Fluorescence properties of humic acid interaction products with s-triazine and bipyridilium herbicides and their Cu complexes: a multivariate approach

Purpose

Excitation–emission matrices spectroscopy (EEMS) of soil humic acids (HAs) contains large amount of information on their properties, as well as on the dynamics related to their intra- and inter-molecular interactions. The objective of this research was (i) to show that EEMS represents a useful tool to investigate the molecular and mechanistic aspects of HA adsorbing capacity towards atrazine (A) and paraquat (P) with or without Cu²⁺ ions and (ii) to evaluate if additional information on these mechanisms can be obtained by combining EEMS with principal component analysis (PCA).

Materials and methods

HAs have been isolated from soil samples collected in a citrus field at three locations, within the plant rows (HAa); between the rows (HAb) and in a adjacent, control soil (HAc). Interaction products were obtained between each HA and A and P, with or without Cu ions. Elemental analysis and Fourier-transorm Infrared Spectroscopy were applied to support fluorescence data. Fluorescence spectra were recorded on aqueous solutions, and fluorescence intensity (FI) values were normalized using a quinine sulphate standard. PCA analysis was performed using the software STATGRAPHICS Centurion XV.I.

Results and discussion

The EEM spectra of the three unreacted HAs are characterized by the presence of two fluorophores α and β, in the region of longer wavelengths pairs (EEWP). The EEM spectra of the HA-A interaction products are featured by the same two fluorophores, with FI values decreased of about 50%, whereas those of the HA-P interaction products show a unique peak, γ, at intermediate EEWP. Finally, the EEM spectra of HA-A-Cu²⁺ show, with respect to HA-A samples, a blue shift of the α peak with an additional decrease of FI values (about 60%) and the disappearance of the peaks β, whereas those of HA-P-Cu²⁺ feature, in comparison with those of HA-P, a small red-shift of the peak γ. PCA data suggest that Cu ions do not affect the interaction mechanism between HA and P, whereas it appears to exercise a strong influence on interaction between HA and A.

Conclusions

The results obtained indicate that EEMS allows direct measurements of the adsorbing capacity of HA towards atrazine and paraquat. Additional information obtained by PCA analysis show that Cu ions behave like a good antagonist in preventing the formation of ionic bonds between HA and atrazine, whereas Cu is not able to affect the prevalent mechanism of HA interaction with paraquat, which is a charge-transfer bond.

     

Eco-physiological and Antioxidant Responses of Holm Oak (<Emphasis Type="Italic">Quercus ilex</Emphasis> L.) Leaves to Cd and Pb

Plants of the urban environment are exposed to a wide range of pollutants, including heavy metals. This research studies in situ the eco-physiological and antioxidant responses of holm oak (Q. ilex) leaves to Pb and Cd to assess the mechanisms of metal tolerance in this species, widely used as biomonitor. Leaves of plants grown at parks and roadsides were analyzed for photosynthetic activity, Pb and Cd concentration in tissues and cell-free extracts, thiol groups, D1 and Rubisco protein content, ascorbic acid (AsA) amount, and catalase (CAT) activity. The main results evidenced that Cd concentration was higher in leaves collected at the park out from the downtown; whereas Pb was most abundant in leaves sampled at the roadside nearby the highway. Pb in cell-free extracts was higher in park than in roadside leaves. Although Cd in the leaf tissues was twofold lower than Pb, it was more abundant than Pb in cellular extracts deprived of all particulate matter. Leaves responded to different concentration of Cd and Pb modulating some eco-physiological and biochemical traits, roadside leaves showed reduced leaf lamina, higher content of photosynthetic pigments, hydrogen peroxide, and AsA, as well as higher CAT activity compared to park leaves. In the roadside leaves, a stress condition for photosynthetic apparatus can be hypothesized on the basis of the decline of photochemical activity, the increase of NPQ, and the reduction of Rubisco and D1 protein content. The elevated presence of thiol groups in these leaves suggests a possible role of Pb and Cd in activation of antioxidant responses.     

