Insights about levels and sources of organic pollution in an urbanized Amazon estuary (Belém, PA, Northern Brazil).

Amazon aquatic systems have been affected by organic pollution from urbanized regions. This study was conducted to determine the levels, sources, and distribution patterns of 16 polycyclic aromatic hydrocarbons (PAHs) and 6 steroid markers in surficial sediments from an important urbanized Amazon estuarine system (Belém, PA, Northern Brazil). Total PAH concentration (∑PAH) ranged from 878.2 to 9905.7 ng g-1, 3295.2 ng g-1 on average, suggesting a highly contaminated environment. PAH molecular ratios and statistical analysis indicated that PAH originated from a mixture of local sources emissions, mainly related to the combustion of fossil fuels and biomass. Coprostanol levels (maximum concentration = 292.52 ng g-1) could be compared to the mid-range reported in the literature. Studied stations, except for one, presented sterol ratio data indicating organic matter related to untreated sewage. Sterols indicative of sewage contamination showed a correlation with pyrogenic PAH amounts which are transported by the same channels where sewage is discharged.

Combined Reconstruction of the Anterior Cruciate Ligament and Anterolateral Ligament Injury Compared to the Isolated Reconstruction of the Anterior Cruciate Ligament: A Meta-Analysis.

The present research aims to compare the outcomes from the combined reconstruction of the anterior cruciate ligament (ACL) and of the anterolateral ligament (ALL) with the standard isolated ACL reconstruction in patients with chronic ACL injury. To do so, a meta-analysis was carried out to determine whether the combined ACL and ALL reconstruction would lead to a significant improvement in knee function according to the International Knee Documentation Committee (IKDC), the Lysholm test and KT-2000 evaluation scores and lower graft rupture rates in comparison with isolated reconstruction. To identify randomized controlled trials (RCTs) comparing the combined ACL and ALL reconstruction with the isolated ACL reconstruction, papers published between 2010 and 2019 were searched in the MEDLINE, EMBASE, SPORTDiscus, LILACS and Cochrane Central Register of Controlled Trials databases, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) criteria. The stability of the knee joint is only marginally improved with the combined reconstruction of ACL and ALL, and both reconstruction techniques show functional results. The main outcomes sought were patient function and graft stability and rupture rates after ACL reconstruction. Out of the 421 studies identified, 6 were included in our meta-analysis. Study quality (internal validity) was assessed using the Cochrane risk-of-bias tool; in general, the studies included presented moderate-quality evidence. The graft rupture rate was higher in patients undergoing isolated ACL reconstruction (relative risk, 0.22; 95% confidence interval, 0.12 to 0.41; p < 0.00001).

The postharvest ripening of strawberry fruits induced by abscisic acid and sucrose differs from their in vivo ripening.

In this study, we compared the chemical composition of strawberry (Fragaria × ananassa) fruits that were ripened in vivo (attached to plant) to those ripened during postharvest storage. The effects of the application of abscisic acid (ABA) and sucrose on the postharvest ripening were also evaluated. The results suggested that the postharvest ripening process was dependent on the signal triggered by ABA and differed from in vivo ripening, resulting in fruits with altered chemical composition and firmness. The application of sucrose in unripe strawberries resulted in the induction of ripening, which is dependent on ABA and its derivatives. This induction was more pronounced during the first days of storage and associated with the application of mannitol rather than water, suggesting that mannitol negatively regulated the postharvest strawberry ripening. These results provide further insights into the role of ABA and sucrose in the regulation of postharvest ripening of strawberry.

ABA-dependent salt and drought stress improve strawberry fruit quality.

Strawberry crop is very sensitive to osmotic stress conditions. We investigated the effect of the stress induced by mild drought (DS) and salt (SS) stresses, on molecular, physiological, and metabolic processes in the strawberry crop (Fragaria ananassa), cv. Camarosa. The results showed that the fruit yield was not affected. Mild DS and SS also resulted in an increased content of phenolics, anthocyanins, and l-ascorbic acid, and an increased antioxidant activity. These effects were accompanied by increased levels of ABA and its derivatives (phaseic and dehydrophasic acids), alongside the upregulation of several genes involved on their synthesis. Therefore, the results obtained in this study suggest that mild DS and SS improve the functional quality of strawberry fruits through and ABA-dependent mechanism.

Usefulness of in-house PCR methods for hepatitis B virus DNA detection.

