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Book ChapterDOI

The Determination of Collagen and Elastin

01 Jan 1967-Methods of biochemical analysis (Methods Biochem Anal)-Vol. 15, pp 25-76
TL;DR: A critical evaluation of the various methods of quantitating collagen and elastin is concerned, pointing out the pitfalls to be encountered, and recommending the methods suitable for the particular situations that are currently under investigation in many laboratories.
Abstract: Collagen and elastin are two fibrous proteins which occur in varying amounts in practically every tissue. They are particularly abundant in the connective tissues. Collagen, and possibly elastin, is extremely widespread throughout the phylogenetic system. In the last 15 years there has been a widespread interest in these proteins, including physiological and clinico-pathological studies which depend almost exclusively on an accurate determination of the amount of these proteins in various tissues. In a good proportion of these studies, misunderstandings of the limitations of the methods used have reduced the value of the published data. This article is therefore concerned with a critical evaluation of the various methods of quantitating collagen and elastin, pointing out the pitfalls to be encountered, and recommending the methods suitable for the particular situations that are currently under investigation in many laboratories.
Citations
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Journal Article
TL;DR: A life-span study was carried out on longevity, pathologic lesions, growth, lean body mass, and selected aspects of muscle of barrier-maintained SPF Fischer 344 rats fed either ad libitum (Group A) or group A (Group B).
Abstract: A life-span study was carried out on longevity, pathologic lesions, growth, lean body mass, and selected aspects of muscle of barrier-maintained SPF Fischer 344 rats fed either ad libitum (Group A)...

533 citations

Journal ArticleDOI
TL;DR: Food restriction not only increased the mean length of life but also acted to extend life span since more than 60% of the Group R rats lived longer than the longest lived Group A rat.
Abstract: A life-span study was carried out on longevity, pathologic lesions, growth, lean body mass and selected aspects of muscle of barrier-maintained SPF Fischer 344 rats fed either ad libitum (Group A) or 60% of the ad libitum intake (Group R). Food restriction was as effective in prolonging the life of already long-lived SPF rats as previously shown for rats maintained in conventional facilities. Food restriction not only increased the mean length of life but also acted to extend life span since more than 60% of the Group R rats lived longer than the longest lived Group A rat. Renal lesions occurred at an earlier age in Group A rats than in Group R rats and progressed more rapidly. Death of most Group A rats was associated with severe renal lesions while few Group R rats showed such lesions at death. Food restriction was also found to delay or prevent interstitial cell tumors of the testes, bile duct hyperplasia, myocardial fibrosis and myocardial degeneration. Gastrocnemius muscle mass declined in advanced age and food restriction delayed this decline. Interestingly, however, lean body mass did not progressively decline with increasing age but rather decline occurred only after the onset of the terminal disease process.

515 citations

Journal ArticleDOI
TL;DR: This review integrates the biophysical, biomechanical, and biological aspects of interstitial and lymph fluid and its transport in tissue physiology, pathophysiology, and immune regulation.
Abstract: The interstitium describes the fluid, proteins, solutes, and the extracellular matrix (ECM) that comprise the cellular microenvironment in tissues. Its alterations are fundamental to changes in cell function in inflammation, pathogenesis, and cancer. Interstitial fluid (IF) is created by transcapillary filtration and cleared by lymphatic vessels. Herein we discuss the biophysical, biomechanical, and functional implications of IF in normal and pathological tissue states from both fluid balance and cell function perspectives. We also discuss analysis methods to access IF, which enables quantification of the cellular microenvironment; such methods have demonstrated, for example, that there can be dramatic gradients from tissue to plasma during inflammation and that tumor IF is hypoxic and acidic compared with subcutaneous IF and plasma. Accumulated recent data show that IF and its convection through the interstitium and delivery to the lymph nodes have many and diverse biological effects, including in ECM reorganization, cell migration, and capillary morphogenesis as well as in immunity and peripheral tolerance. This review integrates the biophysical, biomechanical, and biological aspects of interstitial and lymph fluid and its transport in tissue physiology, pathophysiology, and immune regulation.

