Acid Adenine Research Articles

Qualitative and Quantitative Evaluation of Active Constituents in Callus of Lavandula angustifolia plant in Vitro

This study was conducted to describe a protocol for the callus establishing culture of Lavandula angustifolia plant and estimating their content of volatile oil. The quantity of volatile oil callus tissues was compared with that of leaves production. Callus was induced from leaf explants on Murashige and Skoog medium (MS) supplemented with Naphthalene acetic acid (NAA) and Benzyl adenine (BA) in different concentrations. Maximum callus fresh weight was obtained in the combination of 10 mg/L BA and 3 mg/L NAA which reached 18 g after four weeks. The results of this work showed that the quantity of volatile oil from the highest fresh weight callus was 6 ml compared with quantity of 18g of leaves which gave 0.5 ml. Volatile oil of leaf and callus extracts were analyzed using gas chromatography mass spectrometry method (GC-MS) which showed linoleic acid (56.61%) and oleic acid (57.93%) as main components.

In Vitro Propagation of Artemisia japonica

ABSTRACTAn effective protocol for in vitro propagation was developed for Artemisia japonica using nodal explants. The efficiency of the explants to induce the multiple shoots on Murashige and Skoog (MS) medium supplemented with 2.5, 5.0, 7.5, and 10 µM of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-napthalene acetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), 6-benzylaminopurine (BAP), thidiazuron (TDZ), kinetin (Kn), and 2-isopentenyl adenine (2iP) was tested. A maximum of 20 axillary shoots per explant were induced on MS medium supplemented with 10 µM BAP. Shoots were rooted on MS medium supplemented with IBA (10 µM). The resulting plantlets were hardened in a mixture of cocopeat and perlite (1:1). Cytological analysis revealed that regenerated plants had 2n = 18 chromosomes, which was same as that of the mother plant.

CBIO-02KYNURENINE 3-MONOOXYGENASE INHIBITION SYNERGIZES WITH THE ALKYLATING CHEMOTHERAPY TEMOZOLOMIDE TO ATTENUATE PROLIFERATION IN HUMAN GLIOBLASTOMA CELLS

BACKGROUND: Activation of the kynurenine pathway (KP) in cancer cells suppresses anti-tumor immunity, resulting partly from KP metabolite effects. The KP is the principal route of tryptophan metabolism leading to the production of metabolites, including kynurenine (KYN), anthralinic acid (AA) and nicotinamide adenine dinucleotide (NAD+). Kynurenine 3-monooxygenase (KMO) is a critical enzyme in the KP directly responsible for the production of the immunosuppressive metabolites, 3-hydroxyanthranilic acid (3-HAA) and 3-hydroxykynurenine (3-HK). Inhibition of one of the enzymes that degrades tryptophan in the KP, indoleamine 2,3-dioxygenase (IDO-1), suppresses cancer formation in animal models. However, no studies have evaluated the therapeutic potential of KMO inhibitors in brain cancer. METHODS: The effects of the KMO inhibitor, Ro 61-8048 (Ro), and the chemotherapeutic, temozolomide (TMZ), on glioblastoma (GBM) cell proliferation was evaluated using the MTS assay and GBM cell lines, U87, T98G and U138. KP metabolites were quantified using LC-MS/MS and KMO protein expression was evaluated using western blot. RESULTS: GBM cell lines expressed the KMO protein constitutively and following stimulation with the IDO-1 inducer, IFN-γ. GBM cell lines also produced the KP immunosuppressive metabolites, AA and 3-HK and increased KYN production concomitant with tryptophan catabolism following IFN-γ stimulation. KMO inhibition with Ro caused a dose-dependent decrease in GBM cell line proliferation. Ro also synergized with TMZ to decrease U87 cell proliferation more effectively than Ro treatment alone when cells were stimulated with IFN-γ. CONCLUSIONS: These findings suggest that KMO may play a significant role in GBM cell proliferation and highlight KMO inhibition as a potential new therapy for GBM. Clinically, KMO inhibition may shunt KP metabolism away from the immunosuppressive 3-HK and 3-HAA, restoring anti-tumor immunity while reducing the capacity for malignant cells to produce NAD + , which is necessary for cellular energy production, DNA repair and proliferation.

Improved spectrophotometric human interferon-gamma bioassay

Interferon gamma (IFNγ) is a cytokine involved in many anti-viral and immunoregulatory processes. One of the major mechanisms through which IFNγ exerts these effects is by inducing expression of indoleamine 2,3 dioxygenase-1 (IDO1), an enzyme that catalyses the first, rate-limiting step of the kynurenine pathway. In this pathway, tryptophan can be catabolised to many products, including picolinic acid and nicotinamide adenine dinucleotide. However, in endothelial cells, the pathway ends at the production of kynurenine. This is due to little or no expression of enzymes that metabolise kynurenine. Production of kynurenine has been used as an indicator of human IDO1 activity, and hence as an hIDO1 bioassay. Due to IFNγ's ability to induce IDO1 expression, kynurenine production can also be a measure of human IFNγ (hIFNγ) bioactivity. Previously, the levels of hIFNγ have been commonly determined by anti-viral assays, high performance liquid chromatography and ELISA. Apart from their technical complexity, these assays are costly and only the anti-viral assay measures bioactive IFNγ. Here, we report the development of an improved IFNγ spectrophotometric bioassay using a human brain endothelial cell line (HBEC 5i). The method is sensitive, easy to perform and cost efficient.

