Interaction of abscisic acid & indole acetic acid on transcription.
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3H-Indole acetic acid bound to cytosol proteins of embryonic chicken liver in vitro to an average capacity of at least 20 pmoles/mg binding protein. The auxin-binding protein complexes could be resolved into 4 major zones by anion exchange chromatography; they sedimented at 3-7 S in sucrose density gradients, and were also heterogenous in agarose gel electrophoresis.
The cotton root rot fungus, Phymatotrichum omnivorum, synthesized indole acetic acid (IAA) in culture medium containing tryptophan as the only source of nitrogen. Synthesis of IAA was not affected by illumination, and maximum amounts of the auxin were produced at 30 days. The optimum hydrogen ion concentration and temperature for IAA production were 6.5 and 28 degrees C, respectively. Indole ethanol (IEA) was also detected in tryptophan media, primarily during the active growth of the organism.
The polyamide layer technique for the chromatographic separation of dimethylaminonaphthalene sulphonyl amino acids has been adapted to the qualitative analysis of amino acids in media before and after the growth of micro-organisms. The method has been used to study the amino acids metabolized by cultures of proteolytic clostridia growing in a medium consisting of an acid hydrolysate of casein as a source of amino acids and small amounts of yeast extract and trypticase as sources of growth factors. The chromatograms of the media after growth showed which amino acids were used and which new amino acids were produced. Clostridium botulinum type F (proteolytic), C. ghoni, C. mangenoti and C. putrificum were found to reduce proline to 5-aminovaleric acid and to produce 2-aminobutyric acid, properties they shared with C. sporogenes and C. sticklandii. C. botulinum type G and C. subterminale used glycine, lysine, serine, and arginine but in contrast to C. sticklandii they neither reduced proline to 5-aminovaleric acid nor produced 2-aminobutyric acid. Both organisms oxidized phenylalanine, tyrosine and tryptophan to phenylacetic acid, p-hydroxyphenyl acetic acid and indole acetic acid respectively. C. lituseburense and C. scatologenes used serine, threonine and arginine and produced 2-amino butyric acid and ornithine. C. lentoputrescens, C. limosum and C. malenomenatum resembled C. tetanomorphum by using glutamic acid and tyrosine. The chromatograms always showed the physiological group to which an organism belonged and in some cases were characteristic of the species.
The weak luminescence that accompanies the linoleate-lipoxygenase reaction was greatly enhanced by the addition of indole analogues, and especially indole acetic acid. The main emitting species in the indole acetic acid-linoleate-lipoxygenase system was analysed spectrophotometrically in the visible region and ascribed to the transition of excited indole acetate in triplet state to its ground state. Such an excited indole acetate could be generated by transfer of energy from the excited CO2 and excited carbonyl (generated by the linoleate-lipoxygenase reaction) to indole acetate in the ground state, but not by cleavage of the dioxetane analog (positions 2 and 3 on the indole ring).
Indole-3-acetic acid (IAA) is a tryptophan-derived gut microbial metabolite with reported anti-inflammatory activities, but the organisms and anaerobic pathways that support robust production remain unclear. Screening 206 human gut bacterial isolates by LC-MS revealed that IAA production is rare: only five strains exceeded the limit of quantitation, and high-capacity production was confined to the acetogens Blautia hydrogenotrophica and Intestinibacter bartlettii. Across growth conditions, IAA was a minor product that rose alongside carbohydrate-sensitive, OFOR-linked catabolism of multiple amino acids, generating abundant branched-chain and aromatic organic acids. In gnotobiotic mice mono-colonized with I. bartlettii, these metabolites were produced in vivo but showed distinct host handling, with branched-chain fatty acids largely extracted between portal and peripheral plasma, whereas aromatic acids and their glycine conjugates appeared in plasma and urine. Genomic analyzes and heterologous enzyme assays identified expanded repertoires of 2-oxoacid:ferredoxin oxidoreductases (OFORs) with activities spanning pyruvate/oxaloacetate, branched-chain, and aromatic 2-oxoacids, including indolepyruvate conversion to indoleacetyl-CoA, a putative intermediate en route to IAA. Finally, position-specific 13C tracing showed that CO2 released during amino acid oxidation is reassimilated into acetate via reductive acetogenesis, indicating that gut acetogens can maintain redox balance without fermenting partner strains. Together, these findings show that high IAA output is restricted to select gut acetogens and linked to a broader OFOR-driven anaerobic metabolism that generates additional metabolites that are absorbed by the host.
These studies evaluated the teratogenic potential of indole-3-acetic acid (IAA), a naturally occurring plant hormone, in CF-1 mice and Sprague-Dawley rats. Mice were given 5, 50, 200, or 500 mg IAA/kg/day by gavage on days 7 through 15 of gestation. Rats were given 50, 200, or 500 mg IAA/kg/day by gavage on days 7 through 15 of gestation. IAA was teratogenic in mice and rats at 500 mg/kg/day; cleft palate was induced in both species at this dose level. In mice, other malformations including exencephaly, ablepharia, dilated cerebral ventricles, and crooked tail were also observed. Mice given 500 mg/kg of IAA gained less than control mice during gestation; no evidence of maternal toxicity was observed in rats. IAA did not cause fetal resorptions in either species and was not teratogenic at dose levels below 500 mg/kg.
