indigo-carmine and indirubin

Topics: Color; Coloring Agents; Indigo Carmine; Indoles

Topics: Animals; Antineoplastic Agents; Cell Line, Tumor; Female; Glycosides; Glycosylation; Humans; Indigo Carmine; Indoles; Isomerism; Male; Mice; Neoplasms

Accessed to literatures at home and abroad, we introduced the process of indigo naturalis transforming from dyestuff industry into pharmaceutical industry. It is affirmed that the precursors of indigo and indirubin are isatan A, isatan B, isatan C and indicant. Meanwhile, author clarified the mechanism of transformation for these precursors transforming into indigo and indirubin. And we summarized methods of determination for these precursors. In summary, these references provide us accordance of study on processing principle of Indigo naturalis, and lay the foundation for technics of making indigo and indirubin entering into modern industry.

Topics: Indigo Carmine; Indoles; Oxidation-Reduction; Pigmentation; Plants; Temperature

Purple urine bag syndrome (PUBS) is a rare occurrence, in which the patient has a purple-colored urine bag following urinary catheterization for hours to days. Most of authors believe it is a mixture of indigo (blue) and indirubin (red) that becomes purple. Previous study showed that PUBS occurred predominantly in chronically catheterized, constipated women. We collected 10 elderly patients with PUBS in two nursing homes. The first two cases were identified by chart review in 1987 and 2003, and then later eight cases (42.1%) were collected among 19 urinary catheterized elderly in the period between January 2007 and June 2007. In the present report, PUBS probably can occur in any patients with the right elements, namely urinary tract infection (UTI) with bacteria possessing these enzymes, diet with enough tryptophan, and being catheterized. Associations with bed-bound state, Alzheimer's, or dementia from other causes are reflections of the state of such patients who are at higher risk for UTI, and hence PUBS occurred. Although we presented PUBS as a harmless problem, prevention and control of the nosocomial catheter-associated UTIs (CAUTIs) has become very important in the new patient-centered medical era. Thus, we should decrease the duration of catheterization, improve catheter care, and deploy technological advances designed for prevention, especially in the elderly cared for in nursing homes.

Topics: Aged; Aged, 80 and over; Bacteria; Catheters, Indwelling; Cross Infection; Female; Humans; Indigo Carmine; Indoles; Male; Nursing Homes; Syndrome; Urinary Catheterization; Urinary Tract Infections

Although the color of indigo is strongly dependent on its environment, it is blue in most commonly encountered situations. Indigo's absorption at such long wavelengths for such a small molecule is unique, and I provide here an overview of the concepts advanced to account for this feature. A traditional valence-bond approach may be used to provide a reasonable qualitative explanation. A more rigorous, quantitative explanation is provided by molecular orbital methods of varying degrees of sophistication and several explanations have been proposed based on these models. Commonly, it is suggested that the important structural unit in determining color is based on the cross-conjugated "H-chromophore" concept. A second closely related explanation describes it as two symmetrically coupled merocyanine chains. Another proposal suggests that the basic chromophore may be interpreted as the aza analogue of two coupled anti aromatic-cyclopentadienyl ions. PiSYSTEM, a commercially available quantum mechanics program, has been used to provide a successful quantitative account of the colors of indigo and indirubin, a red isomer.

Topics: Color; Coloring Agents; Indigo Carmine; Indoles

The cytochrome P450 (P450) enzymes involved in drug metabolism are among the most versatile biological catalysts known. A small number of discrete forms of human P450 are capable of catalyzing the monooxygenation of a practically unlimited variety of xenobiotic substrates, with each enzyme showing a more or less wide and overlapping substrate range. This versatility makes P450s ideally suited as starting materials for engineering designer catalysts for industrial applications. In the course of heterologous expression of P450s in bacteria, we observed the unexpected formation of blue pigments. Although this was initially assumed to be an artifact, subsequent work led to the discovery of a new function of P450s in intermediary metabolism and toxicology, new screens for protein engineering, and potential applications in the dye and horticulture industries.

Topics: Animals; Aryl Hydrocarbon Hydroxylases; Biotechnology; Cytochrome P-450 CYP2A6; Cytochrome P-450 Enzyme System; Escherichia coli; Humans; Indigo Carmine; Indoles; Mixed Function Oxygenases; Models, Chemical; Plants, Genetically Modified; Recombinant Proteins

Topics: Animals; Anti-Inflammatory Agents; Colitis; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Indigo Carmine; Mice; Reactive Oxygen Species

Indigo (IND) and indirubin (INB) have demonstrated a synergistic effect in treating ulcerative colitis at a ratio of 7.5:1. However, the colon mucus layer, a critical physiological barrier against external threats, is also a biological barrier, limiting the potential for effective drug delivery to the lamina propria for regulating inflammatory cells. Inspired by the potential of Hyaluronic acid (HA), to enhance cellular uptake by inflammatory cells, and Pluronic® F127 (F127), known for overcoming the mucus barrier, this study innovatively developed INB/IND nanosuspensions by co-modifying with F127 and HA. Moreover, inulin serves a dual purpose as a spray protective agent and a regulator of intestinal flora. Therefore, it was incorporated into INB/IND nanosuspensions for subsequent spray drying, resulting in the preparation of INB/IND nanocrystals (INB/IND-NC). The mucus penetration of INB/IND-NC was 24.30 times that of the control group. Besides, INB/IND-NC exhibited enhanced cellular uptake properties proximately twice that of Raw INB/IND. Importantly, INB/IND-NC exhibited improved therapeutic efficacy in DSS-induced mice by regulating the expression of cytokines, regulating immune responses via downregulating the expression of macrophages, neutrophils, and dendritic cells and maintaining intestinal flora homeostasis. Our study provides a new perspective for applying natural products for treating inflammatory diseases.

Topics: Animals; Colitis; Colitis, Ulcerative; Colon; Dextran Sulfate; Disease Models, Animal; Gastrointestinal Microbiome; Hyaluronic Acid; Indigo Carmine; Inulin; Mice; Mice, Inbred C57BL; Nanoparticles

Topics: Aged, 80 and over; Anti-Bacterial Agents; Bacteria; Color; Humans; Indigo Carmine; Indoles; Male; Urinary Tract Infections

Natural plant dyes have been developed and used across many traditional societies worldwide. The blue pigment indigo has seen widespread usage across South America, Egypt, Europe, India and China for thousands of years, mainly extracted from indigo-rich plants. The utilization and genetic engineering of indigo in industries and ethnobotanical studies on the effects of cultural selection on plant domestication are limited due to lack of relevant genetic and genomic information of dye plants. Strobilanthes cusia (Acanthaceae) is a typical indigo-rich plant important to diverse ethnic cultures in many regions of Asia. Here we present a chromosome-scale genome for S. cusia with a genome size of approximately 865 Mb. About 79% of the sequences were identified as repetitive sequences and 32 148 protein-coding genes were annotated. Metabolic analysis showed that the main indigoid pigments (indican, indigo and indirubin) were mainly synthesized in the leaves and stems of S. cusia. Transcriptomic analysis revealed that the expression level of genes encoding metabolic enzymes such as monooxygenase, uridine diphosphate-glycosyltransferase and β-glucosidase were significantly changed in leaves and stems compared with root tissues, implying their participation in indigo biosynthesis. We found that several gene families involved in indigo biosynthesis had undergone an expansion in number, with functional differentiation likely facilitating indigo biosynthesis in S. cusia. This study provides insight into the physiological and molecular bases of indigo biosynthesis, as well as providing genomic data that provide the basis for further study of S. cusia cultivation by Asia's traditional textile producers.

Topics: Acanthaceae; Chromosomes, Plant; Evolution, Molecular; Gene Expression Profiling; Genome, Plant; Indigo Carmine; Indoles; Plant Leaves; Plant Stems; Plants, Medicinal

This paper presents a comparative study on natural indigo and indirubin in terms of molecular structures and spectral properties by using both computational and experimental methods. The spectral properties were analyzed with Fourier transform infrared (FTIR), Raman, UV-Visible, and fluorescence techniques. The density functional theory (DFT) method with B3LYP using 6-311G(d,p) basis set was utilized to obtain their optimized geometric structures and calculate the molecular electrostatic potential, frontier molecular orbitals, FTIR, and Raman spectra. The single-excitation configuration interaction (CIS), time-dependent density functional theory (TD-DFT), and polarization continuum model (PCM) were used to optimize the excited state structure and calculate the UV-Visible absorption and fluorescence spectra of the two molecules at B3LYP/6-311G(d,p) level. The results showed that all computational spectra agreed well with the experimental results. It was found that the same vibrational mode presents a lower frequency in indigo than that in indirubin. The frontier molecular orbital analysis demonstrated that the UV-Visible absorption and fluorescence bands of indigo and indirubin are mainly derived from π → π* transition. The results also implied that the indigo molecule is more conjugated and planar than indirubin, thereby exhibiting a longer maximum absorption wavelength and stronger fluorescence peak.

