Cartap hydrolysis relative to its action at the insect nicotinic channel

The insecticide cartap is the bis(thiocarbamate) derivative of 2-(dimethylamino)propane-1,3-dithiol, which on oxidation forms the natural toxicant nereistoxin (NTX) [4-(dimethylamino)-1,2-dithiolane]. Both cartap and NTX are ion channel blockers of the nicotinic acetylcholine receptor (nAChR). Cartap was originally proposed to act only after metabolic conversion to NTX and later suggested to block directly without activation. The present study uses a new approach to differentiate these hypotheses, that is, pH effects on channel-blocking activity and hydrolysis rates. As controls, mecamylamine (the classic channel blocker) and NTX are stable and similar in channel-blocking potency ([(3)H]thienylcyclohexylpiperidine binding assay, honeybee nAChR) at pH 6.1-8.4. In contrast, cartap is >200-fold more effective at pH 7.4 than at pH 6.1, indicating that it undergoes hydrolytic activation. Cartap slowly hydrolyzes to cartap monothiol at pH 6.1 but quickly forms the dithiol and some NTX at pH 7.4. The relationship between potency and hydrolysis products at various pH ranges suggests that cartap dithiol is the most plausible blocking agent.     

Hydroxymethanesulphinate salts and related decomposition products in wastewater

Hydroxymethanesulphinate (HMS) and its decomposition products such as HCHO, Zn and sulphites can be determined quantitatively in each other's presence by a polarographic method. In the alkaline medium it is possible to have well defined and separated waves relative to the oxidation of HMS ion and to the reduction of HCHO and zincate ion. The quantitative determination of these compounds is carried out with standard addition methods since the concentration-current relationship is linear. The SO₂ is determined in HClO₄ 0.1 M and in the same sample it is possible to determine even HMS, HCHO and Zn after addition of NaOH up to 0.1 M. Moreover, a method of indirect determination of HMS is herein described. This is based on the determination of HCHO released by means of a weak oxidation of HMS which is stoichiometrically proportional to the total HMS. The method is sensitive and is made specific for such types of wastewater which come from textile industries.     

In vitro effect of classical swine fever virus on a porcine aortic endothelial cell line

The effect of classical swine fever (CSF) virus on some phenotypic and functional features of an established porcine aortic endothelial cell (AOC) line was investigated. AOC cells show most of the characteristics of primary endothelial cells, avoiding the alterations and senescence that these cells undergo after a few passages in culture. AOC cells were susceptible to CSF virus infection to a high degree, reaching 90% of CSF virus positive cells after 24 h of infection; however as with other porcine susceptible cells, no cytopathic effect could be observed. In these conditions none of the surface molecules studied, including SLA-II MHC antigens, adhesion or co-stimulatory molecules, were altered by virus infection after 24 or 48 h. Functionally CSF virus infection induced a decrease in the pro-coagulant activity of the AOC cells, determined by the increase in the clot formation time shown by the lysates of these cells. This contrasts with the increase observed in the expression of mRNA corresponding to IL-1 alpha and IL-6, two proinflammatory and pro-coagulant cytokines, in CSF virus-infected AOC cells.     

Influence of the species and geographical location on volatile composition of Spanish oak wood (Quercus petraea Liebl. and Quercus robur L.)

The influence of the species and geographical origin on the volatile composition of wood samples from 80 Spanish oaks (55 Quercus petraea Liebl. and 25 Quercus robur L.) has been studied. Oak volatile components were isolated by simultaneous distillation-extraction and analyzed by gas chromatography-mass spectrometry. cis- and trans-beta-methyl-gamma-octalactones were the main constituents, the cis stereoisomer being predominant. Other important volatile components were furfural, 5-methylfurfural, guaiacol, eugenol, vanillin, or syringaldehyde. The main differences were established between species, Quercus petraea being significantly richer in volatile compounds than Quercus robur; however, the variability found among trees was high. Differences among geographical provenances were much less important than those found between species.     

