Gas exchange and osmotic adjustment in cotton cultivars subjected to severe salt stress.

Salinity is harmful to crops when the concentration of soluble salts overcomes the salinity threshold of the crop, causing osmotic stress and limitations in plant growth. In this scenario, adopting tolerant cultivars is the most adequate strategy to minimize agricultural losses. However, the inheritance of tolerance depends on the genotype. From this perspective, this study assessed the tolerance to severe salt stress in 11 cotton cultivars based on gas exchange parameters and the free proline content. The cultivars were grown in a greenhouse and subjected to 34 days of saline irrigation (10 dS m-1), starting 45 days after seedling emergence (B1 phase). Plant growth was monitored weekly until the end of the salt stress period. The treatments consisted of a combination of two factors: eleven cultivars associated with two electrical conductivity levels of irrigation water (ECw: 0.3 and 10.0 dS m-1). The experimental design was in randomized blocks in a 11 × 2 factorial arrangement with three replications (66 plots), with the experimental unit consisting of one plant per plot. Salinity impacted plant growth, being reflected on the gas exchange and free proline data of most cultivars. However, BRS 286, FMT 705, BRS 416, and BRS Acácia, and CNPA 7MH withstood the effects of stress and osmotically adjusted to the salt stress conditions, thus minimizing the damage to growth. Those cultivars are the most indicated for improvement programs aiming at tolerance to salt stress based on the results found in this research.

Calcium pyruvate in the attenuation of the water deficit on the agro-industrial quality of ratoon sugarcane.

Sugarcane is one of the largest agricultural commodities when considering the export volume and the number of jobs generated. Sugarcane production in the Brazilian Northeast region is generally low due to several factors, including the irregular rainfall distribution, which highlights the importance of studies aimed at mitigating the deleterious effects of water stress. In this scenario, this study aimed to evaluate calcium pyruvate as a water deficit attenuator on the agro-industrial quality of sugarcane in the second cycle of cultivation. The experiment was conducted out under greenhouse conditions of the Federal University of Campina Grande, where five sugarcane commercial genotypes tested (G1- RB863129, G2- RB92579, G3- RB962962, G4- RB021754, and G5- RB041443) and three irrigation management strategies (E1- full irrigation, E2- water deficit with application of 30 mM of calcium pyruvate, and E3- water deficit without calcium pyruvate application), distributed in randomized blocks in 5 × 3 factorial arrangement with three replications. The RB021754 genotype under water deficit and without foliar application of calcium pyruvate increased the fiber content (13.2%) and the sugarcane moist cake weight (143.5 g). The effects of water deficit in sugarcane genotypes are attenuated by the exogenous application of 30 mM of calcium pyruvate, with benefits on the polarized sucrose content, apparent sucrose content of the juice, soluble solids content, purity, corrected cane POL, total recoverable sugars, and stem mass in relation to plants under water deficit without calcium pyruvate application.

Salicylic acid does not relieve salt stress on gas exchange, chlorophyll fluorescence, and hydroponic melon growth.

The objective of this study was to evaluate the salicylic acid applications in attenuating the harmful effects of saline nutrient solution on the physiology and growth of 'Gaúcho' melon cultivated in the NFT hydroponic system. The experiment was conducted in a greenhouse, in Pombal-PB, Brazil. The cultivation system used was the Nutrient Film Technique - NFT hydroponics. A completely randomized split-plot design was used, with the plot referring to four levels of salinity in the nutrient solution - ECns (2.1 control, 3.2, 4.3, and 5.4 dS m-1) and the sub-plot four concentrations of salicylic acid (SA) (0, 1.5, 3.0, and 4.5 mM), applied via foliar spray, with six replications. Nutrient solution of 4.3 and 5.4 dS m-1 electrical conductivity promotes higher maximum and variable fluorescence, respectively. The stomatal conductance, transpiration, stem diameter, main branch length, leaf dry mass, and stem dry mass of 'Gaúcho' melon plants decrease with the increase in salinity of the nutrient solution. Salicylic acid increases the initial fluorescence and the main branch length of 'Gaúcho' melon plants in hydroponic cultivation. Salicylic acid at a concentration of 1.5 to 4.5 mM did not attenuate the effects of salt stress on the internal CO2 concentration, CO2 assimilation rate, and root dry mass of 'Gaúcho' melon plants.

Pyruvic acid as attenuator of water deficit in cotton plants varying the phenological stage.