The Acinetobacter calcoaceticus-baumannii complex includes high-priority, multidrug-resistant pathogens for which novel antibiotics are urgently needed. Many bacterial strains from this complex harbor a so-called iac gene cluster that codes for the catabolism of indole-3-acetic acid (IAA). Here, we demonstrate that possession and expression of iac genes represent an Achilles' heel for Acinetobacter species, which can be exploited to suppress bacterial growth by treatment with IAA and its analog 5-chloro-IAA.IMPORTANCEAcinetobacter baumannii is a deadly bacterial pathogen and one of the leading causes of hospital-acquired infections worldwide. It is also known for its resistance to many antibiotics currently available. In this study, we show that Acinetobacter bacteria choke on a mixture of IAA and 5-chloro-IAA, offering a path to the discovery and development of a novel drug treatment.
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Blood levels of serotonin (5-HT) and urinary excretion of 5-hydroxyindole-acetic acid (5-HIAA), tryptamine (T), and indole-acetic acid (IAA) were determined in 39 cases of systemic scleroderma and in 7 cases of very severe cutaneous scleroderma. The T/IAA ratio was normal and serotonin elevated in mild and rather severe acrosclerosis alike, i.e. in systemic scleroderma with pronounced vascular involvement. The T/IAA ratio was increased and serotonin normal in severe acroscleroderma, representing an intermediate form or transition to diffuse scleroderma, in severe diffuse scleroderma, and in severe cutaneous scleroderma. The T/IAA ratio and serotonin level were both elevated in a few cases of systemic scleroderma and in severe generalized morphea with concomitant vascular changes. The findings suggest impaired monoamine oxidase (MAO) activity in scleroderma with consequent accumulation in the organism of biogenic amines derived from tryptophan. An increased T/IAA ratio seems to be of prognostic significance in scleroderma, suggesting an adverse course of the disease.
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Five male patients, complaining of daytime sleep attacks, and hav ing a history of automatic behavior, were recorded continously by polygraph for 108 h under two different protocols: (1ents were given specific tests to perform during Phase 1. Six normal males were run as controls during Phase . At the end of the recordings, patients had measurements of homovanillic acid (HVA) and 5 Hydroxy-indol Acetic Acid (5HIAA) in CSF before and after Probenecid testing. Patients appeared to present an abnormal sleep structure, with lack of Stage 3 and 4 NREM sleep, but had no significant increase in total sleep time during 24 h compared to normals. They also had repetitive very short periods of "micro-sleep" which account for their automatic behavior and were responsible for their great decrement in performance compared to normal controls. No CSF HVA and 5HIAA abnormality could be detected. This syndrome, which greatly impairs the social life and working ability of the patients, may be more related to an impairment of the "wakefulness" structures than to a dysfunction of the "sleep" structures.
The effect of indomethacin 3 mg/kg on levels of homovanillic acid (HVA), 4-hydroxy-3-methoxy phenyl ethylene glycol (HMPG) and 5-hydroxy indol acetic acid (5HIAA) was studied in rat striatum and olfactory tubercle with and without pretreatment with morphine 10 mg/kg. Indomethacin caused a small decrease in resting levels of HVA in striatum but not in olfactory tubercle. No effects were seen on resting or morphine induced changes in the levels of these monoamine metabolites. Likewise indomethacin 20 mg/kg failed to alter the elevation of HVA induced by chlorpromazine 15 mg/kg or the decrease of HVA induced by apomorphine (1--10 mg/kg) in the rat striatum. Our results do not support a major role for endogenous prostaglandins in the modulation of monoamine neurotransmission in the rat brain.
Peroxidation of endogenous lipid by liver microsomes, coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and measured as thiobarbituric acid reactive materials, is markedly stimulated in the presence of indomethacin [1-(p-chlorobenzyl)-5-methoxy-2-methyl-3-indole acetic acid] (0.1--1.0 mM). Concurrently, indomethacin enhances the lipolysis of membrane phospholipid containing arachidonic acid but has no effect on the rate of O2 uptake in these samples. The system generates a rapidly developed chemiliminescense (CL), the intensity and rate of development of which are related to indomethacin concentration. The microsomal CL generated in the presence of indomethacin is distinct from the previously reported CL in that the time required for maximum intensity development is a matter of seconds (20--180) rather than hours. The enhanced CL is believed to be due to an energy transfer reaction whereby a high energy species transfers energy to the indomethacin molecule, which, in turn, decays via chemiluminescense. An enhanced chemiluminescense was also observed when indomethacin was added to a lipoxidase system and superoxide generating system (axanthine oxidase). Based on inhibitor studies, the rapidly developed chemiluminescense of the microsomal system requires cytochrome P-450 in addition to NADPH and coordinated iron ions. The results indicate that the CL is related to neither hydroxyl free radical nor superoxide anion formation.
