration of all walls and vaults.
The vertical geometry and the wall’s elevation of the Upper Church is very different from the Lower Church. As said, the Upper Church design expresses the introduction of the gothic design canons, inspired by the gothic examples in the French and English Normandy and the Scholastic church fathers. The design implies a predominant ad triangulum geometry and the application of the different types of pointed arches as indicated by Villard de Honnecourt [6].
The arithmetic numbering (conversion of metric unit: Roman foot = 0,296 m)
The numeric quantities in both churches reflect a large collection out of the mentioned sources from pagan antiquity, the Holy Bible, the Christian Scholastics, and the practical need for simple and rounded numbers. It starts with the choice of the horizontal as well as the vertical modulus in both churches, equal 40 rf (a number appearing more than 100 times in the Holy Bible and also the modulus of King Solomon’s Temple); the overal inside length (from apse to entrance) of the sepulchral church signs 200 rf (=5x40), the inside width of the transept signs 100 rf, i.e. a single and a double hecatompedon (the length of the mayor antique Greek temple); the inside length of the extended Upper Church signs 250 rf; the free height in the Lower Church from the pavement to the top of the transversal arch signs 30 rf and 33 rf to the vaults keystone signs; the piers’ impost in the Upper Church signs also 33 rf; the height of the vaults keystone signs 60 rf or a 1:2 proportion regarding the Lower Church height. In the Lower Church one finds the omnipresence of number three in different combinations, in the Upper Church, one finds similar arithmetic combinations of number five in dimensions (e.g. length 250 rf = 2x5x5x5) and in structural elaboration (e.g. five-lobate wall-piers). The number five cult and the ad quadratum design is largely visible in the Upper Church east façade, which could serve as the tangible synthesis of a rich and most interesting intangible program (Figure 7g-h).
Maybe the best known and most enigmatic medieval building with regard to architectural design is the castle built mid XIII°century near the site of a former small Our Lady’s convent in Andria (Puglia, Italy), commissioned by Frederic II (1194–1250), the then Holy Roman Emperor. It illustrates the amalgamation of European (Romanesque), classic antiquity (Roman) and Islamic design traditions, clearly combined with Christian semantics about life, rebirth, cosmic structure and the role of Jerusalem as religious and geographic center of the flat terrestrial world. Indeed, the octagon was since Babylonian times the main symbol for rebirth and eternal life (cfr. The supreme goddess Isthar was imaged by a octagonal star); the Pythagoreans loved the number eight for being the first prime ‘regenerating’ three times (8 = 2x2x2), and also the Christians used the octagonal plan for the baptistery, as this is the building where men get spiritual rebirth. In metaphysics, the number eight was the sum of three and five, two numbers with particular semantic meaning, and the octagon was associated with the person of Christ, as He was part of a tripartite God but also human being, using the five human senses (3 + 5 = 8), reborn the third day after death.
In the case of Castle del Monte, we propose a more tangible explanation for the triple octagonal design (i.e. the inner courtyard, the outside perimeter and eight octagonal edge-towers). As emerges from the plan and the geometry, the octagonal design seems a quasi-imitation, of the ca. 600 years earlier built Islamic octagonal ‘Dome of the Rock’ built on top of Mount Moriah in Jerusalem. This ‘Dome of the Rock’ as well as the adjacently located smaller ‘Dome of the Chain’ were at Frederick’s time converted into a Christian ‘Templum Dominum’, considered the geographic center of the world and also the most holy place for all three monotheist religions (Islam, Judaic and Christian). The ‘bare stone platform’, the center of the Dome of the Rock, was covered with the most extraordinary intangible content as it should have been the place where Abram sacrificed(Gen. 22), where Jacob dreamed (Gen. 28,11),where Solomon’s Temple had his main altar (1Kings 6; 2 Cron.), and from where Mohammed ascended to heaven. Notwithstanding the Muslim origin and property of this Dome, it seems quite plausible that Frederick II, also crowned King of Jerusalem since 1229, was inspired by this unique symbolic content, and that he ordered the construction of an octagonal ‘center of the world’- interpretation in his South Italian territories. Although the origin as a pagan (= Islamic) building, it was seen as inalienable part of the Christian heritage, and the imitation of similar octagonal building, substituting Solomon’s Temple, was the best way to prove this towards his subjects. On top of this, it could be seen as a sign of obligingness from an illuminated and open minded Emperor towards the Muslim society (who had many Arabic scientists at his court) (Figure 8d).
