Efficient Synthesis of 8-Thiosubstituted Guanine Derivatives as Potential Tools for Biochemical and Biological Studies

Abstract

A method for the selective introduction of the N²-(dimethylamino)methylene group into 8-thio-9-(2-hydroxyethoxymethyl)guanine (1) has been developed. The effect of the N²-amidine protection on the S-alkylation of 1 was studied.     

A Convenient Method for the Synthesis of 2-(β-D-Glycopyranosylthio) Pyridines

Abstract

Treatment of piperidinium salts of dihydropyridinethiolates 3 with glycosyl bromides 4 in dry acetone provides a convenient and high yielding synthesis of 1,4-dihydro-3-cyanopyridine thioglycosides 5. The structures of 5 were confirmed by oxidation as well as by ¹H NMR and ¹³C NMR spectral analysis.     

Base-Labile Group Protected Biotinphosphoramidite Reagents for Solid Phase Biotinylation of Oligonucleotides

Abstract

A general and rapid method is described for the synthesis of N¹ -base labile group protected biotinphosphoramidite reagents, useful for the synthesis of biotinylated oligonucleotides in automated DNA synthesizer.     

An Improved Post-Synthetic Substitution Approach for Synthesis of Oligodeoxynucleotides Containing Labile 4-Substituted Thymines

Abstract

A simple procedure is described for the preparation of a versatile oligodeoxynucleotide which contains 4-phenylthiothymidine. This versatile oligomer has been successfully used for synthesis of oligonucleotides containing labile 5-methyl-N⁴, N⁴-ethanocytosine (7) or 4-azido-5-methyl-2-pyrimidinone-1-β-(2′-deoxyriboside) (8).     

Conformational Aspects of Differences in Requirements for Oxytocin and Vasopressin Receptors

Abstract

Conformational energy calculations were carried out on three non-peptide antagonists of oxytocin and vasopressin: penicilide (compound 1; selective for oxytocin receptors), 1- {1-[4-(3-acetylaminopropoxy (benzoyl]-4-piperidyl}-3,4-dihydro-2(1H)-quinoline (compound 2; selective for vasopressin V₁ receptors) and 5-dimethylamino-1-{(2-methylbenzylamino)-benzoyl}-2,3,4,5–tetrahydro-1H-benzapine (compound 3; selective for vasopressin V₂ receptors). The obtained low-energy conformations of compound 1 were compared with low-energy conformations of oxytocin (OT) and low-energy conformations of compounds 2 and 3 were compared with low-energy conformations of arginine vasopressin (AVP). It was found that the affinity of the non-peptide antagonists and their selectivity for vasopressin and oxytocin receptors is probably connected with mimicking the aromatic rings of the Tyr² and the Phe³ residues of AVP in the case of compounds 2 and 3 and with mimicking the Tyr² residue and the Ile³ or Leu⁸ residues of OT by the outer benzene ring and the isobutyl group of compound 1. Application of the results in the design of more potent non-peptide antagonists of OT and VP is also discussed.     

Redefining the dose of the entomopathogenic fungus Metarhizium brunneum (Ascomycota,Hypocreales) to increase Fe bioavailability and promote plant growth in calcareous and sandy soils

Background and aims

Plants differ in their ability to use different nitrogen (N) chemical forms, these differences can be related to their ecology and drive community structure. The capacity to uptake intact organic N has been observed in plants of several ecosystems. However, soil organic N uptake by Mediterranean plants is unknown despite organic N being abundant in Mediterranean ecosystems. We compare the uptake of different N forms in two widespread coexisting Mediterranean forest trees with contrasting ecophysiological characteristics: Quercus ilex and Pinus halepensis.

Methods

To estimate root uptake rate of each N form we used equimolar solutions (1 mM N) of ¹⁵NO₃ ^?, ¹⁵NH₄ ⁺ and ¹⁵N-¹³C glycine.

Results

NH₄ ⁺ and glycine were taken up at a similar rate, but faster than NO₃ ^? in both species. Intact dual labeled glycine was found in both species, demonstrating that both species can absorb intact organic N.