The lack of water during crop growth causes damage to any production system, especially when it occurs during the initial establishment or beginning of the reproductive stage. Although cotton can be properly managed in regions with water limitation, its yield is affected at different levels according to the genetics of the cultivar adopted. Exogenous application of some organic components has shown a stress-mitigating effect and can be a valuable procedure to enhance the yield of water stress-sensitive cultivars. The objective of this work was to evaluate the benefits of exogenous application of pyruvic acid (100 µM) in cotton plants under water deficit varying the phenological stage of the crop. The experiment was conducted in a greenhouse, where the plants were grown in pots and subjected to seven days of water suspension, initiated individually in stages V2 and B1. Each pot contained two plants. The treatments adopted were: T1 - control, T2 - water suppression; and T3 - water suppression + pyruvate application. The design was randomized blocks in a factorial scheme (3 × 3) with three replicates. The reductions in gas exchange and growth of the cultivars BRS Seridó, CNPA 7MH and FM 966 were more significant in the reproductive stage, especially for FM 966, which was more sensitive. Pyruvate application reduced the effects of water suppression on boll production by 31% in BRS Seridó and 34% in CNPA 7MH and FM 966.

Salicylic acid does not mitigate salt stress on the morphophysiology and production of hydroponic melon.

The excess of salts present in the water can limit the hydroponic cultivation of melon in semi-arid regions of the Brazilian Northeast, making it necessary to use strategies that allow the use of these waters. Among these strategies, the use of elicitor substances stands out, such as salicylic acid. In this context, this study aimed to evaluate the effect of foliar application of salicylic acid in mitigating the harmful effects of salt stress on the morphophysiology and production of 'Gaúcho' melon cultivated in a hydroponic system. A completely randomized design was adopted in a split-plot scheme, with four levels of electrical conductivity of the nutrient solution - ECsn (2.1, 3.2, 4.3, and 5.4 dS m-1) considered the plots and four salicylic acid concentrations - SA (0, 1.5, 3.0, and 4.5 mM), the subplots, with six replications. The foliar application of salicylic acid concentrations did not mitigate the deleterious effects of salt stress on the morphophysiology and yield of melon grown in hydroponic system. The concentration of 4.5 mM of salicylic acid intensified the harmful effects of the salinity of the nutrient solution on gas exchange and fresh weight of hydroponic melon.

Can Bradyrhizobium strains inoculation reduce water deficit effects on peanuts?

Drought is one of the environmental factors that most affects peanut cultivation in semi-arid regions, resulting in economic losses to growers. However, growth promoting bacteria are able to reduce water deficit damage in some plant species. In this context, this study aimed to evaluate the interaction of Bradyrhizobium strains reducing water stress effects on peanut genotypes by antioxidant enzymes activities, leaf gas exchanges and vegetative growth, as well as to determine the taxonomic positioning of strain ESA 123. The 16S rRNA gene of ESA 123 was amplified by PCR and sequenced by dideoxy Sanger sequencing method. An experiment was performed in greenhouse with three peanut genotypes (BRS Havana, CNPA 76 AM and 2012-4), two Bradyrhizobium strains (SEMIA 6144 and ESA 123), a mineral source of N and an absolute control (without N) under two water regimes (with and without irrigation). Seeds of peanut were sown and the plants were grown until 30 days after emergence. On the 20th day, the water deficit plants group had their irrigation suspended for 10 days. At in silico analyzes, ESA 123 presented 98.97% similarity with the type strain of B. kavangense. Leaf gas exchange was affected by water deficit; as well as alteration of antioxidant activities and reduction of vegetative growth variables. However, some plants inoculated with SEMIA 6144 and ESA 123 strains presented lower reductions and increment of some evaluated variables, mainly the ones inoculated with the ESA 123 strain, Bradyrhizobium sp. from the semi-arid region of Northeast Brazil. This data suggests beneficial effects of the peanut-Bradyrhizobium interaction in a water stress condition, specially with the ESA 123 strain.

ADME studies and preliminary safety pharmacology of LDT5, a lead compound for the treatment of benign prostatic hyperplasia.

This study aimed to estimate the absorption, distribution, metabolism and excretion (ADME) properties and safety of LDT5, a lead compound for oral treatment of benign prostatic hyperplasia that has previously been characterized as a multi-target antagonist of α1A-, α1D-adrenoceptors and 5-HT1A receptors. The preclinical characterization of this compound comprised the evaluation of its in vitro properties, including plasma, microsomal and hepatocytes stability, cytochrome P450 metabolism and inhibition, plasma protein binding, and permeability using MDCK-MDR1 cells. De-risking and preliminary safety pharmacology assays were performed through screening of 44 off-target receptors and in vivo tests in mice (rota-rod and single dose toxicity). LDT5 is stable in rat and human plasma, human liver microsomes and hepatocytes, but unstable in rat liver microsomes and hepatocytes (half-life of 11 min). LDT5 is highly permeable across the MDCK-MDR1 monolayer (Papp ∼32×10-6 cm/s), indicating good intestinal absorption and putative brain penetration. LDT5 is not extensively protein-bound and is a substrate of human CYP2D6 and CYP2C19 but not of CYP3A4 (half-life >60 min), and did not significantly influence the activities of any of the human cytochrome P450 isoforms screened. LDT5 was considered safe albeit new studies are necessary to rule out putative central adverse effects through D2, 5-HT1A and 5-HT2B receptors, after chronic use. This work highlights the drug-likeness properties of LDT5 and supports its further preclinical development.