Anti-chemokine activities of ixodid ticks depend on tick species,developmental stage,and duration of feeding

Ixodid ticks require comparatively large bloodmeals for their development and survival. Blood-feeding elicits signaling events in the host leading to wound healing responses (hemostasis, inflammation, and tissue repair) and immunity. Bioactive molecules present in tick saliva sabotage these host responses at several levels. One of them is neutralization of cellular communication by binding of specific saliva molecules to cytokines that have important roles in innate and adaptive immunity. Chemokines are a subset of cytokines having chemotactic activities. We show anti-chemokine activities in salivary gland extracts (SGE) of adult Rhipicephalus appendiculatus ticks against human chemokines CXCL8, CCL2, CCL3, CCL5, and CCL11. At comparable protein concentrations, male Ixodes ricinus SGE showed activity against all the chemokines; SGE of female I. ricinus had comparatively lower levels of activity against all the chemokines but no detectable activity against CCL5 and CCL11. However, when the equivalent of a single pair of salivary glands was tested, male I. ricinus showed little or no activity against CCL3 and CCL5. No fundamental differences in activity were observed against mouse compared with human chemokines. A comparison with previously published data for Dermacentor reticulatus and Amblyomma variegatum indicates that the level of anti-cytokine activity depends on the species, developmental stage (adult or nymph), and amount of SGE used, as well as on the number of days the tick has been feeding.     

Qualitative Comparison of 0.27T, 1.5T,and 3T Magnetic Resonance Images of the Normal Equine Foot

Magnetic resonance (MR) imaging has become an important diagnostic tool in the investigation of foot pain in horses. The aim of this study was to qualitatively compare ex-vivo MR images of the same equine feet obtained at three magnetic field strengths: 0.27, 1.5, and 3 tesla (T). Ten cadaver feet were used. All feet were imaged with two high-field systems (3T, 1.5T) and with a low-field (LF) system at 0.27 T designed for standing horses. Images were acquired using similar pulse sequences in all 3 MR units. MR images were subjectively evaluated by three independent experienced image analysts for image quality and clarity of visualization of individual anatomical structures using a four-point grading scale. The images from all of the examinations were considered to be of diagnostic value except for the hoof capsule where substantial artifacts were present in LF images with distortion and loss of signal at the dorsal/distal aspect of the hoof capsule in LF images. Anatomical structure scoring values of images obtained at 3T and 1.5T were significantly greater than scores of images obtained at 0.27T. Scores for images obtained at 3T were significantly higher than those for images obtained at 1.5T. Mean score differences between 1.5T and 3T were higher for cartilage of the distal interphalangeal joint and for the ungular cartilages.     

Effect of living cells of microalgae or their extracts on soil enzyme activities

The enzymatic activity of the soil and potential plant growth can both be affected by the addition of different substances, such as biostimulants. The final objective of this work was to evaluate the effect of Chlorella vulgaris, Scenedesmus quadricauda or their extracts applied directly into the soil, by monitoring the complex soil-microorganism-plant system. Experiments were conducted in pots using agricultural soil and tomato plants in a growth chamber at 25 ± 2 °C illuminated by artificial light with a 16-h photoperiod. The soil biological activity was analyzed by monitoring fluorescein diacetate hydrolysis, dehydrogenases, acid and alkaline phosphomonoesterase and urease activities. Dry weight and chlorophyll index values of tomato plants were also investigated. Results showed that both the microalgae and their extracts positively affected soil biological activity by increasing values of the biochemical index of potential soil fertility (Mw) both in cultivated and uncultivated soils. The highest Mw value was reached in cultivated soil treated for 11 days with the lower concentration of S. quadricauda extract (Mw 9.47). All treatments significantly increased the growth of tomato plants with respect to untreated soil. These results are very promising with a view to improving soil biological activity as well as increasing plant growth.     

Biotechnological production of vitamin B2-enriched bread and pasta

Lactic acid bacteria (LAB) were obtained from durum wheat flour samples and screened for roseoflavin-resistant variants to isolate natural riboflavin-overproducing strains. Two riboflavin-overproducing strains of Lactobacillus plantarum isolated as described above were used for the preparation of bread (by means of sourdough fermentation) and pasta (using a prefermentation step) to enhance their vitamin B2 content. Pasta was produced from a monovarietal semolina obtained from the durum wheat cultivar PR22D89 and, for experimental purposes, from a commercial remilled semolina. Several samples were collected during the pasta-making process (dough, extruded, dried, and cooked pasta) and tested for their riboflavin content by a high-performance liquid chromatography method. The applied approaches resulted in a considerable increase of vitamin B2 content (about 2- and 3-fold increases in pasta and bread, respectively), thus representing a convenient and efficient food-grade biotechnological application for the production of vitamin B2-enriched bread and pasta. This methodology may be extended to a wide range of cereal-based foods, feed, and beverages. Additionally, this work exemplifies the production of a functional food by a novel biotechnological exploitation of LAB in pasta-making.     

