Factors affecting the efficacy of non-fumigant nematicides for controlling root-knot nematodes

Second-stage juveniles (J2) and egg masses of root-knot nematodes as well as root debris heavily infected by the latter were exposed for different periods of time to six different doses of the nematicides cadusafos and fenamiphos. The efficacy of the nematicides increased significantly with increasing exposure time. Both nematicides were more effective against J2, although they could not provide acceptable control of J2 inside egg masses or heavily galled root debris. The effect of different application strategies on the efficacy and persistence of certain nematicides was also assessed in a field study. Cadusafos, fenamiphos, fosthiazate and oxamyl were applied in field micro-plots either as a single full dose at the time of crop establishment or as multiple reduced-rate applications at 14-day intervals throughout the cropping period, and their efficacy and persistence were determined using bioassays and analytical studies. Fosthiazate was the most efficient nematicide studied, and this was mainly attributed to its long soil persistence. Oxamyl also provided adequate nematode control for the first 48-56 days after its application, regardless of the application method used and its relatively rapid field dissipation. Fenamiphos and cadusafos failed to provide adequate nematode control, although cadusafos was the most persistent of the nematicides tested. The failure of fenamiphos to provide adequate nematode control was mainly attributed to its rapid degradation by soil micro-organisms, which were stimulated after its repeated low-rate application at 14-day intervals. In contrast cadusafos failure was attributed to the inability of the nematicide to reduce nematode populations even at relatively high concentrations in soil.     

Saffron as a source of novel acetylcholinesterase inhibitors: molecular docking and in vitro enzymatic studies

Inhibitors of acetylcholine breakdown by acetylcholinesterase (AChE) constitute the main therapeutic modality for Alzheimer's disease. In the search for natural products with inhibitory action on AChE, this study investigated the activity of saffron extract and its constituents by in vitro enzymatic and molecular docking studies. Saffron has been used in traditional medicine against Alzheimer's disease. Saffron extract showed moderate AChE inhibitory activity (up to 30%), but IC(50) values of crocetin, dimethylcrocetin, and safranal were 96.33, 107.1, and 21.09 μM, respectively. Kinetic analysis showed mixed-type inhibition, which was verified by in silico docking studies. Safranal interacts only with the binding site of the AChE, but crocetin and dimethylcrocetin bind simultaneously to the catalytic and peripheral anionic sites. These results reinforce previous findings about the beneficial action of saffron against Alzheimer's disease and may be of value for the development of novel therapeutic agents based on carotenoid-based dual binding inhibitors.     

Lipophilic Caffeic Acid Derivatives Protect Cells against H(2)O(2)-Induced DNA Damage by Chelating Intracellular Labile Iron

Naturally occurring cinnamic acid derivatives are ubiquitously distributed in the plant kingdom, and it has been proposed that their consumption contributes to the maintenance of human health. However, the molecular mechanisms underlying their health keeping effects remain unknown. In the present investigation, we evaluated the capacity of several cinnamic acid derivatives (trans-cinnamic, p-coumaric, caffeic and ferulic acids, as well as caffeic acid-methyl and -propyl esters) to protect cells from oxidative stress-induced DNA damage. It was observed that effective protection was based on the ability of each compound to (i) reach the intracellular space and (ii) chelate intracellular "labile" iron. These results support the notion that numerous lipophilic iron chelating compounds, present abundantly in plant-derived diet components, may protect cells in conditions of oxidative stress and in this way be important contributors toward maintenance of human health.     

Responses of cucumber grown in recirculating nutrient solution to gradual Mn and Zn accumulation in the root zone owing to excessive supply via the irrigation water

