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Open pit mining can cause loss in different ecosystems, including damage to habitats of rare and endemic species. Understanding the biology of these species is fundamental for their conservation, and to assist in decision-making. Sporobolus multiramosus is an annual grass endemic to the Amazon canga ecosystems, which comprise rocky outcrop vegetation covering one of the world's largest iron ore reserves. Here, we evaluated whether nitric oxide aids S. multiramosus in coping with water shortages and examined the physiological processes behind these adaptations. nitric oxide application improved the water status, photosynthetic efficiency, biomass production, and seed production and germination of S. multiramosus under water deficit conditions. These enhancements were accompanied by adjustments in leaf and root anatomy, including changes in stomata density and size and root endodermis thickness and vascular cylinder diameter. Proteomic analysis revealed that nitric oxide promoted the activation of several proteins involved in the response to environmental stress and flower and fruit development. Overall, the results suggest that exogenous nitric oxide has the potential to enhance the growth and productivity of S. multiramosus . Enhancements in seed productivity have significant implications for conservation initiatives and can be applied to seed production areas, particularly for the restoration of native ecosystems.

MicroRNAs (miRNAs) are short non-coding RNA sequences with the ability to inhibit the expression of a target mRNA at the post-transcriptional level, acting as modulators of both the degenerative and regenerative processes. Therefore, these molecules constitute a potential source of novel therapeutic tools. In this study, we investigated the miRNA expression profile that presented in enthesis tissue upon injury. For this, a rodent enthesis injury model was developed by creating a defect at a rat's patellar enthesis. Following injury, explants were collected on days 1 ( n = 10) and 10 ( n = 10). Contra lateral samples ( n = 10) were harvested to be used for normalization. The expression of miRNAs was investigated using a "Fibrosis" pathway-focused miScript qPCR array. Later, target prediction for the aberrantly expressed miRNAs was performed by means of the Ingenuity Pathway Analysis, and the expression of mRNA targets relevant for enthesis healing was confirmed using qPCRs. Additionally, the protein expression levels of collagens I, II, III, and X were investigated using Western blotting. The mRNA expression pattern of EGR1 , COL2A1 , RUNX2 , SMAD1 , and SMAD3 in the injured samples indicated their possible regulation by their respective targeting miRNA, which included miR-16, -17, -100, -124, -133a, -155 and -182. Furthermore, the protein levels of collagens I and II were reduced directly after the injury (i.e., day 1) and increased 10 days post-injury, while collagens III and X showed the opposite pattern of expression.

The use of multiphasic scaffolds to treat injured tendon-to-bone entheses has shown promising results in vitro. Here, we used two versions of a biphasic silk fibroin scaffold to treat an enthesis defect created in a rat patellar model in vivo. One version presented a mixed transition between the bony and the tendon end of the construct (S-MT) while this transition was abrupt in the second version (S-AT). At 12 weeks after surgery, the S-MT scaffold promoted better healing of the injured enthesis, with minimal undesired ossification of the insertion area. The expression of tenogenic and chondrogenic markers was sustained for longer in the S-MT-treated group and the tangent modulus of the S-MT-treated samples was similar to the native tissue at 12 weeks while that of the S-AT-treated enthesis was lower. Our study highlights the important role of the transition zone of multiphasic scaffolds in the treatment of complex interphase tissues such as the tendon-to-bone enthesis.