Topics: Density Functional Theory; Indigo Carmine; Indoles; Models, Chemical; Models, Molecular; Molecular Structure

Purple urine bag syndrome (PUBS) is a rare condition characterised by urine discolouration. The management of PUBS remains controversial. Four females (mean age 84.5±9.7 years) with palliative conditions (two cancer and two non-cancer cases) presenting PUBS were identified. Urine bags were changed in all cases. Urinary catheters were changed in three cases. Oral antibiotics were prescribed in two cases and used in one case. Urine discolouration was resolved in all cases. One patient (without antibiotic treatment) died on day 5 after presentation of PUBS. Three patients (one out of three cases used oral antibiotics) were clinically stable after the management of PUBS. There was no recurrence of PUBS. Caring for patients with PUBS should be based on clinical decisions, patient status and the goals of care. Palliative care teams should focus on the prevention of PUBS by shortening the duration of catheterisation and minimising modifiable risk factors for this condition.

Topics: Aged; Aged, 80 and over; Anti-Bacterial Agents; Female; Humans; Indigo Carmine; Indoles; Palliative Care; Treatment Outcome; Urinary Catheterization; Urinary Tract Infections

Baphicacanthus cusia (Nees) Bremek (B. cusia) is an effective herb for the treatment of acute promyelocytic leukemia and psoriasis in traditional Chinese medicine. Methyl jasmonate (MeJA) is a well-known signaling phytohormone that triggers gene expression in secondary metabolism. Currently, MeJA-mediated biosynthesis of indigo and indirubin in B. cusia is not well understood. In this study, we analyzed the content of indigo and indirubin in leaf and root tissues of B. cusia with high-performance liquid chromatography and measured photosynthetic characteristics of leaves treated by MeJA using FluorCam6 Fluorometer and chlorophyll fluorescence using the portable photosynthesis system CIRAS-2. We performed de novo RNA-seq of B. cusia leaf and root transcriptional profiles to investigate differentially expressed genes (DEGs) in response to exogenous MeJA application. The amount of indigo in MeJA-treated leaves were higher than that in controled leaves (p = 0.004), and the amounts of indigo in treated roots was higher than that in controlled roots (p = 0.048); Chlorophyll fluorescence of leaves treated with MeJA were significantly decreased. Leaves treated with MeJA showed lower photosynthetic rate compared to the control in the absence of MeJA. Functional annotation of DEGs showed the DEGs related to growth and development processes were down-regulated in the treated leaves, while most of the unigenes involved in the defense response were up-regulated in treated roots. This coincided with the effects of MeJA on photosynthetic characteristics and chlorophyll fluorescence. The qRT-PCR results showed that MeJA appears to down-regulate the gene expression of tryptophan synthase β-subunits (trpA-β) in leaves but increased the gene expression of anthranilate synthase (trp 3) in roots responsible for increased indigo content. The results showed that MeJA suppressed leaf photosynthesis for B. cusia and this growth-defense trade-off may contribute to the improved adaptability of B. cusia in changing environments.

Topics: Acanthaceae; Acetates; Adaptation, Physiological; Biosynthetic Pathways; China; Cyclopentanes; Gene Expression Profiling; Gene Expression Regulation, Plant; Indigo Carmine; Indoles; Oxylipins; Photosynthesis; Plant Breeding; Plant Growth Regulators; Plant Leaves; Plant Roots; Plants, Medicinal; Tryptophan

Topics: Double-Blind Method; Drugs, Chinese Herbal; Ecosystem; Humans; Indigo Carmine; Indoles; Malassezia; Metabolome; Psoriasis

The rinds of surface-ripened cheeses have expected aesthetic properties, including distinct colors, that contribute to overall quality and consumer acceptance. Atypical rind pigments are frequently reported in small-scale cheese production, but the causes of these color defects are largely unknown. We provide a potential microbial explanation for a striking purple rind defect in a surface-ripened cheese. A cheese producer in the United States reported to us several batches of a raw-milk washed-rind cheese with a distinctly purple rind. We isolated a Proteus species from samples with purple rind defect, but not from samples with typical rind pigments, suggesting that this strain of Proteus could be causing the defect. When provided tryptophan, a precursor in the indigo and indirubin biosynthesis pathway, the isolated strain of Proteus secreted purple-red pigments. A Psychrobacter species isolated from both purple and normal rinds also secreted purple-red pigments. Using thin-layer chromatography and liquid chromatography-mass spectrometry, we confirmed that these bacteria produced indigo and indirubin from tryptophan just as closely related bacteria make these compounds in purple urine bag syndrome in medical settings. Experimental cheese communities with or without Proteus and Psychrobacter confirmed that these Proteobacteria cause purple pigmentation of cheese rinds. Reports of purple rinds in two other cheeses from Europe and the observation of pigment production by Proteus and Psychrobacter strains isolated from other cheese rinds suggest that purple rind defect has the potential to be widespread in surface-ripened cheeses.

Topics: Animals; Cattle; Cheese; Color; Indigo Carmine; Indoles; Milk; Pigments, Biological; Proteus; Psychrobacter; Tryptophan

This research aimed to develop an FTIR-based method for rapid and low-cost integrated quality assessment of organic-inorganic composite herbs, which are kinds of herbs composed of both organic and inorganic active ingredients or matrix components. A two-step quality assessment route was designed and verified using the example of Indigo Naturalis (

Topics: Algorithms; Calcium; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Medicine, Chinese Traditional; Plant Extracts; Quality Control; Spectroscopy, Fourier Transform Infrared

Topics: Aged; Catheter-Related Infections; Color; Female; Humans; Indigo Carmine; Indoles; Klebsiella Infections; Klebsiella pneumoniae; Stroke; Urinary Catheterization; Urinary Tract Infections

Terpenoid natural products comprise a wide range of molecular architectures that typically result from C-C bond formations catalysed by classical type I/II terpene cyclases. However, the molecular diversity of biologically active terpenoids is substantially increased by fully unrelated, non-canonical terpenoid cyclases. Their evolutionary origin has remained enigmatic. Here we report the in vitro reconstitution of an unusual flavin-dependent bacterial indoloterpenoid cyclase, XiaF, together with a designated flavoenzyme-reductase (XiaP) that mediates a key step in xiamycin biosynthesis. The crystal structure of XiaF with bound FADH

Topics: Bacteria; Bacterial Proteins; Cyclization; Flavin-Adenine Dinucleotide; Hydroxylation; Inactivation, Metabolic; Indigo Carmine; Indoles; Lyases; Molecular Structure; Phylogeny; Terpenes; Xenobiotics

Purple urine bag syndrome is a rare, benign phenomenon of bacterial colonization. The syndrome affects mainly women and is usually asymp- tomatic. Factors influencing the development of purple urine bag syndrome stem from the pathophysiology of indigo production by bac- teria, and from extrinsic fac- tors.. This case report de- scribes a woman with a complex history placing her at higher risk for developing this syndrome. However, with conservative management she did not develop any complications from this rare syndrome.. Although mainly benign, those popu- lations particularly susceptible to this phenomenon are intrinsically at risk for further complications such as sepsis and altered mental status and must be managed accordingly.

Topics: Female; Humans; Indigo Carmine; Indoles; Middle Aged; Nephrotomy; Tryptophan; Urinary Tract Infections; Urine

The phenylacetone monooxygenase, isolated from Thermobifida fusca, mainly catalyzes Baeyer-Villiger oxidation reaction towards aromatic compounds. Met446 plays a vital role in catalytic promiscuity, based on the structure and function of phenylacetone monooxygenase. Mutation in Met446 locus can offer enzyme new catalytic feature to activate C-H bond, oxidizing indole to finally generate indigo and indirubin, but the yield was only 1.89 mg/L. In order to further improve the biosynthesis efficiency of the whole-cell catalyst, metabolic engineering was applied to change glucose metabolism pathway of Escherichia coli. Blocking glucose isomerase gene pgi led to pentose phosphate pathway instead of the glycolytic pathway to become the major metabolic pathways of glucose, which provided more cofactor NADPH needed in enzymatic oxidation of indole. Engineering the host E. coli led to synthesis of indigo and indirubin efficiency further increased to 25 mg/L. Combination of protein and metabolic engineering to design efficient whole-cell catalysts not only improves the synthesis of indigo and indirubin, but also provides a novel strategy for whole-cell catalyst development.

Topics: Escherichia coli; Glucose; Indigo Carmine; Indoles; Industrial Microbiology; Metabolic Engineering; Metabolic Networks and Pathways; Protein Engineering

Topics: Color; Female; Humans; Indigo Carmine; Indoles; Middle Aged; Syndrome; Tryptophan; Urinary Catheterization; Urine

Topics: Aged; Catheters, Indwelling; Escherichia coli Infections; Female; Humans; Indican; Indigo Carmine; Indoles; Polyvinyl Chloride; Urinary Catheterization; Urinary Tract Infections

To examine the role of a gene encoding flavin-containing monooxygenase (cFMO) from Corynebacterium glutamicum ATCC13032 when cloned and expressed in Escherichia coli for the production of indigo pigments.. The blue pigments produced by recombinant E. coli were identified as indigo and indirubin. The cFMO was purified as a fused form with maltose-binding protein (MBP). The enzyme was optimal at 25 °C and pH 8. From absorption spectrum analysis, the cFMO was classified as a flavoprotein. FMO activity was strongly inhibited by 1 mM Cu(2+) and recovered by adding 1-10 mM EDTA. The enzyme catalyzed the oxidation of TMA, thiourea, and cysteamine, but not glutathione or cysteine. MBP-cFMO had an indole oxygenase activity through oxygenation of indole to indoxyl. The recombinant E. coli produced 685 mg indigo l(-1) and 103 mg indirubin l(-1) from 2.5 g L-tryptophan l(-1).. The results suggest the cFMO can be used for the microbial production of both indigo and indirubin.