A standard and a high manganese (Mn) level (10 and 160 μM) were combined with a standard and a high zinc (Zn) level (4 and 64 μM) in the nutrient solution supplied to cucumber in closed‐cycle hydroponic units to compensate for nutrient uptake. The concentrations of all nutrients except Mn and Zn were identical in all treatments. The objectives of the experiment were to establish critical Zn and Mn levels in both nutrient solutions and leaves of cucumber grown hydroponically, to assess the impact of gradual Zn and/or Mn accumulation in the external solution on nutrient uptake and gas exchange, and to find whether Mn and Zn have additive effects when the levels of both ions are excessively high in the root zone. The first symptoms of Mn and Zn toxicity appeared when the concentrations of Mn and Zn in the leaves of cucumber reached 900 and 450 mg kg^–1 in the dry weight, respectively. Excessively high Mn or/and Zn concentrations in the leaves reduced the fruit biomass production due to decreases in the number of fruits per plant, as well as the net assimilation rate, stomatal conductance, and transpiration rate, but increased the intercellular CO₂ levels. Initially, the Mn or Zn concentrations in the recirculating nutrient solution increased rapidly but gradually stabilized to maximal levels, while the corresponding concentrations in the leaves constantly increased until the end of the experiment. The uptake of Mg, Ca, Fe, and Cu was negatively affected, while that of K and P remained unaffected by the external Mn and Zn levels. The combination of high Mn and Zn seems to have no additive effects on the parameters investigated.     

Application and fate of Cyromazine in a closed-cycle hydroponic cultivation of bean (Phaseolus vulgaris L.)

The fate of cyromazine applied via the nutrient solution (20, 40, and 60 mg of active ingredient per plant) in a closed-cycle soilless cultivation of bean with zero discharge of effluents was traced in both the recycled drainage solution and the plant tissues for 99 days. The insecticide was applied once, 15 days after planting (16 days prior to the first harvest). In addition to cyromazine, the residues of melamine, its metabolite, in the drainage solution and plant tissues were also regularly determined during the 99 days. The two higher application doses induced toxicity symptoms on the leaves of the bean plant. The maximum cyromazine levels were measured 8 days after application in the drainage solution (17-46 mg l(-1)), 16 days in the roots (1.1-2.4 mg kg(-1) fresh weight [f. wt.]) and the vegetative shoot (4.5-9.5 mg kg(-1) f. wt.), and 24 days after application in the pods (2.6-4.1 mg kg(-1) f. wt.). However, the cyromazine residues in pods were clearly below the maximum acceptable levels for bean. The half-life of cyromazine in the drainage solution ranged from 16 to 19 days for the three doses. The melamine residues in the drainage solution and in the roots reached a concentration peak 16 days after cyromazine application, whereas in the vegetative shoot and the pods they were constantly increasing over the 99 days after application. Nevertheless, the melamine residues were constantly much lower than those of cyromazine, although on the last sampling day (99) they tended toward convergence. Cyromazine proved to be highly persistent, as indicated by the remarkably high residues measured in both the drainage solution and the plant tissues, even 99 days after application. Nevertheless, the application of cyromazine via the nutrient solution to beans grown in closed-cycle hydroponic systems at doses not exceeding 20 mg per plant seems to be safe with respect to both phytotoxicity and residue levels in the edible pods.     

Oscillatory dynamics of Cdc42 GTPase in the control of polarized growth

Cells promote polarized growth by activation of Rho-family protein Cdc42 at the cell membrane. We combined experiments and modeling to study bipolar growth initiation in fission yeast. Concentrations of a fluorescent marker for active Cdc42, Cdc42 protein, Cdc42-activator Scd1, and scaffold protein Scd2 exhibited anticorrelated fluctuations and oscillations with a 5-minute average period at polarized cell tips. These dynamics indicate competition for active Cdc42 or its regulators and the presence of positive and delayed negative feedbacks. Cdc42 oscillations and spatial distribution were sensitive to the amounts of Cdc42-activator Gef1 and to the activity of Cdc42-dependent kinase Pak1, a negative regulator. Feedbacks regulating Cdc42 oscillations and spatial self-organization appear to provide a flexible mechanism for fission yeast cells to explore polarization states and to control their morphology.     

Effect of solution conductivity on the volatile constituents of Origanum dictamnus L. in nutrient film culture

The chemical composition of the essential oils obtained from leaves and bracts of hydroponically cultivated Origanum dictamnus L. (Cretan dittany), growing under various electrical conductivity (EC) levels (2.0, 4.0, and 6.0 mS/cm), was studied, using the nutrient film technique (NFT). The analysis of the essential oil content was achieved by GC-MS technique, and totals of 41 and 38 different compounds were identified in both cases of large-leaved and narrow-leaved samples of leaves and bracts, respectively. Differences in the composition content and of the percentage of each of the constituents in the two studied samples (i.e., large-leaved and narrow-leaved) and within the essential oils of leaves and bracts in both samples were observed. Carvacrol and p-cymene were identified as the main constituents in all essential oils, whereas thymoquinone was found in higher percentage in the essential oils of large-leaved than in narrow-leaved plants. The results obtained from GC-MS analysis were submitted to chemometric analysis, and a phenotypic similarity of the essential oils of narrow-leaved O. dictamnus was observed, whereas the essential oils of large-leaved O. dictamnus showed two separate subgroups.     

