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The anti-leishmania effects of HIV peptidase inhibitors (PIs) have been widely reported; however, the biochemical target and mode of action are still a matter of controversy in Leishmania parasites. Considering the possibility that HIV-PIs induce lipid accumulation in Leishmania amazonensis, we analysed the effects of lopinavir on the lipid metabolism of L. amazonensis promastigotes. To this end, parasites were treated with lopinavir at different concentrations and analysed by fluorescence microscopy and spectrofluorimetry, using a fluorescent lipophilic marker. Then, the cellular ultrastructure of treated and control parasites was analysed by transmission electron microscopy (TEM), and the lipid composition was investigated by thin-layer chromatography (TLC). Finally, the sterol content was assayed by gas chromatography–mass spectrometry (GC/MS). TEM analysis revealed an increased number of lipid inclusions in lopinavir-treated cells, which was accompanied by an increase in the lipophilic content, in a dose-dependent manner. TLC and GC–MS analysis revealed a marked increase of cholesterol-esters and cholesterol. In conclusion, lopinavir-induced lipid accumulation and affected lipid composition in L. amazonensis in a concentration–response manner. These data contribute to a better understanding of the possible mechanisms of action of this HIV-PI in L. amazonensis promastigotes. The concerted action of lopinavir on this and other cellular processes, such as the direct inhibition of an aspartyl peptidase, may be responsible for the arrested development of the parasite.
The conservation of pineapple in urban backyards is an innovative strategy that aims to involve city residents. A program of this nature requires careful planning and monitoring because of the involvement of people who do not have technical knowledge. This paper reports the implementation and evaluation of 30 gardens in urban backyards in Cabaceiras do Paraguaçú, Bahia, Brazil, to establish the parameters to allow creating a model for pineapple germplasm conservation cells with the collaboration of urban residents. A questionnaire was applied in two steps to people interested in participating, from which it was possible to choose and evaluate a general profile of the participants. Thirty pineapple accessions from the Active Germplasm Bank (AGB) of the Embrapa were selected for testing in the gardens. Two production cycles were considered, during which quantitative and qualitative traits of the plants and fruits were evaluated. The data were compared with the characteristics of the same accessions in the Pineapple AGB and were analysed by mixed principal component analysis. With respect to adequate maintenance, 20 gardens were well cared for until the end of the assessments, five were reasonably well tended and five were lost, due to home remodelling or lack of care by the guardians. Despite the loss of the five gardens, no accession was totally lost, thanks to the experimental design with three plants of each accession in three different gardens. The plants preserved in the gardens did not differ from those maintained by the AGB, demonstrating the effectiveness of this conservation strategy.
Intrauterine growth restriction (IUGR) can induce deleterious changes in the modulatory ability of the vascular endothelium, contributing to an increased risk of developing cardiovascular diseases in the long term. However, the mechanisms involved are not fully understood. Emerging evidence has suggested the potential role of endothelial progenitor cells (EPCs) in vascular health and repair. Therefore, we aimed to evaluate the effects of IUGR on vascular reactivity and EPCs derived from the peripheral blood (PB) and bone marrow (BM) in vitro. Pregnant Wistar rats were fed an ad libitum diet (control group) or 50% of the ad libitum diet (restricted group) throughout gestation. We determined vascular reactivity, nitric oxide (NO) concentration, and endothelial nitric oxide synthase (eNOS) protein expression by evaluating the thoracic aorta of adult male offspring from both groups (aged: 19–20 weeks). Moreover, the amount, functional capacity, and senescence of EPCs were assessed in vitro. Our results indicated that IUGR reduced vasodilation via acetylcholine in aorta rings, decreased NO levels, and increased eNOS phosphorylation at Thr495. The amount of EPCs was similar between both groups; however, IUGR decreased the functional capacity of EPCs from the PB and BM. Furthermore, the senescence process was accelerated in BM-derived EPCs from IUGR rats. In summary, our findings demonstrated the deleterious changes in EPCs from IUGR rats, such as reduced EPC function and accelerated senescence in vitro. These findings may contribute towards elucidating the possible mechanisms involved in endothelial dysfunction induced by fetal programming.
