Apoptosis of pancreatic cancer BXPC-3 cells induced by indole-3-acetic acid in combination with horseradish peroxidase

AIM： To explore the mechanisms underlying the apoptosis of human pancreatic cancer BXPC-3 cells induced by indole-3-acetic acid （IAA） in combination with horseradish peroxidase （HRP）. METHODS： BXPC-3 cells derived from human pancreatic cancer were exposed to 40 or 80 μmol/L IAA and 1.2 μg/mL HRP at different times. Then, Mn- assay was used to detect the cell proliferation. Flow cytometry was performed to analyze cell cycle. Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling assay was used to detect apoptosis. 2,7-Dichlorofluorescin diacetate uptake was measured by confocal microscopy to determine free radicals. Level of malondialdehyde （MDA） and activity of superoxide dismutase （SOD） were measured by biochemical methods. RESULTS： IAA/HRP initiated growth inhibition of BXPC-3 cells in a dose- and time-dependent manner. Flow cytometry revealed that the cells treated for 48 h were arrested at G1/G0. After exposure to 80 μmol/L IAA plus 1.2 μg/mL HRP for 72 h, the apoptosis rate increased to 72.5‰, which was nine times that of control. Content of MDA and activity of SOD increased respectively after treatment compared to control. Meanwhile, IAA/HRP stimulated the formation of free radicals. CONCLUSION： The combination of IAA and HRP can inhibit the growth of human pancreatic cancer BXPC-3 cells in vitro by inducing apoptosis.

Synthesis,Spectroscopic Characterization,Thermogravimetric and Biological Activity Evaluation of Te(Ⅳ),Se(Ⅳ),V(Ⅲ),Nb(Ⅴ),Ta(Ⅴ)Complexes With Indole-3-Acetic Acid Plant Hormone Ligand

Te(Ⅳ),Se(Ⅳ),V(Ⅲ),Nb(Ⅴ)and Ta(Ⅴ)complexes of indole-3-acetic acid(IAAH)ligand were synthesized,characterized by elemental analysis and various spectroscopic techniques like,IR,1H-NMR,X-ray powder diffraction,UV-Visible,thermogravimetry analysis,magnetic measurements,molar conductance and surface morphology using SEM.All the synthesized complexes of IAAH ligand have 1∶2 stoichiometry of the types[Te(IAA)2(NH 3)2]·2Cl(Ⅰ),[Se(IAA)2(NH 3)2]·2Cl(Ⅱ),[V(IAA)2(NH 3)(Cl)](Ⅲ),[Nb(IAA)2(Cl)3](Ⅳ),and[Ta(IAA)2(Cl)3](Ⅴ).Spectral analysis indicates octahedral geometry for the Te(Ⅳ),Se(Ⅳ)and V(Ⅲ)complexes,whereas both Nb(Ⅴ)and Ta(Ⅴ)have a seven-coordination.The bonding sites are the oxygen atoms of carboxylate group for the deprotonated indole-3-acetic acid(IAA)ligand.The thermogravimetry analysis studies gave evidence for the presence of other coordinated molecules(Cl or NH 3)in the composition of IAA complexes,which were further supported by IR and micro analytical measurements.The higher molar conductance data of tellurium and selenium(Ⅳ)complexes reveal that these chelates are electrolytes,while low conductivity values for the vanadium(Ⅲ),niobium and tantalum(Ⅴ)chelates indicated a non-electrolytes.To test the antibacterial property of the five complexes in this study,four bacterial strains Klebsiella(G-),Escherichia coli(G-),Staphylococcus aureus(G+)and Staphylococcus epidermidis(G+)were used in the investigation.The effects of the five complexes in the cytotoxicity of Caco-2 and Mcf-7 human cancer cell lines were studied Neutral red uptake assay for the estimation of cell viability/cytotoxicity protocol.

THE APPLICATION OF SEROTONIN AND 5-HVDROXYINDOLE-3-ACETIC ACID IN THE DIAGNOSIS OF APUDOMA

In our report, the values of whole blood serotonin (5-HT) in 87 apudoma patients (diagnosed by operation and pathology) were summarized. Among them, the levels of urinary 5-hydroxyindole-3-acetic acid (5-HIAA) of 44 patients were also tested. The results showed that both parametres of apudoma patients were higher than those of non-apudoma, post-operative patients of apudoma as well as the normal. The increasing extent of the levels of whole blood 5-HT and urinary 5-HIAA in small intestinal carcihoid was the most obvious one but that of rectum was not. The referable diagnostic values suggested were: 5-HT>130 ng ml, 5-HIAA>30 mg/ 24 hours.

