In vitro antitumor activity of methotrexate via pH-sensitive chitosan nanoparticles

Nanoparticles with pH-sensitive behavior may enhance the success of chemotherapy in many cancers by efficient intracellular drug delivery. Here, we investigated the effect of a bioactive surfactant with pH-sensitive properties on the antitumor activity and intracellular behavior of methotrexate-loaded chitosan nanoparticles (MTX-CS-NPs). NPs were prepared using a modified ionotropic complexation process, in which was included the surfactant derived from Nα,Nε-dioctanoyl lysine with an inorganic lithium counterion. The pH-sensitive behavior of NPs allowed accelerated release of MTX in an acidic medium, as well as membrane-lytic pH-dependent activity, which facilitated the cytosolic delivery of endocytosed materials. Moreover, our results clearly proved that MTX-CSNPs were more active against the tumor HeLa and MCF-7 cell lines than the free drug. The feasibilty of using NPs to target acidic tumor extracellular pH was also shown, as cytotoxicity against cancer cells was greater in a mildly acidic environment. Finally, the combined physicochemical and pH-sensitive properties of NPs generally allowed the entrapped drug to induce greater cell cycle arrest and apoptotic effects. Therefore, our overall results suggest that pH-sensitive MTX-CS-NPs could be potentially useful as a carrier system for tumor and intracellular drug delivery in cancer therapy.

Membrane-destabilizing activity of pH-responsive cationic lysine-based surfactants: role of charge position and alkyl chain length

Many strategies for treating diseases require the delivery of drugs into the cell cytoplasm following internalization within endosomal vesicles. Thus, compounds triggered by low pH to disrupt membranes and release endosomal contents into the cytosol are of particular interest. Here, we report novel cationic lysine-based surfactants (hydrochloride salts of N¿- and N¿-acyl lysine methyl ester) that differ in the position of the positive charge and the length of the alkyl chain. Amino acid-based surfactants could be promising novel biomaterials in drug delivery systems, given their biocompatible properties and low cytotoxic potential. We examined their ability to disrupt the cell membrane in a range of pH values, concentrations and incubation times, using a standard hemolysis assay as a model of endosomal membranes. Furthermore, we addressed the mechanism of surfactant-mediated membrane destabilization, including the effects of each surfactant on erythrocyte morphology as a function of pH. We found that only surfactants with the positive charge on the ¿-amino group of lysine showed pH-sensitive hemolytic activity and improved kinetics within the endosomal pH range, indicating that the positive charge position is critical for pH-responsive behavior. Moreover, our results showed that an increase in the alkyl chain length from 14 to 16 carbon atoms was associated with a lower ability to disrupt cell membranes. Knowledge on modulating surfactant-lipid bilayer interactions may help us to develop more efficient biocompatible amino acid-based drug delivery devices.

pH Sensitive surfactants from lysine: assessment of their cytotoxicity and environmental behavior

The toxicity and environmental behavior of new pH-sensitive surfactants from lysine are presented. Three different chemical structures are studied: surfactants with one amino acid and one alkyl chain, surfactants with two amino acids on the polar head and one alkyl chain, and gemini surfactants. The pH sensitivity of these compounds can be tuned by modifying their chemical structures. Cytotoxicity has been evaluated using erythrocytes and fibroblast cells. The toxic effects against these cells depend on the hydrophobicity of the molecules as well as their cationic charge density. The effect of hydrophobicity and cationic charge density on toxicity is different for each type of cells. For erythrocytes, the toxicity increases as hydrophobicity and charge density increases. Nevertheless, for fibroblasts cationic charge density affects cytotoxicity in the opposite way: the higher charge density, the lower the toxicity. The effect of the pH on hemolysis has been evaluated in detail. The aquatic toxicity was established using Daphnia magna. All surfactants yielded EC50 values considerably higher than that reported for cationic surfactants based on quaternary ammonium groups. Finally, their biodegradability was evaluated using the CO2 headspace test (ISO 14593). These lysine derivatives showed high levels of biodegradation under aerobic conditions and can be classified as"readily biodegradable compounds".