ADME studies and preliminary safety pharmacology of LDT5, a lead compound for the treatment of benign prostatic hyperplasia

This study aimed to estimate the absorption, distribution, metabolism and excretion (ADME) properties and safety of LDT5, a lead compound for oral treatment of benign prostatic hyperplasia that has previously been characterized as a multi-target antagonist of α1A-, α1D-adrenoceptors and 5-HT1A receptors. The preclinical characterization of this compound comprised the evaluation of its in vitro properties, including plasma, microsomal and hepatocytes stability, cytochrome P450 metabolism and inhibition, plasma protein binding, and permeability using MDCK-MDR1 cells. De-risking and preliminary safety pharmacology assays were performed through screening of 44 off-target receptors and in vivo tests in mice (rota-rod and single dose toxicity). LDT5 is stable in rat and human plasma, human liver microsomes and hepatocytes, but unstable in rat liver microsomes and hepatocytes (half-life of 11 min). LDT5 is highly permeable across the MDCK-MDR1 monolayer (Papp ∼32×10-6 cm/s), indicating good intestinal absorption and putative brain penetration. LDT5 is not extensively protein-bound and is a substrate of human CYP2D6 and CYP2C19 but not of CYP3A4 (half-life >60 min), and did not significantly influence the activities of any of the human cytochrome P450 isoforms screened. LDT5 was considered safe albeit new studies are necessary to rule out putative central adverse effects through D2, 5-HT1A and 5-HT2B receptors, after chronic use. This work highlights the drug-likeness properties of LDT5 and supports its further preclinical development.

Atividade analgésica e antiedematogênica de Polygala paniculata L. (Polygalaceae) selvagem e obtida por micropropagação / Analgesic and antiedematogenic activities of wild and micropropagated Polygala paniculata L (Polygalaceae)

Polygala paniculata L. (Polygalaceae) é uma erva que ocorre em todas as regiões do Brasil. No presente trabalho, foram avaliadas as atividades analgésica, através do teste da placa quente, da retirada de cauda e da formalina, e antiedematogênica, através do teste do edema de orelha induzido por óleo de cróton, dos extratos etanólicos obtidos das partes aéreas de Polygala paniculata selvagem e cultivadas por micropropagação. A aplicação oral do extrato etanólico de Polygala paniculata apresentou atividade analgésica, em ratos, tanto em testes de dor induzida por agentes térmicos (testes da placa quente e de retirada da cauda) quanto por agentes químicos (teste da formalina), de modo que os melhores resultados foram obtidos na dose de 400 mg/kg. Também foi observada redução na formação de edema de orelha induzida pela aplicação de óleo de cróton. Os efeitos provocados pelos extratos obtidos a partir das plantas cultivadas in vitro foram menos pronunciados que aqueles produzidos pelos extratos das plantas selvagens, embora ambos tenham sido significativos. Estes resultados sugerem que o extrato etanólico de Polygala paniculata possui atividades analgésica e antiedematogênica.

The ethanolic extracts of Polygala paniculata L. (Polygalaceae), wich is a herbaceous plant widely distributed all over Brazil, were tested for their analgesic effects using hot plate, tail flick and formalin test models, and for their antiedematogenic effects using croton oil induced ear oedema. The ethanolic extracts obtained from wild and micropropagated plants produced analgesic effects against thermal and chemical induced pain. The highest results were observed at the dose of 400 mg/kg. The inhibition of ear oedema in mice was also observed after treatment with ethanolic extract of Polygala paniculata. The effects produced by micropropagated plants were lower than wild plants, whereas both had produced significant effects. These results suggest that the ethanolic extracts from wild and micropropagated Polygala paniculata possess analgesic and antiedematogenic effects.

Anti-inflammatory and analgesic activity of Bouchea fluminensis.