The Difference of Sensitivity between BXPC-3 and K562 Cells by Treatments with Combination of Indole-3-acetic Acid and Horseradish Peroxidase

The difference of sensitivity to indole- 3-acetic acid （ IAA ） combined with horseradish peroxidase （HRP） in K562 and BXPC- 3 cells was investigated. The cell proliferation was determined by MTF assay. The cell cycle and apoptosis of K562 and BXPC-3 cells were examined by a fluorescence flow cytometer （FCM） and terminal deoxynacleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） respectively. The experimental results show that IAA and HRP could inhibit BXPC- 3 cell proliferation greatly compared with K562 cell during the first 48 h . The cell cycle was arrested predominantly at G2/ M phase in K562 and BXPC- 3 cells. The cell apoptosis of K562 and BXPC- 3 was induced by IAA/ HRP. There was a significant difference between the two cell lines since BXPC-3 cells were more sensitive than K562 cells by treatments with combination of IAA and HRP.

Sulfur nutrient availability regulates root elongation by affecting root indole-3-acetic acid levels and the stem cell niche

Sulfur is an essential macronutrient for plants with numerous biological functions. However, the influence of sulfur nutrient availability on the regulation of root development remains largely unknown. Here, we report the response of Arabidopsis thaliana L. root development and growth to different levels of sulfate, demonstrating that low sulfate levels promote the primary root elongation. By using various reporter lines, we examined in vivo IAA level and distribution, cell division,and root meristem in response to different sulfate levels.Meanwhile the dynamic changes of in vivo cysteine, glutathione,and IAA levels were measured. Root cysteine, glutathione, and IAA levels are positively correlated with external sulfate levels in the physiological range, which eventually affect root system architecture. Low sulfate levels also downregulate the genes involved in auxin biosynthesis and transport, and elevate the accumulation of PLT1 and PLT2. This study suggests that sulfate level affects the primary root elongation by regulating the endogenous auxin level and root stem cell niche maintenance.

One-step co-electrodeposition of graphene oxide doped poly(hydroxymethylated-3,4-ethylenedioxythiophene) film and its electrochemical studies of indole-3-acetic acid

A novel graphene oxide （GO） doped poly（hydroxymethylated-3,4-ethylenedioxythiophene） （PEDOTM） film has been achieved via one-step co-electrodeposition and utilized for electrochemical studies of indole-3-acetic acid （IAA）. The incorporation of GO into PEDOTM film facilitated the electrocatalytic activity and exhibited a favorable interaction between the PEDOTM/GO film and the phytohormone during the oxidation of IAA. Under optimized conditions, differential pulse voltammetry and square wave voltammetry were used for the quantitative analysis of IAA, respectively, each exhibiting a wide linearity range from 0.6 μmol L-1 to 10 μmol L-1 and 0.05 μmol L-1 to 40 μmol L-1, good sensitivity with a low detection Iimit of 0.087 μmol L-1 and 0.033μmol L T, respectively, as well as good stability. With the notable advantages of a green, sensitive method, expeditious response and facile operation, the as-prepared PEDOTM/GO organic-inorganic composite film provides a promising platform for electrochemical studies of IAA.

Amperometric Determination of lndole-3-acetic Acid Based on Platinum Nanowires and Carbon Nanotubes

Platinum nanowire （PtNW） can be grown by electrodeposition in polycarbonate membrane, with the average diameter of the nanowires about 250 nm. The PtNW and multiwalled carbon nanotubes （CNT） are then dispersed into chitosan （CHIT） solution. The resulting PtNW-CNT-CHIT material brings new capabilities for electrochemical devices by using the synergistic action of the electrocatalytic activity of PtNW and CNT. By dropping the PtNW-CNT-CHIT film onto the glassy carbon （GO） electrode surface, and after evaporation an amperometric sensor for the determination of indole-3-acetic acid （IAA） was developed. The oxidation current of IAA increased significantly at the PtNW-CNT-CHIT film coated GC electrode, in contrast to that at the CNT-CHIT modified GC. The linear response of the sensor is from 50 ng/ml to 50 μg/ml with a detection limit of 25 ng/mL.

