elastin and Lathyrism

Topics: Aerosols; alpha 1-Antitrypsin Deficiency; Animals; Blood Platelets; Cadmium; Collagen; Disease Models, Animal; Elastin; Emphysema; Forecasting; Hexanes; Lathyrism; Macrophages; Mice; Mice, Mutant Strains; Microscopy, Electron; Neutrophils; Nutrition Disorders; Oxygen; Pancreatic Elastase; Papain; Physical Exertion; Pulmonary Fibrosis; Pulmonary Gas Exchange; Pulmonary Heart Disease; Respiration; Trypsin

Topics: Aminopropionitrile; Animals; Ascorbic Acid; Collagen; Connective Tissue; Copper; Cross-Linking Reagents; Diet; Dietary Proteins; Drug Interactions; Elastin; Energy Intake; Extracellular Matrix; Flavonoids; Genes; Homocysteine; Lathyrism; Manganese; Monoamine Oxidase Inhibitors; Nutritional Physiological Phenomena; Penicillamine; Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase; Procollagen-Proline Dioxygenase; Protein Biosynthesis; Protein-Lysine 6-Oxidase; Proteins; Proteoglycans; Pyridoxine; Riboflavin; Thiamine; Vitamin A; Vitamin D; Vitamin E; Vitamin K; Vitamins; Zinc

It is apparent that significant progress has been made in our understanding of the biosynthesis, modifications, and maturation of collagen and elastin. We now recognize and partially understand special reactions involved in hydroxylations within the cell and complex cross-linking processes occurring outside the cell. Recent experiments (191) have shown that in human diploid fibroblast cultures of limited doubling potential (191) the hydroxylation of collagen prolyl residues appears to be "age" or passage-level dependent. With increasing passage level of these cultures, both the ascorbate requirements and the extent of collagen hydroxylation decrease. "Young" cell cultures have a strong requirement for complete hydroxylation and without ascorbate there is only about 50% of the normal level. "Middle-aged" cultures show higher hydroxylation without and full hydroxylation with ascorbate, whereas "old" (or cultures close to "senescence") are incapable of full hydroxylation with or without ascorbic acid. Although the overall system may show some deterioration with increasing passage levels, it appears that with increasing passage levels other components in the cell replace the ascorbate dependence of the hydroxylase system to a greater exten. In some ways, aging WI-38 cultures begin to resemble some transformed cells in their biochemical reactions, although they continue to remain diploid and eventually lose the ability to replicate. It is not yet known whether old animals can produce collagen, which may now be underhydroxylated, perhaps contributing to certain senescent changes. Careful examination of the hydroxylation index of collagen produced in organoid cultures of tissue biopsies as a function of donor age might be informative, particularly if one looks at the quality of collagen by employing collagenase and other proteolytic digests with collagen (191). One could comare the levels of frequent and characteristic peptide triplet sequences such as Gly-Pro-Hyp to Gly-Pro-Pro, Gly-Ala-Hyp to Gly-Ala-Pro, or Gly-Pro-Hyl to Gly-Pro-Lys and others for evaluation of hydroxylation throughout the entire molecule or at selected sequences.

Topics: Amino Acid Sequence; Animals; Antibody Specificity; Ascorbic Acid; Collagen; Connective Tissue; Copper; Ehlers-Danlos Syndrome; Elastin; Epitopes; Homocystinuria; Humans; Hydralazine; Lathyrism; Marfan Syndrome; Molecular Conformation; Platelet Aggregation; Procollagen-Proline Dioxygenase; Skin Diseases; Syndrome

Topics: Abnormalities, Drug-Induced; Amino Acid Oxidoreductases; Aminopropionitrile; Aneurysm; Animals; Aortic Aneurysm; Blood Vessels; Bone and Bones; Bone Diseases; Cleft Palate; Collagen; Connective Tissue; Elastin; Fetus; Heart Defects, Congenital; Humans; Kyphosis; Lathyrism; Lysine; Neurologic Manifestations; Nitriles; Poultry Diseases; Rodent Diseases; Spinal Cord; Spinal Cord Diseases; Stomach; Turkeys