Topics: Cloning, Molecular; Copper; Corynebacterium glutamicum; Enzyme Inhibitors; Enzyme Stability; Escherichia coli; Gene Expression; Hydrogen-Ion Concentration; Indigo Carmine; Indoles; Oxygenases; Recombinant Proteins; Spectrum Analysis; Temperature

A change in the colour of urine is always of clinical significance, and a source of concern for the patient and his physician. Among the different urine colours observed, purple is the least common. Although purple discolouration of a catheter and a urine bag is an uncommon finding, it was reported in the literature as early as 1978, by Barlow and Dickson. We present a unique case of purple urine bag syndrome in a patient with bilateral nephrostomy tubes (NT) and associated urine bags (UB) with only the left nephrostomy tube and urine bag exhibiting the purple colour, which resolved with a course of appropriate antibiotics eradicating the causative bacterial pathogen, and change of NT and UB.

Topics: Aged, 80 and over; Anti-Bacterial Agents; Catheters, Indwelling; Color; Humans; Indigo Carmine; Indoles; Male; Nephrostomy, Percutaneous; Pseudomonas aeruginosa; Syndrome; Urinary Catheterization; Urinary Tract Infections; Urine

DNA family shuffling is a powerful method of directed evolution applied for the generation of novel enzymes with the aim of improving their existing features or creating completely new enzyme properties. This method of evolution in vitro requires parental sequences containing a high level of sequence similarity, such as is found in family members of cytochrome P450 enzymes. Cytochromes P450 (P450s or CYPs) are capable of catalyzing a variety of chemical reactions and generating a wide range of products including dye production (e.g., pigments indigo and indirubin). Application of the method of DNA family shuffling described here has enabled us to create novel P450 enzymes and to further extend the capacity of P450 to oxidize indole, leading to pigment formation.

Topics: Cloning, Molecular; Coloring Agents; Cytochrome P-450 Enzyme System; DNA Shuffling; Gene Library; Humans; Indigo Carmine; Indoles; Nucleic Acid Amplification Techniques

The hydrophilicity of the normal decoction pieces (NDP) of Indigo Naturalis is not good, therefore, it is not suit for decoctions. In this paper, powder modification technology is used and some NDP and alcohol are ground together in the vibromill to prepare the hydrophilic decoction pieces (HDP) of Indigo Naturalis. Initially, the properties of NDP, ultrafine decoction pieces (UDP) and HDP are compared, the hydrophilicity of UDP was promoted slightly, that of HDP is promoted dramatically. Then, three batches of Indigo Naturalis are prepared to HDP separately, but there is no obvious difference in the contact angle. Furthermore, the size distribution, surface area and micro-shape of HDP are bigger than that of UDP and smaller than NDP. The contents of indigo and indirubin in three decoction pieces are the same, as well as the species of inorganic substance, although there is a little difference in the proportion of five inorganic substances. The fact suggests the change of physical state and the qualitative and quantitative change of organism and inorganic substances are not the main factors to influence the hydrophilicity. In addition, hydroxyl, methylene and methyl can be identified at the wavenumber of 3 356 cm(-1) and 1 461 cm(-1) in infrared spectrum; the content of alcohol in HDP is 0.67% measured by gas chromatogram. The stability of HDP in the heating condition is studied, the fact suggests the hydrophilic effect of HDP at 40 degrees C is relatively stable. All above research suggests that the alcohol is the main factor to influence the hydrophilicity and maybe the intermolecular force which fixed alcohol molecule on the surface of Indigo Naturalis is the basic principle to produce the hydrophilicity.

Topics: Acanthaceae; Alcohols; Hydrophobic and Hydrophilic Interactions; Indigo Carmine; Indoles; Isatis; Microscopy, Electron, Scanning; Particle Size; Plants, Medicinal; Polygonum; Powders; Surface Properties; Technology, Pharmaceutical; X-Ray Diffraction

To provide a reference evidence for choosing salt-resistent and drought-resistent varieties of Isatis indigotica Fort... The tissue culture seedlings of different Isatis indigotica tetraploids were used as plant materials. The diploid was used as CK, which were cultured for 30 days. 2 g/L NaCl and 15% PEG-6000 were used as stress treatment respectively for 12 hours, then poured out the solution, the materials were continuously cultured for 7 days. The contents of indirubin and indigotin were determined by HPLC.. The contents of indigotin and indirubin of DB3, DB5 and DB12 with PEG treatment and DB2, DB3 and DB12 with NaCl treatment were increased.. DB3 and DB12 can be further studied as reproduction materials.

Topics: Chromatography, High Pressure Liquid; Indigo Carmine; Indoles; Isatis; Plant Roots; Polyethylene Glycols; Quality Control; Seedlings; Sodium Chloride; Stress, Physiological; Tetraploidy; Water

Topics: Aged; Bacteriuria; Color; Female; Humans; Incidental Findings; Indigo Carmine; Indoles; Polyvinyls; Syndrome; Urinary Catheterization; Urinary Tract Infections

Topical indigo naturalis ointment is clinically proved to be an effective therapy for plaque-type psoriasis. Indirubin, as the active component of indigo naturalis, inhibits cell proliferation of epidermal keratinocytes. However, the detailed underlying mechanism is not fully understood.. To further investigate the anti-proliferating effects of indigo naturalis and indirubin on epidermal keratinocytes.. The decreased expression of CDC25B in indigo naturalis- or indirubin-treated epidermal keratinocytes, as revealed by cDNA microarray analysis, was studied. The CDC25B expression was examined under different serum concentrations and compared between primary and immortalized keratinocytes. The activation of EGFR and the effect of EGF on the cell proliferation and CDC25B expression were also investigated in epidermal keratinocytes. RT/real-time PCR and western blot method were used to analyze the CDC25B expression at the mRNA and protein levels, respectively.. Indigo naturalis and indirubin were confirmed to down-regulate CDC25B expression significantly at both the mRNA and protein levels. The growth-dependent expression of CDC25B was demonstrated by the increased expression in serum-stimulated and immortalized keratinocytes. The activation of EGF receptor, known to be highly expressed in psoriatic lesions, was inhibited by indigo naturalis or indirubin. The cell proliferation and CDC25B expression of epidermal keratinocytes were induced by EGF alone and confirmed to be inhibited by indigo naturalis or indirubin.. Except being a common therapeutic target in various cancers, CDC25B also plays an important role in the hyper-proliferation of epidermal keratinocytes which can be suppressed by anti-psoriatic drug indigo naturalis and its component, indirubin.

Topics: cdc25 Phosphatases; Cell Proliferation; DNA, Complementary; Dose-Response Relationship, Drug; Epidermal Cells; Epidermal Growth Factor; Epidermis; ErbB Receptors; Gene Expression Regulation; Humans; Indigo Carmine; Indoles; Keratinocytes; Oligonucleotide Array Sequence Analysis; Plant Extracts; Psoriasis; Skin; Time Factors; Transfection

This bibliography lists and provides commentary on publications describing the history, preparation, chemistry and applications of indigo and related dyes. The dates of the publications are mostly, but not exclusively, later than the previously published indigo bibliography. Note that commentary by the author is placed after the bibliographic information and is set in italics.

Topics: Coloring Agents; Indigo Carmine; Indoles; Plants

A simple and low-cost method based on matrix solid-phase dispersion (MSPD) extraction, HPLC separation, and diode array detection has been developed for the determination of indigo and indirubin in Folium isatidis. The experimental parameters that may affect the MSPD method, including dispersing sorbent, ratio of dispersing sorbent to sample, elution solvent, and volume of the elution solvent were examined and optimized. The optimized conditions were determined to be that C18 was used as dispersing sorbent, the ratio of C18 to sample mass was selected to be 4:1, and 10 mL of N,N-dimethyl formamide was used as elution solvent. The highest extraction yields of the two compounds were obtained under the optimized conditions. The method showed good linearity (r > 0.9995) and precision (RSD < 3.0%) for indigo and indirubin, with the limits of detection of 18 and 22.5 ng/mL, respectively. The recoveries were in the range of 90.33-100.74% with RSD values ranging from 1.7 to 3.6%. Comparing to ultrasonic and Soxhlet methods, the proposed MSPD procedure was more convenient and less time-consuming with reduced requirements on sample and solvent amounts. The proposed procedure was applied to analyzed three real samples that were collected from different localities.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Isatis; Molecular Structure; Solid Phase Extraction

To overcome the problem of poor aqueous solubility and bioavailability of indirubin-3-oximes, the compounds were modified by attaching a quaternary ammonium group at the oxime moiety. Exploring the prodrug concept, an oxime ester with acetyl-l-carnitine was prepared, and the rate of its hydrolysis was investigated to assess its suitability for clinical administration. In addition, the cytotoxic potency of new stable oxime ethers with a choline moiety and their influence on the cell cycle were tested in human cancer cell lines.