Lateral root elongation in maize is related to auxin synthesis and transportation mediated by N metabolism under a mixed NO_(3)^(–) and NH_(4)^(+) supply

A mixed nitrate (NO_(3)^(–)) and ammonium (NH_(4)^(+)) supply can promote root growth in maize (Zea mays),however,the changes in root morphology and the related physiological mechanism under different N forms are still unclear.Here,maize seedlings were grown hydroponically with three N supplied in three different forms (NO_(3)^(–)only,75/25 NO_(3)^(–)/NH_(4)^(+)and NH_(4)^(+)only).Compared with sole NO_(3)^(–)or NH_(4)^(+),the mixed N supply increased the total root length of maize but did not affect the number of axial roots.The main reason was the increased total lateral root length,while the average lateral root (LR) length in each axle was only slightly increased.In addition,the average LR density of 2nd whorl crown root under mixed N was also increased.Compared with sole nitrate,mixed N could improve the N metabolism of roots (such as the N influx rate,nitrate reductase (NR) and glutamine synthase (GS)enzyme activities and total amino content of the roots).Experiments with exogenously added NR and GS inhibitors suggested that the increase in the average LR length under mixed N was related to the process of N assimilation,and whether the NR mediated NO synthesis participates in this process needs further exploration.Meanwhile,an investigation of the changes in root-shoot ratio and carbon (C) concentration showed that C transportation from shoots to roots may not be the key factor in mediating lateral root elongation,and the changes in the sugar concentration in roots further proved this conclusion.Furthermore,the synthesis and transportation of auxin in axial roots may play a key role in lateral root elongation,in which the expression of ZmPIN1B and ZmPIN9 may be involved in this pathway.This study preliminarily clarified the changes in root morphology and explored the possible physiological mechanism under a mixed N supply in maize,which may provide some theoretical basis for the cultivation of crop varieties with high N efficiency.

Theoretical Studies on the Aminolysis of Ester: Effects of the Catalysis and Substituent to the Reaction of Methyl Indole-3-acetate with Ammonia

A comprehensive exploration of the aminolysis mechanism for methyl indole-3-acetate with ammonia is carried out by employing the B3 LYP/6-311++G(d,p), M06-2 X/6-311++G(d,p) and MP2/6-311++G(d,p)//M06-2 X/6-311++G(d,p) levels. Two alterative reaction channels of the concerted and addition/elimination stepwise processes including the uncatalyzed, base-catalyzed reactions are taken into consideration. Subsequently, the substituent effects and solvent effects in methanol are also evaluated at the M06-2 X/6-311++G(d,p) level. The calculated results indicate that the calculated values of M06-2 X level are quite close to those of MP2, the stepwise pathway has more advantages to the concerted one for all of the reaction processes and the catalyst facilitates the proton migration and decreases the energy barriers as well. It is shown that the most preferred mechanism is the based-catalyzed stepwise process, the substituent of NH2 group slightly accelerates all the aminolysis reaction processes, and the solvent effect does not remarkably change the mechanism of the reaction.

Endogenous nitric oxide and hydrogen peroxide detection in indole-3-butyric acid-induced adventitious root formation in Camellia sinensis

Nitric oxide（NO）and hydrogen peroxide（H2O2）are essential signaling molecules with key roles in auxin induced adventitious root formation in many plants.However,whether they are the sole determinants for adventitious root formation is worth further study.In this study,endogenous NO and H2O2 were monitored in tea cutting with or without indole-3-butyric acid（IBA）treatment by using the fluorescent probes diaminofluorescein diacetate（DAF-2DA）and 2＇,7＇-dichlorodihydrofluorescein diacetate（DCF-DA）,respectively.The overproduction of NO and H2O2 was detected in the rooting parts of tea cuttings treated with or without IBA.But little NO and H2O2 was detected before the initiation phase of tea cuttings even with IBA treatment indicating that they might be not directly induced by IBA.Further carbon and nitrogen analysis found that the overproduction of NO and H2O2 were coincident with the consumption of soluble sugars and the assimilation of nitrogen.These results suggest that rooting phases should be taken into consideration with the hypothesis that auxin induces adventitious root formation via NO-and H2O2-dependent pathways and sink establishment might be a prerequisite for NO and H2O2 mediated adventitious root formation.

