Structure-based drug design studies of the interactions ofent-kaurane diterpenes derived from Wedelia paludosa with the Plasmodium falciparumsarco/endoplasmic reticulum Ca2+-ATPase PfATP6

Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosawere tested in silico against the Plasmodium falciparumCa2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.

Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors

Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients.

Effects of the encapsulation of usnic acid into liposomes and interactions with antituberculous agents against multidrug-resistant tuberculosis clinical isolates

Mycobacterium tuberculosis (Mtb) has acquired resistance and consequently the antibiotic therapeutic options available against this microorganism are limited. In this scenario, the use of usnic acid (UA), a natural compound, encapsulated into liposomes is proposed as a new approach in multidrug-resistant tuberculosis (MDR-TB) therapy. Thus the aim of this study was to evaluate the effect of the encapsulation of UA into liposomes, as well as its combination with antituberculous agents such as rifampicin (RIF) and isoniazid (INH) against MDR-TB clinical isolates. The in vitro antimycobacterial activity of UA-loaded liposomes (UA-Lipo) against MDR-TB was assessed by the microdilution method. The in vitro interaction of UA with antituberculous agents was carried out using checkerboard method. Minimal inhibitory concentration values were 31.25 and 0.98 µg/mL for UA and UA-Lipo, respectively. The results exhibited a synergistic interaction between RIF and UA [fractional inhibitory concentration index (FICI) = 0.31] or UA-Lipo (FICI = 0.28). Regarding INH, the combination of UA or UA-Lipo revealed no marked effect (FICI = 1.30-2.50). The UA-Lipo may be used as a dosage form to improve the antimycobacterial activity of RIF, a first-line drug for the treatment of infections caused by Mtb.

Meanings attributed by family members in pediatrics regarding their interactions with nursing professionals

Abstract OBJECTIVE Understanding the meanings attributed by family caregivers of children in hospital environments about their interactions with nursing professionals. METHODS This qualitative study used Symbolic Interactionism as a theoretical reference and Grounded Theory as the methodological framework. It was carried out in a Pediatrics Center in southern Brazil, in the first half of 2013. Participants were 15 family caregivers of hospitalized children. Data were collected through interviews and submitted to open and axial analysis. RESULTS Interactions with the nursing team enable family to trust or distrust in the provided child care and to positively evaluate the care received. CONCLUSION Interactions between family members and the nursing team contribute to the significance attributed by the family to the nursing care received by the child. Nurses should be aware of the attitudes of the nursing team regarding the child and their family, prioritizing humanized care.

Interactions between magnesium, calcium, and aluminum on soybean root elongation

Alleviation of Al rhizotoxicity by Ca and Mg can differ among species and genotypes. Root elongation of soybean [Glycine max (L.) Merr.] line N93-S-179 and cvs. Young and Ransom exposed to varying concentrations of Al, Ca and Mg were compared in two experiments using a vertically split root system. Roots extending from a surface compartment with limed soil grew for 12 days into a subsurface compartment with nutrient solution treatments maintained at pH 4.6 with either 0 or 15 µmol L-1 Al. Calcium and Mg concentrations in treatments ranging from 0 to 20 mmol L-1. Although an adequate supply of Mg was provided in the surface soil compartment for soybean top growth, an inclusion of Mg was necessary in the subsurface solutions to promote root elongation in both the presence and absence of Al. In the absence of Al in the subsurface solution, tap root length increased by 74 % and lateral root length tripled when Mg in the solutions was increased from 0 to either 2 or 10 mmol L-1. In the presence of 15 µmol L-1 Al, additions of 2 or 10 mmol L-1 Mg increased tap root length fourfold and lateral root length by a factor of 65. This high efficacy of Mg may have masked differences in Al tolerance between genotypes N93 and Young. Magnesium was more effective than Ca in alleviating Al rhizotoxicity, and its ameliorative properties could not be accounted for by estimated electrostatic changes in root membrane potential and Al3+ activity at the root surface. The physiological mechanisms of Mg alleviation of Al injury in roots, however, are not known.

Determinação de Fe, Zn, Cu e Mn extraídos do solo por diferentes extratores e dosados por espectrofotometria de emissão ótica em plasma induzido e espectrofotometria de absorção atômica

Para aumentar a precisão nas análises químicas de fertilidade do solo e dosar simultaneamente vários elementos, alguns laboratórios vêm optando pelo uso da técnica da espectrofotometria de emissão ótica em plasma induzido (ICP), em detrimento da técnica da espectrofotometria de absorção atômica (EAA), hoje comumente utilizada nos laboratórios de análise de solos. Este trabalho, além de comparar as duas técnicas de dosagem quanto à precisão, à reprodutibilidade e à magnitude dos teores dos micronutrientes Fe, Zn, Cu e Mn, extraídos por Mehlich-1, Mehlich-3 e DTPA-TEA, objetivou, também, selecionar os comprimentos de onda que apresentam menores interferências espectrais no ICP. Foram utilizadas 36 amostras (0 a 0,2 m) de solos coletadas nos Estados de Minas Gerais e Bahia, com ampla variação nos teores de micronutrientes, sendo selecionados três solos para definir os comprimentos de onda do ICP e avaliar a precisão e a reprodutibilidade dos métodos de dosagem. Os comprimentos de onda com menores interferências espectrais no ICP foram: 259,939 nm para Fe em Mehlich-1 e DTPA-TEA e 234,349 nm em Mehlich-3; 213,857 nm para Zn e 324,752 nm para Cu nos três extratores; e 259,372 nm para Mn em Mehlich-1 e DTPA-TEA e 260,568 nm em Mehlich-3. Tanto o ICP quanto o EAA foram precisos e reprodutíveis nas dosagens de Fe e Mn, sendo o ICP, em virtude do seu menor limite de detecção, mais preciso e reprodutível nas dosagens de Zn e Cu. Os métodos de dosagem diferiram estatisticamente (p < 0,01) pelo teste de identidade aplicado, para as dosagens de Fe, Zn, Cu e Mn, utilizando Mehlich-1, Mehlich-3 e DTPA-TEA, comprometendo assim a interpretação dos resultados gerados pelo ICP, com base nos níveis críticos gerados a partir do EAA.

Hydraulic conductivity and soil-sewage sludge interactions

One of the main problems faced by humanity is pollution caused by residues resulting from the production and use of goods, e.g, sewage sludge. Among the various alternatives for its disposal, the agricultural use seems promising. The purpose of this study was to evaluate the hydraulic conductivity and interaction of soil with sandy-silty texture, classified as Spodosols, from the Experimental Station Itapirema - IPA, in Goiana, state of Pernambuco, in mixtures with sewage sludge from the Mangueira Sewage Treatment Station, in the city of Recife, Pernambuco at rates of 25, 50 and 75 Mg ha-1. Tests were conducted to let water percolate the natural saturated soil and soil-sludge mixtures to characterize their physical, chemical, and microstructural properties as well as hydraulic conductivity. Statistical data analysis showed that the presence of sewage sludge in soils leads to an increase of the < 0.005 mm fraction, reduction in real specific weight and variation in optimum moisture content from 11.60 to 12.90 % and apparent specific dry weight from 17.10 and 17.50 kN m-3. In the sludge-soil mixture, the quartz grains were covered by sludge and filling of the empty soil macropores between grains. There were changes in the chemical characteristics of soil and effluent due to sewage sludge addition and a small decrease in hydraulic conductivity. The results indicate the possibility that soil acidity influenced the concentrations of the elements found in the leachate, showing higher levels at higher sludge doses. It can be concluded that the leaching degree of potentially toxic elements from the sewage sludge treatments does not harm the environment.