Early Archaic human skeletal remains found in a burial context in Lapa do
Santo in east-central Brazil provide a rare glimpse into the lives of
hunter-gatherer communities in South America, including their rituals for
dealing with the dead. These included the reduction of the body by means of
mutilation, defleshing, tooth removal, exposure to fire and possibly
cannibalism, followed by the secondary burial of the remains according to
strict rules. In a later period, pits were filled with disarticulated bones
of a single individual without signs of body manipulation, demonstrating
that the region was inhabited by dynamic groups in constant transformation
over a period of centuries.
Soybean oil (SBO) is rich in polyunsaturated fatty acids (FA) and rumen bypass of SBO can contribute to increase the polyunsaturated FA proportion in milk fat. Citrus pulp (CPP) is a source of antioxidants but there is little information on the effects of CP administration on milk properties. This study was performed to determine the role of rumen microorganisms in the transfer of antioxidants from CPP into milk when cows receive SBO as a source of polyunsaturated FA. Four ruminally fistulated lactating Holstein cows were assigned to a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments: (1) SBO administered in the rumen; (2) SBO infused in the abomasum; (3) SBO + CPP administered in the rumen; and (4) SBO + CPP infused in the abomasum. Product and site of administration had no effect on yield of milk components. Concentrations of total polyphenols and flavonoids, reducing power and production of conjugated diene (CD) hydroperoxides in milk were not affected by products, but infusion in the abomasum compared with administration in the rumen increased production of CD. Milk fat FA profile was not affected by products. However, cows infused in the abomasum compared with those administered in the rumen showed lower proportions of short-chain and monounsaturated FA and higher proportions of polyunsaturated, omega 3 and omega 6 FA in milk fat, which resulted in enhanced health-promoting index of milk. Administration of SBO and CPP (0·2 + 1·0 kg/d) in the rumen or the abomasum resulted in similar milk antioxidant properties, thus suggesting that the rumen microbes have little involvement in the metabolism of antioxidants from CPP.
A retrospective space–time permutation model with non-Euclidean distance criteria was applied within a high-complexity hospital setting to quantitatively explore cluster patterns of 273 patients infected with or colonized by carbapenemase-producing Klebsiella pneumoniae during 4 years. Results were compared to standard nosocomial active-surveillance methods. Two clusters were identified in the period, suggesting that space–time strategies for cluster quantification within confined environments may be useful.
The West Midlands was the first English region to report sustained community transmission during the ‘containment’ phase of the influenza A(H1N1)pdm09 pandemic in England. To describe the epidemiological experience in the region, West Midlands and national datasets containing laboratory-confirmed A(H1N1)pdm09 virus cases in the region during the ‘containment’ phase were analysed. The region accounts for about 10·5% of England's population, but reported about 42% of all laboratory-confirmed cases. Altogether 3063 cases were reported, with an incidence rate of 56/100 000 population. School-associated cases accounted for 25% of cases. Those aged <20 years, South Asian ethnic groups, and residents of urban and socioeconomically deprived areas were disproportionately affected. Imported cases accounted for 1% of known exposures. Regional R0 central estimates between 1·41 and 1·43 were obtained. The West Midlands experience suggests that interpretation of transmission rates may be affected by complex interactions within and between sub-populations in the region.