Topics: Acetylcarnitine; Antineoplastic Agents; Cations; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Survival; Humans; Hydrolysis; Indigo Carmine; Indoles; Oximes; Solubility; Water

Attempts at identification of mollusc species producing Tyrian purple from archaeological material are usually done with high performance liquid chromatography in the reversed-phase system, but the peaks obtained are often wide and asymmetric. This is due to the low solubility of the indigoids and their brominated derivatives in the mobile phase, especially 6,6'-dibromoindigotin, which is soluble in only few, particular solvents. Our study focused on improving both symmetry and peak height for more precise quantification. The influence of various factors was evaluated: stationary phase characteristics, mobile phase composition, elution gradient parameters and temperature on the peak shape of the main components of Tyrian purple. The best results were obtained using highly retentive, but moderately bonded ODS stationary phases (about 2.8 μmol m(-2)), percolated with gradient of acetonitrile with acidified aqueous mobile phases (0.1% strong acid) at elevated temperatures (70°C). The upper quantification limit for 6,6'-dibromoindigotin was improved by over 350%, between standard and optimised systems. Using them, the detection and quantification of trace Tyrian purple components (less than 0.15%) aside from major indigoids becomes possible. Consequently, for the first time, the new analogues of brominated and unbrominated indirubins were found in the shellfish purple from Hexaplex trunculus.

Topics: Acetonitriles; Bromine; Chromatography, High Pressure Liquid; Indigo Carmine; Indoles; Solubility; Spectrophotometry, Ultraviolet; Temperature

Topics: Acute Kidney Injury; Adenocarcinoma; Aged; Biotransformation; Color; Female; Humans; Hydrogen-Ion Concentration; Indigo Carmine; Indoles; Klebsiella Infections; Klebsiella pneumoniae; Ovarian Neoplasms; Peritoneal Neoplasms; Proteus Infections; Proteus mirabilis; Tryptophan; Urinary Tract Infections; Urine

Different harvest times of Isatidis Folium had a significant effect on the yield and the quality of Isatidis Radix and Isatidis Folium. The harvest could increase the yield of Isatidis Folium, but reduce the yield of Isatidis Radix, the quality of Isatidis Radix and Isatidis Folium. One, two and three harvests of the Isatidis Folium reduced the yield of Isatidis Radix as 18.3%, 58.6%, 67.4% and increased the yield of the Folium as 107.3%, 86.3% and 116.6%. Ethanol-soluble extract of Isatidis Radix was 42.50%, 42.24%, 31.77%, which were 1.19%, 1.79%, 26.13% lower than those of the control, respectively. The water-soluble extract, indirubin, indigo content reduced with increase of the harvest times. Indirubin contents with two or three times harvests were higher than that of the control, but the content of water-soluble extract, ethanol-soluble extract, indigo were lower than those of the control.

Topics: Agriculture; Desert Climate; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Isatis; Plant Leaves; Plant Roots; Quality Control; Time Factors

Oxygenases are useful for the production of many industrially important molecules. Screening of an effluent treatment plant (ETP) sludge metagenomic library identified two clones encoding proteins, B1 and B2, with similarity to putative flavin monooxygenases from Mesorhizobium loti and Sphingomonas wittichi, respectively. The deduced amino acid sequences show only 20% identity, but both have a paired Rossman fold and a flavin monooxygenase (FMO) motif. B1 and B2 appear to be members of the flavin-containing monooxygenase and the Baeyer-Villiger monooxygenases subfamilies, respectively. When expressed in Escherichia coli, the two clones produced activities that oxidized indole to a mixture of indigo and indirubin pigments. These results suggest that B1 and B2 have potential as a biocatalyst in indigo/indirubin production.

Topics: Amino Acid Sequence; Biotransformation; Clone Cells; Gene Library; Indigo Carmine; Indoles; Metagenomics; Mixed Function Oxygenases; Molecular Sequence Data; Phylogeny; Pigments, Biological; Sewage; Waste Disposal, Fluid; Water Purification

To screen the factors that affect indirubin-generated significantly in the process of preparing indigo naturalis, optimize level combination and determine the optimum technology for indirubin-generated.. Using concentration of indirubin (mg x g(-1)) that generated by fresh leaf as an index, Plackett-Burman design, Box-Behnken design response surface analysis as the statistical method, we screened the significantly influencing factors and the optimal level combination.. The soaking and making indirubin process in preparing indigo naturalis was identified as the wax is not removed before immersion with immersion pH 7, solvent volume-leaf weight (mL: g)15, soaked not avoided light, soaking 48 h, temperature 60 degrees C, ventilation time of 180 min, and added ammonia water to adjust pH to 10.5.. The soaking and making indirubin process in preparing indigo naturalis is optimized systematically. It clarify the various factors on the impact of the active ingredient indirubin which controlled by industrialized production become reality in the process of preparing indigo naturalis, at the same time, it lay the foundation for processing principle of indigo naturalis.

Topics: Control Groups; Dose-Response Relationship, Drug; Hydrogen-Ion Concentration; Indigo Carmine; Indigofera; Indoles; Myanmar; Plant Leaves

A proposal for a European Pharmacopoeia monograph concerning Indigo naturalis has recently been published, whereby the indigo (1) and indirubin (2) content should be determined by HPLC-UV. This method was tested, but problems were seen with the dosage of indigo due to poor solubility. A quantitative assay for indigo based on (1)H-NMR was developed as an alternative. The HPLC and qNMR assays were compared with eight Indigo naturalis samples. The HPLC assay consistently gave much lower indigo concentrations because solubility was the limiting factor in sample preparation. In one sample, sucrose was identified by (1)H-NMR as an organic additive. Simple wet chemistry assays for undeclared additives such as sugars and starch were tested with artificially spiked Indigo naturalis samples to establish their limits of detection, and sulfate ash determinations were carried out in view of a better assessment of Indigo naturalis in a future European monograph.

Topics: Chromatography, High Pressure Liquid; Indigo Carmine; Indigofera; Indoles; Magnetic Resonance Spectroscopy; Reference Standards

Indigo naturalis has shown efficacy in treating psoriasis in our previous clinical studies.. To investigate the potential effect of indigo naturalis on regulating keratinocyte proliferation and differentiation.. Skin samples from six patients were analyzed for proliferating cell nuclear antigen (PCNA) and involucrin expression by immunohistochemical staining. In addition, indigo naturalis extracts from 10 to 500 microg/ml were added to cultured keratinocytes and cell viability determined. Real-time RT-PCR, Western blotting analysis and indirect immunofluorescent labeling were used to investigate the messenger (m)RNA and protein expressions of PCNA and involucrin. Finally, high-performance liquid chromatography (HPLC) was used to identify major components of indigo naturalis and their in vitro effects compared.. Immunohistochemical results demonstrated decreased PCNA and increased involucrin in psoriatic lesions after indigo naturalis treatment. Cultured keratinocytes decreased after indigo naturalis treatment, while G(0)/G(1) arrest was observed to dose-dependently increase. Staining revealed decreased PCNA-stained nuclei and increased cytosolic involucrin in treated keratinocytes. Decreased PCNA and increased involucrin at both the mRNA and protein levels were confirmed. Both major components, indirubin and indigo, could cause G(0)/G(1) phase arrest; however, only indirubin modulated the expressions of PCNA and involucrin similar to indigo naturalis.. Together, these findings indicate that the anti-psoriatic effects of indigo naturalis are mediated, at least in part, by modulating the proliferation and differentiation of keratinocytes, with indirubin as the major active component.

Topics: Cell Cycle; Cell Survival; Cells, Cultured; Dermatologic Agents; Down-Regulation; Gene Expression; Humans; Indigo Carmine; Indigofera; Indoles; Keratinocytes; Phytotherapy; Plant Extracts; Proliferating Cell Nuclear Antigen; Protein Precursors; Psoriasis

Purple urine bag syndrome is a rare disorder where the plastic urinary catheter bag and tubing turn purple. The discolouration is due to the presence of indigo and indirubin pigments which are metabolites of tryptophan. It is associated with urinary tract infection. Bacteria that produce sulphatase and phosphatase are involved in the formation of these pigments. Purple urine bag syndrome is associated with higher morbidity and mortality, compared to urinary tract infection without this phenomenon. We present a case report of this rare phenomenon occurring in a 68-year-old woman.