Conclusions

Despite their ecological differences, both species had similar preference for N forms suggesting no fundamental niche complementarity for N uptake. The higher preference for NH₄ ⁺ and glycine over NO₃ ^? possibly reflects adaptation to the differing proportions of N forms in Mediterranean soils.

     

Interactive effects of chromate and arsenate on their uptake and speciation in Pteris ensiformis

Background and aims

Arsenate (AsV) and chromate (CrVI) inhibit each other’s uptake and translocation in As-hyperaccumulator Pteris vittata. In the present study, we extended the research to As-sensitive plant Pteris ensiformis to better understand the mechanism of their interactions.

Methods

Plants were exposed to 0, 0.75 or 7.5 mg L^?1 AsV and 0, 0.52, or 5.2 mg L^?1 CrVI for 7 d in hydroponics. Arsenic and Cr speciation were determined in nutrient solutions and plant biomass.

Results

P. ensiformis accumulated high levels of As and Cr in the rhizomes and roots with low levels in the fronds. However, P. ensiformis was much more effective in taking up Cr than As, as much more Cr was accumulated in the roots (306–6015 vs. 87–642 mg kg^?1). AsV and CrVI increased each other’s uptake in the rhizomes and roots when co-present. The AsV and CrVI taken up by P. ensiformis were reduced to arsenite (AsIII) and chromite (CrIII), possibly serving as detoxification mechanism.

Conclusions

Uptake of As and Cr induced oxidative stress as indicated by increased lipid peroxidation and electrical conductivity. Arsenic and Cr increased each other’s uptake by P. ensiformis.

     

High-performance liquid chromatographic profile and 1H quantitative nuclear magnetic resonance analyses for quality control of a Xinjiang licorice extract

ABSTRACT

Various pharmacological properties of Xinjiang licorice flavonoids have been reported recently. We have investigated constituents corresponding to distinct peaks on the high-performance liquid chromatography (HPLC) profile of a flavonoid-rich extract from licorice, and identified 13 flavonoids, including licochalcone A (1), licochalcone B (3), glabrone (4), and echinatin (5), by isolating them and then performing high-resolution electrospray ionization mass spectrometry and ¹H nuclear magnetic resonance (NMR) spectral analyses. We then applied the ¹H quantitative NMR (qNMR) method for analysis of major flavonoids, 1 and 3–5 in the extract. The ¹H qNMR results were supported by ¹³C NMR analysis. The results demonstrated the utility of the combination of HPLC profiling and qNMR analyses for quality control of Xinjiang licorice. Additionally, we observed a moderate inhibitory effect of the most abundant constituent, licochalcone A (1), on acetylcholine esterase activity, suggesting utility as a seed for drug development.     

Hints on Research for Bird-watchers

Capsule Fledgling Golden Eagles in northern Sweden preferred clearcuts and other open forest habitats, as well as steep slopes.

Aims To study the post-fledging habitat use and ranging behaviour of juvenile Golden Eagles on their natal territories.

Methods Fourteen juvenile Golden Eagles in northern Sweden were marked with GPS transmitters and tracked until they left their natal territory.

Results Eagles fledged at the end of July–beginning of August and remained on their natal territories until October–early November. Fledged eagles' home range size before flying south was on average 41?km². Juvenile eagles showed a preference for clearcuts, coniferous forest on lichen-covered bedrock and edges between clearcuts and forest, whilst all other habitat types were used less than expected. The eagles showed a preference for steep slopes, in particular south-facing ones, whilst north-facing slopes were used less than expected.

Conclusion Golden Eagles' preference for clearcuts and steep slopes can be used in the planning and management of ‘eagle friendly’ wind farms.     

