Implications of nighttime temperature on metamitron impacts on the photosynthetic machinery functioning of Malus x domestica Borkh.

Metamitron (MET) is a fruitlet thinning compound for apple trees, needing better understanding of its action on leaf energy metabolism, depending on nighttime temperature. A trial under environmental controlled conditions was set with 'Golden Reinders' potted trees, under 25/7.5 and 25/15 °C (diurnal/nighttime temperature), with (MET, 247.5 ppm) or without (CTR) application, and considering the monitoring of photosynthetic and respiration components from day 1 (D1) to 14 (D14). Net photosynthesis (Pn) decline promoted by MET after D1 was not stomatal related. Instead, non-stomatal constraints, reflected on the photosynthetic capacity (Amax), included a clear photosystem (PS) II inhibition (but barely of PSI), as shown by severe reductions in thylakoid electron transport at PSII level, maximal (Fv/Fm) and actual (Fv'/Fm') PSII photochemical efficiencies, estimate of quantum yield of linear electron transport (Y(II)), and the rise in PSII photoinhibition status (Fs/Fm' and PIChr) and uncontrolled energy dissipation (Y(NO)). To Pn inhibition also contributed the impact in RuBisCO along the entire experiment, regardless of night temperature, here reported for the first time. Globally, MET impact on the photosynthetic parameters was usually greater under 7.5 °C, with maximal impacts between D4 and D7, probably associated to a less active metabolism at lower temperature. Cellular energy metabolism was further impaired under 7.5 °C, through moderate inhibition of NADH-dependent malate dehydrogenase (MDH) and pyruvate kinase (PK) enzymes involved in respiration, in contrast with the increase of dark respiration in MET 7.5 until D7. The lower impact on PK and MDH under 15 °C and a likely global higher active metabolism at that temperature would agree with the lowest sucrose levels in MET 15 at D4 and D7. Our findings showed that MET alters the cell energy machinery in a temperature dependent manner, affecting the sucrose balance mainly at 15 °C, justifying the observed greater thinning potential.

Evaluation of human papillomavirus self-collection offered by community health workers at home visits among under-screened women in Brazil.

OBJECTIVE: To explore the acceptability of high-risk human papillomavirus self-testing, involving community health workers, for never/under-screened Brazilian women. Cervical cancer is the most common cause of cancer-related death among adult women in a large number of low-income and lower-middle-income countries, where it remains a major public health problem. High-risk human papillomavirus persistence is required for the development of cervical neoplasia. METHODS: The target population was all women aged 30+ from the list of families available in healthcare centre data, who had never been screened or were not screened in the previous 3 years (under-screened women), and who were living in the 17 cities included in this study. RESULTS: Of the 377 women included, 16.9% (n = 64) had never had a pap smear. Of all samples included in the study, 97.1% (n = 366) were considered adequate for evaluation, as 2.9% (n = 11) were considered invalid for all high-risk human papillomavirus types. Analysing these 366 samples, 9.6% (n = 35) of the women were infected by at least one high-risk human papillomavirus type and 90.4% (n = 331) had no infection with any high-risk type of the virus. CONCLUSIONS: Vaginal self-sampling is an adequate strategy to improve the effectiveness of the cervical cancer program by increasing screening in a high-risk group.

Long-Term Assessment of Air Quality and Identification of Aerosol Sources at Setúbal, Portugal.

Understanding air pollution in urban areas is crucial to identify mitigation actions that may improve air quality and, consequently, minimize human exposure to air pollutants and their impact. This study aimed to assess the temporal evolution of the air quality in the city of Setúbal (Portugal) during a time period of 10 years (2003-2012), by evaluating seasonal trends of air pollutants (PM10, PM2.5, O3, NO, NO2 and NOx) measured in nine monitoring stations. In order to identify emission sources of particulate matter, PM2.5 and PM2.5-10 were characterized in two different areas (urban traffic and industrial) in winter and summer and, afterwards, source apportionment was performed by means of Positive Matrix Factorization. Overall, the air quality has been improving over the years with a decreasing trend of air pollutant concentration, with the exception of O3. Despite this improvement, levels of PM10, O3 and nitrogen oxides still do not fully comply with the requirements of European legislation, as well as with the guideline values of the World Health Organization (WHO). The main anthropogenic sources contributing to local PM levels were traffic, industry and wood burning, which should be addressed by specific mitigation measures in order to minimize their impact on the local air quality.