Topics: Aged; Antidepressive Agents, Second-Generation; Escherichia coli Infections; Fatal Outcome; Female; Humans; Indigo Carmine; Indoles; Tryptophan; Urinary Catheterization; Urinary Tract Infections

Topics: Catheters, Indwelling; Diagnosis, Differential; Humans; Indigo Carmine; Indoles; Male; Middle Aged; Protein Kinase Inhibitors; Proteus Infections; Proteus mirabilis; Syndrome; Tryptophan; Urinalysis; Urinary Catheterization; Urinary Tract Infections

Purple urine bag syndrome (PUBS) is considered as an uncommon phenomenon. This cohort study aimed to figure out the causative factors and the potential risks of PUBS.. We recruited 84 patients with long-term urinary catheterization from seven nursing institutions and home care centre, then compared the differences of demographic, clinical, urine test and bacteriological data between the patients with and without PUBS.. The 2-month period prevalence of PUBS in chronic catheterized patients was 16.7%. PUBS-affecting patients had significantly higher proportion of female gender (P = 0.034), self oral intake (P = 0.036) and eating self-prepared food (P = 0.007). Otherwise, no statistical associations were found. Five different bacterial species were isolated from the urine of PUBS-affecting patients. No sequelae were found after forward follow-up for 6 months. We further discuss the associations between certain factors and PUBS according to literature review and the results of our study.. PUBS is not as rare as we thought before. The causative factors of PUBS have not been clearly characterized. It may be the combination of several factors that cause the PUBS. Female gender and food content were found to be associated factors of PUBS in our study. Asymptomatic PUBS is unnecessary to be treated by antibiotics.

Topics: Aged; Aged, 80 and over; Color; Constipation; Female; Humans; Hydrogen-Ion Concentration; Indigo Carmine; Indoles; Male; Middle Aged; Syndrome; Urinary Catheterization; Urinary Tract Infections

Halogenated and polycyclic aromatic hydrocarbons, exogenous ligands of the aryl hydrocarbon receptor (AhR), cause various toxicological effects through the transformation of the AhR. In this study, we investigated the antagonistic effects of indigoids on the transformation in addition to their agonistic ones. In a cell-free system, indigoids induced the transformation dose-dependently, but suppressed the transformation induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin and the binding of 3-methylcholanthrene to the AhR. In mouse hepatoma Hepa-1c1c7 cells, indigoids, especially indirubin, suppressed the transformation and expression of CYP1A1 by inhibiting the translocation of AhR into the nucleus. When orally administered to mice at 10mg/kg BW/day for three successive days, indigoids did not induce AhR transformation and expression of the CYP1A subfamily in the liver, while indirubin and indigo upregulated quinone reductase activity. These results indicate that indigoids are able to bind to the AhR as ligands and exhibit antagonistic effects at lower concentrations in mammalian cells.

Topics: Animals; Carcinoma, Hepatocellular; Cell Line, Tumor; Dose-Response Relationship, Drug; Indigo Carmine; Indoles; Liver; Mice; Piperidones; Receptors, Aryl Hydrocarbon

Cytochrome P450 BM-3 with the mutations A74G, F87V, and L188Q could catalyze indole to produce indigo and indirubin. To further enhance this capability, site-directed and random mutageneses on the monooxygenase domain of P450 BM-3 mutant (A74G/F87V/L188Q; 3X) were performed. The mutant libraries created by error-prone polymerase chain reaction were screened using a colorimetric colony-based method on agar plates followed by a spectroscopic assay involving in absorption of indigo at 670 nm and NADPH at 340 nm in microtiter plate. Three mutants (K434R/3X, E435D/3X, and D168N/ A225V/K440N/3X) exhibited higher hydroxylation activity toward indole in comparison to parent enzyme. Moreover, using saturation site-directed mutagenesis at amino acid positions 168, 225, 434, 435, and 440, two P450 BM-3 variants (D168H/3X, E435T/3X) with an up to sixfold increase in catalytic efficiency (kcat/Km) were identified, and the mutant D168H/3X acquired higher regioselectivity resulting in more indigo (dimerized 3hydroxy-indole) compared to parent mutant (93 vs 72%).

Topics: Bacillus megaterium; Bacterial Proteins; Base Sequence; Catalytic Domain; Cytochrome P-450 Enzyme System; Directed Molecular Evolution; DNA, Bacterial; Hydroxylation; Indigo Carmine; Indoles; Kinetics; Mixed Function Oxygenases; Models, Molecular; Mutagenesis; Mutagenesis, Site-Directed; NADPH-Ferrihemoprotein Reductase; Protein Conformation; Recombinant Proteins

A rapid, selective, and sensitive LC-APCI-MS method is developed in this study for detecting and analyzing tryptanthrin, indigo, and indirubin in daqingye and banlangen, which are, respectively, the leaves and roots of Isatis indigotica and Strobilanthes cusia in traditional Chinese medicine. The detection of the three active components is linear in concentrations ranging from 100 to 1500 ng/mL, the squared correlation coefficient is higher than 0.996, the precision as measured by the relative standard deviation is no larger than 9.5%, and the recovery is greater than 86.6%. The analysis of the 21 banlangen samples led to considerably different conclusions on the contents of tryptanthrin, indigo, and indirubin in fresh leaves versus those in dried leaves. These results should shed some light on future plant selection and breeding. Compared with the traditional TLC and HPLC-UV methods, the new LC-APCI-MS approach has proven to be an optimal tool for detecting and analyzing the three marker compounds in the Chinese herbal medicines of daqingye and banlangen.

Topics: Calibration; Chromatography, High Pressure Liquid; Coloring Agents; Drugs, Chinese Herbal; Indicators and Reagents; Indigo Carmine; Indoles; Isatis; Mass Spectrometry; Plant Leaves; Plant Roots; Quinazolines; Reference Standards; Reproducibility of Results

An HPLC method has been developed and validated for the quantification of the pharmacologically active principles tryptanthrin (1), 1,3-dihydro-3-[(4-hydroxy-3,5-dimethoxyphenyl)methylene]-2 H-indol-2-one (indolinone) (3), indirubin (4), alpha-linolenic acid (2), and indigo (5), an isomer of indirubin, in extracts from the traditional anti-inflammatory plant Isatis tinctoria (woad). The chromatographic separation was performed on a C-18 column with a linear gradient of acetonitrile in water containing 0.1% formic acid. The method combines UV and electrospray MS detection in the positive ion mode for the detection of the alkaloids, with a switch to the negative mode for the analysis of alpha-linolenic acid. The method was applied to the analysis of woad extracts obtained by supercritical fluid (SFE) CO2 extraction, and by pressurized liquid extraction (PLE) with dichloromethane and methanol, respectively. While the highest concentration of alpha-linolenic acid was found in the SFE extract (7.43%), the concentrations of tryptanthrin , indolinone, indirubin and indigo were the highest in the dichloromethane extract (0.30, 0.035, 2.48 and 0.84%, respectively). Compound 3 was not detected in the methanolic extract and only traces of compounds 1, 4 and 5 and low amount of alpha-linolenic acid (0.39%) were present in this extract.

Topics: alpha-Linolenic Acid; Animals; Anti-Inflammatory Agents; Chromatography, High Pressure Liquid; Indigo Carmine; Indoles; Isatis; Mass Spectrometry; Models, Animal; Plant Extracts; Plant Leaves; Quinazolines; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Ultraviolet

The roots and leaves of Isatis indigotica, named 'Ban-Lan-Gen' and 'Da-Qing-Ye', respectively, are widely used for the treatment of influenza, viral pneumonia, mumps, pharyngitis, and hepatitis. The indoxyl derivatives detected in the roots and leaves of I. indigotica have been reported to be biologically active. In the present study, a liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method was developed to determine indican, isatin, indirubin and indigotin in the roots and leaves of I. indigotica. The method has been validated for linearity, precision and accuracy. Using multiple reaction monitoring (MRM), the limits of detection (LODs) were determined as 0.004 ng for indican, 0.01 ng for isatin, 0.01 ng for indrubin and 0.03 ng for indigotin, while the limits of quantitation (LOQs) were 0.015 ng for indican, 0.04 ng for isatin, 0.04 ng for indirubin and 0.1 ng for indigotin. Compared with previously reported methods, the current method is more rapid, selective and sensitive. This is the first report of the LC/MS/MS determination of indican, isatin, indirubin and indigotin.

Topics: Chromatography, High Pressure Liquid; Indican; Indigo Carmine; Indoles; Isatin; Isatis; Plant Extracts; Plant Leaves; Plant Roots; Spectrometry, Mass, Electrospray Ionization

Topics: Aged; Bacteriuria; Humans; Indican; Indigo Carmine; Indoles; Male; Pseudomonas aeruginosa; Urinary Catheterization; Urine

Discoloration of urine is not uncommonly encountered in clinical practice and may indicate a significant pathology. However, the majority of instances are benign and occur as the result of trauma to the urological system during procedures or ingestions of substances such as medication or food. Purple discoloration of a urinary catheter bag is rare and can be alarming to both patients and healthcare workers. This phenomenon is known as the purple urine bag syndrome. It is associated with urinary tract infections occurring in catheterized patients, generally elderly females with significant comorbidities and constipation. The urine is usually alkaline. Gram-negative bacteria that produce sulfatase and phosphatase are involved in the formation of pigment, indirubin and indigo. Tryptophan metabolism is involved in the pathogenesis. We present two cases of this rare and interesting phenomenon and discuss the underlying pathogenesis.