O6-(4-Nitrophenyl)inosine and -Guanosine as Chromogenic Substrates for Adenosine Deaminase

Abstract

O⁶-(4-Nitrophenyl)inosine (la), O⁶ -(4-nitrophenyl)guanosine (1c) and O⁶ -(4-methylumbelliferonyl)inosine (2) were obtained by reaction of 6-chloro-9-(β-D-ribofuranosyl)purine (3a) or 2-amino-6-chloro-9-(β-D-ribofuranosyl)purine (3c) with sodium salts of 4-nitrophenol or 4-methylumbelliferone in N,N-dimethylformamide. Similarly, 6-chloro-9-(β-D-2,3-isopropylideneribofuranosyl)purine (3b) was transformed to 2′,3′-O-isopropylidene-O⁶-(4-nitrophenyl)inosine (1b). Deprotection of 1b with CF₃COOH gave compound la and O⁶ -(4-nitrophenyl)hypoxanthine (4). Compounds 1a and 1c are substrates for adenosine deaminase releasing 4-nitrophenol which is readily detected visually or spectrophotomemcally. Rate and extent of hydrolysis of la are significantly increased in the presence of purine nucleoside phosphorylase but xanthine oxidase has no influence. A potential fluorogenic analogue 2 is not a substrate for adenosine deaminase.

     

Homo-C-Nucleoside Analogs† II. Synthesis and Anomeric Configuration of 4-(2,5-Anhydro-D-Gluco-PEntitol-1-YL)-2-Phenyl2H-1,2,3-Triazole††

Abstract

Treatment of 4-(D-gluco-pentitol-l-y1)-2-pheny1–2H-1,2,3-triazole (1) with p-toluenesulfonyl chloride in pyridine solution, afforded the homo-C-nucleoside analog, 4-(2,5-anhydro-D-gluco-pentitol-1-yl)-2-phenyl-2H-1,2,3-triazole (2) as well as its partial p-toluenesulfonyl derivative (3). 4-(5-Chloro-5-deoxy-D-gluco-pentitol-1-yl)-2-phenyl-2H-1,2,3-triazole (8), was isolated as a byproduct from the reaction. The structure and anomeric configuration of 2 was determined by acylation, ¹H, ¹³C NMR, and NOE, spectroscopy as well as mass spectrometry.

     

Enantiospecific Synthesis of Carbocyclic Aminoimidazole Carboxamide Ribonucleotide (C-AICAR), Succinoaminoimidazole Carboxamide Ribonucleotide (C-SAICAR), and a New Intermediate for SAICAR Analogs

Abstract

The (-)-enantiomer of the carbocyclic analogs of aminoimidazole carboxamide ribonucleotide (C-AICAR¹, 7), and succinoaminoimidazole carboxamide ribonucleotide (C-SAICAR, 14) have been prepared. En route, a new intermediate (19) for the preparation of SAICAR analogs was developed.     

Novel 2.6-Disubstituted Adenosine Analogues: Potential Agonists for Adenosine Receptors

Abstract

Synthesis of novel 2-lodo- and 2-oxo- N⁶?cyclosubstituted adenoslne analogues with potential adenoslne A₁ or A₂ agonist activity is described. The procedure for the 2-oxo compounds represents a new and general synthetic approach to isoguanoslne analogues.     

Aflatoxin B1 and DNA Adducts. Proposed Model for Surface Noncovalent and Covalent Complexes with N(7) of Guanine. II.

Abstract

An alternate model for surface noncovalent and surface covalent binding of aflatoxin B₁ to N(7) of guanine in DNA is proposed. This model considers the out-of-plane motions of C(8) of aflatoxin B₁ in those interactions.

The covalent intercalated fit of aflatoxin B₁ into DNA arises from steric adjustments made by DNA at the covalent intercalation site as well as local strain in the bond angles about N(7) of guanine and C(8) of aflatoxin B₁. The bond angle about N(7) deviates modestly from the sp² value toward the sp³ value.

This study suggests that the surface covalent aflatoxin B₁ -DNA complex serves only a minor role in aflatoxin's precarcinogenic interaction with DNA and is a likely correctable error.     

In vitro and ex vivo biofilms of dermatophytes: a new panorama for the study of antifungal drugs

Abstract

This study describes an ex vivo model that creates an environment for dermatophyte biofilm growth, with features that resemble those of in vivo conditions, designing a new panorama for the study of antifungal susceptibility. Regarding planktonic susceptibility, MIC ranges were 0.125-1?µg ml^?1 for griseofulvin and 0.000097-0.25?µg ml^?1 for itraconazole and terbinafine. sMIC50 ranges were 2->512?µg ml^?1 for griseofulvin and 0.25->64?µg ml^?1 for itraconazole and terbinafine. CLSM images demonstrated a reduction in the amount of cells within the biofilm, but hyphae and conidia were still observed and biofilm biomass was maintained. SEM analysis demonstrated a retraction in the biofilm matrix, but fungal structures and water channels were preserved. These results show that ex vivo biofilms are more tolerant to antifungal drugs than in vitro biofilms, suggesting that environmental and nutritional conditions created by this ex vivo model favor biofilm growth and robustness, and hence drug tolerance.     

