Biochemistry

"One of the most sinister features of DDT and related chemicals is the way they are passed on from one organism to another through all the links of the s. For example, fields of are dusted with DDT; meal is later prepared from the alfalfa and fed to hens, the hens lay eggs which contain DDT. Or the hay, containing residues of 7 to 8 parts per million, may be fed to cows. The DDT will turn up in the milk in the amount of about 3 parts per million, but in butter made from this milk the concentration may run to 65 parts per million. Through such a process of transfer, what started out as a very small amount of DDT may end as a heavy concentration."

"The robin was twitching, tremoring, convulsing uncontrollably, and peeping occasionally. The student handed the bird to me, and in a few minutes it was dead in my hands. It was April 23, 1963, and I was in my laboratory at in , when the student walked in with the bird. A week earlier the s of Hanover had been sprayed with the insecticide DDT to control the spread of by s."

"As an information specialist, Carson was in daily contact with wildlife biologists in those Fish and Wildlife divisions concerned with predator and pest control. They, along with the biologists at in nearby , were alarmed by some of the the early test results of the new synthetic pesticide dichlorodiphenyl-tricholorethane, known as DDT. Carson was particularly well informed on the progress of this research because her government mentor, friend, and former supervisor Elmer Higgins was collaborating with noted biological Clarence Cottam on a series of research reports on the impact of DDT on fish and other wildlife. These reports came to Carson's desk for editing, and their conclusions were the subject of debate and discussion around the office."

"A mosquito was heard to complain That a chemist had poisoned her brain The cause of her sorrow Was para-dichloro- Diphenyltrichloroethane."

"In the late 1960s, two chemists began to argue head to head in newspapers, journals, and courtrooms about a pressing question inspired by Rachel Carson's Silent Spring: ... namely, should the pesticide dichlorodiphenyltrichloroethane (DDT) be banned in the U.S.? The protracted debate between Stony Brook University professor Charles Wurster and University of California at Berkeley professor initially concerned the pesticide's effects on nontarget wildlife, including robins, hawks, salmon, and crabs. As the debate gained momentum and visibility, however, it became clear that it concerned DDT's effects on another species as well: humans. Jukes argued that few technological breakthroughs had done as much as DDT to save human lives, by protecting millions from malaria, typhus, and starvation. Wurster, on the other hand, maintained that DDT threatened the lives of millions more by destroying the ecosystems on which humanity depended for its well-being. Wurster argued that DDT's effects on wildlife were a harbinger of what was to come for humankind. Significantly, he also denounced the chemical as a pervasive likely carcinogen."

"Collagen is the primary component of the extracellular matrix in the human body. It has proved challenging to fabricate collagen scaffolds capable of replicating the structure and function of tissues and organs. We present a method to 3D-bioprint collagen using freeform reversible embedding of suspended hydrogels (FRESH) to engineer components of the human heart at various scales, from capillaries to the full organ. Control of pH-driven gelation provides 20-micrometer filament resolution, a porous microstructure that enables rapid cellular infiltration and microvascularization, and mechanical strength for fabrication and perfusion of multiscale vasculature and tri-leaflet valves. We found that FRESH 3D-bioprinted hearts accurately reproduce patient-specific anatomical structure as determined by micro–computed tomography. Cardiac ventricles printed with human cardiomyocytes showed synchronized contractions, directional action potential propagation, and wall thickening up to 14% during peak systole."

"The normal microbial colonization of sites in the body's tissues by certain bacteria requires that the bacteria first bind to extracellular secreted constituents, cell-surface membranes, or cell matrixes. This study examines two interactions of a variety of bacteria with the cell matrix noncollagenous proteins fibronectin and laminin and with basement membrane (Type IV) collagen. Adherence of bacteria to matrix proteins coated on tissue culture wells was examined with the use of radiolabeled bacteria. Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus sanguis bound well to fibronectin, laminin, and Type IV collagen, whereas a variety of gram-negative organisms did not bind. The interaction of soluble laminin, fibronectin, and Type IV collagen with bacteria was monitored by nephelometry with the use of a platelet aggregometer. S. aureus aggregated in response to fibronectin, laminin, or Type IV collagen. In contrast, gram-negative organisms did not aggregate with these proteins. It appears that fibronectin, laminin, and Type IV collagen can bind and aggregate certain gram-positive bacteria, and this binding is dependent on the surface characteristics of the organism. These adhesion molecules may play a role in the normal colonization of sites by microorganisms and in invasion during infections."