Effect of the Developmental Stage of Microspores, Growth Regulator and Medium type on Callus indication from Broad bean Vicia faba Anthers Culture

anthers were taken from bean flower buds of the variety Aquadlce and microspore developmental stages were determined according to flower bud sizes. Anthers were cultured on modified Schenk and Hildebrandt medium (SH) supplemented with EDTA ferric monosodium salt to the SH medium which was taken from formulation of Murashige and Skoog (MS), casein hydrolysate 400 mg/l, some amino acids, and different combinations of growth regulators; kinetin and naphthalene acetic acid and 2, 4- dichlorophenoxyacetic acid (Kin + NAA + 2, 4-D) and benzyl adenine (BA + NAA + 2,4-D) in different concentrations. The combination (BA + NAA + 2, 4-D) used with flower bud sizes (7.1-8) mm, containing uninucleate microspores, gave better callus induction (50%). It was also found that the concentrations [1.0, 2.0] mg/l of ascorbic acid were better to prevent the accumulation of phenols in the medium than the concentration [0.0] mg/l. The cytological analysis revealed that the number of chromosomes in callus induced from flower bud sizes (2.5-6.4) mm contained diploid (2n=12 chromosomes), whereas chromosomes in callus induced from flower bud sizes (6.5-8.0) mm contained haploid (n=6 chromosomes) with the presence of some diploid cells (12 chromosomes).

Production of cycloartane triterpene glycosides in shoot cultures of Astragalus sieberi DC.

Shoot tips of the germinated seeds of Astragalus sieberi DC. were cultured on MS-medium supplemented with 0.1 mg L−1 each of naphthalene acetic acid (NAA) and benzyl adenine (BA) for establishment of shoot cultures. The effect of different concentrations of growth regulators on saponin content was studied and optimised. Saponin content was monitored by HPLC analysis. The most appropriate growth regulator with which to produce the highest content of saponins in shoot cultures was 0.5 mg L−1 each of NAA and BA. The isolated cycloastragenol-3-O-glucoside was identified by spectral analysis and compared with an authentic sample.

Some Novel Adenosine Mimics: Synthesis and Anticancer Potential against Cervical Cancer caused by Human Papilloma Virus

Two novel adenosine analogs, viz. 9-(1′-β-D-arabinofuranosyl)-6-nitro-1,3-dideazapurine or Ara-NDDP (1) and 9-(5′-deoxy-5′-S-(propionic acid) (1′-β-D-ribofuranosyl) adenine or SAH analog (2), indigenously synthesized, have been found to be potential anticancer agents against cervical cancer caused by human papilloma virus.

Plant growth regulators cannot be used to alter significantly the commercial harvest date of Boronia heterophylla F. Muell. (Rutaceae)

Harvest of the cut fiower crop, Boronia heterophylla F. Muell., is restricted to a 2-week period in early spring. Extension of the fiowering period by plant growth regulators was examined in plants growing in the field under standard, commercial production conditions. Flowering time, and hence harvest date, were delayed by 6{9 days following the application of gibberellic acid and benzyl adenine (GA4+7+BA). High concentrations of GA4+7+BA (40, 80 mg/L) delayed flowering longer than 20 mg/L, but reduced flower number. BA alone (4 applications of 20, 50, or 100 mg/L) reduced flower number and did not aflect harvest date. GA3 (4 applications of 20 mg/L) prevented flowering. Paclobutrazol (100 mg/L) reduced flower number and delayed flowering by 4 days. The timing of applications at the commencement of flower-promoting conditions, when plants had been exposed to 10-20 nights of temperatures <10˚C (mid autumn), appeared to be critical. A delay of harvest of 6 days is a 40% extension of the harvest period but this may not be sufficient for commercial production.

Carbaryl as a Component in Integrated Crop Management of Apple

The economic viability of most northeastern apple orchards depends upon the production of a high proportion of large, well-colored fruit of high quality. Adjustment of fruit load, or thinning, is an integral part of achieving this production goal. Although the effects of feeding damage caused by leafhoppers is controversial, many orchardists believe that this damage, particularly early in the season, is detrimental to production, and they apply insecticide treatments to maintain infestations below a threshold level. Carbaryl has been long recognized as a fruit-thinning agent, as well as a broad-spectrum insecticide. Field experiments were conducted during 1995 and 1996 to examine the horticultural benefits of carbaryl when used in combination with the plant growth regulators naphthaleneacetic acid and 6-benzyl adenine, and to assess the efficacy of carbaryl against an indigenous leafhopper complex when applied during a typical thinning window. In a majority of cases, the addition of carbaryl to naphthaleneacetic acid or 6-benzyl adenine significantly enhanced fruit thinning and increased fruit size. Two thinning rates of carbaryl with or without naphthaleneacetic acid significantly reduced early-season infestations of leafhopper nymphs. The highest rate suppressed nymphs and adults for the entire season and allowed lower leaf damage ratings. Results show that carbaryl in combination with other chemical thinners is beneficial in the management of crop load, and at the same time effects suppression of the leafhopper complex. In orchards that do not rely upon natural control of phytophagous mites, this unique horticultural chemical fits well into integrated crop management programs on apple.

