The influence of N-alkyl chains in benzoyl-thiourea derivatives on urease inhibition: Soil studies and biophysical and theoretical investigations on the mechanism of interaction.

Ureases are enzymes produced by fungi, plants, and bacteria associated with agricultural and clinical problems. The urea hydrolysis in NH3 and CO2 leads to the loss of N-urea fertilizers in soils and changes the human stomach microenvironment, favoring the colonization of H. pylori. In this sense, it is necessary to evaluate potential enzyme inhibitors to mitigate the effects of their activities and respond to scientific and market demands to produce fertilizers with enhanced efficiency. Thus, biophysical and theoretical studies were carried out to evaluate the influence of the N-alkyl chain in benzoyl-thiourea derivatives on urease enzyme inhibition. A screening based on IC50, binding constants, and theoretical studies demonstrated that BTU1 without the N-alkyl chain (R = H) was more active than other compounds, so the magnitude of the interaction was determined as BTU1 > BTU2 > BTU3 > BTU4 > BTU5, corresponding to progressively increased chain length. Thus, BTU1 was selected for interaction and soil application essays. The binding constants (Kb) for the supramolecular urease-BTU1 complex ranged from 7.95 to 5.71 × 103 M-1 at different temperatures (22, 30, and 38 °C), indicating that the preferential forces responsible for the stabilization of the complex are hydrogen bonds and van der Waals forces (ΔH = -15.84 kJ mol-1 and ΔS = -36.61 J mol-1 K-1). Theoretical and experimental results (thermodynamics, synchronous fluorescence, and competition assay) agree and indicate that BTU1 is a mixed inhibitor. Finally, urease inhibition was evaluated in the four soil samples, where BTU1 was as efficient as NBPT (based on ANOVA two-way and Tukey test with 95% confidence), with an average inhibition of 20% of urease activity. Thus, the biophysics and theoretical studies are strategies for evaluating potential inhibitors and showed that increasing the N-alkyl chain in benzoyl-thiourea derivatives did not favor urease inhibition.

Understanding the multifunctionality of pyroligneous acid from waste biomass and the potential applications in agriculture.

Efforts have been directed to the development of environmentally friendly processes and manufacturing of green products, use of renewable energy and more sustainable agricultural practices. Pyroligneous acid (PA) is a byproduct of biomass pyrolysis that consists of a complex mixture of bioactive substances. The complexity and richness of PA composition have opened a window for PA application in agriculture and mitigation of environmental pollution. This review brings a brief historical on the use of PA and regulatory policies adopted in Brazil, China, Japan and Thailand for PA application in agriculture. The composition and stability of PAs of several origins are presented, together with a discussion of the use of PA to boost plant growth and crop productivity, remove toxic metals from soil, inhibit soil ureases, mitigate the emission of greenhouse gases, control phytopathogen proliferation and weed dissemination. A great variety of biomass types are reported as feedstock to produce PA with distinct chemically diverse and active substances at wide-ranging concentrations. PA has been shown to successfully improve farming practices in a more sustainable fashion. The disclosure of the mechanisms of action that drive the PA's effects, together with the pursue of safety and efficacy data in a case-by-case way to address toxicity and shelf stability, will be valuable to expand the use of PA worldwide for food production.

Interplay between gasotransmitters and potassium is a K+ey factor during plant response to abiotic stress.

Carbon monoxide (CO), nitric oxide (NO) and hydrogen sulfide (H2S) are gasotransmitters known for their roles in plant response to (a)biotic stresses. The crosstalk between these gasotransmitters and potassium ions (K+) has received considerable attention in recent years, particularly due to the dual role of K+ as an essential mineral nutrient and a promoter of plant tolerance to abiotic stress. This review brings together what it is known about the interplay among NO, CO, H2S and K+ in plants with focus on the response to high salinity. Some findings obtained for plants under water deficit and metal stress are also presented and discussed since both abiotic stresses share similarities with salt stress. The molecular targets of the gasotransmitters NO, CO and H2S in root and guard cells that drive plant tolerance to salt stress are highlighted as well.

Does oxidative stress determine the thermal limits of the regeneration niche of Vriesea friburgensis and Alcantarea imperialis (Bromeliaceae) seedlings?