"Anti-laminin antibodies were sought for in the serum of workers exposed to mercury vapour (Hg, n = 58), lead (Pb, n = 38) or cadmium (Cd, n = 47). Thirty-one workers removed from Cd exposure for an average of eight years were also examined. Compared with control workers matched for age and socio-economic status, the prevalence of circulating anti-laminin antibodies was not increased in workers exposed to Hg (mean duration of exposure: 7.9 years and mean urinary excretion of Hg: 72 μg/g creatinine) nor in those exposed to Pb (mean duration of exposure: 10.6 years and mean Pb levels in blood: 535 μg/l). In contrast, anti-laminin antibodies were significantly more prevalent in Cd-exposed workers whose urinary Cd exceeded 20 μg/g creatinine. This observation was made in both currently exposed workers and in workers removed from Cd exposure (mean duration of exposure: 9.4 and 24.6 years and mean urinary Cd: 7.8 and 13.4 μg/g creatinine respectively). These autoantibodies were found in Cd workers with normal renal function as well as in those with increased proteinuria."

"The major glycoprotein component of animal cell basement membranes, laminin, is involved in a variety of cellular activities, including cell adhesion, differentiation, and mito- genesis, that are mediated by the interaction of laminin with specific cell-surfacereceptors. A laminin-binding protein with an apparent molecular mass of 68 to 72 kD was first char- acterized in mammalian tumor cells and considered as “the laminin receptor” (Liotta et al., 1986; Wewer et al., 1986). Severa1 putative cDNA clones encoding this protein have been isolated from mammals (Yow et al., 1988; Rao et al., 1989; Van den Ouweland et al., 1989; Grosso et al., 1991). A11 the clones contained an open reading frame coding for a highly conserved polypeptide with a calculated molecular mass of 33 kD. Independently, a cDNA encoding an identical polypeptide was isolated from mouse tumor cells (Makrides et al., 1988), but the expressed protein, named factor p40, was shown to be a component of the translation machinery (Auth and Brawerman, 1992). Recently, DNA-deduced amino acid sequences exhibiting homology with the previ- ously characterized 33-kD ”laminin receptor” were identified from hydra (Keppel and Schaller, 1991), Drosophila (M.B. Melnick, T.B. Chou, and N. Perrimon, accession No. M90422), and yeast (J. Miles and T.G. Formosa, accession No. M88277)."

"Extracellular matrix protein laminin binds specifically to yeast forms of Paracoccidioides brasiliensis and enhances adhesion of the fungus to the surface of epithelial Madin-Darby canine kidney cells in vitro. Immunoblotting of fungal extracts showed that the gp43 glycoprotein is responsible for adhesion. This was confirmed by binding assays using purified gp43, with a Kd of 3.7 nM. The coating of P. brasiliensis yeast forms with laminin before injection into hamster testicles enhanced the fungus virulence, resulting in a faster and more severe granulomatous disease. These results indicate that interaction of fungi with extracellular matrix elements may constitute a basis for the evolution of fungal infection toward regional spreading and dissemination."

"Pseudomonas aeruginosa is a major human pathogen known to infect tissues that have been previously damaged in some way. In wounded human respiratory tissues, P. aeruginosa cells were found attached to exposed basement membranes following epithelial denudation, suggesting that the affinity for extracellular matrix proteins may account for the bacterium's opportunistic character. By using microtiter wells coated with different P. aeruginosa strains, we demonstrated that laminin binds to both colonizing bacterial strains, isolated from asymptomatic carriers, and strains isolated from infected patients. Binding of soluble laminin to piliated P. aeruginosa PAK and to the nonpiliated isogenic mutant PAK/p—was shown to be saturable. Binding of laminin to the piliated PAK strain was not different from binding to the nonpiliated PAK/p—strain but was significantly higher than binding to the avirulent, nonpiliated PAK-N1 rpoN mutant. By transmission electron microscopy, we localized the laminin-binding sites on a loose material in the outermost layer of the bacteria. Western immunoblotting results suggested that 57- and 59-kDa nonpilus adhesins from the microbial outer membranes account for the binding of P. aeruginosa to laminin. We speculate that bacterial affinity for laminin may be of biological significance in the pathogenesis of P. aeruginosa infection of injured tissues."

