Talisin is a storage protein from
seeds that presents lectin-like and peptidase inhibitor properties. These characteristics suggest that talisin plays a role in the plant defense process, making it a ...multifunctional protein. This work aimed to investigate the effects of chronic intake of talisin on fifth instar larvae of
, considered the main insect pest of maize and the cause of substantial economic losses in several other crops. The chronic intake of talisin presented antinutritional effects on the larvae, reducing their weight and prolonging the total development time of the insects. In addition, talisin-fed larvae also showed a significant reduction in the activity of trypsin-like enzymes. Midgut histology analysis of talisin-fed larvae showed alterations in the intestinal epithelium and rupture of the peritrophic membrane, possibly causing an increase of aminopeptidase activity in the midgut lumen. Talisin also proved to be resistant to degradation by the digestive enzymes of
. The transcription profile of trypsin, chymotrypsin and aminopeptidase genes was also analyzed through qPCR technique. Talisin intake resulted in differential expression of at least two genes from each of these classes of enzymes. Molecular docking studies indicated a higher affinity of talisin for the less expressed enzymes.
Chagas disease (CD), which is caused by Trypanosoma cruzi and was discovered more than 100 years ago, remains the leading cause of death from parasitic diseases in the Americas. As a curative ...treatment is only available for the acute phase of CD, the search for new therapeutic options is urgent. In this study, nitroazole and azole compounds were synthesized and underwent molecular modeling, anti-T. cruzi evaluations and nitroreductase enzymatic assays. The compounds were designed as possible inhibitors of ergosterol biosynthesis and/or as substrates of nitroreductase enzymes. The in vitro evaluation against T. cruzi clearly showed that nitrotriazole compounds are significantly more potent than nitroimidazoles and triazoles. When their carbonyls were reduced to hydroxyl groups, the compounds showed a significant increase in activity. In addition, these substances showed potential for action via nitroreductase activation, as the substances were metabolized at higher rates than benznidazole (BZN), a reference drug against CD. Among the compounds, 1-(2,4-difluorophenyl)-2-(3-nitro-1H-1,2,4-triazol-1-yl)ethanol (8) is the most potent and selective of the series, with an IC50 of 0.39 µM and selectivity index of 3077; compared to BZN, 8 is 4-fold more potent and 2-fold more selective. Moreover, this compound was not mutagenic at any of the concentrations evaluated, exhibited a favorable in silico ADMET profile and showed a low potential for hepatotoxicity, as evidenced by the high values of CC50 in HepG2 cells. Furthermore, compared to BZN, derivative 8 showed a higher rate of conversion by nitroreductase and was metabolized three times more quickly when both compounds were tested at a concentration of 50 µM. The results obtained by the enzymatic evaluation and molecular docking studies suggest that, as planned, nitroazole derivatives may utilize the nitroreductase metabolism pathway as their main mechanism of action against Trypanosoma cruzi. In summary, we have successfully identified and characterized new nitrotriazole analogs, demonstrating their potential as promising candidates for the development of Chagas disease drug candidates that function via nitroreductase activation, are considerably selective and show no mutagenic potential.
Quality of life (QL) assessments of children with incapacitating diseases, such as cerebral palsy (CP), have often been conducted with the help of the representatives of a child, making QL assessment ...more subjective. The Autoquestionnaire Qualité de Vie Enfant Imagé (AUQEI) is a QL assessment designed for children to self-report-it uses images to facilitate the reporting process.
evaluate the psychometric properties of AUQEI when responses are given by children with CP.
Children aged 4 to 12 years (45 with CP and 45 healthy children) gave responses to the questionnaire. The data quality, reliability and validity were assessed. The data loss rate ranged from 8.8% to 46.7%, and was highest for the "autonomy" factor. No floor or ceiling effect was detected. The success rate for reliability of the internal consistency of the items was less than 80% for the "autonomy" factor. Cronbach's alpha coefficient was 0.71 for the instrument and less than 0.5 for the factors. All the factors had a success rate of greater than 80% for the discriminating validity of the items. The factors did not have correlations between each other, thus indicating adequate discriminating validity. Convergent validity was tested and a significant correlation was demonstrated only between the AUQEI "functioning" factor and the Child Health Questionnaire--50-Item (CHQ-PF50) physical summary score (r = 0.31, p = 0.042). The AUQEI scores did not have correlations with the gross motor function scores (p>0.05) as expected for divergent validity. Regarding construct validity, the total AUQEI score obtained by the CP group was lower (median: 47.3) than that of the healthy group (median: 51.0) (p<0.01).