Association between physical properties of cervical mucus and ovulation rate in superovulated cows

Possible associations between certain physical properties of cervical mucus (CM) and ovulation rate were studied in 21 superovulated Holstein cows. In CM samples collected at the beginning of estrus (0 h) and in 4 h intervals for the following 24 h, the pH, the spinnbarkeit (spinability), and the crystallization value were measured. Blood samples, collected at the same time points with CM samples, were assessed for progesterone and estradiol concentrations. At 48 h the number of ovulated follicles was counted by transrectal ultrasonography and the animals were allotted into 2 groups according to the occurrence of at least one (group A, n = 16) or no (group B, n = 5) ovulations. The pH was lower (P < 0.05) at 8 h (7.00 + 0.24) in group A compared with group B (7.55 + 0.12). In group A, spinnbarkeit was significantly lower at 0 h and 20 h, and higher at 8 h and 16 h compared with group B (0 h: 2.50 + 0.82 versus 6.95 + 0.41; 20 h: 3.00 + 1.89 versus 5.38 + 0.94; 8 h: 7.00 + 0.87 versus 2.75 + 0.43; 16 h: 7.00 + 1.41 versus 4.30 + 0.71, for groups A versus B, respectively). Crystallization was significantly lower at 4 h (2.00 + 0.63) and 20 h (1.50 + 0.82) in group A compared with group B (3.13 + 0.32 at 4 h and 3.00 + 0.41 at 20 h). Progesterone at all time points, and estradiol at 16 h, 20 h, and 24 h were lower (P < 0.05) in group A than in group B. The pH, crystallization, estradiol, and progesterone differed (P < 0.05) within one group, while sbk differed within both groups.Our results imply that during the periovulatory period, steadily low progesterone concentrations trigger alterations of certain CM characteristics, while extremely high estradiol concentration could prevent the occurrence of these alterations.     

A minimally invasive method for clinical trans‐diaphragmatic pressure measurement in anaesthetized dogs

ObjectiveTo measure trans-diaphragmatic pressures, as an indication of diaphragmatic contractility, in anaesthetized dogs breathing normally, or during inspiratory obstruction (Mueller’s manoeuvre) in order to assess if the method is practicable for use in clinical circumstances.Study designPilot study.AnimalsTwenty eight client-owned dogs, ASA I or II, 1–10 years old, 5–30 kg bodyweight, which required anaesthesia for surgery, and were to be positioned in lateral recumbency.MethodsFollowing a standardized regimen of premedication and anaesthetic induction, anaesthesia was stabilized and maintained with isoflurane. Two commercially available balloon catheters were introduced orally, and advanced, one into the stomach and one into the mid-third of the oesophagus. Oesophageal and gastric pressures were measured from these catheters, and trans-diaphragmatic pressure (P_di) calculated and recorded continuously. At three separate time points during anaesthesia, for one breath, inspiration was obstructed (Mueller’s manoeuvre) and P_di was measured.ResultsPlacement of the catheters in the stomach was not easy, and failed in three cases. In five dogs, their size resulted in failure of correct placement of catheters. Good traces of all three pressures reflecting respiratory cycles were obtained from 20 dogs. During normal spontaneous breathing (mean ± SD [range]) P_di was 5 ± 2.1 (3–10) mmHg. During Mueller’s manoeuvre, P_di was 14.6 ± 4.5 (9–21) mmHg. Abnormal waveforms were seen included cardiac oscillations (five dogs), inadequate intra-gastric pressure tracing (one dog), deflections with a double peak (one dog), and multiple artifacts when there was increased heart rate and tachypnoea (two dogs in response to surgery).Conclusions and clinical relevanceMeasurement of trans-diaphragmatic pressure with balloon catheters was practicable in suitably sized dogs anaesthetized for clinical purposes and might be a useful tool in the assessment of diaphragmatic function. A range of catheters are required if the technique is to work in all dogs.