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Urinary delta amino levulinic acid (ALA) in the cow, dog and cat.

The level of delta amino levulinic acid (ALA) was determined in the urine of normal cows and dogs, and in the urine of dogs, cows and cats with a variety of diseases. ALA was measured in the urine of two steers and two dogs experimentally poisoned with lead. Experimental lead poisoning was followed by increased excretion of ALA in the urine. Higher levels of ALA were present in the urine of sick dogs and cats than in the urine of sick cows.

Amino Acid Metabolism, Inborn Errors↗

Nest liquid resources of several cavity nesting bees in the genus Centris and the identification of a preservative, levulinic acid.

Twig-nesting species of bees in the genus Centris including C. bicornuta, C. analis, C. vittata, and C. nitida, found in the dry forest of Guanacaste Province of Costa Rica, provision their nests with pollen and nectar, rather than pollen and oil as reported for other Centris species. The liquid contents of the nests of these four species were found to contain sugars including 66-75% fructose, 25-33% glucose, and a trace of sucrose. The sugar concentration averaged 47.2%, slightly higher than most flower nectars. No tri-, di-, or monoglycerides, the main components of the flower oil of Byrsonima crassifolia, were detected in the nest provisions. Although these four Centris species are also known to collect oil from B. crassifolia, the oil appears to be used for activities other than nest provisioning. The liquid nest contents did have a slight goat-like odor, suggesting the presence of short-chain fatty acids, and were found to contain a small amount (less than 1%) of three fatty acids. Two of these, butanoic and octanoic acid, were found in trace amounts and are responsible for the goat-like odor. A third was identified as levulinic acid, which made up about 99% of the nest fatty acid contents. This fatty acid had little odor, but may be important as a fungicidal agent. Attempts to determine the source of the fatty acids, were not successful.

Animals↗

5-Chloro[1,4-13C]levulinic acid modification of mammalian and bacterial porphobilinogen synthase suggests an active site containing two Zn(II).

5-Chloro[1,4-13C]levulinic acid ([1,4-13C]CLA) is an active site-directed inactivator of porphobilinogen synthase (PBGS). PBGS asymmetrically condenses two molecules of 5-aminolevulinic acid (ALA) which are called A-side ALA and P-side ALA in reference to their fates as the acetyl and propionyl halves of the product. [1,4-13C]CLA modifies bovine PBGS at the A-side ALA binding site. The C4 chemical shift indicates an intact keto moiety; the C1 chemical shift indicates a deprotonated carboxyl group. In contrast, [1,4-13C]CLA modification of Escherichia coli PBGS is heterogeneous and occurs preferentially at the P-side ALA binding site. The C1 chemical shifts indicate substantially deprotonated carboxylic acid groups. For one of four observed forms of [1,4-13C]CLA-modified E. coli PBGS, an analog of the P-site Schiff base is found. Bovine and E. coli PBGS contain two different zincs, ZnA and ZnB. Past results placed ZnA near A-side ALA. [1,4-13C]CLA modifies E. coli PBGS at Cys119 or Cys129, which is part of a four-cysteine cluster implicated in binding ZnB. This result places ZnB near P-side ALA. E. coli PBGS binds a third type of divalent metal, MgC or MnC, which is found to have no significant effect on the 13C NMR spectrum of the [1,4-13C]CLA-modified protein.

Amino Acid Sequence↗

Inhibition of 5-amino levulinic acid dehydratase activity by arsenic in excised etiolated maize leaf segments during greening.

In vivo as well as in vitro supply of sodium arsenate inhibited the 5-Amino levulinic acid dehydratase (5-aminolevulinate-hydrolyase EC 4.2.1.24, ALAD) activity in excised etiolated maize leaf segments during greening. The percent inhibition of enzyme activity by arsenate (As) was reduced by the supply of KNO3, but it was increased by the glutamine and GSH. Various inhibitors, such as, chloramphenicol, cycloheximide and LA, decreased the % inhibition of enzyme activity by As. The % inhibition of enzyme activity was also reduced by in vivo supply of DTNB. The enzyme activity was reduced substantially by in vitro inclusion of LA, both in the absence and presence of As. In vitro inclusion of DTNB and GSH inhibited the enzyme activity extracted from leaf segments treated without arsenate (-As enzyme) and caused respectively no effect and stimulatory effect on arsenate treated enzyme (+As enzyme). Increasing concentration of ALA during assay increased the activity of -As enzyme and +As enzyme to different extent, but double reciprocal plots for both the enzymes were biphasic and yielded distinct S0.5 values for the two enzymes (-As enzyme, 40 micromol/L and +As enzyme, 145 micromol/L) at lower concentration range of ALA only. It is suggested that As inhibits ALAD activity in greening maize leaf segments by affecting its thiol groups and/or binding of ALA to the enzyme.