Induction of callus from axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) and subsequent plantlet regeneration

Axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) cultured on half strength Murashige-Skoog medium (HMS) containing taro extract (HMSTE) and 2, 4, 5-trichlorophenoxyacetic acid produce a compact, hard, slow growing callus which is not very active morphogenetically and produces only a few plantlets. When cultured on HMSTE plus 5 mg 1(-1) each of naphthaleneacetic acid and benzyl adenine (HMSNB) the buds produce a fast growing, friable and morphogenetically active callus. Meristematic regions form on the friable callus after 30 days on HMSNB. If transferred to HMSTE at this point the callus gives rise to plantlets. Addition of taro extract to the media is required for the culture of buds, induction of callus and plantlet regeneration.

Biochemical and functional features of blood plates in myeloproliferative diseases

The aim of this work was to study the relationship between changes in blood platelet sialic acid levels and adenine nucleotide pool regulation with the parameters of platelet hemostasis (adhesive-aggregation and retractile capacity of blood plates) in chronic myeloleukemia and idiopathic myelofibrosis. The studies were performed on platelets of healthy subjects (20), patients with chronic myeloleukemia (14) and idiopathic myelofibrosis (11) with advanced clinical manifestations.

High-performance liquid chromatographic determination of water-soluble vitamins in intravenous fluids. I. Ascorbic acid (AsA) and flavin adenine dinucleotide (FAD)

High-performance liquid chromatographic determination of water-soluble vitamins in intravenous fluids. I. Ascorbic acid (AsA) and flavin adenine dinucleotide (FAD)

Synthesis and polymerization of nucleic acid base substituted L-lysine

Poly-l-lysines containing nucleic acid bases, that is, adenine, thymine, uracil and theophylline were synthesized. The nucleic acid base substituted l-lysine derivatives were prepared from α,N-Cbz-l-lysine and the p-nitrophenyl esters of the carboxyethyl derivatives of the nucleic acid bases. The polymers were obtained from these l-lysine derivatives by the NCA method.

Enhancement by Low Temperatures of the Anaerobic Induction of Cocklebur Seed Germination

Non-dormant, upper cocklebur (Xanthium pensylvanicum Wallr.) seeds, incapable of germinating under ordinary conditions, can germinate when previously subjected to anaerobiosis; this has been termed the anaerobic induction of seed germination. Aerobic presoaking of the seeds was also required for successful anaerobic induction, and exerted two counter-acting effects on seed germination. When the time period of aerobic presoaking was sufficiently prolonged, the increasing duration of an anaerobic treatment resulted in proportional increase of germination potential but, when it was short, the effect of the anaerobiosis was saturated in a few days. Prolonging the aerobic presoaking period caused less response of the seed to the anaerobic induction, suggesting the development of some germination-inhibiting system during the aerobic presoaking period. This system could not develop in the absence of O2 or at low temperature. Thus, low temperature during prolonged presoaking produced a maximal response to anaerobic induction. Various germination stimulants, CO2, ethylene, gibberellic acid and benzyl adenine, did not significantly alter the effects of the presoaking.

Relations between fatty acid activation and adenine nucleotide translocase systems in mitochondria

Experiments with freshly isolated rat liver mitochondria and mitochondria aged by two different methods showed that the activity of the mitochondrial fatty acid activation system can lead to inhibition of adenine nucleotide (AN) transport through the inner mitochondrial membrane. Inhibition of AN translocase was abolished by preincubating the mitochondria with carnitine. A special feature distinguishing inhibition of AN translocase by aged mitochondria is that these mitochondrial membranes contain fatty acids in sufficient concentration to cause inhibition on the addition of CoA and ATP. The results suggest a role for acyl-CoA formed by mitochondrial acyl-CoA synthetase in the regulation of AN transport in the mitochondrion.

Interaction of amino acids, N-acetyl amino acid esters, thymine and adenine with sephadex LH-20 gel

N-Acetyl aromatic amino acid esters, tryptophan, adenine and thymine show strong retention in Sephadex LH-20 gel in an aqueous phase. The decreased retention in 6 M urea, the absence of retention in absolute methanol and the increased retention at higher temperatures in the aqueous phase indicate that hydrophobic interaction is responsible for the observed retention of the amino acids and their esters in the gel. Adenine was found to be retained by polar interaction in the gel. The increased retention of the solutes in the presence of different electrolytes suggests that the lyotropic effect is more important than the ionic strength effect. The relevance of the results obtained with amino acids and esters to the conformational aspects of proteins in aqueous solution is discussed.