Organic and Nitrogen Fertilization of Soil under ‘Syrah’ Grapevine: Effects on Soil Chemical Properties and Nitrate Concentration

ABSTRACT Viticulture is an activity of great social and economic importance in the lower-middle region of the São Francisco River valley in northeastern Brazil. In this region, the fertility of soils under vineyards is generally poor. To assess the effects of organic and nitrogen fertilization on chemical properties and nitrate concentrations in an Argissolo Vermelho-Amarelo (Typic Plinthustalf), a field experiment was carried out in Petrolina, Pernambuco, on Syrah grapevines. Treatments consisted of two rates of organic fertilizer (0 and 30 m3 ha-1) and five N rates (0, 10, 20, 40, and 80 kg ha-1), in a randomized block design arranged in split plots, with five replications. The organic fertilizer levels represented the main plots and the N levels, the subplots. The source of N was urea and the source of organic fertilizer was goat manure. Irrigation was applied through a drip system and N by fertigation. At the end of the third growing season, soil chemical properties were determined and nitrate concentration in the soil solution (extracted by porous cups) was determined. Organic fertilization increased organic matter, pH, EC, P, K, Ca, Mg, Mn, sum of bases, base saturation, and CEC, but decreased exchangeable Cu concentration in the soil by complexation of Cu in the organic matter. Organic fertilization raised the nitrate concentration in the 0.20-0.40 m soil layer, making it leachable. Nitrate concentration in the soil increased as N rates increased, up to more than 300 mg kg-1 in soil and nearly 800 mg L-1 in the soil solution, becoming prone to leaching losses.

Hemodynamic effects of sodium bicarbonate in critically ill neonates.

OBJECTIVE: To analyze the cardiovascular effects of sodium bicarbonate in neonates with metabolic acidosis. DESIGN: Prospective, open, non-randomized, before-after intervention study with hemodynamic measurements performed before and 1, 5, 10, 20, and 30 min after bicarbonate administration. SETTING: Neonatal intensive care unit, tertiary care center. PATIENTS: Sequential sample of 16 paralysed and mechanically ventilated newborn infants with a metabolic acidosis (pH < 7.25 in premature and < 7.30 in term infants, base deficit > -8). INTERVENTION: An 8.4% sodium bicarbonate solution diluted 1:1 with water (final osmolality of 1000 mOsm/l) was administered in two equal portions at a rate of 0.5 mmol/min. The dose in mmol was calculated using the formula "base deficit x body weight (kg) x 1/3 x 1/2". MEASUREMENTS AND RESULTS: Sodium bicarbonate induced a significant but transient rise in pulsed Doppler cardiac output (CO) (+27.7%), aortic blood flow velocity (+15.3%), systolic blood pressure (BP) (+9.3%), (+14.6%), transcutaneous carbon dioxide pressure (PtcCO2) (+11.8%), and transcutaneous oxygen pressure (PtcO2) (+8%). In spite of the PaCO2 elevation, pH significantly improved (from a mean of 7.24 to 7.30), and the base deficit decreased (-39.3%). Calculated systemic vascular resistance (SVR) (-10.7%) and diastolic BP (-11.7%) decreased significantly, while PaO2 and heart rate (HR) did not change. Central venous pressure (CVP) (+6.5%) increased only slightly. By 30 min after bicarbonate administration all hemodynamic parameters, with the exception of the diastolic BP, had returned to baseline. CONCLUSION: Sodium bicarbonate in neonates with metabolic acidosis induces an increase in contractility and a reduction in afterload.

Liming effects on growth of native woody species from Brazilian Savannah

The liming effects on the growth of fifteen woody species of Brazil were evaluated under glasshouse conditions. The species used belong to different ecologic groups, namely: pioneer, secondary and climax trees. The soil treatments consisted in the absence of liming (-LIM) and liming sufficient to reach soil pH 6.0 (+LIM). In general, the pioneer and secondary species presented higher responses in total dry matter production (TDM) to soil liming, whereas the TDM of the climax species were not affected by the soil treatments. Thus, the ranking of species in relation to soil acidity tolerance ranged from highly sensitive to highly tolerant. The pioneer and secondary species growing in limed soil (+LIM) showed higher calcium (Ca), magnesium (Mg) and phosphorus (P) contents, and, at the same time lower Ca, Mg utilization efficiency (CaUE and MgUE respectively), whereas the P utilization (PUE) was higher. In contrast, the Ca, Mg and P content in the climax species were only slightly affected by the soil liming. In general the climax species were less efficient in the CaUE and MgUE than the pioneer and secondary species.