The predicted environmental changes may be detrimental to initial seedling growth, particularly the expected increase in air temperature. We therefore investigated the thermal limits for growth and development of Vriesea friburgensis and Alcantarea imperialis seedlings in the context of oxidative stress. The optimal temperatures for the growth of V. friburgensis and A. imperialis were 25 and 25-30 °C, respectively. Extreme temperatures (15, 30, or 35 °C) induced oxidative stress in both species with significant accumulation of hydrogen peroxide (H2O2) and nitric oxide (NO). Under oxidative stress, the amount of chlorophyll decreased in both species, more prominently in V. friburgensis, while carotenoid levels dramatically increased in A. imperialis. Notably, the activities of superoxide dismutase, catalase (CAT), and ascorbate peroxidase increased in A. imperialis at extreme temperatures. Similar results were observed for V. friburgensis; however, the activity of CAT remained unaffected regardless of temperature. Seedlings of A. imperialis survived at a wider range of temperatures than V. friburgensis, which had greater than 40% mortality when growing at 30 °C. Overall, precise control of cellular H2O2 and NO levels takes place during the establishment of A. imperialis seedlings, allowing the species to cope with relatively high temperatures. The thermal limits of the fundamental niches of the species investigated, determined based on the ability of seedlings to cope with oxidative stress, were distinct from the realized niches of these species. The results suggest that recruitment success is dependent on the ability of seedlings to handle extreme temperature-triggered oxidative stress, which limits the regeneration niche.

Highly functionalized piperidines: Free radical scavenging, anticancer activity, DNA interaction and correlation with biological activity.

Twenty-five piperidines were studied as potential radical scavengers and antitumor agents. Quantitative interaction of compounds with ctDNA using spectroscopic techniques was also evaluated. Our results demonstrate that the evaluated piperidines possesses different abilities to scavenge the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the anion radical superoxide (•O2-). The piperidine 19 was the most potent radical DPPH scavenger, while the most effective to •O2- scavenger was piperidine 10. In general, U251, MCF7, NCI/ADR-RES, NCI-H460 and HT29 cells were least sensitive to the tested compounds and all compounds were considerably more toxic to the studied cancer cell lines than to the normal cell line HaCaT. The binding mode of the compounds and ctDNA was preferably via intercalation. In addition, these results were confirmed based on theoretical studies. Finally, a linear and exponential correlation between interaction constant (Kb) and GI50 for several human cancer cell was observed.

Physiological traits and antioxidant metabolism of leaves of tropical woody species challenged with cement dust.

Tropical woody species occurring in limestone outcrops are frequently exposed to particulate material from cement factories. The effects of 60-day cement dust exposure on physiological traits and enzymatic antioxidant system of young plant leaves of Guazuma ulmifolia Lam., Myracrodruon urundeuva Allemão and Trichilia hirta L. were investigated. Cement dust (2.5 or 5mgcm-2) was applied to the leaf surface or soil or both (leaf plus soil) and plants were maintained at greenhouse. Cement dust barely affected the mineral nutrient levels, except for iron whose content was decreased in leaves/leaflets of all species studied. The incident light was partly blocked in cement dust-treated leaves, regardless of the plant species, causing a decrease in the photosynthetic pigments in M. urundeuva. The chlorophyll b content, however, increased in G. ulmifolia and T. hirta leaves upon cement dust treatment. The potential quantum yield of photosystem II in challenged leaves of G. ulmifolia was 3.8% lower than that of control plants, while such trait remained unaffected in the leaves of the other species. No changes in leaf stomatal conductance and antioxidant enzymes activities were observed, except for M. urundeuva, which experienced a 31% increment in the superoxide dismutase activity upon 5mgcm-2 cement dust (leaf plus soil treatment), when compared with control plants. Overall, the mild changes caused by cement dust in the in physiological and biochemical traits of the species studied indicate that such species might be eligible for further studies of revegetation in fields impacted by cement factories.

Salinity-induced accumulation of endogenous H2S and NO is associated with modulation of the antioxidant and redox defense systems in Nicotiana tabacum L. cv. Havana.

Salinity is one of the abiotic factors that most affect crop growth and production. This study focused on the effect of high salinity on the endogenous levels of the signaling molecules hydrogen sulfite (H2S) and nitric oxide (NO) in Nicotiana tabacum leaves and the extent of these for the biochemically-driven plant tolerance to such abiotic stress. The NaCl treatment for 10days led to an expressive augment of H2S and NO levels. This increase was correlated with the raise of l-Cys and l-Arg and the induction of l-cysteine desulfhydrase, cyanoalanine synthase, cysteine synthase, nitrate reductase and arginase, enzymes known to be involved in the biosynthesis of H2S or NO. The enzymatic antioxidant system (superoxide dismutase and catalase activity) was boosted and the non-enzymatic antioxidant glutathione was intensively oxidized in leaves upon stress allowing plants to cope with oxidative stress. Lower stomatal conductance was observed in stressed plants in comparison with control ones. Moreover, the high activity of antioxidant enzymes and high rate of glutathione oxidation following salt stress were considerably decreased upon NO or H2S scavenging. Thus, increment in NO and H2S levels and their interplay, along with metabolic and physiological changes, contributed to tobacco survival to extreme salinity conditions.