Topics: Aging; Amino Acids; Aneurysm; Animals; Arteriosclerosis; Brain Diseases; Copper; Deficiency Diseases; Elastic Tissue; Elastin; Endocardial Fibroelastosis; Growth Disorders; Hair; Humans; Inflammation; Lathyrism; Marfan Syndrome; Metabolic Diseases; Respiratory Tract Diseases; Skin Diseases

Topics: Aging; Aldehydes; Animals; Bone Diseases; Collagen; Connective Tissue; Elastin; Lathyrism; Lysine; Peptide Biosynthesis; Pyridinium Compounds

Topics: Collagen; Collagen Diseases; Connective Tissue; Elastin; Humans; Lathyrism

Topics: Age Factors; Aminopropionitrile; Animals; Arteriosclerosis; Chemical Phenomena; Chemistry, Physical; Elastin; Lathyrism; Leucine; Molecular Weight; Protein Binding

Weanling Sprague-Dawley rats received beta-aminopropionitrile (BAPN) and/or hypertensive treatment, namely, heminephrectomy and administration of deoxycorticosterone acetate-NaCl. The BAPN-treated rats (lathyritic rats) died of dissecting aneurysm, and the victims with hypertensive treatment was greater in number and died earlier than those without the treatment, indicating that the rise of blood pressure promoted the onset of dissecting aneurysm. The elastic architecture of the ascending aortic media was examined not only by transmission electron microscopy with tannic acid stain and/or toluidine blue O stain, but also by scanning electron microscopy after hot formic acid treatment, and the area of interlaminar elastic fibers were morphometrically analyzed by a point counting method using transmission electron microscopic photographs. In the lathyritic rats, interlaminar elastic fibers showed a significant reduction compared with the control rats, and elastic fibers tended to become round-shaped and were frequently spotted with glycosaminoglycan, which suggest a disturbance of elastogenesis. On the other hand, elastic laminae were not disrupted and smooth muscle cells were well preserved. These results suggest that the alteration of the elastic architecture causes an unstable connection between each elastic lamina, and is related to the initiation and the progression of dissecting aneurysm.

Topics: Aminopropionitrile; Animals; Aorta; Aortic Aneurysm; Aortic Dissection; Blood Pressure; Body Weight; Elastic Tissue; Elastin; Glycosaminoglycans; Hypertension; Lathyrism; Male; Rats; Rats, Inbred Strains

We evaluated the effect of beta-aminopropionitrile (beta APN) on the nitrogen dioxide (NO2) animal model of emphysema. Rats maintained on a beta APN-supplemented or a regular diet were exposed to 30 ppm NO2 for intervals ranging from 1 to 8 wk. Emphysema development was assessed by histologic evaluation and by changes in lung volume and mean linear intercept values. Evidence of pathologic changes were also documented by clinical and radiographic findings of osteolathyrism. The induction of centriacinar emphysema was attributed specifically to NO2 exposure. Neither the severity of the emphysema nor the time course of its development was altered by the beta APN-supplemented diet. These findings are in marked contrast to those observed with the exogenous elastase model of the disease, and they suggest that elastin synthesis and repair may not modulate elastin destruction in the NO2 model of emphysema.

Topics: Aminopropionitrile; Analysis of Variance; Animals; Diet; Disease Models, Animal; Elastin; Germ-Free Life; Lathyrism; Lung; Male; Nitrogen Dioxide; Organ Size; Pulmonary Emphysema; Rats; Rats, Inbred Strains; Time Factors

These experiments were designed to determine if local differences in elastic properties or wall structure contributed to the development of aortic rupture in turkeys. Static elastic properties were measured from pressure-volume curves of cylindrical segments of upper and lower thoracic aorta and innominate artery in two groups of lathyritic birds and two age-matched control groups. The biggest changes were in the young lathyritic group that was on 0.04% beta-aminoproprionitrile at 3 weeks of age and continued for 2, 3, or 4 weeks. Much smaller changes were seen in the older group that started on 0.05% beta-aminoproprionitrile at 6 weeks of age for 10 days and then 0.02% beta-aminoproprionitrile for the next 9-12 weeks. The biggest changes were seen in the abdominal aorta, which ruptured spontaneously in 50% of the young lathyritic group. The birds had a marked decrease in the number of elastin layers per unit wall thickness. Analysis of the elastic diagrams showed that the elastance of both elastin and collagen were altered in the young lathyritic group. Rupture appeared to follow partial dissection of plaques, but true longitudinal dissection did not occur.