Dried leaves extract from Bouchea fluminensis was assessed in anti-inflammatory (mouse paw edema) and analgesic models (acetic acid-induced writhings and hot plate). Oral pretreatment of animals with a crude mixture (IG) and a purified mixture of ursolic, oleanolic and micromeric acids (IG-59) at doses ranging from 1 to 30 mg/kg, significantly inhibited carrageenin-induced edema formation. At the same doses, IG and IG-59 also exhibited peripheral and central analgesic activity. It seems that B. fluminensis triterpenes develop their analgesic effect through central opioid receptors, due to the observation that naloxone reverted analgesic activity on the hot plate model.

Ação de extratos do açaí (Euterpe oleracea Mart. ) sobre a produção de óxido nítrico em células RAW 264.7

Açaí (Euterpe oleracea Mart.), é uma palmeira tropical muito apreciada por sua beleza e valor nutricional. Estudos químicos revelaram a presença de ácidos graxos e esteróides. No presente trabalho foi avaliada a ação de extratos obtidos dos frutos e flores sobre a produção de óxido nítrico (ON), molécula que apresenta várias atividades fisiológicas, tais como vasodilatação, neurotransmissão, além de atividades tumoricidas e citotóxicas. Células Raw 264,7 estimuladas com lipopolissacarídeo bacteriano (LPS, 100 ng/ml) e interferon-alfa (IFN-alfa, 10 U/ml) produziram grande quantidade de óxido nítrico (35 μM) quando comparadas com as células não estimuladas (3 μM). Os extratos com hexano, diclorometano, acetato de etila e n-butanol apresentaram alta capacidade de inibição em células ativadas com LPS e IFN-alfa, de acordo com a concentração, sendo que na concentração mais alta ocorreu uma inibição de quase 100 por cento. Também avaliamos se o efeito inibitório seria devido a seqüestro do radical livre (ON), através do uso do SNAP (um doador de ON). Somente o extrato em acetato de etila mostrou atividade sequestrante. Esforços estão sendo empregados na tentativa de compreender os possíveis mecanismos associados ao efeito inibitório destes extratos.

Açaí (Euterpe oleracea Mart.) is a tropical palm tree appreciated for its attractive beauty and for nutritional purposes. Chemical studies have revealed the presence of fatty acids and steroids. In the present work, it has been tested the action of the extracts obtained from the fruits and flowers on the nitric oxide (NO) production, a very important molecule with a lot of physiological rules such as vasodilatation, neurotransmission, tumoricidal and cytotoxic activity. Cells RAW 264.7 stimulated with bacterial lipopolysaccharide (LPS, 100 ng/ml) and interferonalpha (IFN-alpha, 10 U/ml) produce large amounts of nitric oxide (35 μM) when compared with non-stimulated cells (3μM). The hexane, dichloromethane, ethyl acetate and n-butanol extracts have shown high inhibition capacity, concentration-dependent in the cells activated with LPS and IFN-alpha, and the highest concentration has promoted almost 100 percent of inhibition. We also have tested if the inhibitory effect was due to a scavenger action using a NO donor, the SNAP. Only the ethyl acetate extract has shown significant scavenger action. At this moment an effort is going on to try to understand the possible mechanisms associated to the inhibition of those extracts.

Role of nitric oxide and superoxide in Giardia lamblia killing.

Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (approximately 60% dead trophozoites). The effect was inhibited (> 90%) by an NO synthase inhibitor (200 microM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors, S-nitroso-acetyl-penicillamine (SNAP) and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 microM, respectively). A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentrations up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (approximately 35% dead trophozoites at 1 mM). The mixtures of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoites killing and this effect is not mediated by peroxynitrite.

Role of nitric oxide and superoxide in Giardia lamblia killing

Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (~60 per cent dead trophozoites). This effect was inhibited (>90 per cent) by an NO synthase inhibitor (200 muM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors S-nitroso-acetyl-penicillamine(SNAP)and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 muM, respectively). A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentration up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (~35 per cent dead trophozoites at 1 mM). The mixture of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoite killing and this effect is not mediated by peroxynitrite.

Depolymerization of macrophage microfilaments prevents induction and inhibits activity of nitric oxide synthase.