OPTIMAS-DW: A comprehensive transcriptomics,metabolomics, ionomics,proteomics and phenomics data resource for maize

Background

The wild barley Hordeum chilense fulfills some requirements for being a useful tool to investigate the endosperm yellow pigment content (YPC) in the Triticeae including its diploid constitution, the availability of genetic resources (addition and deletion stocks and a high density genetic map) and, especially, its high seed YPC not silenced in tritordeums (amphiploids derived from H. chilense and wheat). Thus, the aim of this work was to test the utility of the H. chilense genome for investigating the YPC in the Triticeae.

Results

Twelve genes related to endosperm carotenoid content and/or YPC in grasses (Dxr, Hdr [synonym ispH], Ggpps1, Psy2, Psy3, Pds, Zds, e-Lcy, b-Lcy, Hyd3, Ccd1 and Ppo1) were identified, and mapped in H. chilense using rice genes to identify orthologs from barley, wheat, sorghum and maize. Macrocolinearity studies revealed that gene positions were in agreement in H. vulgare and H. chilense. Additionally, three main regions associated with YPC were identified in chromosomes 2H^ch, 3H^ch and 7H^ch in H. chilense, the former being the most significant one.

Conclusions

The results obtained are consistent with previous findings in wheat and suggest that Ggpps1, Zds and Hyd3 on chromosome 2H^ch may be considered candidate genes in wheat for further studies in YPC improvement. Considering the syntenic location of carotenoid genes in H. chilense, we have concluded that the H^ch genome may constitute a valuable tool for YPC studies in the Triticeae.

     

Impact of the TaMATE1B gene on above and below-ground growth of durum wheat grown on an acid and Al3+-toxic soil

Background and aims

Subsoil acidity with a high aluminium (Al³⁺) soil content inhibits root growth and proliferation of durum wheat (tetraploid AABB, Triticum turgidum) leading to poor nutrient and water uptake. This study evaluated the impact of Al³⁺-tolerantTaMATE1B allele on root and shoot traits of durum wheat grown in an acidic soil with a high Al³⁺concentration.

Methods

Two durum wheat lines, Jandaroi–TaMATE1B with the TaMATE1B gene introgressed from Al³⁺-tolerant bread wheat and Jandaroi–null (a sister line lacking the Al³⁺-tolerant TaMATE1B allele), were grown in rhizoboxes in a glasshouse. We mapped root growth and proliferation over time and measured shoot traits and grain yield.

Results

Introgression of the Al³⁺-tolerant TaMATE1B allele into durum wheat enabled root growth and proliferation below 0.25 m of the soil profile, where the soil pH was low (4.1, CaCl₂ extract) with high Al³⁺ content (16.5 mg kg⁻¹), and increased total root length and biomass at 42 days after sowing (DAS; Z₃₃) by 38.3 and 22%, respectively, relative to the Jandaroi–null. Differences in root growth between the two lines were apparent from tillering stage (Z₃₃) and by 50% anthesis (Z₆₄), respectively. Jandaroi–TaMATE1B had 69.2% greater root biomass, 76.2% greater root length, 5.89% greater leaf area and 18% greater shoot biomass than Jandaroi–null at 50% anthesis (Z₆₄). Time to anthesis and physiological maturity was delayed 6–7 days in Jandaroi–TaMATE1B, compared to Jandaroi–null. Jandaroi–TaMATE1B tended to have relatively greater, but not significantly different, shoot biomass, grain yield and yield components than Jandaroi–null.

Conclusions

Introgression of the Al³⁺-tolerant TaMATE1B allele into durum wheat enabled root growth and proliferation down an acidic soil profile with a high Al³⁺ concentration. We assume that in the field where plants need to acquire water at depth differences in above-ground parameters would be amplified.