Andria, Castel del Monte: (a) General aerial view. (b, c) squared and circle geometry of the ground floor (photo, drawings: Pinterest). (d) Jeruzalem, ‘Dome of the Rock’ and the adjacent, at east, small ‘Dome of the Chain’, both with octogonal plan,located in the center of the Temple Mount platform at Jeruzalem (Wikipedia).
There also might have been a second motive in play. Political history learns about the quasi permanent conflict between Frederick II and the then Pope Gregory IX. This last one had started in 1228 the before cited Sepulchral Basilica for S. Francis in Assisi, including also some small residence at the papal service, mainly for devotional reasons but also to consolidate papal political power in Central Italy. As the Castle del Monte was built in the same years (i.e. before 1240–1249) as the S. Francis Basilica at Assisi (built 1228–1253)), this Castle del Monte might also have been meant as a imperial answer in virtual confrontation with the papal project in Assisi. This hypothesis of ours can explain the evident link in the design with the biblical structure in Jerusalem and changes the Castle from a hunting refuge, as presented in literature, into a political and religious statement, expressed by this most unusual triple octagonal castle design. (One should note that the actual rectangular Al-aqsa mosque as well as the ‘Dome of the Rock’- shrine has been damaged and rebuilt several times, but the mayor geometry of the ‘Dome’ has been preserved).
Many hypothesis on the semantics of this exceptional format have been proposed, but still today, there are more questions than answers [24]. In most literature, it is presented as a ‘hunting’ castle, or part of a greater military defense chain of castles by Frederick II, but none of this hypothesis make sense in this totally isolated location on a ca. 500 m high top. As said, it seems more a political sign against Pope Gregory IX (building the S. Francis basilica on top of the Assisi’s Collis Inferiore), and a confirmation of the emperor’s personal contribution as the leader of the 6th crusade, bringing the Jerusalem holy places under Christian control. The plausible intention of goodwill towards the Muslim world, the probable and significant astrologic input in the design of the castle, and the most probable sacral, social and political message gives this monument a complex and multilayered content, as intriguing as his design mixes the geometric and the arithmetic canons of Europe and the Mediterranean Basin.
The design draws two concentric octagons, with an open courtyard at the center and eight small octagonal towers at each corner of the external perimeter. Based on a possible (but not sure) metric unit of the ‘imperial foot’ of 0,52 m, the side of the overall circumscribed octagon signs (again) 40 feet and the ca.54,60 m diameter of the circle circumscription is to convert into the number 54,60: 0,52 = 105 feet, which indicates the particular arithmetic sequence of 105 = 1x3x5x7 i.e. the product of the first uneven prime numbers. The plan design is strictly squared in all particularities (Figure 8b,c). Each of the two floors is divided in eight trapezium-form rooms, covered by a central cross-rib square vault and two triangular side vaults. The parallel with the ‘Dome of the Rock ‘-temple is evident as also this one is designed as a double concentric octagon signing the same overall diameter of 105 feet or ca. 55 m, what supports the hypothesis about a conscious imitation in Andria. A further connection with Islamic architecture is given by the slim-line corner-towers reminding the typology of the 4th century square towers alongside the byzantine defense wall round Constantinople, although the octagonal towers at the Andria-castle do not seem too much for defense as well as for semantic reasons.