"Retinal explants from embryonic or adult mice were placed on laminin or merosin substrates and the outgrowth of optic fibers was assayed under serum-free conditions. Both substrates strongly promoted outgrowth. A blocking antibody to the β1/β3 integrin subunits completely blocked laminin-dependent growth of embryonic optic fibers but had no detectable effect on adult fibers. Similarly, a blocking antibody against the main neurite-promoting region within the globular domain of the E8 fragment of laminin inhibited growth of embryonic fibers but had no effect on adult optic fibers. The β1 integrin subunit was identified immunohistochemically on both embryonic and adult fibers. These findings indicate that adult fibers have lost the β1 function which dominates laminin-dependent growth in embryonic fibers but express a receptor for laminin-dependent growth that is not detectable in embryonic fibers. These findings suggest that there are intrinsic differences between embryonic and adult optic fibers that may have implications for regenerative failure in the central nervous system of adult mammals."

"Adhesion of Aspergillus fumigatus, the causative agent of human aspergillosis, to the extracellular matrix protein laminin has been previously demonstrated. This study investigated the expression of laminin receptors during swelling of conidia, a step leading to germination and subsequent colonization of tissues. Scanning electron microscopy showed that the laminin binding sites were distributed over the external rodlet layer of resting conidia. During swelling, the characteristic rodlet layer progressively disintegrated and conidia surrounded by a smooth cell wall layer appeared. Flow cytometry using fluorescein isothiocyanate-conjugated laminin demonstrated that expression of laminin receptors at the surface of conidia was swelling dependent. Resting conidia expressed high levels of laminin receptors on their surface. A gradual decrease of laminin binding was then observed as swelling occurred, reaching a minimum for 4-h-swollen conidia. This correlated with a loss of adherence of swollen conidia to laminin immobilized on microtiter plates. Trypsin pretreatment of conidia reduced laminin binding. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ligand blotting with laminin identified in a cell wall extract a major 72-kDa cell wall glycoprotein which binds laminin. Thus, one of the initial events in the host colonization may be the recognition of basement membrane laminin by this 72-kDa cell wall surface component."

"Laminins, heterotrimers of α, β, and γ chains, are prominent constituents of basal laminae (BLs) throughout the body. Previous studies have shown that laminins affect both myogenesis and synaptogenesis in skeletal muscle. Here we have studied the distribution of the 10 known laminin chains in muscle and peripheral nerve, and assayed the ability of several heterotrimers to affect the outgrowth of motor axons. ... we show that motor axons respond in distinct ways to different laminin heterotrimers: they grow freely between laminin 1 (α1β1γ1) and laminin 2, fail to cross from laminin 4 to laminin 1, and stop upon contacting laminin 11. The ability of laminin 11 to serve as a stop signal for growing axons explains, in part, axonal behaviors observed at developing and regenerating synapses in vivo."

"We have previously demonstrated that Staphylococcus aureus, a highly invasive bacteria, presents a 52-kDa surface protein that mediates its binding to laminin. In order to better characterize this receptor, we excised this putative laminin receptor from two-dimensional (2-D) PAGE and used it as antigen for raising a mouse hyperimmune serum which was for screening an S. aureus expression library. A single clone of 0.3 kb was obtained, and its sequence revealed 100% homology with S. aureus α-enolase. Moreover, amino acid sequencing of the 52-kDa protein eluted from the 2-D gel indicated its molecular homology with α−enolase, an enzyme that presents a high evolutionary conservation among species. In parallel, monoclonal antibodies raised against the S. aureus 52-kDa band also recognized yeast α-enolase in western blot analysis. These monoclonal antibodies were also able to promote capture of iodine-labeled bacteria when adsorbed to a solid phase, and this capture was inhibited by the addition of excess rabbit muscle α-enolase. Finally, the cell surface localization of S. aureus α-enolase was further confirmed by flow cytometry. Hence, α-enolase might play a critical role in the pathogenesis of S. aureus by allowing its adherence to laminin-containing extracellular matrix."