Quinolines: Microwave-assisted synthesis and their antifungal, anticancer and radical scavenger properties.

An efficient method for the synthesis of quinolines using microwave irradiation was developed providing 28 quinolines with good yields. The reaction procedures are environmentally friendly, convenient, mild and of easy work-up. Quinolines were evaluated for their antifungal, anticancer and antioxidant properties and exhibited high activities in all tests performed.

Impact of cement dust pollution on Cedrela fissilis Vell. (Meliaceae): A potential bioindicator species.

Considering the impacts caused to vegetation in the vicinity of cement factories, the aim of this study was to evaluate the impacts of cement dust on the structural organization and physiological/biochemical traits of Cedrela fissilis leaflets, a woody species native to tropical America. Plants were exposed to 2.5 or 5 mg cm-2 cement dust applied to the leaf surface, to the soil or simultaneously to the leaf surface and the soil.. Leaves of shoot-treated plants exhibited chlorosis, marginal and inter veins necrosis, diminished thickness, epidermal cells less turgid, cellular collapse, obstructed stomata, senescence, rolling and some abscission. In few cases, individual death was recorded. Cement dust-treated plants also presented decreased amount of photosynthetic pigments and iron (Fe) and increase in calcium (Ca) levels. The cement crust formed in leaves surface blocked from 30 to 50% of the incoming light and reduced the stomatal conductance and the potential quantum yield of photosystem II. Control or soil-treated plants did not exhibit morphophysiological changes throughout the experiment. The activity of superoxide dismutase, catalase and ascorbate peroxidase increased in leaves of plants upon treatment with 2.5 mg cm(-2) cement dust, independent of the site application. Overall, these results indicate that C. fissilis is highly sensitive to cement dust at the initial stage of development.

Leaf structural traits of tropical woody species resistant to cement dust.

Cement industries located nearby limestone outcrops in Brazil have contributed to the coating of cement dust over native plant species. However, little is known about the extent of the response of tropical woody plants to such environmental pollutant particularly during the first stages of plant development and establishment. This work focused on the investigation of possible alterations in leaf structural and ultrastructural traits of 5-month-old Guazuma ulmifolia Lam. (Malvaceae), 6-month-old Myracrodruon urundeuva Allemão (Anacardiaceae), and 9-month-old Trichilia hirta L. (Meliaceae) challenged superficially with cement dust during new leaf development. Leaf surface of plants, the soil or both (leaf plus soil), were treated (or not) for 60 days, under controlled conditions, with cement dust at 2.5 or 5.0 mg cm(-2). After exposure, no significant structural changes were observed in plant leaves. Also, no plant death was recorded by the end of the experiment. There was also some evidence of localized leaf necrosis in G. ulmifolia and T. hirta, leaf curling in M. urundeuva and T. hirta, and bulges formation on epidermal surface of T. hirta, after cement dust contact with plant shoots. All species studied exhibited stomata obliteration while T. hirta, in particular, presented early leaf abscission, changes in cellular relief, and organization and content of midrib cells. No significant ultrastructural alterations were detected under the experimental conditions studied. Indeed, mesophyll cells presented plastids with intact membrane systems. The high plant survival rates, together with mild morphoanatomic traits alterations in leaves, indicate that G. ulmifolia is more resistant to cement dust pollutant, followed by M. urundeuva and T. hirta. Thus, the three plant species are promising for being used to revegetate areas impacted by cement industries activities.

Stressing conditions as tools to boost the biosynthesis of valuable plant natural products.

There is a consensus that plants are great sources of metabolites with a broad variety of functions. This is particularly important because plants cannot run away from environmental conditions that can threat their existence. The numerous biological activities exhibited by plant natural products prompted humanity to use such substances or their derivatives for the treatment and/or prevention of diseases. The more we know the flora around the world the higher is the chance to find new lead compounds for the design of more potent drugs or nutraceuticals. This review first deals with Brazilian flora, contextualizing the most studied medicinal species and related patents. It also describes a compilation of relevant works based on the use of stress conditions to enhance the biosynthesis of valuable metabolites in cell cultures, tissue cultures (hairy roots) and whole plants by using native or crop plants around the world.