514 citations

Journal ArticleDOI
TL;DR: Results showed the SIS remodeled neotendons to be stronger than the musculotendinous origin or the boney insertion by 12 weeks after surgery and to consist of organized collagen-rich connective tissue similar to the contralateral normal tendons.
Abstract: A study was conducted to evaluate the tissue response to a xenogeneic biomaterial when this material was used to repair an experimentally induced Achilles tendon defect in the dog. Twenty dogs had a 1.5 cm segmental defect of the Achilles tendon created surgically which was then repaired with acellular connective tissue derived from porcine small intestinal submucosa (SIS). The animals were sacrificed at 1, 2, 4, 8, 12, 16, 24, and 48 weeks and the neotendons examined for uniaxial longitudinal tensile strength, morphologic appearance, hydroxyproline (collagen) content, and disappearance of the originally implanted SIS material over time. The contralateral normal Achilles tendons served as controls as did four additional dogs that had a 1.5 cm segmental Achilles tendon defect created surgically without subsequent surgical repair with SIS. Results showed the SIS remodeled neotendons to be stronger than the musculotendinous origin or the boney insertion (> 1000 N) by 12 weeks after surgery and to consist of organized collagen-rich connective tissue similar to the contralateral normal tendons. The four dogs in which no SIS was implanted showed inferior strength at the comparable time points of 4, 8, 12, and 16 weeks. Immunohistochemical studies suggest that the SIS biomaterial becomes degraded within the first eight weeks and serves as a temporary scaffold around which the body deposits appropriate and organized connective tissue. SIS is a promising biomaterial worthy of further investigation for orthopedic soft tissue applications.

506 citations

Journal ArticleDOI
TL;DR: Results indicate that FAP depletion inhibits tumor cell proliferation indirectly, increases accumulation of collagen, decreases myofibroblast content, and decreases blood vessel density in tumors, providing proof of principle that targeting stromal cell-mediated modifications of the tumor microenvironment may be an effective approach to treating epithelial-derived solid tumors.
Abstract: Membrane-bound proteases have recently emerged as critical mediators of tumorigenesis, angiogenesis, and metastasis. However, the mechanisms by which they regulate these processes remain unknown. As the cell surface serine protease fibroblast activation protein (FAP) is selectively expressed on tumor-associated fibroblasts and pericytes in epithelial tumors, we set out to investigate the role of FAP in mouse models of epithelial-derived solid tumors. In this study, we demonstrate that genetic deletion and pharmacologic inhibition of FAP inhibited tumor growth in both an endogenous mouse model of lung cancer driven by the K-rasG12D mutant and a mouse model of colon cancer, in which CT26 mouse colon cancer cells were transplanted into immune competent syngeneic mice. Interestingly, growth of only the K-rasG12D-driven lung tumors was also attenuated by inhibition of the closely related protease dipeptidyl peptidase IV (DPPIV). Our results indicate that FAP depletion inhibits tumor cell proliferation indirectly, increases accumulation of collagen, decreases myofibroblast content, and decreases blood vessel density in tumors. These data provide proof of principle that targeting stromal cell-mediated modifications of the tumor microenvironment may be an effective approach to treating epithelial-derived solid tumors.

358 citations

References
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Journal ArticleDOI
TL;DR: Method has been applied to a study of hydroxyproline distribution in cell particulates, tissue fluids, and purified plant and animal proteins, and significant amounts of hydroXYproline were found in crystalline preparations of pepsin, elastase.

3,756 citations

Journal ArticleDOI
TL;DR: A short and simple calorimetric method is here described that is applicable to the determination of hydroxyproline in hydrolysates of 40 to 100 y of collagen with a reproducibility of f2 per cent and an accuracy of f1 per cent as judged by recovery of hydroXYproline from elastin hydrolysate and from an amino acid mixture simulating collagen.

1,249 citations