"Mutations of LAMB2 typically cause autosomal recessive Pierson syndrome, a disorder characterized by congenital nephrotic syndrome, ocular and neurologic abnormalities, but may occasionally be associated with milder or oligosymptomatic disease variants. LAMB2 encodes the basement membrane protein laminin β2 which is incorporated in specific heterotrimeric laminin isoforms and has an expression pattern corresponding to the pattern of organ manifestations in Pierson syndrome. Herein we review all previously reported and several novel LAMB2 mutations in relation to the associated phenotype in patients from 39 unrelated families. The majority of disease-causing LAMB2 mutations are truncating, consistent with the hypothesis that loss of laminin β2 function is the molecular basis of Pierson syndrome. While truncating mutations are distributed across the entire gene, missense mutations are clearly clustered in the N-terminal LN domain, which is important for intermolecular interactions. There is an association of missense mutations and small in frame deletions with a higher mean age at onset of renal disease and with absence of neurologic abnormalities, thus suggesting that at least some of these may represent hypomorphic alleles. Nevertheless, genotype alone does not appear to explain the full range of clinical variability, and therefore hitherto unidentified modifiers are likely to exist."

"Laminins are large molecular weight glycoproteins constituted by the assembly of three disulfide-linked polypeptides, the α, β and γ chains. The human genome encodes 11 genetically distinct laminin chains. Structurally, laminin chains differ by the number, size and organization of a few constitutive domains, endowing the various members of the laminin family with common and unique important functions. In particular, laminins are indispensable building blocks for cellular networks physically bridging the intracellular and extracellular compartments and relaying signals critical for cellular behavior, and for extracellular polymers determining the architecture and the physiology of basement membranes."

"Laminin-211 is a major constituent of the skeletal muscle basement membrane. It stabilizes skeletal muscle and influences signal transduction events from the myomatrix to the muscle cell. Mutations in the gene encoding the α2 chain of laminin-211 lead to congenital muscular dystrophy type 1A (MDC1A), a life-threatening disease characterized by severe hypotonia, progressive muscle weakness, and joint contractures. Common complications include severely impaired motor ability, respiratory failure, and feeding difficulties. Several adequate animal models for laminin-α2 chain deficiency exist and analyses of different MDC1A mouse models have led to a significant improvement in our understanding of MDC1A pathogenesis. Importantly, the animal models have been indispensable tools for the preclinical development of new therapeutic approaches for laminin-α2 chain deficiency, highlighting a number of important disease driving mechanisms that can be targeted by pharmacological approaches."

"Laminins are composed of three polypeptide chains, designated as α, β, and γ. The C-terminal region of laminin heterotrimers, containing coiled-coil regions, short tails, and laminin globular (LG) domains, is necessary and sufficient for binding to integrins, which are the major laminin receptor class. Laminin recognition by integrins critically requires the α chain LG domains and a glutamic acid residue of the γ chain at the third position from the C-terminus. Furthermore, the C-terminal region of the β chain contains a short amino acid sequence that modulates laminin affinity for integrins. Thus, all three of the laminin chains act cooperatively to facilitate integrin binding. Mammals possess 5 α (α1–5), 3 β (β1–3), and 3 γ (γ1–3) chains, combinations of which give rise to 16 distinct laminin isoforms. Each isoform is expressed in a tissue-specific and developmental stage-specific manner, exerting its functions through binding of integrins."