Aminolevulinic Acid↗

Stimulation of Friend erythroleukemic cell cytodifferentiation by 5-amino levulinic acid; porphyrins, cell size, segregation of sialoglycoproteins, and nuclear translocation.

The late events of erythroid differentiation in Friend erythroleukemic cells (FELC), stimulated by 5-amino levulinic acid (5-ALA), were studied. Cultivation of the cells in 5-ALA-enriched media triggered a chain reaction, beginning with an immediate and rapid accumulation of endogenous porphyrins, in particular protoporphyrin and hemin. Incorporation of 14C-ALA was rapid and independent of dimethylsulfoxide (DMSO) induction. In parallel, on the second day of growth, a marked decrease in cell volume was elucidated by flow cytometry. The total protein content was reduced, while Fe uptake and hemoglobin synthesis were increased. The combination of DMSO and 5-ALA produced the most effective induction of the FELC, and the differentiation criteria were the most advanced. The cells exposed to the combined stimulation became loaded with heme and hemoglobin and their generation time was prolonged up to 35 h. Transmission electron microscopy of these treated cells showed a morphological alteration to pearlike cells, associated with a typical nuclear translocation phenomenon and a regional segregation of sialoglycoproteins. An uneven distribution of organelles was revealed; one part of the cell contained numerous ribosomes and the nucleus, while the other part was hemoglobinized, contained mitochondria, and the outer membrane was heavily labeled with ferritin hydrazide, a marker for sialoglycoproteins. The enhanced stimulation of Friend cells by 5-ALA promoted an advanced step of erythroid maturation that has much in common with the late events of normal nuclear extrusion process.

Aminolevulinic Acid↗

The effects of levulinic Acid and 4,6-dioxoheptanoic Acid on the metabolism of etiolated and greening barley leaves.

Application of levulinic acid (LA), a competitive inhibitor of delta-aminolevulinic acid (ALA) dehydratase, to greening plant tissues causes ALA to accumulate at the expense of chlorophyll. 4,6-Dioxoheptanoic acid (DA), which has been reported to be an effective inhibitor of this enzyme in animal systems, has a similar but more powerful effect on ALA and chlorophyll metabolism in greening leaves of Hordeum vulgare L. var. Larker. Both LA and DA also inhibit the uptake of [(14)C]amino acids into etiolated and greening barley leaves and reduce their incorporation into protein. Treatment of etiolated and greening leaves with these compounds results in the inhibition of (14)CO(2) evolution from labeled precursors, including amino and organic acids. Inhibition of (14)CO(2) evolution by these compounds is more effective in greening leaves than in etiolated leaves when [4-(14)C]ALA or [1-(14)C]glutamate are employed as precursors. Both LA and DA also inhibit the uptake and increase the incorporation of (32)Pi into organophosphorus by etiolated barley leaves. These results indicate that LA and DA have more far-reaching effects upon plant metabolism than was previously believed.

Journal Article↗

Sorption of levulinic acid onto weakly basic anion exchangers: equilibrium and kinetic studies.

The equilibrium and dynamics of levulinic acid sorption on two weakly basic anion exchangers, in free base form, in single-component aqueous solutions were investigated. Adsorption isotherms such as Langmuir, Sips, Radke-Prausnitz, and Toth were applied to correlate the experimental data in the temperature range 285-315 K. Modeling results showed that the Toth model is the best one to correlate the sorption isotherms. The resulting Toth equations were used with the Clausius-Clapeyron equation to determine the isosteric heat of sorption. The sorption kinetics was experimentally measured via a completely stirred finite-bath batch experiment under different initial concentrations and at varying solution temperatures. The pseudo-second-order kinetic model and the Elovich equation were used to represent the kinetic data and the equation parameter values were also evaluated. The pseudo-second-order equation cannot simulate the experimental kinetic data, while the Elovich equation fitted the sorption dynamic data very well under all the operating conditions studied. Finally, the apparent activation energy of sorption was also determined.