The exact mechanisms of the relationship between obesity and cardiovascular events are not yet fully understood; however, oxidative stress may be involved. Thus, the aim of the present study was to evaluate the effects of resveratrol and fish oil on catecholamine-induced mortality in obese rats. To begin with, rats were divided into five groups: (1) lean, (2) obese, (3) obese supplemented with resveratrol, (4) obese supplemented with fish oil and (5) obese supplemented with resveratrol and fish oil (n 18 rats per group), for 2 months. After supplementation, the groups were subdivided as with (n 10) and without (n 8) cardiovascular catecholaminergic stress after isoproterenol (60 mg/kg) injection. At 24 h later, the survival rate was analysed. The obese group showed lower survival rates (10 %) when compared with the lean group (70 %). On the other hand, resveratrol (50 %) and fish oil (40 %) increased the survival rate of obese rats (χ2 test, P= 0·019). Biochemical analyses of the myocardium and aorta revealed that obese rats had higher levels of superoxide and oxidative damage to lipids and protein. This was associated with reduced superoxide dismutase and glutathione peroxidase activity in both the myocardium and aorta. The supplementation increased antioxidant enzyme activities and reduced oxidative damage. We also evaluated the nuclear factor-erythroid 2 p45-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 antioxidant pathway. Nrf2 protein levels that were reduced in obese rats were increased by the antioxidant treatment. Taken together, these results showed that resveratrol and fish oil reduce catecholamine-induced mortality in obese rats, partly through the reduction of oxidative stress.
This paper reports the effects of BnSP-7 toxin, a catalytically inactive phospholipase A2 from Bothrops pauloensis snake venom, on Leishmania (Leishmania) amazonensis. BnSP-7 presented activity against promastigote parasite forms both in the MTT assay, with IC50 of 58·7 μg mL−1 of toxin, and a growth curve, inhibiting parasite proliferation 60–70% at concentrations of 50–200 μg mL−1 of toxin 96 h after treatment. Also, the toxin presented effects on amastigotes, reducing parasite viability by 50% at 28·1 μg mL−1 and delaying the amastigote–promastigote differentiation process. Ultrastructural studies showed that BnSP-7 caused severe morphological changes in promastigotes such as mitochondrial swelling, nuclear alteration, vacuolization, acidocalcisomes, multiflagellar aspects and a blebbing effect in the plasma membrane. Finally, BnSP-7 interfered with the infective capacity of promastigotes in murine peritoneal macrophages, causing statistically significant infectivity-index reductions (P < 0·05) of 20–35%. These data suggest that the BnSP-7 toxin is an important tool for the discovery of new parasite targets that can be exploited to develop new drugs for treating leishmaniasis.
In the present work, the interactions between forsterite-91 with distilled water and forsterite-91 with artificial seawater were studied at two pHs (2.0 and 8.0) using different techniques. A large increase in pH was observed for samples incubated at an initially acidic pH (2.0) due to the dissolution of forsterite-91 in distilled water and artificial seawater. Thus, in acidic hydrothermal vents, an increase in the amount of hydrocarbons and magnetite should be expected due to the release of Fe(II). The pHPZC decreased and the pHIEP increased when forsterite-91 was treated with distilled water and artificial seawater. The ions from the artificial seawater had an effect on zeta potential. Scanning electron microscopy (SEM) images and X-ray diffractograms showed halite in the samples of forsterite-91 mixed with artificial seawater. The presence of halite or adsorption of ions on the surface of forsterite-91 could affect the synthesis of magnetite and hydrocarbons in hydrothermal vents, due to a decrease in the dissolution rates of forsterite-91. The dissolution of forsterite-91 yields low concentrations of Fe(III) and Mn(II) as detected by electron paramagnetic resonance (EPR) spectroscopy. Microanalysis of forsterite-91 showed a higher amount of Mn, with an oxidation that was likely not +II, as Mn in supernatant solutions was only detected by EPR spectroscopy after mixing with artificial seawater at pH 2.0. As Fe(III) and Mn(II) are catalyst constituents of magnetite and manganese oxide, respectively, their presence is important for synthesis in hydrothermal vents. Etch pits were observed only in the forsterite-91 sample mixed with distilled water at pH 8.0. Na, Cl, S, Ca and K were detected in the samples mixed with artificial seawater by SEM–EDS. Si, Mg, Fe and Al were detected in almost all supernatant samples due to forsterite-91 dissolution. Cr was not dissolved in the experiments, thus Cr in the mineral could serve as an effective catalyst for Fischer Tropsch Types (FTT) reactions in hydrothermal vent systems. X-ray diffractograms of the original forsterite-91 also showed peaks arising from zeolites and clinochlore. After the samples were treated with artificial seawater, X-ray diffractograms showed the dissolution of zeolite. Experiments should be performed in the natural environment to verify the potential for zeolites to act as a catalyst in hydrothermal vents.
