Association between myocardial mechanical dispersion and ventricular arrhythmogenicity in chagas cardiomyopathy.

Chagas disease is a major health concern in Latin America. Ventricular arrhythmia (VA) is a hallmark of Chagas cardiomyopathy (CCM), associated with worse prognosis. The present study aimed to verify the association between myocardial mechanical dispersion (MD) and ventricular arrhythmogenicity in CCM. In a cross-sectional study, 77 patients (55.8 ± 10.4 years) with CCM were evaluated. Global longitudinal strain (GLS) and MD were assessed by echocardiography, derived from the speckle tracking technique. Myocardial MD was measured from the onset of the Q/R wave on electrocardiogram to the peak longitudinal strain in 16 segments of the left ventricle. Frequency and complexity of ventricular extrasystoles (VES) were assessed by dynamic electrocardiography. The density and complexity of VES and the presence of non-sustained ventricular tachycardias (NSVTs) increase as MD increases. In logistic regression, MD was the only variable associated with the presence of paired VES and ventricular bigeminy. In addition, both MD and GLS were associated with the presence of NSVT (both, p < 0.01), and MD was independently associated with NSVT (OR 1.04, 95% CI 1.004-1.201, p = 0.031). In CCM, MD is associated with a higher density and complexity of VES, including NSVT.

Functional tests associated with sarcopenia in moderate chronic obstructive pulmonary disease.

Objectives: To evaluate the association of physical and functional measures with sarcopenia in moderate chronic obstructive pulmonary disease (COPD) and to establish cutoff points for sarcopenia screening.Methods: The study included COPD with and without sarcopenia, of both sexes who were over 50 years old. Participants were assessed for lung function, body composition, grip strength, Short Physical Performance Battery (SPPB), 5-repetition, 10-repetition and 30-s sit-to-stand tests (5STS, 10STS, and 30STS, respectively). In addition, 6-min walking test, respiratory muscular strength, and physical activity level were tested.Results: The study had 35 participants, 24 men (68.6%) and moderate COPD (51.4%). COPD-sarcopenia showed lower values in lean mass, body fat and body mass alongside lower performance in 10 and 30 STS tests, SPPB and gait speed compared to non-sarcopenic group. The cutoff points with better sensitivity and specificity to identify sarcopenia were 10.88 and 34.14 s, 15 repetitions, and 10 points in the 5STS, 10STS, 30STS, and SPPB, respectively. The comparison of the receiver operating curves evidenced no differences between the functional tests. Only 30STS and SPPB showed acceptable discriminatory power.Conclusion: Functional tests, especially 30STS and SPPB, are simple and affordable tools for screening sarcopenia in COPD with moderate obstruction.

Development of an orange juice in-house reference material and its application to guarantee the quality of vitamin C determination in fruits, juices and fruit pulps.

Reference materials are useful for the quality control of analytical procedures and to evaluate the performance of laboratories. There are few and expensive certified reference materials commercially available for vitamin C or ascorbic acid analysis in food matrices. In this study, the preparation and the suitability assessment of an orange juice in-house reference material (RM) for vitamin C analysis in fruits, juices and in fruit pulps is described. This RM was used for the development and full validation of an HPLC method. The results showed excellent linearity (r(2)=0.9995), good accuracy (96.6-97.3%) and precision, as relative standard deviation, ranged from 0.70% to 3.67%. The in-house RM was homogenous and stable at storage conditions (-80°C) during 12 months. According to our results, this in-house RM is an excellent tool to use in quality control and method verification purposes for vitamin C analysis of fruits, juices and fruit pulps matrices. Furthermore, a stabilization solution with perchloric and metaphosphoric acids was developed which prevents degradation of ascorbic acid for a period of 12 months at -80°C.

Ultra-high pressure LC for astaxanthin determination in shrimp by-products and active food packaging.