     

Strong,Specific, Reversible Binding Ligands for Transfer Rna: Comparison By Fluorescence and Nmr Spectroscopies with Distamycin Binding for a New Structural Class of Ligand

Abstract

Binding data are presented for the interaction with brewer's yeast tRNA^Phc of a new structural family of ligands, symmetrical bis-benzimidazoles. In addition specific perturbations in chemical shifts were detected by 1-dimensional NMR spectroscopy at 400 MHz for some imino and aromatic methyl protons of tRNA^Phc when the tRNA was titrated with distamycin. Competitive displacement of the benzimidazole by added distamycin was followed fluorescence spectroscopy.     

Surface charge engineering of Thermomyces lanuginosus lipase improves enzymatic activity and biodiesel synthesis

Objectives

This study was aimed at engineering charged residues on the surface of Thermomyces lanuginosus lipase (TLL) to obtain TLL variant with elevated performance for industrial applications.

Results

Site-directed mutagenesis of eight charged amino acids on the TLL surface were conducted and substitutions on the negatively charged residues D111, D158, D165, and E239 were identified with elevated specific activities and biodiesel yields. Synergistic effect was not discovered in the double mutants, D111E/D165E and D165E/E239R, when compared with the corresponding single mutants. One TLL mutant, D165E, was identified with increased specific activity (456.60 U/mg), catalytic efficiency (k_cat/K_m: 44.14 s^?1 mM^?1), the highest biodiesel conversion yield (93.56%), and comparable thermostability with that of the TLL.

Conclusions

Our study highlighted the importance of surface charge engineering in improving TLL activity and biodiesel production, and the resulting TLL mutant, D165E, is a promising candidate for biodiesel industry.

     

Plant maintenance and environmental stress. Summarising the effects of contrasting elevation,soil, and latitude on Quercus ilex respiration rates

Aims

Rice (Oryza sativa L.), wheat (Triticum aestivum L.) and common bean (Phaseolus vulgaris L.) are major staple food crops consumed worldwide. Zinc (Zn) deficiency represents a common micronutrient deficiency in human populations, especially in regions of the world where staple food crops are the main source of daily calorie intake. Foliar application of Zn fertilizer has been shown to be effective for enriching food crop grains with Zn to desirable amounts for human nutrition. For promoting adoption of this practice by growers, it is important to know whether foliar Zn fertilizers can be applied along with pesticides to wheat, rice and also common bean grown across different soil and environmental conditions.

Methods

The feasibility of foliar application of zinc sulphate (ZnSO₄.7H₂O) to wheat, rice and common bean in combination with commonly used five fungicides and nine insecticides was investigated under field conditions at the 31 sites-years of seven countries, i.e., China, India, Pakistan, Thailand, Turkey, Brazil and Zambia.

Results

Significant increases in grain yields were observed with foliar Zn/foliar Zn?+?pesticide (5.2–7.7 % of wheat and 1.6–4.2 % of rice) over yields with no Zn treatment. In wheat, as average of all experiments, higher grain Zn concentrations were recorded with foliar Zn alone (41.2 mg kg^?1) and foliar Zn?+?pesticide (38.4 mg kg^?1) as compared to no Zn treatment (28.0 mg kg^?1). Though the magnitude of grain Zn enrichment was lesser in rice than wheat, grain Zn concentrations in brown rice were significantly higher with foliar Zn (24.1 mg kg^?1) and foliar Zn?+?pesticide (23.6 mg kg^?1) than with no Zn (19.1 mg kg^?1). In case of common bean, grain Zn concentration increased from 68 to 78 mg kg^?1 with foliar Zn alone and to 77 mg kg^?1 with foliar Zn applied in combination with pesticides. Thus, grain Zn enrichment with foliar Zn, without or with pesticides, was almost similar in all the tested crops.

Conclusions

The results obtained at the 31 experimental site-years of seven countries revealed that foliar Zn fertilization can be realized in combination with commonly-applied pesticides to contribute Zn biofortification of grains in wheat, rice and common bean. This agronomic approach represents a useful practice for the farmers to alleviate Zn deficiency problem in human populations.