Effect of Different Indole-3-Buteric Acid(IBA)Concentrations and Cutting Types on Plane Trees(Platanus orientalis L.)

Effects of different Indole-3-Buteric Acid （IBA） concentrations （0, 1 000, 1 500 and 2 500 ppm） and curing types （soft wood, semi hard and hard wood） of plane tree were investigated with Randomized Complete Block Design （RCBD） at the Agricultural research station, Charsadda. The maximum survival percentage, plant height （cm）, root length （era）, numbers of roots and root weight per cutting were significantly higher in hard wood cuttings. IBA concentration had no speculative effect on plane tree cuttings. Thus, hard wood cutting was the best choice for plan tree propagation.

Cross-linked lipoic acid nanoparticles with indole-3-methanol loading for the PTEN-me diate d TNBC treatment

We here report a non-gene therapy anti-tumor nanoparticles(I3C@cLANPs)based PTEN by loading indole-3-methanol(I3C)into the cross-linked lipoic acid nanoparticles(cLANPs)for triple-negative breast cancer(TNBC)treatment.I3C is a PTEN-specific natural activator while the poor PTEN-activation effi-ciency impedes its ability to achieve the PTEN-mediated tumor therapy.Due to the structural homology of lipoic acid(LA),the cLANPs hold not only LA-like anticancer activity but also PTEN-activation proper-ties,which would synergistically potentiate the PTEN-activation efficiency.The in vitro and in vivo data showed that PTEN expression in the I3C@cLANPs group was 2.1 and 2.8 times higher than that of I3C,respectively.In antitumor evaluation based on the 4T1 tumor-bearing mice showed a tumor inhibitory rate(TIR)of 78.4%at the I3C usage of 20 mg kg^(–1),54.5%higher than that of I3C alone and 19.7%higher than that of first-line chemotherapy drug Doxorubicin hydrochloride(DOX).Thus,the I3C@cLANPs could address the low activation efficiency in the PTEN-mediated tumor strategy and avoid the risks of gene therapy,holding a good prospect for TNBC and related cancer treatment.

Rooting of Stem Cuttings with Different Indole 3 Butyric Acid (IBA) Treatments and Development of Micropropagation Protocol for <i>Piper betle</i>L. Node Culture

The present study, conducted during 2016 and 2017 seasons, aimed to investigate the effect of IBA on rooting of Piper betle L. stem cuttings (softwood and semi-hardwood). The experiment was undertaken in misting house field 2 UPM using the sand media to determine the adventitious roots initiation and development using the histological method. The cuttings were treated with different IBA concentrations (0, 500, 1000, 1500 and 2000 mg/L). The nodes explants were used in the development of a protocol for in vitro propagation of P. betle L., with different concentrations of Clorox with different times of immersion (20% Clorox 10 minutes, 30% Clorox 10 minutes, 20% Clorox 20 minutes, and 30% 20 minutes). In multiplication of the plantlets, Murashige and Skoog (MS) medium with different concentrations of BAP (0, 0.5, 1.0, 2.0 mg/L) were used to investigate the rooting of the explants. The results indicated that the types of the cuttings were different in the rooting capacity and the length of the roots. Moreover, it was found that in comparison with the control treatment, by a rise in the concentrations of the IBA, there was a significant upsurge in the rooting percentage, the root diameter, and the number of the roots. The results indicated that the types of cutting with 1000, 1500 and 2000 mg/L IBA perform better in the root percentage (100%) in the semi hardwood cuttings. The best results, however, were 2000 mg/L IBA in the semi hardwood cuttings, with the number of the roots to be 35.05, and the fresh weight of the roots to be 3.94 g, the dry weight of the roots to be 0.33 g, the length of the roots to be 391.88 cm, the roots diameter to be 1.21 mm, the surface area of the roots to be 121.83 cm2, and the root volume to be 2.99 cm3. Nonetheless, the optimal concentration of Clorox with the time immersion was 20% with the 20-minute immersion time, which produced a shoot induction percentage of 30% dead explants and a mean number of 70.00 shoots per explant and the optimal concentration of benzylaminopurine (BAP) at 1.0 mg/L. It is of note that a shoot induction percentage of 22.29% and a mean number of 4.1% number of auxiliary bud per treatment. P. betle shoots in MS medium without PGR MS (0.0) yielded a good rooting.