Topics: Aminopropionitrile; Animals; Aorta, Thoracic; Aortic Rupture; Brachiocephalic Trunk; Collagen; Elasticity; Elastin; Lathyrism; Male; Poultry Diseases; Turkeys

Experimental lathyrism was induced by feeding newborn chicks a diet containing 0.2 and 0.4% DL-Penicillamine, with or without CuSO4 (10 mg/Kg diet) and Vitamin B6 (100 mg/Kg diet), or 0.015 and 0.1% beta-aminopropionitrile fumarate (beta-APN). After 7, 15, 25 and 55 days of treatment the animals were killed, the aortas removed and processed for electron microscopy in the presence of markers for proteoglycans, and the elastic fibers were carefully examined. Penicillamine, which prevents the formation of desmosine crosslinks by binding to precursors, induced the production of numerous new elastin fibers which appeared normal from the ultrastructural point of view. It seems, therefore, that at least in chick aortas, desmosine crosslinks are not necessary for the aggregation of tropoelastin molecules into structurally normal fibers. On the contrary, beta-APN, a classical inhibitor of lysyl oxidase, induced the tropoelastin molecules to aggregate into abnormal protuberances on the old fibers. Moreover, the elastin deposited during beta-APN treatment was always permeated by cytochemically revealed proteoglycans, which were never observed after penicillamine treatment. It is speculated that, at least in the system under study, the epsilon-amino groups of tropoelastin molecules may offer the binding sites for matrix proteoglycans until they are removed by lysyl oxidase, and that matrix proteoglycans might play a role in elastin fibrogenesis by preventing spontaneous tropoelastin aggregation in areas far from growing elastin fibers.

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Chickens; Cross-Linking Reagents; Desmosine; Elastin; Glycosaminoglycans; Lathyrism; Microscopy, Electron; Penicillamine; Tropoelastin

By using various cytochemical stains, proteoglycans are shown to be present inside elastic fibers in aortas of beta-aminopropionitrile-induced lathyritic chicks. Depending on the characteristics of the dyes, the shape, size and distribution of the proteoglycan-revealing precipitates are described. The monocationic dye toluidine blue O and the tetracationic dye Alcian blue in the presence of 0.3 M MgCl2 give the most detailed results. With these stains the proteoglycans inside lathyritic elastin appear to be lateral branches of matrix proteoglycans, lying on the external surface of the elastic fibers. A possible general biological significance of elastin-proteoglycan association is briefly discussed.

Topics: Aminopropionitrile; Animals; Aorta; Chickens; Elastic Tissue; Elastin; Glycosaminoglycans; Histocytochemistry; Lathyrism; Proteoglycans

Tropoelastin a was isolated from lathyritic chick aortae by using severe denaturing conditions for the initial extraction. The amino acid composition of this new species of tropoelastin is elastin-like in its high proportion of proline, glycine, alanine and valine. However, it differs from authentic tropoelastin b in containing a higher percentage of polar amino acids and cysteine residues. In addition, the amount of proline hydroxylation is 3 times higher than that found in chick tropoelastin b.

Topics: Amino Acids; Animals; Aorta; Chickens; Chromatography, DEAE-Cellulose; Elastin; Electrophoresis, Polyacrylamide Gel; Lathyrism; Protein Denaturation; Tropoelastin

Ruthenium red and toluidine blue O precipitates were described associated with lathyritic elastic fibers in aortas of chickens treated with beta-aminopropionitrile fumarate (I. Pasquali-Ronchetti, C. Fornieri, I. Castellani, G. M. Bressan, and D. Volpin (1981). Alterations of the connective tissue components induced by beta-aminopropionitrile. Exp. Mol. Pathol. 35, 42-56). In this report evidence is given that these precipitates reveal the presence of proteoglycans, as they are completely removed by 5 M guanidine-HCl incubation and by specific enzymatic digestions. In particular, proteoglycans associated with the poorly cross-linked lathyritic elastin can be removed by testicular hyaluronidase, chondroitinase ABC, heparitinase, and nitrous acid treatments, whereas they are rather resistant to streptococcal hyaluronidase and chondroitinase AC. On the contrary, proteoglycans of the matrix or associated with collagen fibers are particularly sensitive to these latter enzymatic treatments. The conclusion is reached that glycosaminoglycans associated with beta-aminopropionitrile-induced lathyritic elastin (i) are different from those of the matrix or associated with collagen, and (ii) include mainly dermatan and heparan sulfates.