The aim of the present study was to evaluate the performance of three in-house PCR techniques for HBV DNA detection and compare it with commercial quantitative methods to evaluate the usefulness of in-house methods for HBV diagnosis. Three panels of HBsAg reactive sera samples were evaluated: (i) 50 samples were examined using three methods for in-house qualitative PCR and the Cobas Amplicor HBV Monitor Assay; (ii) 87 samples were assayed using in-house semi-nested PCR and the Cobas TaqMan HBV test; (iii) 11 serial samples obtained from 2 HBV-infected individuals were assayed using the Cobas Amplicor HBV test and semi-nested PCR. In panel I, HBV DNA was detected in 44 samples using the Cobas Amplicor HBV test, 42 samples using semi-nested PCR (90% concordance with Cobas Amplicor), 22 samples using PCR for the core gene (63.6% concordance) and 29 samples using single-round PCR for the pre-S/S gene (75% concordance). In panel II, HBV DNA was quantified in 78 of the 87 HBsAg reactive samples using Cobas TaqMan but 52 samples using semi-nested PCR (67.8% concordance). HBV DNA was detected in serial samples until the 17th and 26th week after first donation using in-house semi-nested PCR and the Cobas Amplicor HBV test, respectively. In-house semi-nested PCR presented adequate concordance with commercial methods as an alternative method for HBV molecular diagnosis in low-resource settings.

Carotenoid biosynthetic and catabolic pathways: gene expression and carotenoid content in grains of maize landraces.

Plant carotenoids have been implicated in preventing several age-related diseases, and they also provide vitamin A precursors; therefore, increasing the content of carotenoids in maize grains is of great interest. It is not well understood, however, how the carotenoid biosynthetic pathway is regulated. Fortunately, the maize germplasm exhibits a high degree of genetic diversity that can be exploited for this purpose. Here, the accumulation of carotenoids and the expression of genes from carotenoid metabolic and catabolic pathways were investigated in several maize landraces. The carotenoid content in grains varied from 10.03, in the white variety MC5, to 61.50 µg·g⁻¹, in the yellow-to-orange variety MC3, and the major carotenoids detected were lutein and zeaxanthin. PSY1 (phythoene synthase) expression showed a positive correlation with the total carotenoid content. Additionally, the PSY1 and HYD3 (ferredoxin-dependent di-iron monooxygenase) expression levels were positively correlated with ß-cryptoxanthin and zeaxanthin, while CYP97C (cytochrome P450-type monooxygenase) expression did not correlate with any of the carotenoids. In contrast, ZmCCD1 (carotenoid dioxygenase) was more highly expressed at the beginning of grain development, as well as in the white variety, and its expression was inversely correlated with the accumulation of several carotenoids, suggesting that CCD1 is also an important enzyme to be considered when attempting to improve the carotenoid content in maize. The MC27 and MC1 varieties showed the highest HYD3/CYP97C ratios, suggesting that they are promising candidates for increasing the zeaxanthin content; in contrast, MC14 and MC7 showed low HYD3/CYP97C, suggesting that they may be useful in biofortification efforts aimed at promoting the accumulation of provitamin A. The results of this study demonstrate the use of maize germplasm to provide insight into the regulation of genes involved in the carotenoid pathway, which would thus better enable us to select promising varieties for biofortification efforts.

Selection of reliable reference genes for quantitative real-time polymerase chain reaction studies in maize grains.

KEY MESSAGE: The stability of candidate reference genes was evaluated in maize landrace varieties and during multiple grain developmental stages to evaluate the expression of carotenoid-related genes by RT-qPCR for application to maize biofortification. Vitamin A deficiency affects millions of children worldwide; therefore, increasing the content of vitamin A precursors in maize grains is of interest. The study of the expression of genes involved in the carotenoid biosynthetic pathway in maize grains has provided useful information for metabolic engineering approaches. However, reliable results using real-time quantitative polymerase chain reaction (RT-qPCR) experiments are dependent on the use of the appropriate reference genes. In this study, we utilized geNorm and NormFinder softwares to identify the most stably expressed candidate reference genes in samples from seven stages of grain development and from eight landrace varieties. The results of the analysis performed using geNorm indicated that tubulin (TUB) and actin (ACT) were the most suitable reference genes among all experimental conditions, while glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) showed the least stability. The same result was obtained with the NormFinder software. The minimum number of genes required in each experimental condition to normalize the gene expression data was also determined by geNorm. The expression of phytoene synthase gene (PSY1), the first enzyme in the carotenoid biosynthetic pathway, was overestimated when the least stable candidate gene (GAPDH) was used as the internal control instead of the most stable gene pair (ACT + TUB), thus highlighting the importance of validating reference genes before conducting a RT-qPCR experiment to obtain accurate results. This study is the first survey of the stability of genes for use as reference genes to normalize RT-qPCR data from maize landraces during multiple stages of grain development.