The AUQEI was shown to be a reliable and valid instrument for assessing children with CP when the total score was used. Convergent validity should continue to be tested in future studies.
The study evoluated an in-house Spike Receptor Binding Domain Enzyme-Linked Immunosorbent Assay (RBD-IgG-ELISA) for detecting SARS-CoV-2 IgG antibodies in infected and vaccinated individuals. The ...assay demonstrated a sensitivity of 91%, specificity of 99.25%, and accuracy of 95.13%. Precision and reproducibility were highly consistent. The RBD-IgG-ELISA was able to detect 96.25% of Polymerase chain reaction (PCR) confirmed cases for SARS-CoV-2 infection, demonstrating positive and negative predictive values of 99,18% and 91,69%, respectively. In an epidemiological survey, ELISA, lateral flow immunochromatographic assay (LFIA), and electrochemiluminescence immunoassay (ECLIA) exhibited diagnostic sensitivities of 68.29%, 63.41%, and 70.73%, respectively, along with specificities of 82.93%, 80.49%, and 80.49%, respectively. Agreement between RBD-IgG-ELISA/PCR was moderate (k index 0.512). However, good agreement between different assays (RBD-IgG-ELISA/LFIA k index 0.875, RBD-IgG-ELISA/ECLIA k index 0.901). Test performance on individuals' samples were inferior due to seroconversion time and chronicity. The IgG-RBD-ELISA assay demonstrated its effectiveness in monitoring antibody levels among healthcare professionals, revealing significant differences both before and after the administration of the third vaccine dose, with heightened protection levels observed following the third dose in five Coronavirus disease (COVID-19) vaccine regimens. In conclusion, the RBD-IgG-ELISA exhibits high reproducibility, specificity, and sensitivity, making it a suitable assay validated for serosurveillance and for obtaining information about COVID-19 infections or vaccinations.
•SARS-CoV-2 infection and vaccination typically induce a strong humoral immune response.•RBD-IgG-ELISA exhibits high sensitivity and specificity among controlled settings.•False negatives from the test can be directly linked to the timing of antibody production.•Understanding the IgG antibodies profiles is essential to guide COVID-19 vaccine boosting strategies.
Acute cardiogenic pulmonary edema (ACPE) is a life-threatening condition. OSA may be a modifiable risk factor for ACPE recurrence. This study was designed to evaluate the impact of OSA on the ...incidence of cardiovascular events following ACPE recovery.
Consecutive patients with confirmed ACPE from 3 centers underwent a sleep study following clinical stabilization. OSA was defined as an apnea-hypopnea index (AHI) ≥ 15 events/h. The mean follow-up was 1 year, and the primary outcome was ACPE recurrence.
A total of 104 patients were included in the final analysis; 61% of the patients had OSA. A higher rate of ACPE recurrence (25 vs 6 episodes; P = .01) and a higher incidence of myocardial infarction (15 vs 0 episodes; P = .0004) were observed in patients with OSA than in those without OSA. All 17 deaths occurred in the OSA group (P = .0001). In a Cox proportional hazards regression analysis, OSA was independently associated with ACPE recurrence (hazard ratio HR, 3.3 95% CI, 1.2-8.8; P = .01), incidence of myocardial infarction (HR, 2.3 95% CI, 1.1-9.5; P = .02), cardiovascular death (HR, 5.4 95% CI, 1.4-48.4; P = .004), and total death (HR, 6.5 95% CI, 1.2-64.0; P = .005). When the analysis was limited only to patients with OSA, levels of AHI and hypoxemic burden and rates of sleep-onset ACPE were significantly higher in those who presented with ACPE recurrence or who died than in those who did not experience these events.
OSA is independently associated with higher rates of ACPE recurrence and both fatal and nonfatal cardiovascular events.