However, any further comment on a probable metric modulation or other design parallels are hindered by sufficiently controllable historic and metric information about the initial composition (this last aspect hampered by the lack of net interior dimensions as all inside decorative marble wall-cladding has disappeared, what makes authentic reliable measuring impossible). Further research is needed.
The above mentioned small selection of architectural and structural characteristics illustrate the impact and extraordinary image building capacity of the ancient design procedures and the role of semantics and symbolisms. The Vitruvian maxims on a.o. ordinatio, analogia, symmetria, euritmia [1], as well as the Pythagorean semantics on quantity, harmonic numbering and arithmetic sequences were the guiding criteria in the historic design; this same criteria became universal quality indicators by which also modern designers can be inspired in the choice and the definition about the form and the geometry of their project. Recent E.U. research programs [25] have proved in several occasions the possible input of heritage building analysis by digital modeling; the application of similar techniques and algorithms might be reversible and be introduced in creative new design.
Sustainable architectural construction means not only an economic or utilitarian driven concern but implies a at least as important social and humanitarian assignment. The quality of contemporary projects, and the long-term guarantee for visual and functional quality in the built environment will certainly be enriched by a design which is not looking for eye-catching artistic effects, as this last ones change with the wind, but by a comprehensive design based on the universal human scaled modulation of all times, with a simply legible semantic communication.
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',metaTitle:"Publication Agreement - Chapters",metaDescription:"IN TECH aims to guarantee that original material is published while at the same time giving significant freedom to our authors. For that matter, we uphold a flexible copyright policy meaning that there is no transfer of copyright to the publisher and authors retain exclusive copyright to their work.\n\nWhen submitting a manuscript the Corresponding Author is required to accept the terms and conditions set forth in our Publication Agreement as follows:",metaKeywords:null,canonicalURL:"/page/publication-agreement-chapters",contentRaw:'[{"type":"htmlEditorComponent","content":"The Corresponding Author (acting on behalf of all Authors) and INTECHOPEN LIMITED, incorporated and registered in England and Wales with company number 11086078 and a registered office at 5 Princes Gate Court, London, United Kingdom, SW7 2QJ conclude the following Agreement regarding the publication of a Book Chapter:
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\\n\\nLast updated: 2020-11-27
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The Corresponding Author (acting on behalf of all Authors) and INTECHOPEN LIMITED, incorporated and registered in England and Wales with company number 11086078 and a registered office at 5 Princes Gate Court, London, United Kingdom, SW7 2QJ conclude the following Agreement regarding the publication of a Book Chapter:
\n\n1. DEFINITIONS
\n\nCorresponding Author: The Author of the Chapter who serves as a Signatory to this Agreement. The Corresponding Author acts on behalf of any other Co-Author.
\n\nCo-Author: All other Authors of the Chapter besides the Corresponding Author.
\n\nIntechOpen: IntechOpen Ltd., the Publisher of the Book.
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\n\n2.1 Subject to the following Article, the Corresponding Author grants and shall ensure that each Co-Author grants, to IntechOpen, during the full term of copyright and any extensions or renewals of that term the following:
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\n\nThe Corresponding Author confirms that they (and any Co-Author) are and will remain a member of any applicable licensing and collecting society and any successor to that body responsible for administering royalties for the reprographic reproduction of copyright works.
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\n\n3. CORRESPONDING AUTHOR'S DUTIES
\n\n3.1 When distributing or re-publishing the Chapter, the Corresponding Author agrees to credit the Book in which the Chapter has been published as the source of first publication, as well as IntechOpen. The Corresponding Author warrants that each Co-Author will also credit the Book in which the Chapter has been published as the source of first publication, as well as IntechOpen, when they are distributing or re-publishing the Chapter.
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\n\nThe Corresponding Author shall obtain written informed consent for publication from people who might recognize themselves or be identified by others (e.g. from case reports or photographs).