Topics: Aged; Catheters, Indwelling; Chronic Disease; Constipation; Dehydration; Escherichia coli Infections; Fatal Outcome; Female; Humans; Hydrogen-Ion Concentration; Indigo Carmine; Indoles; Middle Aged; Phosphoric Monoester Hydrolases; Sulfatases; Syndrome; Urinary Catheterization; Urinary Tract Infections

We report a case of an elderly lady with a long-term suprapubic catheter who developed purple discoloration of urine around the same time as she developed intussusception. Purple urine bag syndrome is a benign condition known to be associated with intestinal stasis. However, this association with intussusception should be kept in mind before the patient is reassured.

Topics: Aged; Bacteria; Female; Humans; Indigo Carmine; Indoles; Intussusception; Plastics; Urinary Catheterization; Urine

Topics: Aged, 80 and over; Bacteria; Bacteriuria; Catheters, Indwelling; Color; Female; Humans; Indican; Indigo Carmine; Indoles; Plastics; Urinary Catheterization; Urinary Tract Infections

To establish a method for content determination of indirubin and indigo in Baphicacanthus cusia (Nees) Bremek.. HPLC was used with C18 column, methanol-water (75:25) as mobile phase and detecting wavelength 289 nm.. The range linear curve was 3.875 approximately 23.25 microg/ml (indirubin), 3.05 approximately 18.3 microg/ml (indigo). The average recovery was 96.9%, RSD =3.5%, n =5 (indirubin); 96.9% RSD = 2.8%, n = 5 (indigo).. The method is proper for the quality control of Baphicacanthus cusia (Nees) Bremek.

Topics: Acanthaceae; Chromatography, High Pressure Liquid; Drug Contamination; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Plant Leaves; Plant Roots; Plant Stems; Plants, Medicinal; Powders; Quality Control; Seasons

The aim was to establish a method for extraction and chemical fingerprinting of extracts of Isatis indigotica roots ("Ban-Lan-Gen") and to apply the method developed to 18 Ban-Lan-Gen samples. RP-HPLC with gradient elution was performed on authentic reference standards of powdered I. indigotica roots, indigotin and indirubin purchased from the National Institute for the Control of Pharmaceutical and Biological Products (NICPBP) of China. Eighteen "Ban-Lan-Gen" samples (including the reference powdered herb) were bought from Singapore and different regions in China. Comparisons of the chromatograms showed that the samples can be divided into three groups. The chromatograms of the extracts of five samples were found to be similar to that of the extract of the authentic sample. Eight other samples had similar peaks as the authentic sample but the intensities of the peaks were generally lower, except for the peaks between retention times of 10-40 min. Peaks in these regions were more intense than those found in the extract of the authentic sample. Forty-five characteristic peaks could be found in the extracts of all the above samples. Peaks at retention times 52 and 53 min were determined to be indigotin and indirubin, respectively. The remaining four samples had similar chemical fingerprints to each other but were different from that of the authentic sample. Hierarchical clustering analysis gave similar results as the visual comparison. The RP-HPLC method developed allows simple identification and comparisons of I. indigotica roots. This is the first report of hierarchical clustering analysis of I. indigotica root.

Topics: Chromatography, High Pressure Liquid; Cluster Analysis; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Isatis; Molecular Structure; Plant Extracts; Plant Roots; Powders; Reference Standards; Reproducibility of Results; Technology, Pharmaceutical

Purple urine bag syndrome (PUBS) is an uncommon disorder, in which the plastic disposable urinary catheter bag turns purple or blue following hours or days of urinary catheterization. The purple discoloration results from indirubin dissolved in the plastic mixing with indigo in the urine. Bacteria possessing indoxyl sulfatase degrade indoxyl sulfate into indirubin and indigo. Indoxyl sulfate is derived from the metabolism of tryptophan. PUBS usually occurs in chronic catheterized elderly women who are constipated and poorly ambulant. The clinical course is benign and rarely causes sepsis. This investigation reports a 61-year-old female diabetic patient with end-stage renal disease on maintenance hemodialysis, who had two episodes of blue or purple urine bag discoloration. The urine culture of the first episode yielded Klebsiella pneumoniae, whereas that of the second episode yielded Escherichia coli, Enterococcus faecalis, and Proteus vulgaris. Both episodes resolved following oral antibiotics treatment and placement of new foley catheters. To our knowledge, this is the first recorded case of PUBS in a dialysis patient.

Topics: Bacteria; Color; Diabetic Nephropathies; Female; Fermentation; Humans; Indican; Indigo Carmine; Indoles; Middle Aged; Plastics; Renal Dialysis; Urinary Catheterization

A microbial community analysis of forest soil from Jindong Valley, Korea, revealed that the most abundant rRNA genes were related to Acidobacteria, a major taxon with few cultured representatives. To access the microbial genetic resources of this forest soil, metagenomic libraries were constructed in fosmids, with an average DNA insert size of more than 35 kb. We constructed 80,500 clones from Yuseong and 33,200 clones from Jindong Valley forest soils. The double-agar-layer method allowed us to select two antibacterial clones by screening the constructed libraries using Bacillus subtilis as a target organism. Several clones produced purple or brown colonies. One of the selected antibacterial clones, pJEC5, produced purple colonies. Structural analysis of the purified pigments demonstrated that the metagenomic clone produced both the pigment indirubin and its isomer, indigo blue, resulting in purple colonies. In vitro mutational and subclonal analyses revealed that two open reading frames (ORFs) are responsible for the pigment production and antibacterial activity. The ORFs encode an oxygenase-like protein and a putative transcriptional regulator. Mutations of the gene encoding the oxygenase canceled both pigment production and antibacterial activity, whereas a subclone carrying the two ORFs retained pigment production and antibacterial activity. This finding suggests that these forest soil microbial genes are responsible for producing the pigment with antibacterial activity.

Topics: Bacteria; DNA, Bacterial; Genomic Library; Indigo Carmine; Indoles; Molecular Sequence Data; Open Reading Frames; Phylogeny; Pinus; Polymerase Chain Reaction; RNA, Bacterial; RNA, Ribosomal, 16S; Soil Microbiology; Trees

The human bronchial epithelial cells are the primary sites of influenza virus infection. In this study, the effect of indirubin on the expression of the chemokine regulated on activation, normal T cell expressed and secreted (RANTES) by the influenza virus-infected H292 human epithelial cell line was examined. The expression of RANTES mRNA was analyzed using reverse transcription polymerase chain reaction and the concentration of RANTES production was determined by the enzyme-linked immunosorbent assay. At the non-cytotoxic concentrations, indirubin was found to reduce both the expression and production of RANTES in influenza A/NWS/33-infected H292 cells. Inhibition was also observed in influenza virus B/Lee-infected cells. Significant reduction of the expression of IL-8 was not observed after the infection. Indirubin-3'-oxime, a recently developed derivative with kinase inhibitory activity, also mediates a potent inhibitory effect on the expression of RANTES. The influenza virus infection-induced phosphorylation of the nuclear transcription NF-kB regulatory molecule IkBalpha and the p38 MAP kinase were also found to be inhibited by indirubin-3'-oxime. This finding suggests that indirubin is one of the components in the Chinese medicinal herbs Isatis indigotica and Strobilanthes cusia with immunomodulatory activity on the expression of RANTES.

Topics: Bronchi; Chemokine CCL5; Epithelial Cells; Gene Expression Regulation; Humans; I-kappa B Proteins; Indigo Carmine; Indoles; Interleukin-8; Isatin; Mitogen-Activated Protein Kinases; Orthomyxoviridae; p38 Mitogen-Activated Protein Kinases; RNA, Messenger

Indigo and indirubin have been reported to be present at low levels in human urine. The possibility that indigoids are physiological ligands of the aryl hydrocarbon receptor (AhR) has been suggested by initial studies in yeast, where indirubin was found to be 50 times more potent than 2,3,7,8-tetrachlorodibenzo[p]dioxin (TCDD), and indigo was found to be equipotent. To demonstrate that these indigoids are bona fide agonists in mammalian systems, we employed a number of in vitro and in vivo measures of AhR agonist potency. In a hepatoma cell reporter system, indigo yielded an EC50 of approximately 5x10(-6)M (indirubin 3' -oxime EC50 approximately 5x10(-7)M, indirubin EC50 approximately 1x10(-7)M). A comparison of these EC50 values with that of 2,3,7,8-tetrachlorodibenzofuran (TCDBF) ( approximately 3x10(-9)M) indicated that these compounds are less potent than classic halogenated-dibenzofurans or -dibenzo-p-dioxins. Competitive binding assays for AhR occupancy showed similar IC50 values for indirubin and TCDBF ( approximately 2x10(-9) and 5x10(-9)M), with the IC50 values of indigo and indirubin 3' -oxime being approximately 10-fold higher. When rats were treated with these indigoids in the range of 1.5-50mg/kg, induction of hepatic cytochrome P450 1A1 was detected. Differences in the rank-order of potency observed in vivo and in vitro could, in part, be explained by metabolism. Although their biological potencies are not as high as has been previously suggested, collectively the results show that these indole-derived pigments are agonists of AhR in vivo. The in vivo results suggest that solubility, distribution, and metabolism influence the response to the compounds.