"... We here postulate that basement membrane laminin is the key antigen in driving psoriasis, inducing a T cell-mediated autoimmune response. For laminin to be considered as the key autoantigen in psoriasis, it would be reasonable to expect the following to be demonstrable: (1) that autoantigens are present in psoriatic inflammation; (2) that basement membrane laminin is perturbed in involved and uninvolved skin, and that some of the pathological changes associated with psoriasis could be predicted as a sequel to this; (3) that disruption of the basement membrane is among the earliest events in the evolution of psoriatic lesions; (4) that as streptococcal pharyngitis is the most clearly defined event to trigger or exacerbate psoriasis, then a T cell-mediated autoimmune response to laminin should be anticipated as a potential sequelae to streptococcal pharyngitis; (5) that T cells in psoriasis can be shown to react to peptides with homology to laminin; (6) that HLACw6, as the most closely related gene associated with psoriasis and which is involved in antigen expression, should be preferentially expressed within lesional psoriasis towards the basement membrane, together with other proximal associated immune activity; and (7) that there is some association between antilaminin pemphigoid, a humorally mediated autoimmune disease to skin basement membrane laminin, and psoriasis."

"Integrins play an important role in cell adhesion by linking the cytoskeleton of cells to components in the extracellular matrix. In this capacity, integrins cooperate with different cell surface receptors, including growth factor receptors and G-protein coupled receptors, to regulate intracellular signaling pathways that control cell polarization, spreading, migration, survival, and gene expression. A distinct subfamily of molecules in the integrin family of adhesion receptors is formed by receptors that mediate cell adhesion to laminins, major components of the basement membrane that lie under clusters of cells or surround them, separating them from other cells and/or adjacent connective tissue. During the past decades, many studies have provided evidence for a role of laminin-binding integrins in tumorigenesis, and both tumor-promoting and suppressive activities have been identified."

"Laminin-α2-related congenital muscular dystrophy (LAMA2-CMD) is a devastating neuromuscular disease caused by mutations in the LAMA2 gene. These mutations result in the complete absence or truncated expression of the laminin-α2 chain. The α2-chain is a major component of the laminin-211 and laminin-221 isoforms, the predominant laminin isoforms in healthy adult skeletal muscle. Mutations in this chain result in progressive skeletal muscle degeneration as early as neonatally. Laminin-211/221 is a ligand for muscle cell receptors integrin-α7β1 and α-dystroglycan. LAMA2 mutations are correlated with integrin-α7β1 disruption in skeletal muscle."

"The research on laminin α2 chain-deficient congenital muscular dystrophy (LAMA2-CMD) advanced rapidly in the last few decades, largely due to availability of good mouse models for the disease and a strong interest in preclinical studies from scientists all over the world. These mouse models continue to provide a solid platform for understanding the LAMA2-CMD pathology. In addition, they enable researchers to test laborious, necessary routines, but also the most creative scientific approaches in order to design therapy for this devastating disorder."

"The capacity of pathogenic microorganisms to adhere to host cells and avoid clearance by the host immune system is the initial and most decisive step leading to infections. Bacteria have developed different strategies to attach to diverse host surface structures. One important strategy is the adhesion to extracellular matrix (ECM) proteins (e.g., collagen, fibronectin, laminin) that are highly abundant in connective tissue and basement membranes. Gram-negative bacteria express variable outer membrane proteins (adhesins) to attach to the host and to initiate the process of infection. Understanding the underlying molecular mechanisms of bacterial adhesion is a prerequisite for targeting this interaction by “anti-ligands” to prevent colonization or infection of the host. Future development of such “anti-ligands” (specifically interfering with bacteria-host matrix interactions) might result in the development of a new class of anti-infective drugs for the therapy of infections caused by multidrug-resistant Gram-negative bacteria. This review summarizes our current knowledge about the manifold interactions of adhesins expressed by Gram-negative bacteria with ECM proteins and the use of this information for the generation of novel therapeutic antivirulence strategies."