Nowadays, there is increasing interest in natural antioxidants from food by-products. Astaxanthin is a potent antioxidant and one of the major carotenoids in crustaceans and salmonids. An ultra-high pressure liquid chromatographic method was developed and validated for the determination of astaxanthin in shrimp by-products, and its migration from new packaging materials to food simulants was also studied. The method uses an UPLC® BEH guard-column (2.1 × 5 mm, 1.7 µm particle size) and an UPLC® BEH analytical column (2.1 × 50 mm, 1.7 µm particle size). Chromatographic separation was achieved using a programmed gradient mobile phase consisting of (A) acetonitrile-methanol (containing 0.05 m ammonium acetate)-dichloromethane (75:20:5, v/v/v) and (B) ultrapure water. This method was evaluated with respect to validation parameters such as linearity, precision, limit of detection, limit of quantification and recovery. Low-density polyethylene films were prepared with different amounts of the lipid fraction of fermented shrimp waste by extrusion, and migration was evaluated into food simulants (isooctane and ethanol 95%, v/v). Migration was not detected under the tested conditions.

New nutritional data on traditional foods for European food composition databases.

BACKGROUND/OBJECTIVES: There are many different cultures within Europe, each with its own distinct dietary habits. Traditional foods are the key elements that differentiate the dietary patterns of each country. Unfortunately, in most countries, there is little information on the nutritional composition of such foods. Therefore, there is a need to study traditional foods to preserve these elements of European culture and, if possible, enrich and improve dietary habits across the continent. The Traditional Foods work package within the European Food Information Resource (EuroFIR) project aimed to provide new nutritional data on traditional foods for use in national food composition tables. SUBJECTS/METHODS: A EuroFIR consensus-based method with standardised procedures was applied for the systematic study of traditional foods and recipes in selected European countries. Traditional foods were selected on the basis of the EuroFIR definition of the term 'traditional food' and prioritized according to specific criteria. From the prioritized list, the five traditional foods per country to be investigated were selected to represent a full course meal. Protocols with guidelines for the recording of traditional recipes, the collection, preparation and distribution of laboratory samples, as well as quality requirements for laboratory selection, were developed to establish a common approach for use by all countries for the acquisition of reliable data. RESULTS: The traditional character of the selected foods has been documented and traditional recipes have been recorded. Chemical analyses to determine the nutritional composition of 55 traditional foods were performed and the data were evaluated and fully documented according to EuroFIR standards. Information on food description, the recipe, component identification, sampling plan, sample handling, analytical method and performance was collected for each of the 55 investigated traditional foods. CONCLUSIONS: This common methodology for the systematic study of traditional foods will enable countries to further investigate their traditional foods and to continue to update their national food composition databases and EuroFIR's food databank system.

The role of municipal committees in the development of an integrated urban water policy in Belo Horizonte, Brazil.

This paper discusses the challenges involved in adopting an integrated approach to urban water policies and management, a particularly problematic issue in Brazil due to the incomplete nature of urbanization, defined as the lack of adequate and/or universalized access to infrastructure and services, informal housing and conflicts between environmental protection and social housing needs. In the last two decades strong social movements have influenced urban environmental policies from national to local levels. In Belo Horizonte since 1993, decision-making processes have involved important mechanisms of democratic inclusion, which have contributed to fairer urban policies. A brief discussion of the concept of governance follows, introducing the municipal urban policy within which drainage and sanitation policies have been implemented. This paper presents the constitutional and institutional role of the five municipal committees dealing with water governance issues, as they are important arenas for civil society participation. The main constraints to achieving integrated urban water governance at the local level and the extent to which such policies are able to reduce social inequalities and promote social environmental justice in the use and appropriation of urban water, are discussed. This paper is part of the SWITCH-Sustainable Water Management Improves Tomorrow's Cities' Health-research network.

Detection and differentiation of Xylella fastidiosa strains acquired and retained by glassy-winged sharpshooters (Hemiptera: Cicadellidae) using a mixture of strain-specific primer sets.