Topics: Animals; Binding, Competitive; Cell Line, Tumor; Cytochrome P-450 CYP1A1; Cytosol; Dioxins; Humans; Indigo Carmine; Indoles; Luciferases; Male; Mice; Microsomes, Liver; Oxidation-Reduction; Oximes; Radioligand Assay; Rats; Rats, Sprague-Dawley; Receptors, Aryl Hydrocarbon; Response Elements; Transfection

Indirubin and indigo, which are thought to be natural ligands for aryl hydrocarbon receptor (AhR), showed marked AhR ligand activities in a reporter gene assay using recombinant yeast. Their activities were comparable with or more potent than that of 2,3,7,8-tetrachlorodibenzo-p-dioxin. When indirubin and indigo were administered to mice, ethoxyresorufin-O-dealkylase and methoxyresorufin-O-dealkylase activities in the liver were increased, but subsequently decreased within 2 days. Indirubin was more potent than indigo. Levels of cytochrome P450 1A1/2 proteins and mRNAs in the liver of mice dosed with indirubin were also enhanced. These enhancing effects of indirubin and indigo were not observed in AhR knock-out mice. Ethoxyresorufin-O-dealkylase and methoxyresorufin-O-dealkylase activities in rat hepatocytes and HepG2 cells were enhanced by the addition of indirubin or indigo, but less potently than by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Indigocarmine, a sulfate derivative of indigo, which is used as food additive, did not show these inducing effects on drug-metabolizing enzymes. Our results suggest that indirubin and indigo act as inducers for cytochrome P450 1A1/2 mediated by AhR in mammals in vivo.

Topics: Animals; Cell Line, Tumor; Cytochrome P-450 CYP1A1; Cytochrome P-450 CYP1A2; Cytochrome P-450 Enzyme System; Dose-Response Relationship, Drug; Enzyme Induction; Hepatocytes; Indigo Carmine; Indoles; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Microsomes, Liver; Rats; Rats, Sprague-Dawley; Receptors, Aryl Hydrocarbon; RNA, Messenger; Yeasts

Purple urine bag syndrome (PUBS) was first reported in 1978. PUBS is rare, occurs predominantly in constipated women, chronically catheterized and associated with some bacterial urinary infections that produce sulphatase/phosphatase. The etiology is due to indigo (blue) and indirubin (red) or to their mixture that becomes purple. A chain reaction begins in the gastrointestinal tract with tryptophan as described in the article.

Topics: Catheters, Indwelling; Female; Humans; Indican; Indigo Carmine; Indoles; Middle Aged; Syndrome; Tryptophan; Urinary Catheterization; Urinary Tract Infections; Urine

Two indigoid pigments, red indirubin and blue indigotin, were isolated from the urine of healthy male subjects. The two pigments were assayed on HL60 (human promyelocytic cell line) cells. Preliminary results revealed the antiproliferative effect of the pigments. Indigotin seemed more effective than indirubin.

Topics: Cell Division; Growth Inhibitors; HL-60 Cells; Humans; In Vitro Techniques; Indigo Carmine; Indoles; Male

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes involved in xenobiotic metabolism, cellular proliferation, and differentiation. Numerous xenobiotic and biological compounds are known to interact with AhR, but it remains an orphan receptor, because its physiological ligand is unknown. We identified AhR ligands in human urine using a yeast AhR signaling assay and then characterized their properties. Two ligands, indirubin and indigo, were both present at average concentrations of approximately 0.2 nm in the urine of normal donors. Indirubin was also detected in fetal bovine serum and contributed half of the total AhR ligand activity. The activities of indirubin and indigo were comparable with or more potent than that of the archetypal ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin, in yeast AhR activation assays. We suggest that the endogenous levels and potencies of indirubin and indigo are such that they activate AhR-mediated signaling mechanisms in vivo.

Topics: Genes, Reporter; Humans; Indigo Carmine; Indoles; Ligands; Male; Polychlorinated Dibenzodioxins; Receptors, Aryl Hydrocarbon; Saccharomyces cerevisiae; Signal Transduction

A wild-type naphthalene-degrading strain Pseudomonas putida RKJ1 and two recombinant strains each of Ps. putida and Escherichia coli carrying the genes for naphthalene degradation on a recombinant plasmid pRKJ3, produced indigo and indirubin pigments from indole. Naphthalene, salicylate and IPTG induced cells of naphthalene-degrading recombinant bacteria produced up to two times higher indigo compared with the uninduced cells. The maximum rates of indigo formation by Ps. putida RKJ1, Ps. putida RKJ5/pRKJ3, Ps. putida KT2442/pRKJ3, E. coli TB1/pRKJ3 and E. coli AB1157/pRKJ3 were 0.60, 0.80, 0.60, 1.20 and 1.50 nmol min-1 mg dry biomass-1, respectively, using indole as the substrate. The apparent Km values of indigo formation by these same bacteria were 0.22, 0.15, 0.10, 0. 21 and 0.20 mmol l-1, respectively, again using indole as the substrate. The present study revealed that E. coli AB1157 was the most efficient of the hosts tested for the expression of the plasmid encoded genes (pRKJ3) from the wild-type strain Ps. putida RKJ1. In addition, both recombinant E. coli strains were capable of producing indigo directly from nutrient medium.

Topics: Coloring Agents; DNA, Recombinant; Escherichia coli; Indigo Carmine; Indoles; Naphthalenes; Pseudomonas putida

Indole is a product of tryptophan catabolism by gut bacteria and is absorbed into the body in substantial amounts. The compound is known to be oxidized to indoxyl and excreted in urine as indoxyl (3-hydroxyindole) sulfate. Further oxidation and dimerization of indoxyl leads to the formation of indigoid pigments. We report the definitive identification of the pigments indigo and indirubin as products of human cytochrome P450 (P450)-catalyzed metabolism of indole by visible, (1)H NMR, and mass spectrometry. P450 2A6 was most active in the formation of these two pigments, followed by P450s 2C19 and 2E1. Additional products of indole metabolism were characterized by HPLC/UV and mass spectrometry. Indoxyl (3-hydroxyindole) was observed as a transient product of P450 2A6-mediated metabolism; isatin, 6-hydroxyindole, and dioxindole accumulated at low levels. Oxindole was the predominant product formed by P450s 2A6, 2E1, and 2C19 and was not transformed further. A stable end product was assigned the structure 6H-oxazolo[3,2-a:4, 5-b']diindole by UV, (1)H NMR, and mass spectrometry, and we conclude that P450s can catalyze the oxidative coupling of indoles to form this dimeric conjugate. On the basis of these results, we propose that the P450/NADPH-P450 reductase system can catalyze oxidation of indole to a variety of products.

Topics: Animals; Aryl Hydrocarbon Hydroxylases; Cytochrome P-450 CYP2A6; Cytochrome P-450 Enzyme System; Escherichia coli; Humans; Indigo Carmine; Indoles; Isatin; Male; Microsomes, Liver; Mixed Function Oxygenases; NADH, NADPH Oxidoreductases; NADPH-Ferrihemoprotein Reductase; Oxidation-Reduction; Pigments, Biological; Rats; Rats, Wistar; Recombinant Proteins

Polygonum tinctorium Lour. (P. tinctorium) is known to have the ability to suppress inflammation. We attempted to isolate the active compounds from P. tinctorium based on their inhibitory effects on the production of interferon-gamma, which is a well-known inflammatory cytokine. We thus isolated indirubin, an isomer of indigo. Indirubin exerted its inhibitory effects not only on interferon-gamma production by human myelomonocytic HBL-38 cells but also on interferon-gamma and interleukin-6 production by murine splenocytes with no influence on the proliferation of either cells. Because of its inhibitory activity on interferon-gamma production, we further investigated the effects of indirubin on 2,4, 6-trinitro-l-chlorobenzene (TNCB)-induced delayed-type hypersensitivity as a representative inflammatory reaction. When injected intraperitoneally, indirubin significantly inhibited the ear swelling of TNCB-elicited mice. The amount of interferon-gamma in the culture supernatants of elicited mouse lymphocytes was inhibited by indirubin treatment. These results suggest that indirubin is a compound with anti-inflammatory effects.