We have investigated the relationship between peritoneal murine macrophage cytoskeleton and nitric oxide (NO) synthase (NOS). Activation of the cells with lipopolysaccharide plus interferon-gamma (LI) induced iNOS, detected by nitrite or by labeled L-citrulline production and by a specific antibody against macrophage iNOS. Addition of cytochalasin B (a microfilament-depolymerizing agent) caused a dose-dependent inhibition in NO production by macrophages, whereas colchicine (a microtubule depolymerizing agent) inhibited it only by 20% and not dose-dependently. Addition of cytochalasin B together with LI abolished nitrite and L-citrulline accumulation as well as the amount of iNOS antigen in activated macrophage. Moreover, addition of cytochalasin B 6 or 12 h after stimulus, also decreased the nitrite and L-citrulline production by macrophages although iNOS antigen content by Western blot was the same in the presence or in the absence of cytochalasin B added 12 h after activation. Since cytochalasin B failed to inhibit iNOS activity directly, its inhibitory effects on NO production by macrophages is likely to be indirect, through microfilament network in central regions of cells, but not in filaments seen at pseudopodia or edging processes. Our findings demonstrate that disruption of microfilaments but not of microtubules prevents the iNOS induction process and inhibits its enzymatic activity in activated macrophages.

Inhibition of rat paw oedema and pleurisy by the extract from Mandevilla velutina.

This study reports the oral anti-inflammatory profile of the crude extract (CE) of Mandevilla velutina, a plant which has been previously demonstrated to selectively antagonize bradykinin response of the isolated tissues on rat paw oedema and pleurisy caused by different phlogistic agents. The CE (50 to 200 mg/kg), given 60 min before, inhibited in a dose-dependent manner bradykinin (BK) and cellulose sulphate-induced paw oedema, maximal inhibition of 59% and 65%, respectively. In the same range dose the CE also significantly antagonized pleural exudate and cell infiltration caused by these substances, maximal inhibition of 34% and 46%, respectively. In addition, the CE (100 and 200 mg/kg) also inhibited paw oedema induced by serotonin, PAF-acether and zymosan, maximal inhibition of 55%, 38% and 46%, respectively, but enhanced histamine oedema. However, the CE revealed only partial or no inhibition in pleural exudate caused by these agents. The CE (100 and 200 mg/kg) also inhibited in a dose and time-dependent manner carrageenan-induced paw oedema with a maximal inhibition of 44%, but only partially affected carrageenan-induced pleural exudate. The CE also partially inhibited dextran oedema, but even at a higher dose (400 mg/kg) it failed to interfere with Bothrops Jaracaca-induced paw oedema. The CE inhibited BK and to a lesser extent cellulose sulphate-induced cell migration, but failed to interfere with the differential leukocyte migration in the pleural cavity. These findings provide evidence that the CE from M. velutina, besides antagonizing kinin action, exhibit an oral anti-oedematogenic activity against a variety of phologistic agents, but it was more effective in inhibiting those models where kinins are more involved.

Comparative effects of two potentiating peptides (KPP and BPP9a) on kinin-induced rat paw edema.

We have previously shown that KPP, a kinin potentiating peptide generated by tryptic digestion of human plasma proteins potentiated kinin effects on isolated smooth muscle preparations like guinea-pig ileum with high potency and specificity. We also obtained evidence suggesting that, unlike other potentiating peptides, KPP exerts its effect by a mechanism different from the inhibition of kinin metabolism by angiotensin converting enzyme, neutral endopeptidase and kininase I. Here we show the potentiating effect of KPP and of BPP9a, a potentiator derived from snake venom, towards the rat paw edema induced by bradykinin (BK). Our results show that: a) KPP is 25-fold more active than BPP9a in potentiating rat paw edema elicited by BK: b) like BPP9a, KPP is specific in potentiating kinin-induced edema, being ineffective in potentiating edema induced by histamine or serotonin; and c) DesArg9-BK (DABK) elicits a small edematogenic response which can be potentiated by both KPP and BPP9a.

Differential inhibition by two hetrazepine PAF antagonists of acute inflammation in the mouse.

1. The injection of 100 or 300 micrograms of carrageenin into the mouse paw or pleural cavity produced a delayed inflammatory reaction at 48 h while platelet activating factor (PAF)-induced paw oedema and pleurisy were maximal 30 min after its injection. 2. The PAF antagonist, WEB 2086, failed to inhibit mouse paw oedema and pleurisy induced by PAF, but reduced the first phase of oedema (1-4 h) induced by carrageenin without interfering with the second one (48-72 h). In contrast, another structurally-related PAF antagonist, WEB 2170, inhibited dose-dependently both oedema and pleurisy induced by PAF and by carrageenin (48 h). 3. Repeated injections of PAF into the mouse paw or pleural cavity led to significant autodesensitization. The animals desensitized to PAF and injected with carrageenin also displayed a significantly reduced oedema. 4. Our results suggest that PAF may be involved in the inflammatory response to carrageenin in mice. Furthermore, because the different receptor antagonists displayed distinct effects against PAF itself, different sites for in vivo interaction of PAF are available and are species- and drug-dependent.