"In adult rat testes, the basement membrane is structurally constituted by laminin and collagen chains that lay adjacent to the blood-testis barrier (BTB). It plays a crucial scaffolding role to support spermatogenesis. On the other hand, laminin-333 comprised of laminin-α3/ß3/γ3 at the apical ES (ectoplasmic specialization, a testis-specific cell-cell adherens junction at the Sertoli cell-step 8–19 spermatid interface) expressed by spermatids serves as a unique cell adhesion protein that forms an adhesion complex with α6ß1-integrin expressed by Sertoli cells to support spermiogenesis. Emerging evidence has shown that biologically active fragments are derived from basement membrane and apical ES laminin chains through proteolytic cleavage mediated by matrix metalloproteinase 9 (MMP9) and MMP2, respectively. Two of these laminin bioactive fragments: one from the basement membrane laminin-α2 chain called LG3/4/5-peptide, and one from the apical ES laminin-γ3 chain known as F5-peptide, are potent regulators that modify cell adhesion function at the Sertoli-spermatid interface (i.e., apical ES) but also at the Sertoli cell-cell interface designated basal ES at the blood-testis barrier (BTB) with contrasting effects. These findings not only highlight the physiological significance of these bioactive peptides that create a local regulatory network to support spermatogenesis, they also open a unique area of research."

"Laminin, a non-collagenous glycoprotein present in the brain extracellular matrix, helps to maintain blood–brain barrier (BBB) integrity and regulation. Neuroinflammation can compromise laminin structure and function, increasing BBB permeability. ... We found that laminin may be a good indicator of BBB overall structural integrity, although changes in expression are dependent on the pathologic or experimental model used. In ischemic stroke, permanent vascular damage correlates with increased laminin expression (β and γ subunits), while transient damage correlates with reduced laminin expression (α subunits). Laminin was reduced in traumatic brain injury and cerebral hemorrhage studies but increased in multiple sclerosis and status epilepticus studies. Despite these observations, there is limited knowledge about the role played by different subunits or isoforms (such as 411 or 511) of laminin in maintaining structural architecture of the BBB under neuroinflammation."

"Blood vessels in the central nervous system (CNS) are unique in having high electrical resistance and low permeability, which creates a selective barrier protecting sensitive neural cells within the CNS from potentially harmful components in the blood. The molecular basis of this blood–brain barrier (BBB) is found at the level of endothelial adherens and tight junction protein complexes, extracellular matrix (ECM) components of the vascular basement membrane (BM), and the influence of adjacent pericytes and astrocyte endfeet. Current evidence supports the concept that instructive cues from the BBB ECM are not only important for the development and maturation of CNS blood vessels, but they are also essential for the maintenance of vascular stability and BBB integrity. In this review, we examine the contributions of one of the most abundant ECM proteins, laminin to BBB integrity, and summarize how genetic deletions of different laminin isoforms or their integrin receptors impact BBB development, maturation, and stability."

"Misleading data, false ideas, problems of personal interrelationships occur in much if not all scientific work. Consider, for example, the discovery of the basic structure of collagen, the major protein of tendons, cartilage, and other tissues. The basic fiber of collagen is made of three long chains wound around one another. Its discovery had all the elements that surrounded the discovery of the double helix. The characters are just as colorful and diverse. The facts were just as confused and the false solutions just as misleading. Competion and friendliness also played a part in the story. Yet nobody has written even one book about the race for the triple helix. This is surely because, in a very real sense, collagen is not as important a molecule as DNA. Of course this depends to some extent on what you consider important. Before Alex Rich and I worked (quite by accident, incidentally) on collagen, we tended to be rather patronizing about it. "After all," we said, "there's no collagen in plants." In 1955, after we got interested in the molecule, we found ourselves saying, "Do you realize that one-third of all the protein in your body is collagen?" But however you look at it, DNA is more important than collagen, more central to biology, and more significant for further research. So, as I have said before: It is the molecule that has the glamour, not the scientists."

"Collagen type I is the most abundant protein in mammals. It confers mechanical stability, strength and toughness to a range of tissues from tendons and ligaments, to skin, cornea, bone and dentin. These tissues have quite different mechanical requirements, some need to be elastic or to store mechanical energy and others need to be stiff and tough. This shows the versatility of collagen as a building material. While in some cases (bone and dentin) the stiffness is increased by the inclusion of mineral, the mechanical properties are, in general, adapted by a modification of the hierarchical structure rather than by a different chemical composition. The basic building block of collagen-rich tissues is the collagen fibril, a fiber with 50 to a few hundred nanometer thickness. These fibrils are then assembled to a variety of more complex structures with very different mechanical properties."