Journal Article↗

Effects of levulinic acid on 5-aminolevulinic acid production in heterotrophic cultures of Chlorella regularis YA-603.

To investigate the effects of levulinic acid (LA) on extracellular 5-aminolevulinic acid (ALA) accumulation, a heterotrophic culture of Chlorella regularis YA-603 in basal medium containing 0 to 60 mM of LA was carried out. It was found that the extracellular and total ALA concentrations increased, while the chlorophyll (CHL) concentration was the same above initial LA concentrations of 30 mM, with an increasing initial LA concentration up to 50 mM. In the presence of an initial LA concentration of 60 mM, however, the specific growth rate decreased markedly. This suggested that excessive LA exerted a negative effect on ALA production because it also directly affected cellular growth. The relationship between the initial LA and total ALA concentrations mentioned above indicates that LA could promote the ALA-producing activity. Kinetic analysis revealed that an LA concentration of between 30 to 50 mM in the culture broth gave the maximum specific production rate of ALA ((q(ALA))max). When LA was added repeatedly to maintain this optimum range of LA concentration, (q(ALA))max could be maintained at a relatively high level for a longer period, and the maximum concentration of ALA reached 3.86 mM.

Journal Article↗

Biosynthesis of rat kidney catalase. Double-labeling by [14C]leucine and delta-amino[3H]levulinic acid.

Kidney catalase [EC 1.11.1.6] was doubly labeled by injecting a mixture of [14C]-leucine and delta-amino[3H]levulinic acid (ALA) into normal and allyliso-(2-isopropyl-4-pentenoyl)urea (Sedormid)-treated rats. Shortly after the injection of a tracer dose or the precursors, catalase in microsomes showed the highest specific radioactivities among catalases from various cell fractions. On the other hand, peroxisomal catalase was labeled gradually, reaching a plateau at about 90 min. The patterns of incorporation of both isotopes were similar to those obtained previously with rat live catalase, except that the 3H-radioactivities were much higher. From the results obtained it could be postulated that kidney catalase is synthesized on polysomes and then transferred to peroxisomes, directly or via the soluble phase. The ratio of 3H/14C incorporated was lowest in microsomal catalase, highest in microsomal catalase, highest in peroxisomal enzyme and intermediate in catalase of the soluble fraction. The increment with time was large in the latter two catalases, while the former showed a rather small change. The evidence suggests that nascent catalase contains less heme than the completed molecule; further addition of heme to this intermediate seems to occur in the cytosol, and possibly also in peroxisomes. Administration of a porphyrinogenic drug, (2-isopropyl-4-pentenoyl)urea, produced a remarkable decrease in the incorporation of [3H]ALA into kidney catalase, with no significant effect on that of [14C]leucine.

Aminolevulinic Acid↗

Fluorescence detection of bladder tumors with 5-amino-levulinic acid.

BACKGROUND: Preliminary data suggest that photodiagnosis by fluorescence detection of protoporphyrin IX, a biosynthetic product of the photosensitizer 5-amino-levulinic acid (ALA), is superior to conventional cystoscopy in bladder tumor detection. PATIENTS AND METHODS: We instilled 1.5 g of ALA into the bladders of 52 patients with suspect bladder lesions 1 to 4 hours prior to transurethral resection or biopsy. Red fluorescence was induced by violet-blue light (wavelength 380-450 nm). RESULTS: In 13 patients (25%), significant tumors were detected by fluorescence that were missed on conventional cystoscopy. In the present series, ALA photodetection had a sensitivity of 94.6% (compared with 76% for cystoscopy) and a specificity of 43%. CONCLUSION: Fluorescence diagnosis with ALA may become a standard procedure for bladder tumor detection and resection, especially in endoscopically difficult situations (carcinoma in situ, multifocal tumors, multiple prior resections, or previous drug instillation therapy).

Adult↗

Laser-induced fluorescence studies of normal and malignant tumour tissue of rat following intravenous injection of delta-amino levulinic acid.

