Plant P5C reductase as a new target for aminomethylenebisphosphonates

A series of N-substituted derivatives of aminomethylenebisphosphonic acid were evaluated as potential inhibitors of delta1-pyrroline-5-carboxylate reductase (EC 1.5.1.2), the enzyme that catalyzes the last step in proline biosynthesis, partially purified from Arabidopsis thaliana suspension cultured cells. At millimolar concentrations, three compounds out of 26 were found to interfere with the catalytic mechanism. One of them, namely, 3,5-dichloropyridyl-aminomethylenebisphosphonic acid, retained such inhibitory activity in the micromolar range. Kinetic analyses ruled out the possibility that the inhibition could simply rely upon the chelating properties of bisphosphonates and showed mechanisms of a noncompetitive type against NADH and an uncompetitive type against delta1-pyrroline-5-carboxylic acid, with KI values of 199 +/- 6 and 10.3 +/- 1.5 microM, respectively. A computer-aided docking analysis, performed on the basis of the crystal structure of the enzyme from Streptococcus pyogenes, suggested that this phosphonate may interact with amino acid residues near the binding site of delta1-pyrroline-5-carboxylic acid, thus blocking the substrate in a pocket and preventing its interaction with NADH. Because in higher plants the step catalyzed by delta1-pyrroline-5-carboxylate reductase is shared by all pathways leading to proline synthesis, such a compound may represent a lead structure to be exploited for the design of new substances endowed with herbicidal activity.     

Antioxidant activity and phenolic composition of Lavandin (Lavandula x intermedia Emeric ex Loiseleur) waste

The phenolic content of lavandin waste obtained after the distillation of essential oils for the perfume industry was investigated to find an alternative use for this material. The antioxidant activity of different fractions as well as their total phenolic content were evaluated by different methods. Twenty-three phenolic compounds were identified by liquid chromatography coupled to ionspray mass spectrometry (LC/MS/MS), including phenolic acids, hydroxycinnamoylquinic acid derivatives, glucosides of hydroxycinnamic acids, and flavonoids, none of which have previously been reported in lavandin waste. Some structure-activity relationships were proposed by relating the type of scavenging activity of different fractions with the identified phenolic compounds. Contents of representative phenolic acids of Lamiaceae (chlorogenic and rosmarinic) were evaluated by high performance liquid chromatography-diode array detection (HPLC-DAD) and compared with those of other plant species.     

Chemical characterization of red wine grape (Vitis vinifera and Vitis interspecific hybrids) and pomace phenolic extracts and their biological activity against Streptococcus mutans

Grapes are rich sources of potentially bioactive polyphenols. However, the phenolic content is variable depending on grape variety, and may be modified during vinification. In this study, we examined the chemical composition and biological activity of phenolic extracts prepared from several red wine grape varieties and their fermented byproduct of winemaking (pomace) on some of the virulence properties of Streptococcus mutans a well-known dental pathogen. Grape phenolic extracts were obtained from Vitis vinifera varieties Cabernet Franc and Pinot Noir and Vitis interspecific hybrid varieties Baco Noir and Noiret. The anthocyanins and flavan-3-ols content were highly variable depending on grape variety and type of extract (whole fruit vs fermented pomace). Nevertheless, all grape phenolic extracts remarkably inhibited glucosyltransferases B and C (70-85% inhibition) at concentrations as low as 62.5 microg/mL (P < 0.01). Furthermore, the glycolytic pH-drop by S. mutans cells was inhibited by the grape extracts without affecting the bacterial viability; an effect that can be attributed to partial inhibition of F-ATPase activity (30-65% inhibition at 125 microg/mL; P < 0.01). The biological activity of fermented pomace was either as effective as or significantly better than whole fruit grape extracts. The results showed that grape phenolic extracts, especially from pomace, are highly effective against specific virulence traits of S. mutans despite major differences in their phenolic content.     

Survival,growth and photochemical efficiency of silver fir seedlings produced with different technologies

New Forests - Forest tree seedling production technologies impact reforestation success determined with survival and quality of seedlings. Five Abies alba seedling production technologies were...     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.     

Evaluation of single and comparative intradermal tuberculin tests for tuberculosis eradication in caprine flocks in Castilla y León (Spain)

Goats can act as reservoirs for tuberculosis (TB) infection. The main etiological agents of TB in goats are Mycobacterium caprae and Mycobacterium bovis and they infect also a wide range of domestic and wild animals and humans. Control programmes based mainly on the application of single and comparative intradermal tuberculin (SIT and SCIT respectively) tests are being implemented in certain regions of Spain with a high density of caprine flocks as Castilla y León, including goats with epidemiological relationship with cattle. The aim of this study was to evaluate the performance of the intradermal tests in naturally TB-infected caprine flocks from this region. The study was performed using data from 17,450 goats in 54 different flocks that were classified as TB-infected in the control programmes executed in 2010 and 2011. Data from 1237 goats from 7 dairy flocks depopulated after the first intradermal testing were used to estimate the sensitivity (Se) using bacteriology as the gold-standard. Overall Se of the SIT test using the severe interpretation was 43.9% (CI 95%, 40.4–47.4) and decreased to 38.8% (CI 95%, 35.5–42.3) using the standard interpretation. Overall Se of the SCIT test ranged between 21.3% (CI 95%, 17.6–25.4) and 7% (CI 95%, 4.9–9.8) depending of the interpretation criteria. A significant weak positive correlation was found between age and skin fold thickness (Spearman’s test p < 0.05). Results from this study yielded, in general, low Se values probably due the systematic detection and slaughter of reactors as a consequence of the eradication programme in previous years or the presence of factors that may interfere in the diagnosis. Therefore, these results suggest the necessity of including ancillary diagnostic tools and/or strict interpretation criteria to maximize detection of positive animals in infected settings.     