"The origin of collagen, the dominant structural component of metazoan extracellular matrix, has long been cited as a critical step in the evolution of metazoan multicellularity. While collagens were once thought to be found only in metazoans, scattered reports of collagen domains in Fungi, and more recently in close relatives of metazoans, have called into question whether collagens are truly unique to metazoans. Here, we take advantage of recently sequenced genomes and transcriptomes of diverse holozoans (the clade encompassing metazoans and their close relatives), as well as publicly available proteomes from diverse non-holozoan eukaryotes, to conduct a systematic search for collagen domains across eukaryotic diversity. We find that collagen domains are ubiquitous in choanoflagellates, the sister group of metazoans, and widespread across many other major eukaryotic taxa. Many predicted collagens in non-metazoans are comparable to metazoan collagens in length and proline content. Moreover, most are present in species that also encode putative prolyl 4-hydroxylase domains, suggesting that, like metazoan collagens, they may be stabilized through the hydroxylation of prolines. Fibrillar collagen and collagen IV appear to be unique to metazoans, and we posit that their ability to assemble into superstructures may have contributed to the origin of metazoan multicellularity."

"Collagen is the major component of the tumor microenvironment and participates in cancer fibrosis. Collagen biosynthesis can be regulated by cancer cells through mutated genes, transcription factors, signaling pathways and receptors; furthermore, collagen can influence tumor cell behavior through integrins, discoidin domain receptors, tyrosine kinase receptors, and some signaling pathways. Exosomes and microRNAs are closely associated with collagen in cancer. Hypoxia, which is common in collagen-rich conditions, intensifies cancer progression, and other substances in the extracellular matrix, such as fibronectin, hyaluronic acid, laminin, and matrix metalloproteinases, interact with collagen to influence cancer cell activity. Macrophages, lymphocytes, and fibroblasts play a role with collagen in cancer immunity and progression. Microscopic changes in collagen content within cancer cells and matrix cells and in other molecules ultimately contribute to the mutual feedback loop that influences prognosis, recurrence, and resistance in cancer. Nanoparticles, nanoplatforms, and nanoenzymes exhibit the expected gratifying properties. The pathophysiological functions of collagen in diverse cancers illustrate the dual roles of collagen and provide promising therapeutic options that can be readily translated from bench to bedside. The emerging understanding of the structural properties and functions of collagen in cancer will guide the development of new strategies for anticancer therapy."

"The brewer, who does not untie his belt in warm weather, whose hands do not dry the clay."

"Many of the historical brews were fun to drink. The meads brought on laughter. The gruit ales made people talkative. Dr. Butler's ale induced a peculiar golden heaviness. Some were bottled havoc. Whatever their effect, I was fascinated by the process of brewing a historical text into being. Old brewing recipes are records of how yeasts have etched themselves into human lives and minds over the last few hundred years."

"The maltster, never resting in winter."

"Malting to a very great extent determines the character of the to be fermented, and the fermentation acts upon that wort badly or well, not only in direct relation to the quality and nature of the yeast added, and the conditions of temperature and other surrounding influences of the wort, but in a greater and more important manner according to the nature of that wort itself. The wort is not only the element surrounding the yeast-plant, but it is the food-supply also, and upon the character and quality of the food supplied, the development and well-being of the yeast-crop mainly depend. My own experience has taught me that kiln-drying and exercise almost equally great influences upon the character of the wort, and as the one stage necessarily precedes the other, it must take the first place especially as the influences of the kiln are more permanent than those of the mash-tun."

"To dry pale malt well three things are necessary: 1. It should be loaded... as sound green-malt upon a floor at the thickness rarely, if ever, exceeding seven to eight inches. 2. It must remain unturned at a low temperature until nearly all moisture is removed. 3. Heat must then be applied steadily and freely, and be maintained for a considerable time at a nearly uniform height ranging from 160° to as high as 230° in some cases. With these three rules... vast quantities of malt now daily spoiled in the making, could be manufactured into a thoroughly sound and useful article. A really well designed and good kiln can always ensure the following conditions. Malt completely dried (i.e., deprived of 97 per cent of the total moisture present in green malt). Uniformity of dryness, colour, flavour, and aroma. Economy of working in cost of fuel consumed and labour employed."