Brazilian cattle production is mainly carried out in pastures, and mitigating the livestock's greenhouse gas (GHG) emissions has become an essential prerequisite. The importance of planted forests in ...beef cattle pasture-based integrated systems has been widely recognized in the context of GHG emission mitigation. Adopting the intensification and integration with forest and crops of pasture-based livestock production systems are alternatives to optimizing the soil and tree carbon (C) sequestration; however, research on tropical conditions is still necessary. This long-term trial evaluated the potential of C sequestration (soil + trees) of the forest and the grazing systems with two replicate areas of 3 ha each: 1. continuous stocking rate (CLS) – pasture under continuous stocking; 2. agropastoral or crop-livestock system (AGROP) – pasture under rotational stocking in rotation with corn; 3. agrosilvipastoral or crop-livestock-forestry system (AGROSIL) - similar to AGROP but including eucalyptus; 4. silvipastoral or livestock-forestry system (SIL) - pasture under rotational stocking + eucalyptus and 5. Atlantic Forest (FOR). Soil samples were analysed for texture, C and bulk density in eight soil layers from three to eight trenches for each treatment, then C stock and C accumulation rates were calculated. Data from Eucalyptus urograndis of the SIL and AGROSIL systems with 333 trees. ha−1 during five first years and 166 trees. ha−1 in subsequent years were collected (five- and eight-years old trees) and used to define the wood volume and acquire wood rings. Elemental analyzer CHN performed soil and trees C content. Equations for determining stem volume and tree biomass and, afterward, the C pools of the tree trunks were proposed. The potential of GHG emission mitigation (t CO2 e.. ha−1. year−1) by annual C sequestration (soil or soil + trees) was calculated, including the wood used for the lumber-mill scenario. The participation of the C stock from the 0–30 cm soil layer was between 45 % and 53 % of the total C stock in the 0–100 cm soil layer. Regardless of the elevated C soil accumulation rates, 0.88, 2.54, and 1.74 t C. ha-1. year−1 in the conversion stages CLS → AGROP; AGROP → SIL; AGROP → AGROSIL, respectively, it was not possible to compensate for the negative result of the Forest → CLS conversion (-2.55 t C. ha-1. year−1) because this stage lasted 12 years, while the implementation of AGROSIL and SIL systems had only six years. However, when C tree accumulation rates were considered, the potential of GHG emission mitigation (t CO2 e. ha−1. year−1) by annual C sequestration (soil +trees) was 19.89 to AGROSIL and 18.97 to SIL, including the wood used for the lumber-mill scenario. Based on these results, it can be concluded that C sequestration in trees-pasture integrated systems occurs in layers deeper, with the diverse species contributing in different soil layers; there is a positive effect of improved land-use management, with the use of integrated systems on annual C soil sequestration and the trees consists of an important strategy to improve the GHG mitigation.
This study aimed to validate a scale for assessing acute pain in donkeys. Forty-four adult donkeys underwent castration after sedation with intravenous (IV) xylazine, induction with guaifenesin and ...thiopental IV, local anesthetic block, and maintenance with isoflurane. The scale was constructed from a pilot study with four animals combined with algetic behaviors described for equines. After content validation, the scale was evaluated in 40 other donkeys by three blinded and one reference evaluator, by means of edited videos referring to the preoperative and postoperative periods: before anesthesia, 3–4 h after recovery from anesthesia, 5–6 h after recovery from anesthesia (2 h after analgesia with flunixin—1.1 mg/kg, dipyrone—10 mg/kg, and morphine—0.2 mg/kg) IV, and 24 h after recovery. Content validity, sensitivity, specificity, and responsiveness of behaviors were investigated to refine the scale. Intra- and inter-evaluator reliabilities were investigated by the weighted kappa coefficient, criterion validity by comparing the scale with the visual analog scale (VAS), internal consistency by Cronbach's α coefficient, item-total correlation by the Spearman coefficient, and intervention point for rescue analgesic by the receiver operating characteristics curve and Youden index. The scale showed very good intra-evaluator reliability (0.88–0.96), good to moderate (0.56–0.66) inter-evaluator reliability, responsiveness for all items, good criterion validity vs. VAS (0.75), acceptable internal consistency (0.64), adequate item-total correlation, except for head position and direction, and according to the principal component analysis, good association among items. The accuracy of the point for rescue analgesic was excellent (area under the curve = 0.91). The rescue analgesic score was ≥ 4 of 11 points. The scale can diagnose and quantify acute pain in donkeys submitted to castration, as the instrument is reliable and valid, with a defined intervention analgesic score.