Objective: To evaluate the relationship between crack/cocaine use and domestic violence perpetration from the perspective of substance users' families. A secondary retrospective cross-sectional study, with 3162 family members of crack/cocaine users seeking treatment in the Recomeço Família Programme in São Paulo/Brazil was undertaken. Family members of crack/cocaine users reported that their relatives were more involved in domestic violence such as stealing (money and objects) at home [Odds Ratio Adjusted ORA = 2.17 (CI 95% 1.87; 2.53)], the family gave money to the user to buy drugs [ORA = 1.27 (1.08; 1.48)], and having problems with the judiciary [ORA = 1.48 (CI 95% 1.28; 1.71)]. Relatives of snorted cocaine users reported that there was physical and interpersonal violence, such as fathers being assaulted [ORA = 2.50 (CI 95% 1.08; 5.82)], assaulted someone else [ORA = 1.86 (CI 95% 1.32; 2.60)], threats of violence fights, arguments when the family talk about problematic drug use [ORA = 1.50 (CI 95% 1.13; 1.96)] and threatened some family members [ORA = 1.52 (CI 95% 1.14; 2.04)]. In this sample, there was a connection between crack/cocaine use and the perpetuation of domestic violence, corroborating with important implications for public policies, substance use treatment and prevention of domestic violence interventions.

Background: Overweight and obesity are conditions associated with sedentary lifestyle and accumulation of abdominal fat, determining increased mortality, favoring chronic diseases, and increasing cardiovascular risk. Although the evaluation of body composition and fat distribution are highly relevant, the high cost of the gold standard techniques limits their wide utilization. Therefore, the aim of this work was to explore the relationships between simple anthropometric measures and BIA variables using multivariate linear regression models to estimate body composition and fat distribution in adults., Methods: In this cross-sectional study, sixty-eight adult individuals (20 males and 48 females) were subjected to bioelectrical impedance analysis (BIA), anthropometric measurements (waist circumference (WC), neck circumference (NC), mid-arm circumference (MAC)), allowing the calculation of conicity index (C-index), fat mass/fat-free mass (FM/FFM) ratios, body mass index (BMI) and body shape index (ABSI). Statistical analyzes were performed with the R program. Nonparametric Statistical tests were applied to compare the characteristics of participants of the groups (normal weight, overweight and obese). For qualitative variables, the Fisher's exact test was applied, and for quantitative variables, the paired Wilcoxon signed-rank test. To evaluate the linear association between each pair of variables, the Pearson correlation coefficient was calculated, and Multivariate linear regression models were adjusted using the stepwise variable selection method, with Akaike Information Criterion ( p ≤ 0.05)., Results: BIA variables with the highest correlations with anthropometric measures were total body water (TBW), body fat percentage (BFP), FM, FFM and FM/FFM. The multiple linear regression analysis showed, in general, that the same variables can be estimated through simple anthropometric measures., Conclusions: The assessment of fat distribution in the body is desirable for the diagnosis and definition of obesity severity. However, the high cost of the instruments (dual energy X-ray absorptiometry, hydrostatic weighing, air displacement plethysmography, computed tomography, magnetic resonance) to assess it, favors the use of BMI in the clinical practice. Nevertheless, BMI does not represent a real fat distribution and body fat percentage. This highlights the relevance of the findings of the current study, since simple anthropometric variables can be used to estimate important BIA variables that are related to fat distribution and body composition.

Shiga toxins (Stx) released by Stx-producing E. coli (STEC) are virulence factors that are most closely 3associated with hemolytic uremic syndrome (HUS), a life-threatening complication of intestinal infections by STEC. Stx have to enter into the circulatory system before they are delivered to target organs and cause damage. The presence of Stx in sera could be a risk indicator for HUS development. However, the detection of Stx, particularly Stx2, has been difficult due to the presence of Stx2-binding components in human serum. Here, we report new ELISA-based methods for the detection of Stx1 and Stx2 in human serum and the effect of guanidinium chloride on enhancing the sensitivity for the detection of Stx2. The recovery rate for Stx2 was 62% when Stx2-spiked serum samples were treated with guanidinium chloride at a concentration of 200 mM, in contrast to 17% without guanidinium chloride treatment. The effectiveness of guanidinium chloride treatment for the detection of Stx2 in human serum was validated using sera from STEC-infected patients. Coimmunoprecipitation results indicated a specific physical interaction between Stx2 and the human serum amyloid P component (HuSAP) in human serum samples. Our in vitro study demonstrated that the inhibition from HuSAP alone for the detection of Stx2 was only 20%, much less than 69.6% from human serum at Stx2 level 10 ng/mL, suggesting that there may be other factors that bind Stx2 in human serum. This study indicates that treatment of serum samples with guanidinium chloride may be useful for the early and sensitive detection of Stx2 in sera of STEC-infected patients, so preventive measures can be adopted in a timely manner., Competing Interests: The authors declare no conflicts of interest.