Xylella fastidiosa Wells is a bacterial pathogen that causes a variety of plant diseases, including Pierce's disease (PD) of grapevine, almond leaf scorch, alfalfa dwarf, citrus variegated chlorosis, and oleander leaf scorch (OLS). Numerous strains of this pathogen have been genetically characterized, and several different strains occur in the United States. The dominant vector in southern California is the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The high mobility of this insect, and its use of large numbers of host plant species, provides this vector with ample exposure to multiple strains of X. fastidiosa during its lifetime. To learn more about the ability of this vector to acquire, retain, and transmit multiple strains of the pathogen, we developed a polymerase chain reaction (PCR)-based method to detect and differentiate strains of X. fastidiosa present in individual glassy-winged sharpshooter adults. Insects were sequentially exposed to plants infected with a PD strain in grapevine and an OLS strain in oleander. After sequential exposure, a few insects tested positive for both strains (7%); however, in most cases individuals tested positive for only one strain (29% PD, 41% OLS). In transmission studies, individual adults transmitted either the PD or OLS strain of the pathogen at a rate (39%) similar to that previously reported after exposure to a single strain, but no single individual transmitted both strains of the pathogen. PD and OLS strains of X. fastidiosa remained detectable in glassy-winged sharpshooter, even when insects were fed on a plant species that was not a host of the strain for 1 wk.

Differentiation of Strains of Xylella fastidiosa Infecting Grape, Almonds, and Oleander Using a Multiprimer PCR Assay.

Xylella fastidiosa is a xylem-limited, nutritionally fastidious bacterium that causes several plant diseases including Pierce's disease (PD) in grape and leaf scorch in almond (ALS) and oleander (OLS). OLS strains belong to X. fastidiosa subsp. sandyi, PD strains belong to X. fastidiosa subsp. fastidiosa, and strains from almond designated as ALS strains are of two general types belonging either to X. fastidiosa subsp. multiplex or X. fastidiosa subsp. fastidiosa. The ALS strains assigned to X. fastidiosa subsp. multiplex belong to two different genotypes (ALSI and ALSII) below the subspecies level. The OLS strains do not infect grape or almond. PD strains produce diseases in grape, alfalfa, almond, and some weeds, but they do not infect oleander, oak, peach, or citrus. ALS strains that belong to X. fastidiosa subsp. multiplex do not produce disease on grape. In this study, a relatively simple polymerase chain reaction (PCR) based method was developed to distinguish among PD, OLS, and ALS strains. PCR performed with primers XF1968-L and XF1968-R amplified a 638-bp fragment from OLS strains but not from PD strains or ALS strains that belong to X. fastidiosa subsp. fastidiosa. PCR with primers XF2542-L and XF2542-R amplified a 412-bp fragment from PD strains, but not from OLS strains. PCR with primers ALM1 and ALM2 produced a fragment of 521 bp from strains isolated from almond that belong to X. fastidiosa subsp. multiplex. The combination of the three primer sets allowed the distinction of the two ALS genotypes of X. fastidiosa subsp. multiplex. These results are in agreement with those obtained from analysis of sequences of 16S-23S rDNA intergenic spacer regions sequence analysis and with previous results based on randomly amplified polymorphic DNA analysis.

Discovery and Characterization of Xylella fastidiosa Strains in Southern California Causing Mulberry Leaf Scorch.

Mulberry leaf scorch (MLS), caused by Xylella fastidiosa, is a disease of mulberry trees in the United States that has largely been documented from locations in the eastern and central areas of the country. MLS was recently detected for the first time in white mulberry (Morus alba) trees in southern California. Four MLS-strains were isolated from two locations and confirmed as X. fastidiosa by enzyme-linked immunosorbent assay (ELISA), direct isolation of the pathogen, and use of the X. fastidiosa-specific PCR primers RST31-33. Isolated strains were characterized by the sequencing of their 16S-23S rDNA intergenic spacer regions (ISR) and random amplified polymorphic DNA (RAPD) analysis and subsequent comparison with a previously characterized MLS-strain (Mulberry-VA) and representatives of X. fastidiosa subsp. fastidiosa, X. fastidiosa subsp. multiplex, and X. fastidiosa subsp. sandyi. MLS-strains isolated from California were distinct from strains causing almond leaf scorch, oleander leaf scorch, and Pierce's disease and similar to the Mulberry-VA-strain. The ISR sequences of two MLS-strains, MLS063 and MLS059, were 100% identical to that of the Mulberry-VA sequence, whereas MLS012 and MLS024 were 99.8 and 99.6% identical to the Mulberry-VA-strain and 99.4% identical among themselves. Genomic analysis using RAPD revealed no differences among the four strains. The pathogenicity of one strain, MLS063, was confirmed by inoculation of glasshouse-grown white mulberry plants. Three months after inoculation, the pathogen was recovered from 21 of 25 inoculated plants, and 5 of 25 plants were dead within a year of inoculation. Inoculation of grapevines and oleanders with MLS063 did not result in any disease or recovery of the pathogen up to 1 year later, showing that this strain was not cross-infective to these hosts.