Effects of cortisol on pregnancy rate and corpus luteum function in heifers: an in vivo study

To determine whether glucocorticoids affect the function of the bovine corpus luteum (CL) during the estrous cycle and early pregnancy, we examined the effects of exogenous cortisol or reduced endogenous cortisol on the secretion of progesterone (P4) and on pregnancy rate. In preliminary experiments, doses of cortisol and metyrapone (an inhibitor of cortisol synthesis) were established (n=33). Cortisol in effective doses of 10 mg blocked tumor necrosis factor-induced prostaglandin F(2α) secretion as measured by its metabolite (PGFM) concentrations in the blood. Metyrapone in effective doses of 500 mg increased the P4 concentration. Thus, both reagents were then intravaginally applied in the chosen doses daily from Day 15 to 18 after estrus (Day 0) in noninseminated heifers (n=18) or after artificial insemination (n=36). Pregnancy was confirmed by transrectal ultrasonography between Days 28-30 after insemination. Plasma concentrations of P4 were lower in cortisol-treated heifers than in control heifers on Days 17 and 18 of the estrous cycle (P<0.05). However, the interestrus intervals were not different between control and cortisol-treated animals (P>0.05). Moreover, metyrapone increased P4 and prolonged the CL lifespan in comparison to control animals (P<0.05). Interestingly, in inseminated heifers, cortisol increased the pregnancy rate (75%) compared with control animals (58%), whereas metyrapone reduced the pregnancy rate to 16.7% (P<0.05). The overall results suggest that cortisol, depending on the physiological status of heifers (pregnant vs. nonpregnant), modulates CL function by influencing P4 secretion. Cortisol may have a positive influence on CL function during early pregnancy, leading to support of embryo implantation and resulting in higher rates of pregnancy in heifers.     

Age effects and climate response in trees: a multi-proxy tree-ring test in old-growth life stages

Aging in trees implies a progressive reduction in the growth rate, related to a shortening of the growing period and changes in the photosynthetic capability and efficiency. These changes may continue during the old-growth life stages following the juvenile phase and are reflected in tree-ring properties such as growth increment, density or stable isotopes. We studied possible climate age effects in time series of several tree-ring parameters (ring width, wood density and stable carbon and oxygen isotopes) of mature individuals from two age groups of Pinus uncinata and P. nigra at two locations in Spain. The aim was to test whether age differences in trees in the old-growth life stages could lead to diverging climate responses. The results show some differences in response to climate between age groups at a monthly level, but most of these divergences are not significant for seasonal climate variables. Regardless of the age group, the main limiting climate factors constrained tree growth equally. Although our findings do not support the idea of an age-dependent response to climate that may lead to inaccurate climate reconstructions, further studies using tree-ring density and stable isotope series are urgently needed to verify the current results.     

Wheat Bran AX Properties and Choice of Xylanase Affect Enzymic Production of Wheat Bran‐Derived Arabinoxylan‐Oligosaccharides

Wheat bran‐derived arabinoxylan‐oligosaccharides (AXOS) recently have been shown to potentially exert prebiotic effects. In this study, 15 bran samples obtained by milling different wheat cultivars were treated with xylanases from Hypocrea jecorina (XHJ), Aspergillus aculeatus (XAA), and Pseudoalteromonas haloplanktis (XPH) to assess the effect of bran source and xylanase properties on the AXOS yield and structure. The total arabinoxylan (AX) extraction yield was higher with XHJ (8.2–10.7%) and XAA (8.2–10.8%) than with XPH (6.9–9.5%). Irrespective of the enzyme, a significant negative correlation was observed between extraction yield and arabinose to xylose (A/X) ratio of bran AX (r = –0.7), but not between yield and bran AX level. The A/X ratio of the extracted material was 0.27–0.34 for all bran samples and all enzymes, which combined with yield data and microscopic analysis, indicated primary hydrolysis of aleurone and nucellar epidermis AX. The average degree of polymerization (avDP) of the extracted AX was very low for all enzymes (2–3), owing to the release of high levels of monomeric arabinose and xylose. The release of these monosaccharides could be ascribed to 1) the activity of wheat bran‐associated enzymes (arabinofuranosidases and xylosidases); 2) the hydrolytic properties of the xylanases themselves; and 3) the presence of xylosidases as contaminations in enzyme preparation, in that order of importance. Heat treatment of bran before xylanase treatment significantly decreased the levels of monomeric arabinose and xylose in the extract, without affecting the extraction yield, resulting in a higher avDP of 3–7, thus yielding true AXOS. Overall, for AXOS production, wheat cultivars with a low bran A/X ratio of the AX are preferable as starting materials, and inactivation of bran‐associated enzymes before incubation is desirable. The XHJ xylanase was the best enzyme for wheat bran‐derived AXOS production.     