BACKGROUND: Subacute ruminal acidosis (SARA) is a digestive disorder that often severely jeopardizes the health and lactation performance of ruminants fed a high-energy diet. Different dairy ruminants exhibit varying degrees of inflammation accompanied by variations in the rumen microbiota when SARA occurs. Our understanding of the occurrence of SARA and varying degrees of rumen epithelial inflammation is lacking. Hence, we performed rumen metagenomic, metagenome-assembled genome and metabolomic analyses, with transcriptome and single-nucleus RNA sequence analyses, to explore the microbial mechanism of SARA occurrence and different degrees of inflammation. RESULTS: A total of 36 goats fed two diets with gradually increasing levels of rumen-degradable starch (RDS) were included in this study, and SARA goats fed 70% concentrate diets supplemented with whole corn (HGW-SARA) and SARA goats fed 70% concentrate diets supplemented with crushed corn (HGC-SARA) were identified. Moreover, 11 goats fed a control basal diet, named LGW-CON, were also included. Compared with those in the LGW-CON group, the rumen fermentation capacity was enhanced, accompanied by ruminal epithelial and systemic inflammation, in goats from HGW-SARA and HGC-SARA. Between them, HGC-SARA goats presented less inflammation. Notably, the ruminal inflammation-related pathways were increased only in the HGW-SARA group but not in the HGC-SARA group. Metagenomic analysis revealed that the β diversity of SARA goats was significantly different from that of LGW-CON goats. Ruminococcus significantly increased in both SARA groups, whereas Prevotella and Bacteroidales significantly decreased, which was accompanied by a decrease in cellulose and hemicellulose enzymes and an increase in lysozymes and lipopolysaccharide synthesis enzymes. Multi-omics analysis of the ruminal contents and tissues suggested that epithelial inflammation was caused by disturbed ruminal microbiome-induced Th17 cell differentiation and IL-17 signalling pathway activation. Comparative analyses between the HGW-SARA and HGC-SARA groups highlighted the importance of Selenomonas and Bifidobacterium, as well as bacterial tryptophan metabolism, in the production of 3-indoleacetic acid, which mitigated ruminal epithelial inflammation by modulating Th17 cells and inhibiting IL-17 signalling. Ruminal microbiota transplantation from HGW-SARA goats to healthy dairy goats and mice revealed the role of microbes in epithelial inflammation. Additionally, 3-indoleacetic acid supplementation reduced rumen inflammation and the IL-17 concentration in the serum, improved VFAs absorption, and enhanced milk production. CONCLUSIONS: This study unveiled that after SARA was induced by high-concentrate feeding, the rumen homeostasis was disrupted, and rumen fiber degradation capacity of dairy goats decreased, but the LPS synthesis capacity increased, and inflammation of the rumen epithelium was observed. However, the ruminal microbial species from the Bifidobacterium and Selenomonas genera and bacterial 3-indole acetic acid are pivotal in mitigating ruminal epithelial inflammation during SARA in dairy goats. This could potentially be attributed to the modulation of ruminal Th17 cell proportions and the inhibition of IL-17 signalling pathways. Video Abstract.
Peroxidation of endogenous lipid by rat liver microsomes, coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) was markedly stimulated in the presence of indomethacin [1-(p-chlorobenzyl)-5-methoxy-2-methyl-3-indole acetic acid], a potent anti-inflammatory drug. This system also generated a rapidly developed chemiluminescense (CL), the intensity and rate of development of which were related to indomethacin concentration and the amount of peroxidation in the sample. Microsomes from rats fed a liquid diet based on Sego diet drink (Pet Incorporated) failed to chemiluminesce with or without added indomethacin. Supplementation of the Sego diet with a polyunsaturated fatty acid (C18:3) did not restore the chemiluminescense. In vitro and in vivo studies indicate that vitamin E (a component of the Sego liquid diet) inhibited the microsomal chemiluminescense reaction in a dose related manner.
Immature rats can be induced to ovulate with pregnant mare serum (PMS) as long as they weigh over 60 g. In the rats weighing less than 60 g 5-hydroxytryptamine (5HT)-60 micrograms/rat intraventricularly or 2 micrograms/rat in the paraventricular nucleus (PVN)-stimulated ovulation. Injections into several other hypothalamic nuclei were ineffective. In rats weighing greater than 60 g, lesions of the PVN or injection of 10 micrograms/rat p-chlorophenylalanine into the PVN inhibited ovulation. The hypothalamic levels of 5HT and 5-hydroxy indole acetic acid (5HIAA) were generally lower in the afternoon and evening in the less than 60 g when compared with the greater than 60 g rats. It is possible that increased 5HT activity is required to stimulate ovulation and that its site of action is the PVN.