Characterization of atmospheric aerosol (PM10 and PM2.5) from a medium sized city in São Paulo state, Brazil.

Air pollution causes deleterious effects on human health with aerosols being among the most polluting agents. The objective of this work is the characterization of the PM2.5 and PM10 aerosol mass in the atmosphere. The methods of analysis include WD-XRF and EDS. Data were correlated with meteorological information and air mass trajectories (model HYSPLIT) by multivariate analysis. A morphological structural analysis was also carried out to identify the probable sources of atmospheric aerosols in the city of São José do Rio Preto, Brazil. The mean mass concentration values obtained were 24.54 µg/m3 for PM10, above the WHO annual standard value of 20 µg/m3 and 10.88 µg/m3 for PM2.5 whose WHO recommended limit is 10 µg/m3. WD-XRF analysis of the samples revealed Si and Al as major components of the coarse fraction. In the fine fraction, the major elements were Al and S. The SEM-FEG characterization allowed identifying the morphology of the particles in agglomerates, ellipsoids and filaments in the PM10, besides spherical in the PM2.5. The analysis by EDS corroborated WD-XRF results, identifying the crustal elements, aluminosilicates and elements of anthropogenic origin in the coarse fraction. For the fine fraction crustal elements were also identified; aluminosilicates, black carbon and spherical particles (C and O) originating from combustion processes were predominant. The use of multivariate analysis to correlate air mass trajectories with the results of the morpho-structural characterization of the particulate matter allowed confirmation of the complex composition of the particles resulting from the combination of both local and long-distance sources.

Impact of recovery correction or subjecting calibrators to sample preparation on measurement uncertainty: PAH determinations in waters.

The decision on the fitness of a measurement for its intended use and the interpretation of an analytical result requires the assessment of the measurement uncertainty. Frequently, the determination of analytes in complex matrices involves demanding sample preparations in which analyte losses are observed. These losses should be considered when reporting the results, which can be corrected for low recovery by taking the mean recovery observed in the analysis of reference items (e.g. spiked samples) or, alternatively, by subjecting calibrators to the same pre-treatment performed on the samples. In these cases, neat (NC) or adjusted (AC) calibrators are used, respectively. The way analyte losses are handled impacts on the measurement uncertainty. The top-down evaluation of the measurement uncertainty involves combining precision, trueness and additional uncertainty components. The trueness component is quantified by pooling various analyte recovery determinations. This work assesses and compares the uncertainty of polycyclic aromatic hydrocarbons (PAHs) measurements in water based on HPLC-FD calibrations with NC or AC. The trueness component is estimated by pooling mean recoveries observed from the analysis of different spiked samples to which mean recovery uncertainty and degrees of freedom are used to estimate a weighted mean recovery and respective uncertainty. The performance of measurements based on NC and AC are associated with equivalent uncertainty except when large analyte losses are observed, namely in the determination of Naphtalene. In this case, the processing of AC reduces the expanded relative uncertainty from 9.9% to 3.5%. The evaluated expanded uncertainty ranged from 3.5% to 12% of the measured value.

Adaptation of Alpha-Papillomavirus over Millennia.

Papillomaviruses (PVs) are a group of small DNA viruses that, with around 350 million years of evolution, acquired the capacity of infecting a broad range of vertebrates, including humans. To date, more than 300 PV types have been isolated. Viruses that have a long common evolutionary history with their host typically cause unapparent infections. However, in some Alpha-PV infections, lesions become apparent and may cause benign proliferative disorders or even malignant proliferative lesions of the cervix, vulva, vagina, anus, penis, and oropharynx. The incongruence observed between the topology of the phylogenetic tree of Alpha-PVs and that of their hosts suggests that virus-host codivergence is not the only evolutionary force that has driven the progression of PVs. The integration of the precursors of E5, E6, and E7 on the genome of the ancestral Alpha-PV was important and made the colonization of new niches and the emergence of carcinogenic types possible.

Racial/Ethnic Disparities in Cervical Cancer Screening and Outcomes.