This study investigated the synthesis of goethite under conditions resembling those of the prebiotic Earth. The artificial seawater used contains all the major elements as well as amino acids (α-Ala, β-Ala, Gly, Cys, AIB) that could be found on the prebiotic Earth. The spectroscopic methods (FT-IR, EPR, Raman), scanning electron microscopy (SEM) and X-ray diffraction showed that in any condition Gly and Cys favoured the formation of goethite, artificial seawater plus β-Ala and distilled water plus AIB favoured the formation of hematite and for the other synthesis a mixture of goethite and hematite were obtained. Thus in general no protein amino acids (β-Ala, AIB) favoured the formation of hematite. As shown by surface enhanced Raman spectroscopy (SERS) spectra the interaction between Cys and Fe3+ of goethite is very complex, involving decomposition of Cys producing sulphur, as well as interaction of carboxylic group with Fe3+. SERS spectra also showed that amino/CN and C-CH3 groups of α-Ala are interacting with Fe3+ of goethite. For the other samples the shifting of several bands was observed. However, it was not possible to say which amino acid groups are interacting with Fe3+. The pH at point of zero charge of goethites increased with artificial seawater and decreased with amino acids. SEM images showed when only goethite was synthesized the images of the samples were acicular and when only hematite was synthesized the images of the samples were spherical. SEM images for the synthesis of goethite with Cys were spherical crystal aggregates with radiating acicular crystals. The highest resonance line intensities were obtained for the samples where only hematite was obtained. Electron paramagnetic resonance (EPR) and Mössbauer spectra showed for the synthesis of goethite with artificial seawater an isomorphic substitution of iron by seawater cations. Mössbauer spectra also showed that for the synthesis goethite in distilled water plus Gly only goethite was synthesized and in artificial seawater plus Cys a doublet due to interaction of iron with artificial seawater/Cys was observed. It should be pointed out that EPR spectroscopy did not show the interaction of iron with artificial seawater/Cys.
The present paper demonstrates the development of interspecific hybrids between Manihot esculenta Crantz ssp. esculenta (Mee) and M. esculenta Crantz ssp. flabellifolia (Mef) and paternity analysis using microsatellite markers [simple sequence repeat (SSR)]. Three Mef accessions (FLA005, FLA025V and FLA029V) were used for crosses with varieties of Mee: Saracura, Aipim Bravo, COL 1725, Aipim Rosa, Abóbora, Paraná and PER334. The paternity of the interspecific hybrids was investigated using 24 SSRs. The observed heterozygosity (Ho), polymorphic information content (PIC), probability of identity (PI) and paternity exclusion (PE) were evaluated. The rate of breeding success varied from 17 to 92%, and an average of two pollinations were required for each generated hybrid plant. The Ho value ranged from 0·11 to 0·92, and the PIC value ranged from 0·12 to 0·59. The uneven distribution of allele frequencies was accompanied by a high PI average (0·56). However, the combined PE for 21 loci was 0·99, which allows for the determination of the paternity of the hybrids with good discriminatory power. Of the 74 hybrids evaluated, 0·82 had their paternity confirmed using microsatellite markers. Discriminant analysis of principal components (DAPC) indicated the presence of eight clusters, of which, one was composed of only Mef varieties and the supposed hybrid Fla52Sar-H7, which was a product of apomixis. The parent Mee and hybrids were allocated in the other seven clusters. The data obtained demonstrate that SSR markers can be routinely used in breeding programmes to verify the paternity of interspecific crosses of cassava.