BACKGROUND AND OBJECTIVE: Laser-induced fluorescence was studied in normal and tumour tissue of rat after intravenous injection of delta-amino levulinic acid (ALA). The aim of the study was to investigate the protoporphyrin IX accumulation in different tissue types in rat after systemically administered ALA. STUDY DESIGN/MATERIAL AND METHODS: A malignant rat tumour and normal tissue from 13 different organs were investigated in eight rats. The rats were injected with two different ALA doses, 30 and 90 mg/kg b.w., and the investigations were performed at 10, 30 and 240 min after the injection. The fluorescence was recorded utilising an optical fibre based fluorosensor at 405 nm excitation. RESULTS: Fluorescence spectra were recorded in the 400-750 nm wavelength region including the dual-peaked PpIX fluorescence at about 635 and 705 nm, and the tissue autofluorescence peaking at about 500 nm. The maximum tumour build-up of PpIX was achieved already in less than 1 hr after ALA injection. The fluorescence demarcation between tumour and surrounding tissue was a factor of 7-8:1 after 30 min and decreased for longer retention times. The accumulation in 13 different organs was investigated and a particularly high PpIX build-up was found in stomach and intestine. CONCLUSIONS: Fluorescence detection following i.v. injection of ALA provides attractive diagnostics for the experimental tumour used, indicating clinical usefulness.

Aminolevulinic Acid↗

Interaction between blood lead concentration and delta-amino-levulinic acid dehydratase gene polymorphisms increases the odds of essential tremor.

Blood lead (BPb) concentrations are elevated in essential tremor (ET) cases. The delta-amino-levulinic acid dehydratase (ALAD) gene codes for ALAD, the principal enzyme involved in lead kinetics. Carriers of the ALAD-2 allele may be more susceptible to lead toxicity. The objective of this study was to test, using a case-control design, whether an interaction between BPb concentration and ALAD allele status increases the odds of ET. Mean log BPb concentration was significantly higher in 63 cases than 101 controls (0.47+/-0.26 vs. 0.35+/-0.25 microg/dl). Eighteen (28.6%) cases vs. 17 (16.8%) controls had an ALAD-2 allele (OR=1.98; 95% CI=0.93--4.21). In an adjusted logistic regression analysis, the interaction term (ALAD allele status x log BPb concentration) was associated with increased odds for ET. In stratified analyses, log BPb concentration was not associated with odds of ET in individuals with two ALAD-1 alleles (OR=2.69; 95% CI=0.61--11.82), but in individuals with an ALAD-2 allele, BPb concentration was significantly associated with odds of ET (OR=80.29; 95% CI=3.08--2,096.36). There was an interaction between BPb concentration and ALAD allele status; the odds of ET were greatly elevated in individuals with both an ALAD-2 allele and an elevated BPb concentration. The presence of increased circulating BPb concentrations along with a greater potential for lead toxicity (ALAD-2 allele) could result in greater cerebellar damage, thereby increasing the risk of developing tremor.

Adult↗

Heavy metal levels and delta-amino-levulinic acid dehydrase levels in peripheral polyneuropathy.

On the basis of assay of heavy metals in whole blood (lead, cadimum, chromium, copper, nickel and manganese) and delta-amino-levulinic-acid dehydrase (ALA-D) and ALA in urine in a normal Danish population, the levels of these clinco-chemical factors were assayed in 23 patients with peripheral neuropathy of unknown etiology. All patients studied showed electro-physiological sign of denervation and/or reduced motor or sensoric nerve conduction velocity. Cadmium and manganese were never found to be increased. In all but four patients, an increase of one or more heavy metals was found. Ten patients showed raised levels of two or more metals, the dominant metal being lead (10 cases), nine patients showed increased in chromium. A significant corrleation was found between increasing lead levels and decreasing ALA-D activity. Although normal concentrations of manganese were found, correlation analysis revealed a significant correlation between increased manganese and decreased ALA-D. The raised values of heavy metals could not be traced to occupational or other exposure to heavy metals and the increased values were not related to tobacco consumption. The findings are discussed in relation to known data on neuropathy and the results seem to indicate a multifactorial patholgenesis of the disease. Among factors contributing to the precipitation of the syndrome may be raised levels of heavy metals.

Adult↗