Effects of long-term irrigation with treated wastewater. Part I: Evolution of soil physico-chemical properties

The long-term impact of irrigation on a Mediterranean sandy soil irrigated with Treated wastewater (TWW) since 1980 was evaluated. The main soil properties (CEC, pH, size distribution, exchangeable cations and chloride, hydraulic conductivity) as well as the organic matter and Cu, Cr and Pb speciation in an irrigated soil and a non-irrigated control soil at various soil depths were monitored and compared during a 2 years experiment. In this first part, the evolution of the physico-chemical soil properties was described. The irrigation with TWW was beneficial with regard to water and nutrient supplying. All the exchangeable cations other than K(+) were higher in the irrigated soil than in the reference one. A part of the exchangeable cations was not fixed on the exchange complex but stored as labile salts or in concentrated soil solution. Despite the very sandy soil texture, both saturated and unsaturated hydraulic conductivity exhibited a significant diminution in the irrigated soil, but remained high enough to allow water percolation during rainy periods and subsequent leaching of accumulated salts, preventing soil salinization. In the irrigated soil, exchangeable sodium percentage (ESP) exhibited high values (20% on average) and the soil organic C was lower than in the reference. No significant effect was noticed on soil mineralogical composition due to irrigation. (C) 2010 Published by Elsevier Ltd.

Effects of fire on soil nitrogen dynamics and microbial biomass in savannas of Central Brazil

The objective of this work was to study the effects of fire on net N mineralization and soil microbial biomass in burned and unburned cerrado stricto sensu sites. The study was carried out from April 1998 to April 2000. The pH values were significantly higher in the burned site while soil moisture content was significantly higher in the unburned site (P<0.05). The soil C/N ratio was 22/1 and the available NO3-N ranged between 1.5 and 2.8 mg kg-¹ dry weight. However, the NH4-N concentration ranged between 3 and 34 mg kg-1 dry weight in the burned site and between 3 and 22 mg kg-1 dry weight in the unburned site. The NH4-N increased after fire, but no significant changes were observed for NO3-N (P<0.05). The NO3-N accumulation occurred in short periods during the rainy season. The rates of net N mineralization increased during the rainy season while reductions in soil microbial biomass were observed at both sites. This suggested that the peak in microbial activities occurred with the first rain events, with an initial net immobilization followed by net mineralization. Both sites presented the same pattern for mineralization/immobilization, however, the amount of inorganic-N cycled annually in unburned site was 14.7 kg ha-1 per year while the burned site presented only 3.8 kg ha-¹ of inorganic-N, one year after the burning.

Remaining phosphorus and sodium fluoride pH in soils with different clay contents and clay mineralogies

The remaining phosphorus (Prem) has been used for estimating the phosphorus buffer capacity (PBC) of soils of some Brazilian regions. Furthermore, the remaining phosphorus can also be used for estimating P, S and Zn soil critical levels determined with PBC-sensible extractants and for defining P and S levels to be used not only in P and S adsorption studies but also for the establishment of P and S response curves. The objective of this work was to evaluate the effects of soil clay content and clay mineralogy on Prem and its relationship with pH values measured in saturated NaF solution (pH NaF). Ammonium-oxalate-extractable aluminum exerts the major impacts on both Prem and pH NaF, which, in turn, are less dependent on soil clay content. Although Prem and pH NaF have consistent correlation, the former has a soil-PBC discriminatory capacity much greater than pH NaF.

Modulatory effects of acid-sensing ion channels on action potential generation in hippocampal neurons.