The aim of this study was to investigate white matter (WM) abnormalities in obsessive-compulsive disorder (OCD) and its relationship to severity of obsessive-compulsive symptoms.
Conventional and diffusion tensor imaging were acquired in nine patients with OCD and nine gender- and age-matched healthy volunteers. Changes in fractional anisotropy (FA) and mean diffusivity (MD) were investigated using selected regions of interest (ROIs) analyses and whole brain tract-based spatial statistic analyses. A priori ROIs were placed bilaterally in internal capsule (IC), superior longitudinal fascicule (SLF), cingulate bundle (CB), and corpus calosum (CC).
ROIs analyses showed that, as compared to healthy volunteers, patients with OCD exhibited reduced FA values bilaterally in regions of the posterior limb of the IC and in the SLF and increased MD values bilaterally in the posterior limb of the IC, in the left CB, and in the splenium of CC. Voxelwise analysis showed that, as compared to controls, patients with OCD exhibited reduced FA and increased MD in regions of the cortical spinal tract (genu and posterior limb of internal capsule and corona radiata) and the SLF. Severity of OCD correlated with WM alterations in different brain regions, ie, the left (rho=0.70 [MD]) and right (rho=0.70 [MD]) anterior limb of the IC, the left (rho=0.97 [MD]) and right SLF (rho=0.81 [MD]), and the genu of CC (rho=0.66 [MD]; rho=-0.69 [FA]).
Our findings support the involvement of different WM tracts in OCD and suggest that greater impairment in WM integrity is associated with increased severity of OCD symptoms.
To determine whether an educational programme aimed at discouraging students from drinking sugar-sweetened beverages could prevent excessive weight gain.
Forty-seven classes in twenty-two schools were randomised as intervention or control.
Participants were 1140, 9–12-year-old fourth graders (435 in the intervention group and 608 in the control group). Sugar-sweetened beverages and juice intake were measured through one 24 h recall at baseline and another at the end of the trial. The main outcome was the change in BMI (BMI = weight (kg)/height (m2)), measured at the beginning and at the end of the school year. Intention-to-treat analysis was performed taking into account the cluster (classes) effect.
A statistically significant decrease in the daily consumption of carbonated drinks in the intervention compared to control (mean difference = −56 ml; 95 % CI −119, −7 ml) was followed by a non-significant overall reduction in BMI, P = 0·33. However, among those students overweight at baseline, the intervention group showed greater BMI reduction (−0·4 kg/m2 compared with −0·2 kg/m2 in the control group (P = 0·11)), and this difference was statistically significant among girls (P = 0·009). Fruit juice consumption was slightly increased in the intervention group (P = 0·08), but not among girls.
Decreasing sugar-sweetened beverages intake significantly reduced BMI among overweight children, and mainly among girls. Efforts to reduce energy intake through liquids need to emphasise overall sweetened beverages and addition of sugar on juices.
Many well-known combinatorial optimization problems can be stated over the set of acyclic orientations
of an undirected graph. For example, acyclic orientations with certain diameter constraints are
closely related to the optimal solutions of the vertex coloring and frequency assignment problems.
In this paper we introduce a linear programming formulation of acyclic orientations
with path constraints, and discuss its use in the solution of the vertex coloring problem and
some versions of the frequency assignment problem. A study of the polytope associated with the
formulation is presented, including proofs of which constraints of the formulation are facet-defining
and the introduction of new classes of valid inequalities.