Plant Hosts of Xylella fastidiosa In and Near Southern California Vineyards.

Xylella fastidiosa is a xylem-limited bacterium that causes Pierce's disease (PD) of grapevines. A variety of plant species found near a severe outbreak of PD in vineyards in the Temecula Valley of California were tested using enzyme-linked immunosorbent assay, culture on media, and polymerase chain reaction to identify potential inoculum sources in the area. Species that consistently tested positive for X. fastidiosa were the known hosts, grape, almond, and oleander, and two new hosts, Spanish broom (Spartium junceum) and wild mustard (Brassica spp). Sequence analysis of the 16S-23S rRNA spacer region found that strains isolated from grapevine, Spanish broom, wild mustard, and almond clustered with previously sequenced PD strains. Thus, these species could serve as sources of inoculum for infection of grapevines and should be removed or monitored for signs of infection. Sequences from oleander isolates from Temecula formed another cluster with a previously published oleander strain sequence. Oleander strains do not infect grapevines and thus do not appear to cause a direct threat to grapevines. Two additional isolates from almond were determined to be genetically different from PD strains, and the ability of these strains to infect grapevine is not known. Greenhouse transmission studies indicate that the glassy-winged sharpshooter was able to transmit a PD strain of X. fastidiosa to Spanish broom, black mustard, and other hosts.

Field trials measuring the effects of ultraviolet-absorbing greenhouse plastic films on insect populations.

Field studies were conducted to compare insect population levels in greenhouse crops covered with plastics that block the transmission of UV light in two wavelength ranges. Crops grown in greenhouses under a plastic that blocked UV light at wavelengths of 380 nm and below had lower numbers of aphids and thrips compared with a plastic that blocked UV light at wavelengths of 360 nm and below. This is consistent with the results found for thrips in previous studies using small, completely enclosed tunnels with no plant material. The effects were not as dramatic in the commercial greenhouses, perhaps due to unfiltered light entering through the open sides of those greenhouses. There was no reduction in greenhouse whitefly, Trialeurodes vaporariorum Westwood, populations under the <380 UV-absorbing plastic compared to <360 nm UV-absorbing plastic in these field trials. This is inconsistent with results found in small, completely enclosed tunnels, where sticky traps caught a significantly higher proportion (95 +/- 2%) of released greenhouse whiteflies inside tunnels covered with <360 nm absorbing plastic compared with the <380 nm absorbing plastic. The results of these studies suggest that the type of greenhouse plastic used in a structure can affect population levels of some insect species, and may be useful tools in developing integrated pest management programs for insect management. The design of the greenhouse and amount of unfiltered light that enters the system appear to be important factors in determining the level of effect.

Seasonal flight activity of two Homalodisca species (Homoptera: Cicadellidae) that spread Xylella fastidiosa in southern California.

Homalodisca coagulata (Say) and Homalodisca lacerta (Fowler) are vectors of a new bacterial disease of oleander in California known as oleander leaf scorch, induced by the bacterium Xylella fastidiosa. H. coagulata also has been implicated in the spread of the strain of X. fastidiosa that induces Pierce's disease of grapevines in California. We monitored the flight activity of H. coagulata and H. lacerta in oleander and citrus by using yellow sticky cards at three southern California locations where outbreaks of oleander leaf scorch have been documented, and where vector compliments are different. Areas sampled included a mesic coastal area (Irvine, CA) that supports predominantly H. coagulata and few H. lacerta, a dry inland location (Palm Desert, CA) that supports predominantly H. lacerta and few H. coagulata, and an intermediate area (Riverside, CA) supporting both Homalodisca species. From November 1996 to October 1999 peak catches of both Homalodisca species occurred during the midsummer at all locations. H. coagulata was trapped in greater numbers in citrus than in oleander at both the Riverside and the Irvine sites. Likewise, H. lacerta in Riverside was more associated with citrus than oleander, yet H. lacerta in Palm Desert was trapped in greater numbers in oleander than citrus.