Invasive cervical cancer disproportionately affects women without sufficient access to care, with higher rates among minority groups in higher-income countries and women in low-resource regions of the world. Many elements contribute to racial/ethnic disparities in the cervical cancer continuum - from screening and diagnosis to treatment and outcome. Sociodemographic factors, access to healthcare, income and education level, and disease stage at diagnosis are closely linked to such inequities. Despite the identification of such elements, racial/ethnic disparities persist, and are widening in several minority subgroups, particularly in older women, who are ineligible for human papillomavirus (HPV) vaccination and are underscreened. Recent studies suggest that racial/ethnic differences in HPV infection exist and may also have a role in observed differences in cervical cancer. In this review, we provide an overview of the current literature on racial disparities in cervical cancer screening, incidence, treatment and outcome to inform future strategies to reduce persistent inequities.

Shared and divergent pathways for flower abscission are triggered by gibberellic acid and carbon starvation in seedless Vitis vinifera L.

BACKGROUND: Abscission is a highly coordinated developmental process by which plants control vegetative and reproductive organs load. Aiming at get new insights on flower abscission regulation, changes in the global transcriptome, metabolome and physiology were analyzed in 'Thompson Seedless' grapevine (Vitis vinifera L.) inflorescences, using gibberellic acid (GAc) spraying and shading as abscission stimuli, applied at bloom. RESULTS: Natural flower drop rates increased from 63.1% in non-treated vines to 83% and 99% in response to GAc and shade treatments, respectively. Both treatments had a broad effect on inflorescences metabolism. Specific impacts from shade included photosynthesis inhibition, associated nutritional stress, carbon/nitrogen imbalance and cell division repression, whereas GAc spraying induced energetic metabolism simultaneously with induction of nucleotide biosynthesis and carbon metabolism, therefore, disclosing alternative mechanisms to regulate abscission. Regarding secondary metabolism, changes in flavonoid metabolism were the most represented metabolic pathways in the samples collected following GAc treatment while phenylpropanoid and stilbenoid related pathways were predominantly affected in the inflorescences by the shade treatment. However, both GAc and shade treated inflorescences revealed also shared pathways, that involved the regulation of putrescine catabolism, the repression of gibberellin biosynthesis, the induction of auxin biosynthesis and the activation of ethylene signaling pathways and antioxidant mechanisms, although often the quantitative changes occurred on specific transcripts and metabolites of the pathways. CONCLUSIONS: Globally, the results suggest that chemical and environmental cues induced contrasting effects on inflorescence metabolism, triggering flower abscission by different mechanisms and pinpointing the participation of novel abscission regulators. Grapevine showed to be considered a valid model to study molecular pathways of flower abscission competence acquisition, noticeably responding to independent stimuli.

Molecular candidates for early-stage flower-to-fruit transition in stenospermocarpic table grape (Vitis vinifera L.) inflorescences ascribed by differential transcriptome and metabolome profiles.

Flower-to-fruit transition depends of nutrient availability and regulation at the molecular level by sugar and hormone signalling crosstalk. However, in most species, the identities of fruit initiation regulators and their targets are largely unknown. To ascertain the main pathways involved in stenospermocarpic table grape fruit set, comprehensive transcriptional and metabolomic analyses were conducted specifically targeting the early phase of this developmental stage in 'Thompson Seedless'. The high-throughput analyses performed disclosed the involvement of 496 differentially expressed genes and 28 differently accumulated metabolites in the sampled inflorescences. Our data show broad transcriptome reprogramming of molecule transporters, globally down-regulating gene expression, and suggest that regulation of sugar- and hormone-mediated pathways determines the downstream activation of berry development. The most affected gene was the SWEET14 sugar transporter. Hormone-related transcription changes were observed associated with increased indole-3-acetic acid, stimulation of ethylene and gibberellin metabolisms and cytokinin degradation, and regulation of MADS-box and AP2-like ethylene-responsive transcription factor expression. Secondary metabolism, the most representative biological process at transcriptome level, was predominantly repressed. The results add to the knowledge of molecular events occurring in grapevine inflorescence fruit set and provide a list of candidates, paving the way for genetic manipulation aimed at model research and plant breeding.