The rising of diseases caused by multidrug‐resistant bacteria has encouraged researchers to explore more antimicrobial substances, as well as chemicals capable of potentiating the action of existing ...ones against multidrug‐resistant bacteria. Anacardium occidentale produces a fruit known as cashew nut, filled with a dark, almost black, caustic, and flammable liquid called cashew nutshell liquid (CNSL). The goal of the study was to evaluate the intrinsic antimicrobial activity of the major compounds present in CNSL, called anacardic acids (AA), as well as their possible modulatory action as an adjuvant of Norfloxacin against a Staphylococcus aureus strain overproducing the NorA efflux pump (SA1199B). Microdilution assays were performed to determine the minimum inhibitory concentration (MIC) of AA against different microbial species. Norfloxacin and Ethidium Bromide (EtBr) resistance modulation assays were performed in the presence or absence of AA against SA1199‐B. AA showed antimicrobial activity against Gram‐positive bacterial strains tested but no activity against Gram‐negative bacteria or yeast strains. At subinhibitory concentration, AA reduced the MIC values for Norfloxacin and EtBr against the SA1199‐B strain. Furthermore, AA increased the intracellular accumulation of EtBr in this NorA overproducer strain, indicating that AA are NorA inhibitors. Docking analysis showed that AA probably modulates Norfloxacin efflux by spatial impediment at the same binding site of NorA.
•High-intensity interval training (HIIT) avoided liver steatosis in high-fat-fed mice.•HIIT (thrice weekly) prevented hyperglycemia and endoplasmic reticulum (ER) stress.•HIIT preserved mitochondrial ...ultrastructure and enhanced beta-oxidation.•HIIT targeted ER and mitochondrial homeostasis to prevent liver steatosis.
Metabolic dysfunction-associated steatotic liver disease (MASLD) has attracted increasing attention from the scientific community because of its severe but silent progression and the lack of specific treatment. Glucolipotoxicity triggers endoplasmic reticulum (ER) stress with decreased beta-oxidation and enhanced lipogenesis, promoting the onset of MASLD, whereas regular physical exercise can prevent MASLD by preserving ER and mitochondrial function. Thus, the hypothesis of this study was that high-intensity interval training (HIIT) could prevent the development of MASLD in high-fat (HF)-fed C57BL/6J mice by maintaining insulin sensitivity, preventing ER stress, and promoting beta-oxidation. Forty male C57BL/6J mice (3 months old) comprised 4 experimental groups: the control (C) diet group, the C diet + HIIT (C-HIIT) group, the HF diet group, and the HF diet + HIIT (HF-HIIT) group. HIIT sessions lasted 12 minutes and were performed 3 times weekly by trained mice. The diet and exercise protocols lasted for 10 weeks. The HIIT protocol prevented weight gain and maintained insulin sensitivity in the HF-HIIT group. A chronic HF diet increased ER stress-related gene and protein expression, but HIIT helped to maintain ER homeostasis, preserve mitochondrial ultrastructure, and maximize beta-oxidation. The increased sirtuin-1/peroxisome proliferator-activated receptor-gamma coactivator 1-alpha expression implies that HIIT enhanced mitochondrial biogenesis and yielded adequate mitochondrial dynamics. High hepatic fibronectin type III domain containing 5/irisin agreed with the antilipogenic and anti-inflammatory effects observed in the HF-HIIT group, reinforcing the antisteatotic effects of HIIT. Thus, we confirmed that practicing HIIT 3 times per week maintained insulin sensitivity, prevented ER stress, and enhanced hepatic beta-oxidation, impeding MASLD development in this mouse model even when consuming high energy intake from saturated fatty acids.
HIIT prevented overweight and liver steatosis by mitochondrial and ER enhancement. Abbreviations: ACOX1, acyl-coenzyme A oxidase 1; Atf4, activating transcription factor 4; ER, endoplasmic reticulum; GRP78, glucose-regulated protein 78; Chop, C/EBP homologous protein; Fgf21, fibroblast growth factor 21; Gadd45, growth arrest and DNA damage-inducible 45; Ppar, peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (Pgc1-a); Xbp-a, X-box binding protein 1. Created with www.biorender.com. Display omitted
Detection of genetic exchange has been a limiting factor to deepen the knowledge on the mechanisms by which Trypanosoma cruzi is able to generate progeny and genetic diversity. Here we show that ...incorporation of halogenated thymidine analogues, followed by immunostaining, is a reliable method not only to detect T. cruzi fused-cell hybrids, but also to quantify their percentage in populations of this parasite. Through this approach, we were able to detect and quantify fused-cell hybrids of T. cruzi clones CL Brener and Y. Given the increased detection of fused-cell hybrids in naturally-occurring hybrid CL Brener strain, which displays increased levels of RAD51 and BRCA2 transcripts, we further investigated the role of Rad51 - a recombinase involved in homologous recombination - in the process of genetic exchange. We also verified that the detection of fused-cell hybrids in T. cruzi overexpressing RAD51 is increased when compared to wild-type cells, suggesting a key role for Rad51 either in the formation or in the stabilization of fused-cell hybrids in this organism.
PDF
