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Experimental lathyrism in the chick embryo. The distribution of beta-aminopropionitrile.

1. C(14)-labeled beta-aminopropionitrile distributed throughout the egg contents within 10 minutes postinjection. By ion exchange chromatography and electrophoretic analysis three major components of the extractable dialyzable radioactive material could be demonstrated, representing at least 80 per cent of the total. The acidic and basic components were identified as beta-aminopropionitrile and cyanoacetic acid, while the fraction isoelectric at pH 5.3, consisting of two components, remained unidentified. 2. Less than 1 molecule of betaAPN per 100 molecules of protein was present in the highly purified extractable lathyritic bone collagen indicating that binding of the lathyrogen is not a factor in collagen extractability. 3. The proximity of betaAPN to collagen in bone is not essential to its extractability. 4. The effect of incubation temperature of the embryo on collagen extractability suggests the involvement of a metabolic process in this phenomenon.

Aminopropionitrile↗

Structural and functional changes in lung tissue of mice fed with beta-aminopropionitrile fumarate, L-3-cis-hydroxyproline, and L-4-cis-hydroxyproline.

Structural and functional changes in lung tissue of mice fed with beta-aminopropionitrile fumarate, L-3-cis-hydroxyproline, and L-4-cis-hydroxyproline. We fed 0.1% solutions of 3-cis-hydroxyproline (3cisHP), 4-cis-hydroxyproline (4cisHP) or beta-aminopropionitrile fumarate (beta APN) to 5-week-old mice for 1 month and studied the effect of each of these substances on the lung function and structure. Compared to control animals the compliance of the respiratory system in the mean was increased by 3-15% in the mice fed beta APN or 3cisHP, and decreased by 2-8% in the mice fed 4cisHP. On electron microscopial examination no overt morphological changes were detectable although as proven by biochemical analysis 3cisHP and 4cisHP were incorporated into the collagen of the mice who received these substances. We conclude that the effect of substances interfering with the normal production of collagen molecules depends on the kind of collagen affected and on the rate of collagen turnover.

Aminopropionitrile↗

Collagen synthesis in capsules surrounding dimethylbenzanthracene-induced rat breast tumors and the effect of pretreatment with beta-aminopropionitrile.

Collagen synthesis is increased over three-fold in capsules surrounding dimethylbenzanthracene-induced rat breast tumors compared to the tumor parenchyma and over six-fold compared to normal breast connective tissue. Increased collagen synthesis is independent of the rate of tumor growth and final tumor size. Pretreatment of animals with beta-aminopropionitrile to inhibit collagen cross-linking caused an 82% decrease in tumor formation and a significant reduction in tumor volume (approximately 0.4 cu cm) compared to controls (approximately 10 cu cm). The four small tumors that did develop in the lathyritic animals had increased collagen synthesis in the interior tumor stroma and reduced collagen synthesis in the tumor capsule. These findings suggest that the collagenous capsule surrounding dimethylbenzanthracene tumors functions as a physical barrier to protect the tumor from the immune system of the host. The apparent antitumor effects of beta-aminopropionitrile may be due to immunopotentiation and/or cytotoxic actions of the drug.

9,10-Dimethyl-1,2-benzanthracene↗

[Production of cyanide in the rat liver from 3-aminopropionitrile].

CN- Production was investigated from a saturated mononitrile, 3-aminopropionitrile, in small liver pieces and liver homogenates. Results show that CN- production is a function of time and of the concentration of active ingredient, with a trend to saturation in both cases. Similarly, cell integrity is not required for the biotransformation of 3-aminopropionitrile to occur. Cofactors such as NADPH must be present to reach high level activities. The effect of free radicals scavengers such as mannitol and DMSO was also studied.

Aminopropionitrile↗

Urinary excretion of beta-aminopropionitrile and cyanoacetic acid.

Human serum did not convert beta-aminopropionitrile (BAPN) to the deaminated, nonlathyrogenic metabolite, cyanoacetic acid (CAA). Instead, its enzymic activity for oxidizing benzylamine was inhibited by BAPN (I50 = 2 X 10(-3) M). BAPN was found in the urine within one hour of oral administration. Oral 250 mg BAPN at 6-hr intervals each day for 21 days resulted in urinary BAPN recoveries approximating 16% of the total dose. BAPN was not detected in urine in specimens collected later than 7 hr after cessation of BAPN dosage. Urinary CAA appeared more slowly than BAPN and increased gradually to approximately three times that of urinary BAPN. After BAPN was discontinued, there was prolonged urinary excretion of BAPN-derived CAA. These along with earlier findings in experimental animals suggest that unexcreted BAPN is sequestered in tissues where it is metabolized to CAA before slowly released.