Effects of urea on the molecules involved in the olfactory signal transduction: a preliminary study on Danio rerio

Among vertebrates, the physiologically uremic Chondrichthyes are the only class which are not presenting the ciliated olfactory receptor neurons in the olfactory neuroepithelium. The only sequenced genome for this class revealed only three olfactory receptor genes and the immunohistochemical detection of G protein alpha subunit typically coupled to the olfactory receptors (Gα_olf) failed in different species. Chronic renal disease can represent a cause of olfactory impairment in human. In this context, our present study focused on investigating potential effects of high urea concentration on the olfactory epithelium of vertebrates. Larvae of the teleost fish Danio rerio were exposed to urea in order to assess the effects on the olfactory signal transduction; in particular on both the olfactory receptors and the Gα_olf. The endocytosis of neutral red dye in the olfactory mucosa was detected in control and urea-exposed larvae. The amount of neutral red dye uptake was used as a marker of binding and internalization of the Gα_olf. The neutral red dye endocytosis was not affected by urea exposure, hence suggesting that the presence of the Gα_olf and their binding to the odorants are not affected by urea treatment, either. The presence and distribution of Gα_olf were investigated in the olfactory epithelium of control and urea-exposed larvae, using a commercial antibody. The immunoreactivity was increased after urea treatment, suggesting an effect of urea on the expression or degradation of this G protein alpha subunit.     

From biology to management of Savi's pine vole (Microtus savii)

Savi's pine vole (Microtus savii) is a rodent species of the Cricetidae family, inhabiting southern European agroecosystems. It is considered to be the main cause of rodent‐attributed damage in Italy. To achieve an effective management, detailed knowledge of this species is needed. However, the available information about this species is fragmentary and incomplete. In this paper, the existing knowledge of Savi's pine vole taxonomy, reproduction, population dynamics, habitat and food preferences is reviewed in order to organise available information and identify priority areas of future research. Some of the changes in farming practices that have occurred in recent decades may have increased the impact of Savi's pine vole populations in crop fields. To manage this pest species effectively, an integrated strategy is recommended (involving habitat management, trapping and, when appropriate, the use of rodenticides). The apparent lack of cyclical population outbreaks and the relatively small litter size and long gestation and interpartum period of this species suggest that it could be more manageable than other vole species, while its strict herbivorous diet, stable population size in open habitats and wide distribution seem to indicate it as an ideal model species for risk assessment studies. © 2015 Society of Chemical Industry     

Treatment of intracranial neoplasia in dogs using higher doses: A randomized controlled trial comparing a boosted to a conventional radiation protocol

BackgroundLocal progression of intracranial tumors can be the consequence of insufficient radiation dose delivered. Dose increases in the brain must be made carefully so as not to risk debilitating adverse effects such as radiation necrosis.HypothesisA new protocol with 10 × 4 Gy + 11% physical dose increase limited to the macroscopic tumor volume results in a clinically better outcome compared to a 10 × 4 Gy protocol.AnimalsFifty‐seven client‐owned dogs with primary intracranial neoplasia.MethodsRandomized controlled trial. Twenty‐eight dogs were assigned to the control protocol (10 × 4 Gy) and 29 to the simultaneous integrated boost (SIB) protocol with 4.45 Gy dose increase. Treatment groups were compared for outcome and signs of toxicity.ResultsMild, transient acute or early‐delayed adverse radiation effects were observed in 5 dogs. Severe late adverse effects were not seen. Between the protocols, no significant differences were found for outcome (intention‐to‐treat analysis): overall time to progression (TTP) was 708 days (95% confidence interval (95% CI) [545,872]), in the control group it was 828 days (95% CI [401,1256]), and in the SIB group 627 days (95% CI [282,973]; P = .07). Median overall survival (OS) was 684 days (95% CI [516,853]), in the control group it was 724 days (95% CI [623,826]), and in the SIB group 557 days (95% CI [95,1020]; P = .47). None of the tested variables was prognostic in terms of outcome.Conclusion and Clinical ImportanceThe dose escalation used with an 11% physical dose increase did not result in better outcome.