Extracellular acidification has been shown to generate action potentials (APs) in several types of neurons. In this study, we investigated the role of acid-sensing ion channels (ASICs) in acid-induced AP generation in brain neurons. ASICs are neuronal Na(+) channels that belong to the epithelial Na(+) channel/degenerin family and are transiently activated by a rapid drop in extracellular pH. We compared the pharmacological and biophysical properties of acid-induced AP generation with those of ASIC currents in cultured hippocampal neurons. Our results show that acid-induced AP generation in these neurons is essentially due to ASIC activation. We demonstrate for the first time that the probability of inducing APs correlates with current entry through ASICs. We also show that ASIC activation in combination with other excitatory stimuli can either facilitate AP generation or inhibit AP bursts, depending on the conditions. ASIC-mediated generation and modulation of APs can be induced by extracellular pH changes from 7.4 to slightly <7. Such local extracellular pH values may be reached by pH fluctuations due to normal neuronal activity. Furthermore, in the plasma membrane, ASICs are localized in close proximity to voltage-gated Na(+) and K(+) channels, providing the conditions necessary for the transduction of local pH changes into electrical signals.

The effects of polycyclic aromatic hydrocarbons and heavy metals on terrestrial annelids in urban soils

The effect of soil contamination by polycyclic aromatic hydrocarbons (PAH) and heavy metals on earthworms and enchytraeids was studied in urban parks, in Brno, Czech Republic. In spring and autumn 2007, annelids were collected and soil samples taken in lawns along transects, at three different distances (1, 5 and 30 m) from streets with heavy traffic. In both seasons, two parks with two transects each were sampled. Earthworms were collected using the electrical octet method. Enchytraeids were extracted by the wet funnel method from soil cores. All collected annelids were counted and identified. Basic chemical parameters and concentrations of 16 PAH, Cd, Cu, Pb, and Zn were analysed from soil from each sampling point. PAH concentrations were rather low, decreasing with the distance from the street in spring but not in autumn. Heavy metal concentrations did not decrease significantly with increasing distance. Annelid densities did not significantly differ between distances, although there was a trend of increase in the number of earthworms with increasing distance. There were no significant correlations between soil content of PAH or heavy metals and earthworm or enchytraeid densities. Earthworm density and biomass were negatively correlated with soil pH; and enchytraeid density was positively correlated with soil phosphorus.

Seed priming and sulfur effects on soybean cell membrane stability and yield in saline soil

The objective of this work was to determine the effects of seed priming and sulfur application on cell membrane characteristics, seedling emergence, chlorophyll content and grain yield of soybean (Glycine max) in saline soil. A complete-block design in 4x3 factorial arrangement with three replicates was used to test four types of seed priming (water, auxin, gibberellin and non-priming) and three levels of sulfate availability (0, 70 and 140 kg ha-1 K2SO4). The soil had a silty loam texture with an electrical conductivity of 3.61 ds m-1, a pH of 8.2 and a saturation percentage of about 46%. Seed priming had significant effects on mean emergence rate (MER), emergence percentage, relative water content (RWC) of leaves, relative chlorophyll content, time of maturity, shoot length and grain yield. The highest values for these variables were observed in the priming treatments, except for the time of maturity. Sulfur application had significant effects on MER, shoot length, RWC, membrane injury index and grain yield. Priming treatments provide greater emergence rates and grain yields and interact sinergicaly with sulfur rates.

pH Sensitive surfactants from lysine: assessment of their cytotoxicity and environmental behavior

The toxicity and environmental behavior of new pH-sensitive surfactants from lysine are presented. Three different chemical structures are studied: surfactants with one amino acid and one alkyl chain, surfactants with two amino acids on the polar head and one alkyl chain, and gemini surfactants. The pH sensitivity of these compounds can be tuned by modifying their chemical structures. Cytotoxicity has been evaluated using erythrocytes and fibroblast cells. The toxic effects against these cells depend on the hydrophobicity of the molecules as well as their cationic charge density. The effect of hydrophobicity and cationic charge density on toxicity is different for each type of cells. For erythrocytes, the toxicity increases as hydrophobicity and charge density increases. Nevertheless, for fibroblasts cationic charge density affects cytotoxicity in the opposite way: the higher charge density, the lower the toxicity. The effect of the pH on hemolysis has been evaluated in detail. The aquatic toxicity was established using Daphnia magna. All surfactants yielded EC50 values considerably higher than that reported for cationic surfactants based on quaternary ammonium groups. Finally, their biodegradability was evaluated using the CO2 headspace test (ISO 14593). These lysine derivatives showed high levels of biodegradation under aerobic conditions and can be classified as"readily biodegradable compounds".