Acetates↗

Comparison of dosage-dependent effects of beta-aminopropionitrile, sodium fluoride, and hydrocortisone on selected physical properties of cortical bone.

BAPN, sodium fluoride, and hydrocortisone are reported to induce altered mineralization states. Three separate sets of experiments, one set for each agent, were performed using male New Zealand white rabbits. In each experiment the rabbits were segregated into groups, each fed a specified weight-determined dose for 13 weeks and then sacrificed. Compact bone from the left femur and tibia were tested for density, composition, sonic velocity, longitudinal elastic modulus, equatorial diffraction spacing of mineralized collagen, diaphyseal cross-sectional area, and relative load stress. beta-Aminopropionitrile (BAPN) induced monotonic degradation of all properties at all dose levels, corresponding to the decreasing density with dosage level. The elastic moduli show a decrease; the equatorial diffraction spacing of the collagen increases. The cross-sectioned diaphysis resembled woven bone. The variability in properties increased with dosage. The total cross-sectional area for a given weight increased, implying that the decreased elastic properties were compensated for by a larger area to support the weight. There was a slight increase in average density and other properties for fluoride-treated rabbits, peaking at 20 mg/kg BW/day. For higher dosages the properties are degraded and the values were much lower at high fluoride dosages than for BAPN. There was no peak for the equatorial diffraction spacing, which increased with dosage. It is inferred that the fluorosed apatite is denser than normal apatitic mineral and therefore has a smaller specific volume. A greater weight fraction of fluorosed mineral has a smaller volume fraction than the equivalent normal apatitic mineral. The bone sections look more normal, except for the porosis. The total cross-sectional area decreases when the bone density increases and then increases as the density falls, again implying that the area required to support body weight depends on the magnitude of the elastic moduli. There was a small change in some of the properties of the bones of the hydrocortisone-treated rabbits, but the effects on others were undetectable within the uncertainty of the procedures. There was no change in the cross-sectional areas of the diaphyses.

Aminopropionitrile↗

Toxic effects of beta-aminopropionitrile treatment on developing chicken skin.

beta-aminopropionitrile (BAPN), a compound that inhibits crosslinking of collagen, has been widely used to examine the function of collagen in developing tissues. We used BAPN to examine the function of collagen during the formation and patterning of feather rudiments in embryonic chicken skin. Organ cultures of skin were treated with different concentrations of BAPN and examined for changes in morphology, histology, and extent of collagen crosslinkage. Increasing concentrations of BAPN led to striped patterns of feather rudiments, increases in feather rudiment diameter, and increases in skin thickness in cultured skin. These changes in morphology were paralleled by a progressive decrease in collagen crosslinkage as the concentration of BAPN was increased. However, these changes were also paralleled by an increase in cell death in the epidermis. Culture of monolayers of separated epidermal and dermal cells in the same concentration range of BAPN that altered feather development in the organ cultures showed that BAPN killed epidermal cells but not dermal cells. Our results suggest that developmental alterations caused by BAPN may, in some cases, be due to acute toxicity to specific cell types, and not simply to inhibition of collagen crosslinkage.

Aminopropionitrile↗

Stimulation of matrix formation in rabbit chondrocyte cultures by ascorbate. 1. Effect of ascorbate analogs and beta-aminopropionitrile.

The most consistent effects of 0.2 mM L-ascorbate on monolayer cultures of rabbit articular chondrocytes were a diversion of incorporated radiosulfate into a pericellular matrix and enhancement of cell proliferation. Only with certain batches of fetal bovine serum (FBS) was there a cell-for-cell increase of proteoglycan synthesis. These actions increased as the cell inoculum rose from 0.5 to 2 x 10(5) cells/T25 flask. Maximal effects of ascorbate and D-isoascorbate were found over a range of 0.05-0.2 mM. L-Dehydroascorbic acid was less effective than either, and no stimulatory action was exerted by L-cysteine, glutathione, dithiothreitol, methylene blue, or phenazine methosulfate. Ascorbate increased the hypro:pro ratio of newly synthesized proteins. beta-Aminopropionitrile (1 mM) reduced the proportion of [3H]hydroxyproline and [35S]O4-proteoglycans in the ascorbate-supplemented matrix 31 and 7%, respectively. In corresponding electronmicrographs, the number of pericellular filaments was reduced. We conclude: (a) Ascorbate has a general anabolic effect on chondrocytes in culture and enhances matrix assembly through mechanisms other than its redox function; (b) deposition of proteoglycans in the matrix is not simply the result of mechanical entrapment by allysine- or hydroxyallysine-derived cross-linking of collagen; and (c) contradictory reports on the subject result from variations in the serum employed, inoculum density, and concentration of ascorbate.