Topics: Adjuvants, Immunologic; Animals; Antibiotics, Antineoplastic; Humans; Hypersensitivity, Delayed; Immunoglobulin E; Indigo Carmine; Indoles; Interferon-gamma; Interleukin-2; Interleukin-4; Interleukin-6; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Molecular Structure; Tumor Cells, Cultured

Indirubin is the active ingredient of Danggui Longhui Wan, a mixture of plants that is used in traditional Chinese medicine to treat chronic diseases. Here we identify indirubin and its analogues as potent inhibitors of cyclin-dependent kinases (CDKs). The crystal structure of CDK2 in complex with indirubin derivatives shows that indirubin interacts with the kinase's ATP-binding site through van der Waals interactions and three hydrogen bonds. Indirubin-3'-monoxime inhibits the proliferation of a large range of cells, mainly through arresting the cells in the G2/M phase of the cell cycle. These results have implications for therapeutic optimization of indigoids.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; CDC2-CDC28 Kinases; Cell Cycle; Cell Division; Cell Line; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinases; Enzyme Inhibitors; HL-60 Cells; Humans; Indigo Carmine; Indoles; Isatin; Jurkat Cells; K562 Cells; Leukemia L1210; Medicine, Chinese Traditional; Mice; Models, Molecular; Molecular Conformation; Protein Conformation; Protein Serine-Threonine Kinases; Recombinant Proteins; Spodoptera; Transfection

The plastic of urinary catheter drainage bags occasionally turns purple hours or days after catheterization and the color becomes increasingly intense the longer the same drainage system is left in place. This phenomenon was first reported in 1978 as "purple urine bag syndrome", and had been known to occur with bacterial infection of the urinary tract with chronic constipation. Chronic constipation is commonly associated with bacterial overgrowth in the bowel in which tryptophan has been converted to indol and yields the high levels of indigo (blue) and indirubin (red) in urinary bags of patients with bacterial infection of the urine, because indigo-producing bacteria have indoxyl phosphatase or sulfatase that can produce indigo and indirubin. We determined the serum levels of amino acids in patients with purple urine bag syndrome. The serum level of tryptophan and valine were significantly reduced in patients with purple urine bag syndrome. This result suggests that absorption of amino acids was affected by disturbances of colonic motility and intestinal bacterial overgrowth.

Topics: Aged; Amino Acids; Catheters, Indwelling; Chronic Disease; Female; Humans; Indigo Carmine; Indoles; Syndrome; Time Factors; Urinary Catheterization; Urinary Tract Infections

The p-cumate-degrading strain Pseudomonas putida F1 and the m- and p-toluate-degrading strain P. putida mt-2 transform indole-2-carboxylate and indole-3-carboxylate to colored products identified here as indigo, indirubin, and isatin. A mechanism by which these products could be formed spontaneously following dioxygenase-catalyzed dihydroxylation of the indolecarboxylates is proposed. Indolecarboxylates were employed as chromogenic substrates for identifying recombinant bacteria carrying genes encoding p-cumate dioxygenase and toluate dioxygenase. Dioxygenase gene-carrying bacteria could be readily distinguished as dark green-blue colonies among other colorless recombinant Escherichia coli colonies on selective agar plates containing either indole-2-carboxylate or indole-3-carboxylate.

Topics: Amino Acids; Carboxylic Acids; Chromogenic Compounds; Cloning, Molecular; Coloring Agents; Indigo Carmine; Indoles; Isatin; Oxygenases; Pseudomonas putida

This paper first suggests that 2,4 (1H,3H) quinazolinedione (QZL) may be used as a quality control standard for the Chinese herbal drugs Daqingye (Folium Strobilanthis et Polygoni) and Banlangen (Radix Strobilanthis) as well as its preparations. QZL was found for the first time from the above mentioned two Chinese herbal drugs which has a representative property. Its content is stable. This made it possible to solve the difficulty of quality control for the preparations "antipyretic and common cold granules" and "Banlangan granules" and provide a reliable assay method to evaluate the quality of the herbal preparations. By changing the wavelength and using gradient elution in HPLC, 3 constituents can be determined through one injection by the present method, thus the time of determination may be shortened and the efficiency of assay increased. The method showed good linearity (r = 0.99998, n = 4) in the range of 0.2-3.2 micrograms/ml of QZL, the average recovery was above 95% with RSD < 1.5% (n = 3). HPLC condition: Column: LiChrosorb RP 18; elution gradient: methanol from 35% to 90% in water solution; determination wavelength: 226 nm and 288 nm.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Indigo Carmine; Indoles; Quality Control; Quinazolines

Pigments produced by Escherichia coli containing a cloned piece of DNA from Rhodococcus sp. ATCC 21145 were extracted in chloroform and separated into blue and pink components. Evidence from TLC, NMR spectroscopy, absorption spectrum analysis and solubility behaviour suggested that the blue pigment was indigo and the pink pigment was indirubin, a structural isomer of indigo. The proposed pathway for pigment production on LB agar involves the conversion of tryptophan to indole by tryptophanase of E. coli and the oxidation of indole to indigo by the product of the cloned Rhodococcus DNA insert.

Topics: Chromatography, Thin Layer; Cloning, Molecular; Culture Media; DNA, Bacterial; Escherichia coli; Genetic Vectors; Indigo Carmine; Indoles; Magnetic Resonance Spectroscopy; Pigments, Biological; Plasmids; Rhodococcus; RNA, Bacterial; Solubility; Transformation, Bacterial; Tryptophan; Tryptophanase

The indigotin and indirubin contents of Radix Isatidis, its aqueous extract and residues, as well as extract with 95%, 50% alcohol were determined by TLC-densitometry. The authors throw doubt upon the current processes of extracting with water and precipitating with alcohol for Radix Isatidis, and indicate that the method of extraction with alcohol can give more indigotin and indirubin.

Topics: Antibiotics, Antineoplastic; Chromatography, Thin Layer; Densitometry; Drugs, Chinese Herbal; Ethanol; Indigo Carmine; Indoles; Technology, Pharmaceutical

Branhamella catarrhalis, an occasional cause of human respiratory infections, unlike most other members of the family Neisseriaceae, produces a butyrate esterase. This is capable of breaking the ester linkage between butyryl groups and carrier molecules. B. caviae and B. ovis, which are rarely encountered in pathological specimens, also produce butyrate esterase. This property can be used as a rapid test in the identification of B. catarrhalis. The recently reported rapid test for butyrate esterase relies on the release of methylumbelliferate, which can be detected only by using UV light after 5 min of incubation. In the rapid test described here, indoxyl is liberated from indoxyl butyrate by butyrate esterase and spontaneously forms indigo in the presence of oxygen. B. catarrhalis can be distinguished from other oxidase-positive, gram-negative cocci after 2.5 min by inoculating the organism onto a filter paper strip containing this compound.

Topics: Carboxylic Ester Hydrolases; Humans; Hydrolysis; Indigo Carmine; Indoles; Moraxella catarrhalis; Time Factors

The urine of a patient who suffered from acute myelomonocytic leukemia was red coloured after administration of mitoxantrone and etoposid. The isolation and spectroscopic identification of the excreted pigments resulted in the chemical structures of indigo and indirubin. The structure elucidation has been carried out by taking use of a two-dimensional technique in high-resolution nuclear magnetic resonance spectroscopy and different mass spectrometric methods, including tandem mass spectrometry. Possible reasons for the formation of the indigoid compounds are discussed.

Topics: Antibiotics, Antineoplastic; Chromatography, Ion Exchange; Humans; Indigo Carmine; Indoles; Leukemia; Magnetic Resonance Spectroscopy; Mass Spectrometry

The etiology of the purple urine bag syndrome (PUBS), in which the urinary catheter bag of some elderly patients develops intense purple coloration, was studied. The purple was found to be a mixture of indirubin dissolved in the plastic and indigo on its surface. Six patients with PUBS were studied, and Providencia stuartii was isolated from the urine of five and Klebsiella pneumoniae was isolated from the urine of one. These strains produced indigo in 7.9 mM indoxyl sulfate-containing agar. One hundred and fifty isolates of 41 species of bacteria were tested for their ability to produce indigo on agar containing indoxyl sulfate, but only 17 of 27 strains of P. stuartii, a single strain of Klebsiella pneumoniae, and Enterobacter agglomerans were positive. All of the indigo-producing bacteria had an indoxyl phosphatase with a pI of 6.4. This enzyme also possessed indoxyl sulfatase activity and was not present in strains that were unable to produce indigo from indoxyl sulfate. We conclude that PUBS results from the decomposition of urinary indoxyl sulfate to indigo and indirubin by bacteria (notably P. stuartii). As elderly catheterized patients often have high urinary indoxyl sulfate levels and colonization of their urinary tract with P. stuartii, the condition is most commonly seen in them.

Topics: Bacteriuria; Color; Humans; Indican; Indigo Carmine; Indoles; Isoelectric Focusing; Klebsiella pneumoniae; Phosphoric Monoester Hydrolases; Proteus; Providencia; Sulfatases; Urinary Catheterization

A purple pigment, phyriaviolin, and a blue pigment, phyriaazulin, have been found in relatively large amounts in the urine of patients suffering from two diverse pathological conditions, porphyria cutanea tarda and Crohn's disease. The two pigments have been characterised by chemical, spectroscopic, and chromatographic studies and identified to be indirubin and indigo (indigotin). Possible reasons for their formation are discussed.

Topics: Chromatography, Thin Layer; Crohn Disease; Humans; Indigo Carmine; Indoles; Magnetic Resonance Spectroscopy; Pigments, Biological; Porphyrias; Skin Diseases

Topics: Chromatography, High Pressure Liquid; Fabaceae; Indigo Carmine; Indoles; Medicine, Chinese Traditional; Phytotherapy; Plants, Medicinal

Topics: Antibiotics, Antineoplastic; Chemical Phenomena; Chemistry; Indigo Carmine; Indoles; Leukemia, Myeloid

Topics: Indigo Carmine; Indoles; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Plants, Medicinal; Spectrophotometry

Topics: Chromatography, Paper; Indigo Carmine; Indoles; Spectrophotometry

Topics: Indigo Carmine; Indoles; Spectrophotometry