Effects of biofilm and refrigeration on acerola postharvest conservation

The objective of this work was to compare the effects of four different concentrations of cassava starch film and storage temperature on shelf life and the quality of a genotype of acerola from the Germplasm Active Bank (GAB) of Federal Rural University of Pernambuco, Brazil. Sound orange-reddish acerola fruits were washed with a chlorine solution (100 mg.L-1 active chlorine) and randomly distributed into different lots. The fruits were dipped for 3 min in a cassava starch suspension with concentrations of 1, 2, 3 and 4% (w/v) and the control without coating, stored at 10ºC (85% RH) and 22ºC (85% RH). The total soluble solids (TSS), pH, titratable acidity (TA) and ascorbic acid (AA) were determined at harvest and regular interval during storage. The use of cassava biofilm at 1% on acerola fruits maintained the highest ascorbic acid content and the temperature of 10ºC extended storage life. The fruits coated with 1 and 2% biofilm could be stored for a period up to 15 days at 10ºC, with acceptable quality characteristics.

pH:n stabiilisuus ditioniittivalkaistulla neutraalilla LWC-prosessilla

Tässä työssä Oy Keskuslaboratoriossa sijaitsevalla kiertovesisimulaattorilla on tutkittiu ditioniittivalkaisun ja hylyn määrän vaikutusta pH:n stabiilisuuteen ja häiriöaineiden määrään neutraalissa LWC-prosessissa. Lisäksi on tutkittu ditioniitin määrän seisokkien ja valkaisuviiveiden vaikutusta. Kirjallisuusosassa tutustutaan ditioniittivalkaisuun ja valkaisun kemiaan sekä kalsiumkarbonaatin liukenemiseen. Lyhyesti selvitetään myös paperikoneen vesikiertoja sekä prosessissa olevia häiriöaineita. Karbonaatin liukenemisen osalta tarkastellaan myös mahdollisuutta mallintaa prosessia. Diplomityön aikana ajettiin yhdeksän simulaattoriajoa, jotka ajettiin kahden tai kolmen ajon sarjoissa. Ajoja ajettiin kolmella eri hylkymäärällä. Ditioniittivalkaisun lisäksi suoritettiin referenssiajoja ilman valkaisua. Lyhyen kierron pH pyrittiin pääosin säätämään seitsemään. Valkaisuviiveiden ja yli-yön kestävien seisokkien vaikutusta tutkittiin myös. Kokeissa todettiin ditioniitin alentavan pH tasoa. Tämä pH:n aleneminen johtaa kalsiumkarbonaatin liukenemiseen kalsiumkarbonaatin pyrkiessä puskuroimaan pH:n laskua. Valkaisureaktiossa syntyy negatiivisesti varauruneita yhdisteitä, jotka lisäävät prosessin anionisuutta. Ditioniitin valkaisuvaikutus näkyi lopputuotteen vaaleuden kasvuna sekä kieroveden värillisten yhdisteiden vähenemisenä. Ditioniittivalkaisu näytti poistavan myös noin 30% ligniinistä. Lipofiilisiin uuteaineisiin ditioniitilla ei ollut vaikutusta. Riittävä ditioniittiannos kokeiden perusteella on korkeintaan 1% ditioniittiä käytetyn mekaanisen massan määrästä. Korkeammalla annostuksella ei ollut vaikutusta vaaleuteen. Valkaisuaikana 30 minuuttia on riittävä. Pidempien valkaisuviiveiden ja seisokkien aikana valkaisussa syntyvän sulfiitin todettiin muuttuvan sulfaatiksi. Hylyn määrän lisääminen nosti pH:ta, varausta, johtokykyä ja liuenneen kalsiumin määrään. Hylyn määrän lisäminen paransi myös optisia ominaisuuksia, mikä johtui osin hylyn sisältämästä kalsiumkarbonaatista ja osin hylyn sisältämästä mekaanisesta massasta, joka oli valkaistua.