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\n\n6.2 IntechOpen has the right to use the Corresponding Author’s and any Co-Author’s names and likeness in connection with scientific dissemination, retrieval, archiving, web hosting and promotion and marketing of the Chapter and has the right to contact the Corresponding Author and any Co-Author until the Chapter is publicly available on any platform owned and/or operated by IntechOpen.
\n\n6.3 IntechOpen is granted the authority to enforce the rights from this Publication Agreement, on behalf of the Corresponding Author and any Co-Author, against third parties (for example in cases of plagiarism or copyright infringements). In respect of any such infringement or suspected infringement of the copyright in the Chapter, IntechOpen shall have absolute discretion in addressing any such infringement which is likely to affect IntechOpen's rights under this Publication Agreement, including issuing and conducting proceedings against the suspected infringer.
\n\n7. MISCELLANEOUS
\n\n7.1 Further Assurance: The Corresponding Author shall and will ensure that any relevant third party (including any Co-Author) shall, execute and deliver whatever further documents or deeds and perform such acts as IntechOpen reasonably requires from time to time for the purpose of giving IntechOpen the full benefit of the provisions of this Publication Agreement.
\n\n7.2 Third Party Rights: A person who is not a party to this Publication Agreement may not enforce any of its provisions under the Contracts (Rights of Third Parties) Act 1999.
\n\n7.3 Entire Agreement: This Publication Agreement constitutes the entire agreement between the parties in relation to its subject matter. It replaces and extinguishes all prior agreements, draft agreements, arrangements, collateral warranties, collateral contracts, statements, assurances, representations and undertakings of any nature made by or on behalf of the parties, whether oral or written, in relation to that subject matter. Each party acknowledges that in entering into this Publication Agreement it has not relied upon any oral or written statements, collateral or other warranties, assurances, representations or undertakings which were made by or on behalf of the other party in relation to the subject matter of this Publication Agreement at any time before its signature (together "Pre-Contractual Statements"), other than those which are set out in this Publication Agreement. Each party hereby waives all rights and remedies which might otherwise be available to it in relation to such Pre-Contractual Statements. Nothing in this clause shall exclude or restrict the liability of either party arising out of its pre-contract fraudulent misrepresentation or fraudulent concealment.
\n\n7.4 Waiver: No failure or delay by a party to exercise any right or remedy provided under this Publication Agreement or by law shall constitute a waiver of that or any other right or remedy, nor shall it preclude or restrict the further exercise of that or any other right or remedy. No single or partial exercise of such right or remedy shall preclude or restrict the further exercise of that or any other right or remedy.
\n\n7.5 Variation: No variation of this Publication Agreement shall be effective unless it is in writing and signed by the parties (or their duly authorized representatives).
\n\n7.6 Severance: If any provision or part-provision of this Publication Agreement is or becomes invalid, illegal or unenforceable, it shall be deemed modified to the minimum extent necessary to make it valid, legal and enforceable. If such modification is not possible, the relevant provision or part-provision shall be deemed deleted.
\n\nAny modification to or deletion of a provision or part-provision under this clause shall not affect the validity and enforceability of the rest of this Publication Agreement.
\n\n7.7 No partnership: Nothing in this Publication Agreement is intended to, or shall be deemed to, establish or create any partnership or joint venture or the relationship of principal and agent or employer and employee between IntechOpen and the Corresponding Author or any Co-Author, nor authorize any party to make or enter into any commitments for or on behalf of any other party.
\n\n7.8 Governing law: This Publication Agreement and any dispute or claim (including non-contractual disputes or claims) arising out of or in connection with it or its subject matter or formation shall be governed by and construed in accordance with the law of England and Wales. The parties submit to the exclusive jurisdiction of the English courts to settle any dispute or claim arising out of or in connection with this Publication Agreement (including any non-contractual disputes or claims).
\n\nLast updated: 2020-11-27
\n\n\n\n
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