Aminopropionitrile↗

Intercellular contacts in the media of the thoracic aorta of rat fetuses treated with beta-aminopropionitrile.

Beta-aminopropionitrile (BAPN) alters the scleroproteins of the arterial media in a way which permits an increase in the stress (force/unit area) applied to smooth muscle cells at a given pressure. Pregnant Sprague-Dawley rats were treated with BAPN and the thoracic aortas of exposed fetuses were studied by electron microscopy and image analysis. Transmission electron microscopy of the media of the thoracic aorta revealed that the types of intercellular contacts seen in the controls and in the BAPN group were intermediate junctions (IJs), nexus junctions (NJs), simple appositions (SAs), and interdigitations (Ids) as in adult vessels. The BAPN-treated animals showed an increased density of IJs, SAs, and Ids when compared with controls but the NJs demonstrated no change. Load-sensitive intercellular contacts have been considered to be the major route by which stress is applied across cells, and the increased number in the BAPN group probably represents a response of the vessel wall to increased tangential tension.

Aminopropionitrile↗

Ultrastructural changes in rat palatal epithelium after beta-aminopropionitrile.

Beta-Aminopropionitrile (BAPN) retarded or suppressed epithelial changes in the medial edge of the palatal process in later stages of gestation in rats. Programmed cell death did not follow the usual pattern, and only a few lysosomes were observed on day 18 of gestation. The sensitivity of the medial epithelium to BAPN appeared to be different in various areas of the palatal epithelium; the epithelium on the anterior region of the palatal process was hypertrophied and keratinized, while posteriorly the medial or neighboring epithelium was very thin and, in neonatal rats, the covering was absent. A basal lamina was distinct in the anterior region and indistinct or fragmented posteriorly. Collagen fibers did not develop adjacent to the basal lamina, and an amorphous material was scattered throughout the mesenchymal tissue. These findings suggest that BAPN decreases the "connecting capacity" between mesenchyme and epithelium, and results in a modification of epithelial changes.

Aminopropionitrile↗

The teratogenic effects of beta-aminopropionitrile in hamsters.

The teratogenicity of beta-aminopropionitrile (beta APN) was evaluated in golden Syrian hamsters; single doses of 1250, 2500, and 3750 mg/kg given on days 7-13 produced fetal mortality, growth retardation and malformations. All time-dose combinations produced gross external abnormalities with the highest rate (92.5%) occurring following day-12 treatment. High frequencies of skeletal abnormalities including deformities of the ribs, fibula, and scapula occurred after days-10-12 treatment. Deformed ribs occurred in 100% of fetuses exposed on day 11 to any of the dosages. The types of abnormality generally resembled those produced by lathyrogens in other species except for exencephaly and encephalocele, which occurred after treatment on day 7. These results indicate that betaAPN is a typical teratogen, capable of producing abnormalities of the central nervous system early in gestation, and may reflect a broader mechanism for its activity than previously suggested.

Abnormalities, Drug-Induced↗

The morphogenesis of beta-aminopropionitrile-induced rib malformation in fetal golden Syrian hamsters.

In order to provide information on the mechanism of beta-aminopropionitrile (betaapn) induced teratogenesis, the pathogenesis of a fetal rib abnormality was studied at relatively short time intervals following maternal treatment with 2,500 mg/kg aqueous betaapn on day 11 of gestation. Histochemical tests of ribs from betaapn-exposed fetuses indicated a slight decrease in the level of glycosaminoglycans but at a time when the defect was already morphologically established. Ultrastructural observations on the chondrocytes of ribs from betaapn-exposed fetuses revealed alterations in mitochondrial structure indicative of a slight cytotoxic effect for the teratogen. The mitochondrial changes were transient, occurring initially at three hours after treatment and lasting for nine hours. Alterations in the size of collagen fibres in the cartilage of the fetal rib were also observed in the offspring of betaapn treated females. The mean diameter of collagen fibres in the ribs of control fetuses increased throughout the course of the study. The mean diameter of fibres in the fetuses of betaapn-exposed females failed to show any increase and was found to be significantly less than controls as early as three hours following maternal administration. The results suggested that the principal factor in the production of the fetal rib deformity was fundamentally the same as that known to affect the adult; namely a defect in the extracellular maturation of collagen.

Abnormalities, Drug-Induced↗

Inhibition of beta-aminopropionitrile (beta APN)-induced skeletal teratogenesis by the flavonoid beta-hydroxyethylrutosides (HR) in hamster fetuses.