Topics: Aminopropionitrile; Animals; Aorta; Chickens; Chondroitinases and Chondroitin Lyases; Collagen; Dermatan Sulfate; Elastin; Glycosaminoglycans; Guanidine; Guanidines; Heparitin Sulfate; Hyaluronoglucosaminidase; Lathyrism; Nitrous Acid; Polysaccharide-Lyases

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Cartilage; Chickens; Elastin; Lathyrism; Lung; Methods; Swine; Tropoelastin

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Chickens; Collagen; Connective Tissue; Elastin; Extracellular Space; Lathyrism; Lung; Skin

Topics: Amino Acids; Animals; Aorta; Chick Embryo; Chickens; Elastin; Lathyrism; Lung; Molecular Weight; Organ Culture Techniques; Organ Specificity; Peptide Fragments; Tropoelastin

Topics: Amino Acids; Aminopropionitrile; Animals; Chemical Phenomena; Chemistry; Collagen; Elastin; Hydrazines; Lathyrism; Male; Oxalates; Penicillamine; Pyridinium Compounds; Rats; Rats, Inbred Strains; Semicarbazides

Tropoelastin was isolated from aortae and auricular cartilage obtained from lathyritic piglets. The two tissue-specific tropoelastins were judged homogeneous by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration on a high-pressure liquid chromatograph. Comparative studies of the tropoelastins were performed. Amino acid analysis revealed that the aortic and cartilage tropoelastins were very similar, if not identical, with the only exception that the cartilage tropoelastin contained more hydroxyproline and less lysine residues, both of which can be attributable to post-translational modifications. Both tropoelastins possess an apparent molecular weight of 70 000 and exhibit similar peptide fragments with limited trypsin cleavage. Antiserum raised to the aortic tropoelastin was used to show immunological identity between the two tissue tropoelastins.

Topics: Amino Acid Sequence; Amino Acids; Aminoacetonitrile; Animals; Aorta; Ear Cartilage; Ear, External; Elastin; Lathyrism; Male; Swine; Tropoelastin

Topics: Aging; Amino Acids; Aminoacetonitrile; Aminocaproates; Aminocaproic Acid; Animals; Chickens; Chromatography, Ion Exchange; Elastin; Electrophoresis, Polyacrylamide Gel; Lathyrism; Molecular Weight; Tropoelastin

Soluble elastin was isolated from lathyritic chick aorta using neutral salt solutions in the presence of beta-amino propionitrile. The effect of a carboxy-methylation step in conjunction with proteolytic inhibitors was investigated. Hydrodynamic (Stokes) radii of soluble elastins were measured by gel filtration and the molecular size and weight distribution in purified fractions are reported.

Topics: Amino Acids; Animals; Aorta; Chickens; Disulfides; Elastin; Iodoacetates; Lathyrism; Molecular Weight; Protein Conformation

Topics: Amino Acid Sequence; Amino Acids; Aminopropionitrile; Animals; Chickens; Circular Dichroism; Collagen; Elastin; Epitopes; Lathyrism; Microscopy, Electron; Models, Biological; Molecular Weight; Oxidation-Reduction; Protein Precursors; Solubility; Tropoelastin

The presence of an enzyme(s) associated with purified tropoelastin has been established. Results indicate that the enzyme(s) remains closely associated with the soluble elastin throughout the entire purification procedure suggesting that it is very tightly bound. Enzymatic activity is optimum through the pH range 7-9 (37 degrees C) and can be inhibited by disodium ethylenediaminetetraacetate, N-ethylmaleimide, sulfite, soybean trypsin inhibitor and human alpha-1-antitrypsin. The fragmentation pattern appears to be specific and reproducible.