Solution structure of plantaricin C, a novel lantibiotic.

Plantaricin C, a bacteriocin produced by a Lactobacillus plantarum strain of dairy origin, is a lantibiotic. One dehydroalanine, one lanthionine and three beta-methyl-lanthionine residues were found in its 27 amino acid sequence. The plantaricin C structure has two parts: the first comprises the six NH2-terminal residues, four of which are lysines, which confer a strong positive charge to this stretch. The amino acids in positions 7 and 27 form the lanthionine bridge, giving a globular conformation to the rest of the molecule. The beta-methyl-lanthionine bridges are established between residues 12-15, 13-18 and 23-26. This central region has a charge distribution compatible with an amphipathic alpha-helix, through which plantaricin C would become inserted into the membrane matrix of sensitive organisms, provoking the opening of pores and leakage of the cytoplasmic content.

Cytochrome c" from the obligate methylotroph Methylophilus methylotrophus, an unexpected homolog of sphaeroides heme protein from the phototroph Rhodobacter sphaeroides.

The complete primary structure of an unusual soluble cytochrome c isolated from the obligate methylotrophic bacterium Methylophilus methylotrophus has been determined to contain 124 amino acids and to have an average molecular mass of 14293.0 Da. The sequence has two unusual features: firstly, the location of the heme-binding cysteines is far downstream from the N-terminus, namely at positions 49 and 52; secondly, an extra pair of cysteine residues is present near the C-terminus. In both respects, cytochrome c" is similar to the oxygen-binding heme protein SHP from the purple phototrophic bacterium Rhodobacter sphaeroides. In contrast to SHP, cytochrome c" changes from low-spin to high-spin upon reduction, due to dissociation of a sixth heme ligand histidine which is identified as His-95 by analogy to the class I cytochromes c. The distance of His-95 from the heme (41 residues) and the presence of certain consensus residues suggests that cytochrome c" is the second example of a variant class I cytochrome c.

Solution structure of Desulfovibrio vulgaris (Hildenborough) ferrocytochrome c3: structural basis for functional cooperativity.

Desulfovibrio vulgaris cytochrome c3 is a 14 kDa tetrahaem cytochrome that plays a central role in energy transduction. The three-dimensional structure of the ferrocytochrome at pH 8.5 was solved through two-dimensional 1H-NMR. The structures were calculated using a large amount of experimental information, which includes upper and lower distance limits as well as dihedral angle restraints. The analysis allows for fast-flipping aromatic residues and flexibility in the haem plane. The structure was determined using 2289 upper and 2390 lower distance limits, 63 restricted ranges for the phi torsion angle, 88 stereospecific assignments out of the 118 stereopairs with non-degenerate chemical shifts (74.6%), and 115 out of the 184 nuclear Overhauser effects to fast-flipping aromatic residues (62.5%), which were pseudo-stereospecifically assigned to one or the other side of the ring. The calculated NMR structures are very well defined, with an average root-mean-square deviation value relative to the mean coordinates of 0.35 A for the backbone atoms and 0.70 A for all heavy-atoms. Comparison of the NMR structures of the ferrocytochrome at pH 8.5 with the available X-ray structure of the ferricytochrome at pH 5.5 reveals that the general fold of the molecule is very similar, but that there are some distinct differences. Calculation of ring current shifts for the residues with significantly different conformations confirms that the NMR structures represent better its solution structure in the reduced form. Some of the localised differences, such as a reorientation of Thr24, are thought to be state-dependent changes that involve alterations in hydrogen bond networks. An important rearrangement in the vicinity of the propionate groups of haem I and involving the covalent linkage of haem II suggests that this is the critical region for the functional cooperativities of this protein.