Flower abscission in Vitis vinifera L. triggered by gibberellic acid and shade discloses differences in the underlying metabolic pathways.

Understanding abscission is both a biological and an agronomic challenge. Flower abscission induced independently by shade and gibberellic acid (GAc) sprays was monitored in grapevine (Vitis vinifera L.) growing under a soilless greenhouse system during two seasonal growing conditions, in an early and late production cycle. Physiological and metabolic changes triggered by each of the two distinct stimuli were determined. Environmental conditions exerted a significant effect on fruit set as showed by the higher natural drop rate recorded in the late production cycle with respect to the early cycle. Shade and GAc treatments increased the percentage of flower drop compared to the control, and at a similar degree, during the late production cycle. The reduction of leaf gas exchanges under shade conditions was not observed in GAc treated vines. The metabolic profile assessed in samples collected during the late cycle differently affected primary and secondary metabolisms and showed that most of the treatment-resulting variations occurred in opposite trends in inflorescences unbalanced in either hormonal or energy deficit abscission-inducing signals. Particularly concerning carbohydrates metabolism, sucrose, glucose, tricarboxylic acid metabolites and intermediates of the raffinose family oligosaccharides pathway were lower in shaded and higher in GAc samples. Altered oxidative stress remediation mechanisms and indolacetic acid (IAA) concentration were identified as abscission signatures common to both stimuli. According to the global analysis performed, we report that grape flower abscission mechanisms triggered by GAc application and C-starvation are not based on the same metabolic pathways.

Designing valid and optimised standard addition calibrations: Application to the determination of anions in seawater.

A strategy for designing valid standard addition calibrations and for optimising their uncertainty is presented. The design of calibrations involves the development of models of the sensitivity and precision of the instrumental signal, in a wide range of analyte concentration (or any other studied quantity), and the definition of sample dilution and standard addition procedures that allow fulfilling the assumptions of the linear unweighted regression model in, typically, a smaller range of standard addition calibrations. Calibrators are prepared by diluting the sample and adding analyte with negligible uncertainty to fit in a concentration range where signals are homoscedastic. The minimisation of the uncertainty is supported on detailed measurement uncertainty models function of the calibrators preparation procedure and of analytical instrumentation performance. The number of collected signals replicates is defined by balancing their impact on the estimated expanded uncertainty, the resources needed and the target (maximum) uncertainty for the intended use of measurements. The calibration design strategy was successfully applied to the determination of the mass concentration (mg L(-1)) of Cl(-), Br(-), NO3(-) and SO4(-2) in seawater by ion chromatography. A target expanded uncertainty of 20% was defined for the determination of Cl(-), NO3(-) and SO4(-2), or 40% for the determination of the smaller mass concentration of Br(-). The developed measurement model produced reliable predictions of the measurement uncertainty from approximate concentration of the analyte in the sample, before its accurate quantification, thus proving optimisation is effective. Predictions are more prone to the variability of the measurement uncertainty estimation if based on low number of calibrators signals. The reported relative expanded uncertainty ranged from 7.1% to 49%.

Multilocus profiling with AFLP, ISSR, and SAMPL.

Molecular markers which sample multiple loci simultaneously, like amplified fragment length polymorphism (AFLP), inter-simple sequence repeats (ISSR), and selective amplification of microsatellite polymorphic loci (SAMPL), produce highly informative fingerprints due to their high effective multiplex ratio and expected heterozygosity. Moreover, these markers can be generated for DNA of any organism without initial investment in primer/probe development or in sequence analyses. The fragments produced can be visualized either by agarose or polyacrylamide gel electrophoresis followed by autoradiography or silver staining or via separation and detection on automatic DNA sequencers. Here, we describe detailed protocols based on the original methods aimed to obtain these markers optimized to be resolved on polyacrylamide gel electrophoresis and detected by silver staining which provides a fast, sensitive, and cost-effective method.

Evaluation of the performance of the determination of anions in the water soluble fraction of atmospheric aerosols.