"[C]ultured yeasts have only been in the winemaking picture within the last century. For thousands of years previously, all wine was fermented on wild (indigenous) yeast. Some old world wineries in France still ferment high-end wines on the local yeast."

"The trouble with being descended from a brewer, no matter how long ago he brewered, or whatever you call it, you're supposed to know all about something you don't give a hoot about."

"Much good beer has doubtless been made from very inferior malt, also vast quantities of so-called good beer are made by men who never even saw a proper mash; but neither of these facts can upset or affect the assertion that it is necessary to pay strict and careful attention to the drying of malt to produce a wort of uniform quality and absolute soundness. By careful working, inferior barley can be made into fairly sound and useful malt. By the employment of considerable skill... a brewer can make very good beer from indifferent malt, but his efforts, if carried back to the , are much more certain and reliable in their effects. No brewer needs to be told how much easier is his work, and more certain in its results, if he has malt well made and soundly dried."

"\frac{d{x}}{d{t}} = k_{3}p = \frac{k_{3}e(a-x)}{a-x+\frac{k_{2}+k_{3}}{k_1}} This is Michaelis and Menten's [1913] equation, (k2 + k3)/k1 representing their constant Ks. […] It may be remarked that with this modification of their theory, Michaelis and Menten's analysis of the effects of the products of the reaction, or other substances which combine with the enzyme, still holds good."

"A healthy, mature human being of normal intelligence may have upwards of 20 million RNA molecules [thought to serve as a ‘filing system’ for memory] in each neuron. . . . An RNA molecule made up of merely 25 links could have any one of a million billion different combinations, . . . In fact, every RNA molecule contains many hundreds of units—not merely 25."

"There is no question, then, that RNA presents a filing system perfectly capable of handling any load of learning and memory which the human being is likely to put upon it—and a billion times more than that quantity, too."

"Molecular biology has also shown that the basic design of the cell system is essentially the same in all living systems on earth from bacteria to mammals. In all organisms the roles of DNA, mRNA and protein are identical. The meaning of the genetic code is also virtually identical in all cells. The size, structure and component design of the protein synthetic machinery is practically the same in all cells. In terms of their basic biochemical design, therefore no living system can be thought of as being primitive or ancestral with respect to any other system, nor is there the slightest empirical hint of an evolutionary sequence among all the incredibly diverse cells on earth."

"It is so efficient that all the information . . . necessary to specify the design of all the species of organisms which have ever existed on the planet . . . could be held in a teaspoon and there would still be room left for all the information in every book ever written."

"The era of personalised medicine - medicine that matches treatment to DNA - has begun."

"DNA neither cares nor knows. DNA just is. And we dance to its music."

"DNA studies do not indicate that separate classifiable subspecies exist within modern humans. While different genes for physical traits such as skin and hair color can be identified between individuals, no consistent patterns of genes across the human genome exist to distinguish one race from another. There also is no genetic basis for divisions of human ethnicity. People who have lived in the same geographic region for many generations may have some alleles in common, but no allele will be found in all members of one population and in no members of any other."

"The way DNA encodes information is analogous to how we arrange the letters of the alphabet into words and phrases with specific meanings. The word rat, for example, evokes a rodent; the words tarand art, which contain the same letters, mean very different things. We can think of nucleotides as a four-letter alphabet. Specific sequences of these four nucleotides encode the information in genes. Many genes provide the blueprints for making proteins, which are the major players in building and maintaining the cell and carrying out its activities."

"Does the Covid-19 vaccine change your DNA? Not directly, no. But yes, thanks to your own immune system, your DNA is a tiny bit altered after you get any vaccine."

"There's no genetic basis for any kind of rigid ethnic or racial classification... I'm always asked is there Greek DNA or an Italian gene, but, of course, there isn't... We're very closely related."