Influence of oral co-administration of a preparation containing calcium and magnesium and food on enrofloxacin pharmacokinetics

The objective of this study has been to determine the influence of food and ions on the pharmacokinetics of enrofloxacin (ENRO) in turkeys, administered per os at a dose of 10 mg/kg of body weight (b.w.). Co-administration of ENRO with ions or with food significantly retarded its absorption, and the interaction was more pronounced when the drug was given together with food. The bioavailability of ENRO was 65.78 ± 7.81% and 47.99 ± 9.48% with ions and food, respectively. The maximum concentration (C_max) in plasma of animals exposed to ions reached 0.87 ± 0.26 μg/ml in a t_max of 2.07 ± 0.76 h; in animals which were fed while medicated, the analogous parameters were 0.36 ± 0.13 μg/ml and 8.06 ± 3.08 h. The PK/PD analysis demonstrated that a decrease in the concentration of ENRO in turkeys’ blood due to the interaction with ions or food might impair the drug's clinical efficacy toward some pathogenic microorganisms in turkeys if a routine dose of 10 mg ENRO/kg b.w. is administered.     

Ectomycorrhizal communities associated with silver fir seedlings (Abies alba Mill.) differ largely in mature silver fir stands and in Scots pine forecrops

Context

The requirement for rebuilding forecrop stands besides replacement of meadow vegetation with forest plants and formation of soil humus is the presence of a compatible ectomycorrhizal (ECM) fungal community.

Aims

This study aims to assess ectomycorrhizal fungi diversity associated with silver fir (Abies alba Mill.) seedlings regenerating in silver fir stands and Scots pine forecrops.

Methods

One-year-old seedlings were sampled in six study sites: three mature fir forests and three pine forests. ECM fungi were identified by polymerase chain reaction amplification and sequencing of the internal transcribed spacer of rDNA.

Results

The mean mycorrhizal colonization exceeded 90 %. Thirty-six ectomycorrhizal taxa were identified in fir stands and 23 in pine forecrops; ten out of these species were common to both stands. The fungal communities were different between study sites (R?=?0.1721, p?=?0.0001). Tomentella stuposa was the only species present at all sites.

Conclusion

Silver fir seedlings in Scots pine forecrops supported smaller ECM fungal communities than communities identified in mature silver fir stands. Nevertheless, fungal colonization of seedling roots was similar in both cases. This suggests that pine stands afforested on formerly arable land bear enough ECM species to allow survival and growth of silver fir seedlings.     

The effects of different types of management,functions, and characteristics of stands in Polish forests on the amount of coarse woody debris

In the modern forestry paradigm, many factors influence the amount of coarse woody debris (CWD). The present paper analyzes the effects of both local (national) programs (special functions of forests) and European programs (Natura 2000 sites), as well as the individual characteristics of forest stands. The study was conducted on 2,752 sampling plots distributed over an area of about 17,500 ha and located in lowland stands having a species composition typical of large areas in central Europe. Natura 2000 areas contained significantly more CWD (8.4 m³/ha) than areas not covered by the program (4.8 m³/ha). However, this is due to the fact that Natura 2000 sites involve well-preserved forest areas, such as nature reserves (26.6 m³/ha). In the managed forests that have been covered by the Natura 2000 program over the past several years, the volume of CWD has not increased. Forests with ecological and social functions differed slightly in the amount of CWD. More CWD occurred in protected animal areas (8.7 m³/ha) than in stands damaged by industry (3.9 m³/ha). Intermediate CWD levels were found in water-protection forests and in forests located around cities and military facilities. In managed forests, the lowest CWD volume was observed in middle-aged stands. The species composition of the stand had little effect on the volume of CWD. Only stands with a predominance of ash and alder had higher CWD levels (13.5 m³/ha). More CWD was found in stands whose species composition did not represent the potential site quality (6.4 m³/ha) than in habitats with the optimum species composition (3.8 m³/ha). CWD volume should be systematically increased taking into consideration local natural conditions. Such efforts should be focused on particularly valuable regions, and especially on Natura 2000 sites, where the threshold values reported from other European forests should be reached. Leaving some trees to die naturally and retaining reasonable amounts of such trees ought to be incorporated into CWD management practice in Poland.     

Are naringenin and quercetin useful chemicals in pest-management strategies?

The effects of two polyphenolic flavonoids (flavanone naringenin and flavonol quercetin) on development, fecundity, and mortality of the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), were determined in vitro, on an artificial diets. Also determined in vitro (DC EPG method), on sucrose–agarose gels, were the effects of flavonoids on the probing and feeding behavior of adult apterae. When added to a liquid diet, higher concentrations of studied flavonoids increased the developmental time, the pre-reproductive period, and mortality and decreased fecundity and the intrinsic rate of natural increase of A. pisum. In most events associated with stylet activity (as indicated by EPG waveform g-C), differences in probing behavior did not statistically differ between the control gel and those with flavonoids; quercetin at 10, 100, and 1,000 µg cm^?3 prolonged the number of gel penetrations; and quercetin only at 10,000 μg cm^?3 prolonged the time the first g-C waveform was observed. Addition of flavonoids to the gels generally reduced passive ingestion from fluids of the gels (EPG waveform g-E2). At higher concentrations (>1,000 µg cm^?3) the flavonoids completely stopped salivation (EPG waveform g-E1) and passive ingestion from fluids of the gels (EPG waveform g-E2). In events associated with active ingestion (EPG waveform g-G), however, differences in feeding behavior did not statistically differ between the control gel and those with flavonoids. The present findings demonstrate detrimental effects of the flavanone naringenin and flavonol on the behavior of the pea aphid. This can be employed in a biotechnological projects for plant breeding resistant to herbivores, including aphids.