Shiga-like toxins (verotoxins) are responsible for the virulence associated with a variety of foodborne bacterial pathogens. Direct detection of toxins requires a specific and sensitive technique. In this study, we describe a mass spectrometry-based method of analyzing the tryptic decapeptides derived from the non-toxic B subunits. A gene encoding a single protein that yields a set of relevant peptides upon digestion with trypsin was designed. The (15)N-labeled protein was prepared by growing the expressing bacteria in minimal medium supplemented with (15)NH₄Cl. Trypsin digestion of the (15)N-labeled protein yields a set of (15)N-labeled peptides for use as internal standards to identify and quantify Shiga or Shiga-like toxins. We determined that this approach can be used to detect, quantify and distinguish among the known Shiga toxins (Stx) and Shiga-like toxins (Stx1 and Stx2) in the low attomole range (per injection) in complex media, including human serum. Furthermore, Stx1a could be detected and distinguished from the newly identified Stx1e in complex media. As new Shiga-like toxins are identified, this approach can be readily modified to detect them. Since intact toxins are digested with trypsin prior to analysis, the handling of intact Shiga toxins is minimized. The analysis can be accomplished within 5 h.

The chemical composition of the volatile oils from five Anacardiaceae species and their activities against Gram positive and negative bacteria were assessed. The peroxidative damage within bacterial cell membranes was determined through the breakdown product malondialdehyde (MDA). The major constituents in Anacardium humile leaves oil were (E)-caryophyllene (31.0%) and α-pinene (22.0%), and in Anacardium occidentale oil they were (E)-caryophyllene (15.4%) and germacrene-D (11.5%). Volatile oil from Astronium fraxinifolium leaves were dominated by (E)-β-ocimene (44.1%) and α-terpinolene (15.2%), whilst the oil from Myracrodruon urundeuva contained an abundance of δ-3-carene (78.8%). However, Schinus terebinthifolius leaves oil collected in March and July presented different chemical compositions. The oils from all species, except the one from A. occidentale, exhibited varying levels of antibacterial activity against Staphylococcus aureus, Bacillus cereus and Escherichia coli. Oil extracted in July from S. terebinthifolius was more active against all bacterial strains than the corresponding oil extracted in March. The high antibacterial activity of the M. urundeuva oil could be ascribed to its high δ-3-carene content. The amounts of MDA generated within bacterial cells indicate that the volatile oils induce lipid peroxidation. The results suggest that one putative mechanism of antibacterial action of these volatile oils is pro-oxidant damage within bacterial cell membrane explaining in part their preservative properties.

This study investigates the seasonal variation of three species of Leptospermum (Myrtaceae) grown in Brazil. The chemical composition of the volatile oils of L. flavescens and L. petersonii did not show any significant seasonal variation in the major components, while for Leptospermum madidum subsp. sativum the levels of major constituents of the volatile oils varied with the harvest season. Major fluctuations in the composition of L. madidum subsp. sativum oil included α-pinene (0-15.2%), β-pinene (0.3-18.5%), α-humulene (0.8-30%), 1,8-cineole (0.4-7.1%) and E-caryophyllene (0.4-11.9%). Levels of β-pinene (0.3-5.6%), terpinen-4-ol (4.7-7.2%) and nerolidol (55.1-67.6%) fluctuated seasonally in the L. flavescens oil. In L. petersonii, changes were noted for geranial (29.8-32.8%), citronellal (26.5-33.9%) and neral (22.7-23.5%). The activity of the volatile oils against the tested bacteria differed, depending on season the oils were obtained. In general, the volatile oils were more active against Gram-positive bacteria.