The knowledge of the mass of particulate matter in air, its chemical composition and emission sources is of relevance for taking decisions concerning air quality management in urban areas. The interpretation of these data is a function of the quality of the measurement results expressed by their uncertainties. This study aimed at developing models of the performance of the determination of anions in the water-soluble fraction of atmospheric aerosols, capable of determining, separately, the contribution of aerosols sampling, extraction of water-soluble fraction of atmospheric aerosols and quantification, by ion chromatography, of anions in the extract. The sampling procedure was assessed from the dispersion of results of duplicate parallel sampling after subtracting the analytical component of this dispersion. These models are used to evaluate the adequacy of the measurement procedure for the determination of urban aerosol composition and to support strategies for reducing measurement uncertainty or cost of analysis. The method performance was studied for the following ranges considering extract dilution up to five times: 0.23-8 µg m(-3) for chloride and nitrate, and 0.093-3.25 µg m(-3) for sulphate. Measurements are fit for the analysis of urban aerosols since the relative expanded measurement uncertainty is smaller than a maximum value of 40%. The percentage contribution of the uncertainty components varies with the analyte and its mass concentration, the major components being 24-93% for the extraction, 43-59% for sampling, 0.2-28% for the interpolation of the sample signal in the calibration curve and 4-8% for air volume measurement. The typical composition of analysed air is: (1.12±0.26) µg m(-3), (1.02±0.30) µg m(-3) and (0.76±0.22) µg m(-3) of chloride, nitrate and sulphate in the water soluble fraction of aerosol, respectively, for a confidence level of approximately 95% considering a coverage factor of 2.

Wavefront analysis and Zernike polynomial decomposition for evaluation of corneal optical quality.

Wavefront-guided excimer laser refractive surgery and new customized intraocular lens and contact lens designs are major clinical applications of corneal wavefront analysis. Other therapeutic applications include corneal disorders, conventional excimer laser refractive surgery, incisional techniques and cataract surgery, corneal transplantation, intrastromal corneal ring segment implantation, and crosslinking therapy. Basic data regarding corneal wavefront aberrations, such as distribution in the population and changes with aging, are essential for understanding the nature of each aberration and correcting it. Corneal aberrometry also improved our comprehension of the optical effects of the aforementioned topics while helping us assess the success of the procedures. Zernike polynomials are representations of the higher- and lower-order aberrations of the cornea, allowing a mathematical approach to their determination. Polynomials are used to model individual components of the wavefront in familiar terms. This article reviews the current knowledge of the wavefront aberrations of the human cornea and analyzes studies in the fields of anterior segment surgery and/or therapy, diseases, and optical quality in the context of this knowledge.

Corneal imaging with slit-scanning and Scheimpflug imaging techniques.

Modern anterior segment imaging techniques, such as slit-scanning and Scheimpflug imaging, greatly improved the field of corneal imaging. Devices such as the Orbscan (Bausch & Lomb Surgical, Inc) and, more recently, the Pentacam (Oculus, Inc) and the Galilei (Ziemer Ophthalmic Systems AG) have brought a whole new range of clinical possibilities and sparked interest in the academic community. These new tomographers create three-dimensional models of the cornea and enable the direct measurement of both the anterior and posterior surface elevations. In this paper, we present a review of some available literature on those systems, regarding their optical principles and their performance in current clinical applications.

Rhinovirus C and respiratory exacerbations in children with cystic fibrosis.

To investigate a possible role for human rhinovirus C in respiratory exacerbations of children with cystic fibrosis, we conducted microbiologic testing on respiratory specimens from 103 such patients in São Paulo, Brazil, during 2006-2007. A significant association was found between the presence of human rhinovirus C and respiratory exacerbations.

HIV type 1 subtype C and CB Pol recombinants prevail at the cities with the highest AIDS prevalence rate in Brazil.

HIV-1 B is predominant in Brazil, but HIV-1 C has increasingly been reported in the south of the country. However, many samples clustering with clade C are actually a recombinant, with a small B segment at RT (CRF31). Samples (209) from the three cities with the highest aids prevalence rate are analyzed. Partial polymerase sequences from HIV RNA made it possible to determine HIV clades and recombination patterns and to identify primary drug resistance mutations (DRMs). The incidence was estimated with a BED assay. HIV-1 C and CRF31 patterns were twice as frequent as clade B at all sites, but the proportion of C and CRF31 patterns was significantly different among sites. The incidence estimate for SC was 2.6 persons-years. Infection in recent or younger cases showed no association with clade C. Surveillance DRM was observed in 8.3% (95% CI 5-13), mostly to NNRTIs. Clade F pol genomes had significantly more primary DRM.