Trypanosoma cruzi is the ethiological agent of Chagas
disease. New compounds are being developed based on the biosynthesis and
function of sterols, because T. cruzi has a requirement for
specific endogenous sterols for growth and survival. Sterol biosynthesis
inhibitors (SBIs) are drugs commonly used against fungal diseases. These
drugs act by depleting essential and specific membrane components
and/or inducing the accumulation of toxic intermediary or lateral
products of the biosynthetic pathway. In this work we present the effects
of WSP488, WSP501, and WSP561, specific inhibitors of
Δ24(25)-sterol methyl transferase, on the ultrastructure of
T. cruzi epimastigotes. All three drugs inhibited parasite
multiplication at low concentrations, with IC50 values of 0.48,
0.44, and 0.48 μM, respectively, and induced marked morphological
changes including (a) blockage of cell division; (b) swelling of the
mitochondrion, with several projections and depressions; (c) swelling of
the perinuclear space; (d) presence of autophagosomes and myelin-like
figures; (e) enlargement of the flagellar pocket and of a cytoplasmic
vacuole located in close association with the flagellar pocket; (f)
detachment of the membrane of the cell body; and (g) formation of a
vesicle at the surface of the parasite between the flagellar pocket and
the cytostome. Our results show that these drugs are potent in
vitro inhibitors of growth of T. cruzi.
We describe the preparation, properties and the comparison of SiO2 films deposited by liquid phase deposition from supersaturated solutions with both silica powder and silicic acid. From each solution the resulting [Si (OH)4]-rich supersaturated solution, when suitably diluted with De-Ionised (DI) water deposits SiO2 onto silicon. Our work was motivated by the search for the optimum condition for device quality oxide growth within a low (<300°C) thermal budget, for large area device fabrication, on inexpensive substrates such as glass and plastic. The net growth of the LPD of the oxide layer is result of competition between deposition and etching; the balance and hence the rate of growth being controlled simply and reproducibly by the addition of de-ionised (DI) water. We have produced device grade SiO2 from the solution at 48°C onto both n and p type Si substrates. Control of the oxide properties is achieved in the amount of DI water added to the super-saturated solution. Oxide layers of optimum quality being obtained by the titration of 60ml DI water to 100ml of supersaturated solution. FTIR, ellipsometery, and C-V/I-V measurements were used to characterise the films deposited from solutions of silica powder and silicic acid. The resistivity of the optimum layers from a saturated solution of silica powder was found to be of the order of ≈1011 Ωcm with breakdown field strengths of the order of 3-5 x 106 V/cm. The resistivity of the optimum layers prepared using silicic acid were much higher (1015 Ω-cm), with breakdown field strengths of the order of (9±1) x 106 V/cm. The results approach those for thermally grown silicon dioxide, suggesting that LPD oxide have the potential of device quality insulator on large area substrate.
In this study the Golgi complex of the epimastigote forms of Trypanosoma cruzi were isolated and characterized. Using well-controlled sonication to rupture the cells and centrifugation on a discontinuous sucrose density gradient, a highly enriched Golgi fraction was obtained. The Golgi fraction contained most of the β-galactosyltransferase (β-Gal transferase) and UDP-N-acetyl-glucosamine: polypeptide-α-N-acetyl-glucosaminyltransferase (O-α-GlcNAc transferase) activities with minimal contamination of other organelles, as observed by enzymatic assays and electron microscopy analysis. To characterize the Golgi from T. cruzi cells further, it was incubated with a monoclonal antibody against a 58 kDa protein involved in the association of the Golgi complex with microtubules in mammalian cells. Immunofluorescence microscopy showed that the 58 kDa protein is localized in the T. cruzi Golgi region, a result confirmed by high resolution scanning electron microscopy immunocytochemistry. Thus, our results show, for the first time, that the β-Gal transferase, the O-α-GlcNAc transferase and the 58 kDa protein are present in the Golgi complex of T. cruzi and are novel biochemical markers which can be used in the characterization of this organelle in T. cruzi.
Uniformly sheared flows have been generated in a high-speed wind tunnel at shear rates higher than previously achieved, in an effort to approach those in the inner turbulent boundary layer. As at lower shear rates, the turbulence structure was found to attain a self-similar state with approximately constant anisotropies and exponential kinetic energy growth. The normal Reynolds stress anisotropies showed no systematic dependence upon the mean shear within the examined range; however, the shear stress anisotropy was significantly lower than the low-shear values, in conformity with boundary layer measurements and direct numerical simulations of homogeneous shear flow.