Physical carbon‐sequestration mechanisms under special consideration of soil wettability

The protective impact of aggregation on microbial degradation through separation has been described frequently, especially for biotically formed aggregates. However, to date little information exists on the effects of organic‐matter (OM) quantity and OM quality on physical protection, i.e., reduced degradability by microorganisms caused by physical factors. In the present paper, we hypothesize that soil wettability, which is significantly influenced by OM, may act as a key factor for OM stabilization as it controls the microbial accessibility for water, nutrients, and oxygen in three‐phase systems like soil. Based on this hypothesis, the first objective is to evaluate new findings on the organization of organo‐mineral complexes at the nanoscale as one of the processes creating water‐repellent coatings on mineral surfaces. The second objective is to quantify the degree of alteration of coated surfaces with regard to water repellence. We introduce a recently developed trial that combines FTIR spectra with contact‐angle data as the link between chemical composition of OM and the physical wetting behavior of soil particles. In addition to characterizing the wetting properties of OM coatings, we discuss the implications of water‐repellent surfaces for different physical protection mechanisms of OM. For typical minerals, the OM loading on mineral surfaces is patchy, whereas OM forms nanoscaled micro‐aggregates together with metal oxides and hydroxides and with layered clay minerals. Such small aggregates may efficiently stabilize OM against microbial decomposition. However, despite the patchy structure of OM coating, we observed a relation between the chemical composition of OM and wettability. A higher hydrophobicity of the OM appears to stabilize the organic C in soil, either caused by a specific reduced biodegradability of OM or indirectly caused by increased aggregate stability. In partly saturated nonaggregated soil, the specific distribution of the pore water appears to further affect the mineralization of OM as a function of wettability. We conclude that the wettability of OM, quantified by the contact angle, links the chemical structure of OM with a bundle of physical soil properties and that reduced wettability results in the stabilization of OM in soils.     

Degradation and persistence of rotenone in soils and influence of temperature variations

The persistence and degradation of rotenone and its primary degradation product 12a beta-hydroxyrotenone in soils were determined under standardized laboratory conditions in the dark at 20 or 10 degrees C and at 40% of water holding capacity. Degradation experiments were carried out on two types of soil collected in southern Italy, a silt clay loam (SCL) and a loamy soil (L). A kinetic model was developed to describe degradation rates of rotenone, taking into account the production, retention, and degradation of the main metabolites. The DT50 values of rotenone and 12a beta-hydroxyrotenone, were 8 and 52 days in SCL soil, and 5 and 23 days in L soil at 20 degrees C, respectively. However, at 10 degrees C a tendency for slower degradation of rotenone and 12a beta-hydroxyrotenone was observed (25 and 118 days in SCL and 21 and 35 days in L soils, respectively). The differences were significant for most data sets. Temperature had a strong effect on degradation; a 10 degrees C increase in temperature resulted in a decrease in the DT50 value by a factor of 3.1 and 2.2 in SCL and of 4.2 and 1.4 in L soils for both rotenone and 12a beta-hydroxyrotenone, respectively. Results show that the degradation rates of both rotenone and 12a beta-hydroxyrotenone were greatly affected by temperature changes and soil physicochemical properties. The degradation reaction fits the two compartment or the multiple compartment model pathways better, which clearly indicates a rather complex rotenone degradation process in soils. Results provide further insights on the rates and the mechanisms of rotenone degradation in soils, aiming to more clearly describe the degradation pathway of chemical residues in the environment.     

Clostridium bifermentans and C. subterminale are associated with kiwifruit vine decline,known as moria,in Italy

Since 2012, a new pathogenic syndrome has frequently been observed in many areas of kiwifruit cultivation in Italy. The main symptoms include an initial withering of the leaves followed by a total and sudden collapse of plants, mainly occurring during summer. The withered leaves fall and the main and secondary feeder roots appear rotten, sometimes showing a reddish-brown discoloration. The disease, that affects both the green and yellow-fleshed cultivars, has been called kiwifruit vine decline and is locally known as moria. The syndrome has been found consistently associated with soil waterlogging, which frequently occurs either after the traditional agronomical practice of irrigating orchards through surface irrigation or after very heavy rainfall. So far, the role played by bacteria in this syndrome has not been investigated. In the present study, Clostridium spp. were isolated from both rotten roots and soils obtained from Italian kiwifruit orchards affected by the syndrome, indicating for the first time that anaerobic bacteria are able to cause damage to woody crops. C. bifermentans and C. subterminale incited symptoms in kiwifruit in both in vivo and in vitro pathogenicity tests. Soil waterlogging seems to potentially favour colonization of kiwifruit roots by anaerobic bacteria, probably because saturation of the soil can facilitate proliferation and persistence of these bacteria during long periods of the vegetative growth of the crop. The occurrence of anaerobic bacteria does not exclude the possibility that other microorganisms can play additional/synergic role(s) in causing the kiwifruit vine decline.