Assignment of the ligand geometry and redox potentials of the trihaem ferricytochrome c3 from Desulfuromonas acetoxidans.

Cytochrome c551.5 is a trihaem cytochrome of the cytochrome c3 family isolated from Desulfuromonas acetoxidans. Although several X-ray structures are available for tetrahaem cytochromes of this family, there is no X-ray structure for trihaem cytochromes. Cytochrome C551.5 was studied in the oxidized form by means of two-dimensional NMR. The pattern of observed interhaem NOESY connectivities is in agreement with the haem core structure previously determined by NMR for the reduced protein [Coutinho, I. B., Turner, D. L., Liu, M. Y., LeGall, J. & Xavier, A. V. (1996) J. Biol. Inorg. Chem. 1, 305-311]. The similarities found between the haem core structure and the amino acid sequence of cytochrome c551.5 and those of tetrahaem cytochromes c3 allows each of the haems to be specifically assigned in the polypeptide sequence, and the attribution of the midpoint redox potentials to the individual haems. This also allows individual redox potentials to be assigned to each haem in the NMR spectrum. The paramagnetic shifts of the 13C resonances of the haem substituents were analyzed in terms of pi molecular orbitals with perturbed D4h symmetry. The parameters of this analysis have been shown to be controlled by the orientation of the axial ligands in several other bis-His-coordinated haems and hence the ligand geometry was deduced for cytochrome C551.5. The structural analogy between the relative haem plane orientations in cytochrome c551.5 and the tetrahaem cytochromes c3 is found to extend to the axial ligands with the largest differences being in the vicinity of the deleted fourth haem, using the numbering of cytochrome c3 haems.

An unusual conformation of the methionine haem ligand in cytochrome cL established by two-dimensional 1H-NMR.

A complete relaxation-matrix analysis of NOESY cross-peak intensities was used to determine the conformation of the methionine ligand to the haem group in two ferrocytochromes cL from Methylophilus methylotrophus and Methylobacterium extorquens, including the configuration at the sulphur. The conformation of the axial methionine is of a type reported only for the cytochromes c5 from Pseudomonas mendocina and Azotobacter vinelandii. Although the conformation of the methionine is unusual, the paramagnetic shifts of the haem methyl proton resonances in the oxidized proteins indicate that the electronic structure of the haem groups is similar to that found in the mitochondrial type of cytochrome c.

Characterization of the haem environment in Methylophilus methylotrophus ferricytochrome c" by 1H-NMR.

Two-dimensional NMR techniques have been used to assign proton resonances in the haem cavity of Methylophilus methylotrophus cytochrome c", a monohaem protein with bis-histidinyl ligation which has been shown to couple electron and proton transfer. All the assignments were made directly for the oxidized paramagnetic form of the cytochrome. Nearly all of the haem protons (90%) and the protons of both axial ligands have been assigned; the side-chain protons from four other residues in the haem pocket have also been identified. The data indicate a highly symmetric unpaired-electron distribution in the haem group, which agrees with a perpendicular orientation of the axial imidazole planes. The two haem propionate groups have contrasting degrees of exposure to the solvent, with the propionate group at position 13 being highly exposed. To obtain information on the dynamics of the haem environment, measurements of the 1H/2H-exchange rates of amide protons located in the haem cavity were performed. The two faces of the haem are found to differ markedly with respect to water accessibility. All of this information, together with additional protein sequencing data, indicates that His52 remains attached upon reduction and that the redox-linked protonation occurs via a channel running through the haem cleft on the opposite face.

Pitfalls in assigning heme axial coordination by EPR. c-Type cytochromes with atypical Met-His ligation.

Different monohemic c-type cytochromes were analyzed by visible, EPR and 1H NMR spectroscopies. While the visible and NMR data show unambiguously that the heme iron has a Met-His heme axial coordination, the EPR data indicate an axial ligand field typical of that for a bis-histidinyl ligation. The validity of the widely used EPR methods for the determination of the heme iron axial coordination, based on the crystal field parameters (tetragonality and rhombicity), is questioned.