Topics: alpha 1-Antitrypsin; Amino Acids; Animals; Aorta; Chickens; Edetic Acid; Elastin; Electrophoresis, Polyacrylamide Gel; Ethylmaleimide; Humans; Lathyrism; Molecular Weight; Peptide Hydrolases; Sulfides; Sulfites; Tropoelastin; Trypsin Inhibitor, Bowman-Birk Soybean

Topics: Animals; Aorta; Cell Line; Chickens; Cross Reactions; Elastin; Immunodiffusion; Immunoelectrophoresis; Lathyrism; Tropoelastin

Topics: Amino Acids; Animals; Aorta; Chickens; Elastin; Electrophoresis, Polyacrylamide Gel; Immunodiffusion; Immunoelectrophoresis; Lathyrism; Molecular Weight

Topics: Animals; Arteriosclerosis; Collagen Diseases; Cutis Laxa; Elastin; Humans; Lathyrism; Models, Chemical; Rabbits; Rats; Structure-Activity Relationship

Tropoelastin was isolated from the aortas of chicks rendered lathyritic by treatment with beta-aminopropionitrile. The soluble elastin was judged homogeneous by sodium dodecyl sulfate polyacrylamide gel electrophoresis and possessed an estimated molecular weight of 70000. Automated sequential analysis revealed that the N-terminal region of the chick tropoelastin is very homologous to tropoelastin isolated from copper-deficient piglets. N-terminal analysis of a trypsin digest of chick tropoelastin showed that tyrosine frequently is found adjacent to lysine residues. This positioning of tyrosine residues may be significant in terms of a possible regulatory role in elastin cross-link formation.

Topics: Amino Acid Sequence; Amino Acids; Animals; Aorta; Chickens; Copper; Elastin; Lathyrism; Molecular Weight; Protein Precursors; Swine; Trypsin

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Benzyl Compounds; Binding Sites; Cattle; Chickens; Chromatography, DEAE-Cellulose; Collagen; Connective Tissue; Copper; Diet; Elastin; Electrophoresis, Disc; Lathyrism; Lysine; Oxidoreductases Acting on CH-NH Group Donors; Protein Binding; Swine; Tendons

The isolation of a salt-soluble homogeneous elastin from the aortas of lathyritic chicks by chromatography on DEAE-cellulose and salt precipitation is described. These new techniques, as well as some previously published by other workers, were evaluated with the help of antiserum raised in sheep against insoluble chick elastin. The purified elastin was very basic and behaved in a predictable manner in coacervation studies. The protein migrated in sodium dodecyl sulphate-polyacrylamide gels as a single band moving slightly faster than pyruvate kinase (mol.wt. 57000).

Topics: Amino Acids; Animals; Aorta; Chemical Precipitation; Chickens; Chromatography, DEAE-Cellulose; Elastin; Hydrogen-Ion Concentration; Lathyrism; Molecular Weight; Pyruvate Kinase; Sheep

Topics: Aminopropionitrile; Animals; Chick Embryo; Elastic Tissue; Elastin; Lathyrism; Microscopy, Electron; Muscle, Smooth

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Chickens; Elastin; Electrophoresis; Iodine Isotopes; Lathyrism

Topics: Acetaldehyde; Amino Acid Oxidoreductases; Aminopropionitrile; Animals; Carbon Isotopes; Collagen; Collagen Diseases; Cyanides; Elastin; Lathyrism; Lysine; Metabolism, Inborn Errors; Protein Binding; Protein Denaturation; Rats

Topics: Abnormalities, Drug-Induced; Aminopropionitrile; Aneurysm; Animals; Arteriosclerosis; Bone Diseases; Collagen; Elastin; Female; Humans; Joint Diseases; Lathyrism; Paralysis; Pregnancy; Wound Healing

Topics: Aging; Aminopropionitrile; Animals; Aorta; Brain; Cartilage; Collagen; Elastin; Glycosaminoglycans; Lathyrism; Liver; Male; Rats; Rats, Inbred Strains; Sulfates; Sulfur Isotopes

Topics: Aminopropionitrile; Animals; Aorta; Cartilage, Articular; Collagen; Elastin; Fumarates; Glycosaminoglycans; Hot Temperature; Lathyrism; Rabbits; Tendons; Tissue Extracts