Synthesis, kinetic studies and molecular modeling of novel tacrine dimers as cholinesterase inhibitors.

This study presents the synthesis of 15 new tacrine dimers as well as the Ki and IC50 results, studies of the kinetic mechanism, and molecular docking analysis of the dimers in relation to the cholinesterases hAChE, hBChE, EeAChE and eqBChE. In addition to spectroscopic characterization, X-ray structure determination was performed for two of the new compounds. These new dimers were found to be mixed nanomolar inhibitors of the evaluated targets with a broad and significant selectivity profile, and these properties are dependent on both the type of the linker and the volume of the hydroacridine alicyclic ring. The results indicate that the aromatic linkers play a significant role in generating specific interactions with the half-gorge region of the catalytic center. Thus, these types of linkers can positively modulate the electronic properties of the tacrine dimers studied with an improvement of their cholinesterase inhibition activity.

Cryptococcus gattii: in vitro susceptibility to photodynamic inactivation.

Cryptococus gattii is an emergent primary human pathogen that causes meningismus, papilledema, high intracranial pressure and focal involvement of the central nervous system in immunocompetent hosts. Prolonged antifungal therapy is the conventional treatment, but it is highly toxic, selects for resistant strains, contributes to therapy failure and has a poor prognosis. Photodynamic inactivation (PDI) offers a promising possibility for the alternative treatment of cryptococcosis. The aim of this study was to test the effectiveness of toluidine blue O (TBO) and light-emitting diode (LED) against C. gattii strains with distinct susceptibility profile to antifungal drugs (amphotericin B: 0.015-1.0 µg mL(-1); itraconazole: 0.015-2 µg mL(-1); fluconazole: 4-64 µg mL(-1)). Using 25 µM (6.76 µg mL(-1)) TBO and LED energy density of 54 J cm(-2) these fungal isolates presented variable susceptibility to PDI. The production of reactive oxygen species (ROS)/peroxynitrite was determined, and the catalase and peroxidase activities were measured. After PDI, high amounts of ROS/peroxynitrite are produced and higher catalase and peroxidase activities could be correlated with a lower susceptibility of C. gattii isolates to PDI. These results indicate that PDI could be an alternative to C. gattii growth inhibition, even of isolates less susceptible to classical antifungal drugs, also pointing to mechanisms related to their variable susceptibility behavior.

Nitric oxide synthase-mediated phytoalexin accumulation in soybean cotyledons in response to the Diaporthe phaseolorum f. sp. meridionalis elicitor.

Phytoalexin biosynthesis is part of the defense mechanism of soybean (Glycine max) plants against attack by the fungus Diaporthe phaseolorum f. sp. meridionalis (Dpm), the causal agent of stem canker disease. The treatment of soybean cotyledons with Dpm elicitor or with sodium nitroprusside (SNP), a nitric oxide (NO) donor, resulted in a high accumulation of phytoalexins. This response did not occur when SNP was replaced by ferricyanide, a structural analog of SNP devoid of the NO moiety. Phytoalexin accumulation induced by the fungal elicitor, but not by SNP, was prevented when cotyledons were pretreated with NO synthase (NOS) inhibitors. The Dpm elicitor also induced NOS activity in soybean tissues proximal to the site of inoculation. The induced NOS activity was Ca(2+)- and NADPH-dependent and was sensitive to the NOS inhibitors N(G)-nitro-L-arginine methyl ester, aminoguanidine, and L-N(6)-(iminoethyl) lysine. NOS activity was not observed in SNP-elicited tissues. An antibody to brain NOS labeled a 166-kD protein in elicited and nonelicited cotyledons. Isoflavones (daidzein and genistein), pterocarpans (glyceollins), and flavones (apigenin and luteolin) were identified after exposure to the elicitor or SNP, although the accumulation of glyceollins and apigenin was limited in SNP-elicited compared with fungal-elicited cotyledons. NOS activity preceded the accumulation of these flavonoids in tissues treated with the Dpm elicitor. The accumulation of these metabolites was faster in SNP-elicited than in fungal-elicited cotyledons. We conclude that the response of soybean cotyledons to Dpm elicitor involves NO formation via a constitutive NOS-like enzyme that triggers the biosynthesis of antimicrobial flavonoids.