Since biochemical studies have shown that flavonoids such as beta-hydroxyethylrutosides (HR) protect against the damage to collagen induced by lathyrogens in adult rats, this compound was given to pregnant hamsters in order to determine its effects on the teratogenicity induced by beta-aminopropionitrile (beta APN). A dose of 2,500 mg/kg of beta APN alone given by gavage on day 11 produced a high frequency (69.5%) of skeletal anomalies in the offspring of hamsters. Administration of HR immediately following beta APN to pregnant animals resulted in a significantly decreased teratogenic response (P less than 0.05). These data provide evidence to support the view that the primary mechanism for the beta APN-induced skeletal dysmorphogenesis is the inhibition of cross linking during the maturation of collagen fibers.

Abnormalities, Drug-Induced↗

[Influence of concomitant treatment with D-penicillamine and beta-aminopropionitrile on mechanical properties of rat connective tissue (author's transl)].

In order to study the influence of D-penicillamine (DPA) and beta-aminopropionitrile (BAPN) as well as of a combination of both substances (proportion by weight 1:1) on the collagen of skin, aorta, and femoral epiphysial cartilage, the mechanical properties of these fibrous organs were examined, after having performed tests on onset and dissipation of action, measured by the decrease of ultimate load of excised strips of skin using the two individual compounds. The action of 320 mg/kg of DPA sets in within 7 days reaching a maximum value after 14 days. After discontinuance of the substance the control values are reached within 15 days. BAPN shows its maximal effect on the 18th test day. After discontinuance of treatment the control value is not reached within 35 days. DPA and BAPN show organo-specific actions: DPA has a considerably stronger influence on the collagen of skin than on the collagen of the aorta; under the present test conditions it does, however, not affect the collagen of the distal femoral epiphysial cartilage. As compared to DPA, BAPN exerts a less pronounced influence on the collagen of the skin, similar influence on the collagen of the aorta, and a strong influence on the collagen of the distal femoral epiphysial cartilage. If both substances are administered together, the organospecific actions of DPA on the one hand and BAPN on the other hand are maintained. It is discussed whether the different chemical compositions of the collagen (Type I--IV) may be connected with the organospecific effects of the test substances.

Aminopropionitrile↗

The polymerization of alpha-aminopropionitrile.

The mixture of alpha-aminopropionitrile and alpha, alpha'-iminodipropionitrile polymerized to solidify almost at the temperature near 0 degrees C during 8 years. The conversions based on decreasing of those reactants were 61 and 98% at 4 and 8 years, respectively. The fractionation of 4 and 8 years product using Sephadex G 10 yielded their predominant amounts in the oligomer and polymer section, respectively. The oligomer section product of 8 years product was analyzed by means of ion exchange chromatography and its trimethylsilyl derivative was also analyzed by means of gas chromatography combined with mass spectrometry. These results identified dialanine and trialanine and their amides and nitriles.

Aminopropionitrile↗

The asymmetric adsorption of alpha-aminopropionitrile on D-quartz.

alpha-Aminopropionitrile was adsorbed on the powdered D-quartz, recovered as N-trifluoro-acetylalanyl-(+)-secondarybutyl ester, and analyzed by means of gas chromatography. The asymmetric result supporting the preferential adsorption of L-antipode was obtained and its significance in chemical evolution has been discussed.

Adsorption↗

Purkinje cell toxicity of beta-aminopropionitrile in the rat.

Compounds causing neurolathyrism are putative aetiological agents in neurodegenerative disorders including amyotrophic lateral sclerosis. beta-Aminopropionitrile (BAPN) is one such compound. We have administered this lathyrogenic agent at a dose of 1 g/kg by the intraperitoneal route in experiments in adult Sprague-Dawley rats during a period of 10 weeks. The rats developed marked kyphoscoliosis, ataxia with paralysis and muscle wasting of the hind limbs. Vacuolation and loss of Purkinje cells developed, but no anterior horn cell degeneration was noted. Immunohistochemical studies of phosphorylated neurofilaments and the 72 kDa heat shock protein were normal and no intraneuronal ubiquitinated inclusions were seen. High-dose intraperitoneal BAPN in the rat causes Purkinje cell changes, but no other central nervous system abnormalities.

Aminopropionitrile↗

2-aminopropionitrile polymer. I. The hydrolyzate of the basic fraction.

The basic fraction of a 2-aminopropionitrile polymer was subjected to acid hydrolysis and was analyzed by means of GC-MS, after trimethylsilylation. The fundamental polymer structural units were alanine, 2,2'-iminodipropionic acid, N-(1-cyanoethyl)alanine, N-ethylalanine, glycine, cyanoglycine, and 5-amino-4-carboxyimidazole residues. The last three units may be derived from hydrogen cyanide. Oligomeric combinations of these units were also detected in the hydrolyzate, due to partial hydrolysis of the polymer.

Acetaldehyde↗