Topics: Aminopropionitrile; Animals; Aorta, Abdominal; Arteries; Body Weight; Collagen; Disease Models, Animal; Edema; Elastin; Female; Ferrocyanides; Haplorhini; Hemorrhage; Hemosiderosis; Iliac Artery; Lathyrism; Macaca; Male; Sternum; Thoracic Arteries

Topics: Aldehydes; Amino Acid Oxidoreductases; Animals; Catalysis; Chelating Agents; Chemical Phenomena; Chemistry; Chick Embryo; Chromatography; Chromatography, Gel; Collagen; Copper; Dialysis; Elastin; Lathyrism; Lysine; Tritium

Topics: Adult; Amino Acids; Chromatography; Chromatography, Paper; Collagen; Dipeptides; Elastin; Eye Abnormalities; Humans; Ion Exchange; Lathyrism; Male; Microscopy, Electron; Proteinuria; Spleen

Topics: Amino Acids; Animals; Aorta, Thoracic; Aortic Aneurysm; Ceruloplasmin; Collagen; Copper; Elastin; Female; Histocytochemistry; Lathyrism; Lysine; Male; Rats

Administration of D-penicillamine and lathyrogens such as beta-amino-propionitrile to animals markedly alters connective tissue by preventing the normal cross-linkage of elastin and collagen. It had been shown that beta-aminopropionitrile blocks the cross-linkage of elastin and collagen by preventing the initial step in cross-linkage: the conversion of lysine in peptide linkage to alpha-amino adipic-delta-semialdehyde. We show that penicillamine acts after the initial step, causing the accumulation of an elastin rich in alpha-amino adipic-delta-semialdehyde.

Topics: Adipates; Amino Acids; Aminopropionitrile; Animals; Aorta; Chick Embryo; Collagen; Connective Tissue; Depression, Chemical; Elastin; Lathyrism; Lysine; Penicillamine; Peptide Biosynthesis

Topics: Adipates; Aldehydes; Amino Acids; Animals; Aorta; Carbon Isotopes; Chick Embryo; Chromatography; Culture Techniques; Elastin; Electrophoresis; Fumarates; Lathyrism; Lysine; Poultry

Topics: Animals; Aorta; Collagen; Elastin; Hexosamines; Histocytochemistry; Lathyrism; Male; Rats

Topics: Alkaline Phosphatase; Animals; Aorta; Aortic Aneurysm; Collagen; Elastin; Fumarates; Glycosaminoglycans; Histocytochemistry; Lathyrism; Male; Microscopy, Electron; Muscle, Smooth; Rats

Elastin fibers in the aortas of control, lathyritic, copper-supplemented, and copper-deficient chicks were examined histochemically and chemically for aldehyde content. Diminished staining for aldehydes was obtained in the fibers from the aortas of lathyritic and copper-deficient chicks. Chemical studies of elastin isolated from the aortas of control and lathyritic chicks showed an apparent loss of lysine residues in control elastin to be associated with an increase in aldehyde content providing evidence that lysine is converted to an aldehyde-containing intermediate during biosynthesis of desmosine and isodesmosine. Approximately 6 aldehyde groups were present for every 1000 amino acids in elastin isolated from the aortas of control animals, while the corresponding number in lathyritic elastin was 4. At least two types of aldehydes, saturated and alpha,beta-unsaturated, appear to be associated with elastin, suggesting the presence of more than one intermediate between lysine and the desmosines.

Topics: Aldehydes; Animals; Aorta; Copper; Deficiency Diseases; Elastin; Histocytochemistry; Lathyrism; Poultry

Topics: Animals; Aorta, Thoracic; Collagen; Elastin; In Vitro Techniques; Lathyrism; Microscopy; Rats

Topics: Aminopropionitrile; Animals; Congenital Abnormalities; Cyanides; Elastin; Female; Fetal Death; Fetal Diseases; Lathyrism; Pregnancy; Rats

Topics: Aminopropionitrile; Aorta; Aortic Aneurysm; Chemical Phenomena; Chemistry; Collagen; Cyanides; Elastin; Histological Techniques; Lathyrism; Pathology; Rats; Research

Topics: Amino Acids; Aminopropionitrile; Animals; Aorta; Chick Embryo; Copper; Culture Techniques; Cyanides; Deficiency Diseases; Elastic Tissue; Elastin; Lathyrism; Lysine; Penicillamine