The auxin transporter OsAUX1 regulates tillering in rice(Oryza sativa)

Tillering is an important agronomic trait of rice(Oryza sativa)that affects the number of effective panicles,thereby affecting yields.The phytohormone auxin plays a key role in tillering.Here we identified the high tillering and semi-dwarf 1(htsd1)mutant with auxin-deficiency root characteristics,such as shortened lateral roots,reduced lateral root density,and enlarged root angles.htsd1 showed reduced sensitivity to auxin,but the external application of indole-3-acetic acid(IAA)inhibited its tillering.We identified the mutated gene in htsd1 as AUXIN1(OsAUX1,LOC_Os01g63770),which encodes an auxin influx transporter.The promoter sequence of OsAUX1 contains many SQUAMOSA PROMOTER BINDING PROTEIN-LIKE(SPL)binding sites,and we demonstrated that SPL7 binds to the OsAUX1 promoter.TEOSINTE BRANCHED1(OsTB1),a key gene that negatively regulates tillering,was significantly downregulated in htsd1.Tillering was enhanced in the OsTB1 knockout mutant,and the external application of IAA inhibited tiller elongation in this mutant.Overexpressing OsTB1 restored the multi-tiller phenotype of htsd1.These results suggest that SPL7 directly binds to the OsAUX1 promoter and regulates tillering in rice by altering OsTB1 expression to modulate auxin signaling.

Differences of Plasma Levels of Tryptophan, Serotonin, 5-Hydroxyindole Acetic Acid, and Kynurenine between Healthy People and Patients of Major Monopolar Depression at Various Age and Gender

Background: It is not well analyzed whether there are differences in plasma levels of tryptophan (TRP) metabolites between healthy control people (HC) and patients of major monopolar depression (MMD). Methods: Ultra high-speed liquid chromatography/mass spectrometry has been used for the simultaneous determination of plasma levels of tryptophan metabolites in depressive patients. Results: There are no significant differences between plasma levels of TRP between HC and MMD. Plasma levels of TRP of HC are higher in young men, young women, old men, and old women in this order. Serotonin (5-HT) levels are higher in MMD than HC. Plasma levels of 5-HIAA of HC are also higher than those of patients of MMD. Plasma levels of kynurenine (KYN) of healthy old men and old women are higher than those of young men and old women. Plasma levels of KYN are higher in old women and young men of MMD than those of HC. Conclusion: Plasma levels of 5-HT are higher in patients of MMD than those of HC, which may suggest that use of drugs inhibiting the 5-HT transportation may increase plasma levels of 5-HT in MMD.

AN AMPEROMETRIC BIOSENSOR BASED ON COVALENT IMMOBILIZATION OF ASCORBATE OXIDASE ON BIOCOMPATIABLE AND LOW-TOXIC POLY(THIOPHENE-3-ACETIC ACID) MATRIX

The biocompatiable and low-toxic poly（thiophene-3-aeetic acid） （PTAA） matrix was successfully electrosynthesized in ionic liquid 1-butyl-3-metbylimidazolium hexafluorophosphate （BmimPF6） in comparison with the electrosynthesis of PTAA matrix in acetonitrile （ACN）. Ascorbate oxidase （AO） was used as a model for the development and application of biosensor. Vitamin C （VC） biosensors were facilely fabricated by the covalent immobilization of AO molecules on PTAA matrices electrosynthesized in ACN containing tetrabutylammonium tetrafluoroborate and BmimPF6, respectively. Electrochemical impedance spectroscopy, scanning electron microscopy and FTIR spectroscopy indicated that AO molecules were covalently immobilized on PTAA matrices. Parameters of the as-obtained biosensors such as working potential, pH and temperature have been optimized. The amperometric biosensor based on PTAA matrix electrosynthesized in BmimPF6 exhibited wider linear range, lower detection limit, higher sensitivity and bioaffinity, and better operational and storage stability than that electrosynthesized in ACN under optimal conditions. The as-obtained biosensor based on PTAA matrix electrosynthesized in BmimPF6 was employed for the detection of VC content in comme：rcial juices, and the result was close to the data given by manufacturers. Excellent results indicate that the PTAA matrix electrosynthesized in ionic liquid is a promising platform for the covalent immobilization of biologically-active species and the development of biosensors.