HIV genotyping among female sex workers in the State of Santa Catarina.

The objective of this study was to investigate the frequency of HIV infection among female sex workers in the port area of Imbituba (State of Santa Catarina), and to identify the viral subtype and its susceptibility to antiretroviral medications. Ninety women were interviewed between December 2003 and February 2004. Six (6.7%) were HIV-positive. Genotyping for HIV, performed on four samples, detected subtype C in three of them, which is predominant in Africa and Asia, and subtype B in one of them, which is prevalent in Brazil, USA and Europe. The results suggest that the Port of Imbituba may be one of the gateways for HIV-1 subtype C to enter Brazil, and for its dissemination to the rest of the country and the Mercosul area, along the highway BR-101. This points towards the need for preventive work to reduce the introduction and dissemination of HIV subtype C in Brazil.

HIV genotyping among female sex workers in the State of Santa Catarina

The objective of this study was to investigate the frequency of HIV infection among female sex workers in the port area of Imbituba (State of Santa Catarina), and to identify the viral subtype and its susceptibility to antiretroviral medications. Ninety women were interviewed between December 2003 and February 2004. Six (6.7 percent) were HIV-positive. Genotyping for HIV, performed on four samples, detected subtype C in three of them, which is predominant in Africa and Asia, and subtype B in one of them, which is prevalent in Brazil, USA and Europe. The results suggest that the Port of Imbituba may be one of the gateways for HIV-1 subtype C to enter Brazil, and for its dissemination to the rest of the country and the Mercosul area, along the highway BR-101. This points towards the need for preventive work to reduce the introduction and dissemination of HIV subtype C in Brazil.

O objetivo deste estudo foi verificar a freqüência da infecção pelo HIV em profissionais do sexo, atuantes em Imbituba (SC), identificar o subtipo viral e a suscetibilidade do vírus aos medicamentos antiretrovirais. De dezembro de 2003 a fevereiro de 2004, foram entrevistadas 90 mulheres, profissionais do sexo, e a freqüência de HIV nessa população foi de 6,7 por cento. O teste de genotipagem para o HIV, realizado em quatro amostras, detectou em três delas o subtipo C, que é predominante na Africa e Asia, e em uma o subtipo B, prevalente no Brasil, EUA e Europa. Os resultados sugerem que o Porto de Imbituba pode ser uma das portas de entrada para do HIV-1 subtipo C no Brasil, e a partir dessa localidade ocorrer sua disseminação para o restante do País e países do Mercosul pela rodovia BR-101. Isto aponta para a necessidade de trabalhos de prevenção, com a finalidade de reduzir a introdução, transmissão e disseminação do HIV subtipo C.

Low prevalence of primary antiretroviral resistance mutations and predominance of HIV-1 clade C at polymerase gene in newly diagnosed individuals from south Brazil.

We describe preliminary molecular characterization of HIV-1 pol from 108 consecutive HIV seropositive users of a Voluntary Counseling and Testing (VCT) site of Porto Alegre city, the major metropolitan area in the south of Brazil. Protease and partial reverse transcriptase regions were retrotranscribed from plasma HIV-1 RNA and sequenced after direct nested PCR. Principal antiretroviral resistance mutations (ARM) were observed in 3% of the samples, two cases with K103N and one with M41L, L210W and T215Y, all in HIV-1 clade B infected men. At protease region, no principal mutations were observed, but polymorphisms at secondary codons were frequent. Contrary to other areas in the country where clade B dominates, HIV-1 clade C genomes predominated in this study (58%), clade B (32%) and clade F1 (3%). Of the genomes clustering in clade C, almost half (43%) had a small clade B segment at reverse transcriptase, forming a sub-cluster within clade C with a similar recombinant structure and carrying new amino acid signatures. Other mosaic genomes were also observed (7%). The low prevalence of resistance mutations is consistent with previous observations at this geographical location but the high frequency of HIV-1 clade C and CB mosaics seems pre-eminent and warns close monitoring.