ELECTROSYNTHESES OF FREE-STANDING POLY(THIOPHENE-3-ACETIC ACID) FILM IN MIXED ELECTROLYTES OF BORON TRIFLUORIDE DIETHYL ETHERATE AND TRIFLUOROACETIC ACID

High quality free-standing poly（thiophene-3-acetic acid） （PTAA）, a water-soluble polythiophene derivative, was uccessfully electrosynthesized in boron trifluoride diethyl etherate （BFEE） ＋ 25% （by volume） trifluoroacetic acid （TFA） at 9wer potential （0.38 V versus Pt）. The carboxyl group makes PTAA highly soluble in water, facilitating its potential pplication as a blue-light-emitting material. PTAA films with conductivity of 7 S cm-1 obtained from this medium showed ,etter redox activity and thermal stability. The structure and morphology of the polymer were studied by UV-Vis, FT-IR, H-NMR spectra and scanning electron microscopy, respectively.

Synthesis, Reactions and Antimicrobial Activity of Some New 3-Substituted Indole Derivatives

Reaction of indole-3-carboxaldehydes 4 with hydrazine derivatives and different substituted acid hydrazides afforded the corresponding hydrazine derivatives 5a-c and acid hydrazide derivatives 7-11 respectively. Condensation of indole-3-carboxaldehydes 4 with phenacyl bromide and thiourea gives 1,3-thiazol-2-amine derivative 18. On the other hand, reaction 4 with 3-acetylchromene-2-one afforded chalcone derivative 19. Compound 4 undergoing Knoevenagel condensation with cyanoacetamide, ethyl cyanoacetate, benzimidazol-2-ylacetonitrile, rhodanine-3-acetic acid, 2,3-dihydropyrimidin-4-one derivative and 2,4-dihydropyrazol-3-one afforded the compounds 20a,b, 22, 23, 27 and 28 respectively. The structure of the newly synthesized compounds has been confirmed by elemental analysis and spectra data. The antimicrobial activities of the some newly synthesized compounds were measured and showed that most of them have high

Effects of Zinc Oxide Particles with Different Sizes on Root Development in Oryza sativa

Given the consistent release of zinc oxide(ZnO)nanoparticles into the environment,it is urgent to study their impact on plants in depth.In this study,grains of rice were treated with two different concentrations of ZnO nanoparticles(NP-ZnO,10 and 100 mg/L),and their bulk counterpart(B-ZnO)were used to evaluate whether ZnO action could depend on particle size.To test this hypothesis,root growth and development assessment,oxidative stress parameters,indole-3-acetic acid(IAA)content and molecules/enzymes involved in IAA metabolism were analyzed.In situ localization of Zn in control and treated roots was also performed.Though Zn was visible inside root cells only following nanoparticle treatment,both materials(NP-ZnO and B-ZnO)were able to affect seedling growth and root morphology,with alteration in the concentration/pattern of localization of oxidative stress markers and with a different action depending on particle size.In addition,only ZnO supplied as bulk material induced a significant increase in both IAA concentration and lateral root density,supporting our hypothesis that bulk particles might enhance lateral root development through the rise of IAA concentration.Apparently,IAA concentration was influenced more by the activity of the catabolic peroxidases than by the protective action of phenols.

Transport and Metabolism of the Endogenous Auxin Precursor Indole-3-Butyric Acid

Plant growth and morphogenesis depend on the levels and distribution of the plant hormone auxin. Plants tightly regulate cellular levels of the active auxin indole-3-acetic acid （IAA） through synthesis, inactivation, and transport. Although the transporters that move IAA into and out of cells are well characterized and play important roles in devel- opment, little is known about the transport of IAA precursors. In this review, we discuss the accumulating evidence sug- gesting that the IAA precursor indole-3-butyric acid （IBA） is transported independently of the characterized IAA transport machinery along with the recent identification of specific IBA efflux carriers and enzymes suggested to metabolize IBA. These studies have revealed important roles for IBA in maintaining IAA levels and distribution within the plant to support normal development.