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Protein cis-regulatory elements (CREs) are regions that modulate the activity of a protein through intramolecular interactions. Kinases, pivotal enzymes in numerous biological processes, often undergo regulatory control via inhibitory interactions in cis. This study delves into the mechanisms of cis regulation in kinases mediated by CREs, employing a combined structural and sequence analysis. To accomplish this, we curated an extensive dataset of kinases featuring annotated CREs, organized into homolog families through multiple sequence alignments. Key molecular attributes, including disorder and secondary structure content, active and ATP-binding sites, post-translational modifications, and disease-associated mutations, were systematically mapped onto all sequences. Additionally, we explored the potential for conformational changes between active and inactive states. Finally, we explored the presence of these kinases within membraneless organelles and elucidated their functional roles therein. CREs display a continuum of structures, ranging from short disordered stretches to fully folded domains. The adaptability demonstrated by CREs in achieving the common goal of kinase inhibition spans from direct autoinhibitory interaction with the active site within the kinase domain, to CREs binding to an alternative site, inducing allosteric regulation revealing distinct types of inhibitory mechanisms, which we exemplify by archetypical representative systems. While this study provides a systematic approach to comprehend kinase CREs, further experimental investigations are imperative to unravel the complexity within distinct kinase families. The insights gleaned from this research lay the foundation for future studies aiming to decipher the molecular basis of kinase dysregulation, and explore potential therapeutic interventions.

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CONTEXT: The negative of the Shannon entropy derivative is proposed to account for electron density contraction as the chemical bonds are breaking and forming during a chemical reaction. We called this property the electron density contraction index, EDC, which allows identifying stages in a reaction that are dominated by electron contraction or expansion. Four different reactions were analyzed to show how the EDC index changes along the reaction coordinate. The results indicate that the rate of change of Shannon entropy is directly related to the rate of change of the electron density at the bond critical points between all the atomic pairs in the molecular systems. It is expected that EDC will complement the detailed analysis of reaction mechanisms that can be performed with the theoretical tools available to date. METHODS: Density functional theory calculations at the B3LYP/6-31G(d,p) level of theory were carried out using Gaussian 16 to analyze the reaction mechanisms of the four reactions studied. The reaction paths were obtained via the intrinsic reaction coordinate method, which served as the reaction coordinate to obtain the reaction force and the EDC profiles in each case. Shannon entropy and electron density at the bond critical points were calculated using the Multiwfn 3.7 package.

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Leishmaniasis is a parasitic neglected tropical disease, affecting 12 million people. Available treatments present several limitations, with an increasing number of resistance cases. In the search for new chemotherapies, the natural product dehydrodieugenol B was used as a scaffold for the synthesis of a series of derivatives, resulting in the discovery of the promising analog [4-(4-(5-allyl-3-methoxy-2-((4-methoxybenzyl)oxy)phenoxy)-3-methoxybenzyl)morpholine, 1]. In this work, we investigated the effect of compound 1 on cell signaling in Leishmania (L.) infantum, culminating in cell death, as well as its immunomodulatory effect in the host cell. Additionally, we performed a pharmacokinetic profile study in an animal model. After treatment, compound 1 induced the alkalinization of acidocalcisomes and concomitant Ca2+ release in the parasite. These events may induce depolarization of the mitochondrial potential, with successive collapse of the bioenergetic system, leading to a reduction of ATP and reactive oxygen species (ROS) levels. The analysis of total proteins and protein profile by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) demonstrated that compound 1 also altered the parasite proteins after treatment. Transmission electron microscopy studies revealed ultrastructural damage to mitochondria; together, these data suggest that compound 1 may promote autophagic cell death. Additionally, compound 1 also induced an immunomodulatory effect in host cells, with a reduction of Th1 and Th2 cytokine response, characterizing an anti-inflammatory compound. The obtained pharmacokinetic profile in rats enhances the potential of the compound, with a mean plasma half-life (T1/2) of 21 h. These data reinforce the potential of compound 1 as a new lead for future efficacy studies.

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Procyanidins (PCs) have emerged as agents with potential antimicrobial and antibiofilm activities, although their mechanisms of action and structure-activity relationships remain poorly understood. This review assessed the potential mechanisms of action and applications of these compounds to explore these aspects. Studies on the antimicrobial properties of PCs suggest that they are involved in osmotic imbalance, DNA interactions and metabolic disruption. Although less studied, their antibiofilm activities include antiadhesive effects and the modulation of mobility and quorum sensing. However, most research has used uncharacterized plant extracts for in vitro assays, limiting the understanding of the structure-activity relationships of PCs and their in vivo mechanisms. Clinical trials on the antimicrobial and antibiofilm properties of PCs have not clarified these issues due to nonstandardized methodologies, inadequate chemical characterization, and the limited number of studies, preventing a consensus and evaluation of the in vivo effects. Additionally, patent analysis revealed that technological developments in the antimicrobial and antibiofilm uses of PCs are concentrated in health care and dental care, but new biotechnological uses are emerging. Therefore, while PCs are promising antimicrobial and antibiofilm compounds, further research into their chemical structures and mechanisms of action is crucial for evidence-based applications in biotechnology and health care.

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Traditionally, the performance of sodium-ion batteries has been predicted based on a single characteristic of the electrodes and its relationship to specific capacity increase. However, recent studies have shown that this hypothesis is incorrect because their performance depends on multiple physical and chemical variables. Due to the above, the present communication shows machine learning as an innovative strategy to predict the performance of functionalized hard carbon anodes prepared from grapefruit peels. In this sense, a three-layer feed-forward Artificial Neural Network (ANN) was designed. The inputs used to feed the ANN were the physicochemical characteristics of the materials, which consisted of mercury intrusion porosimetry data (SHg and average pore), elemental analysis (C, H, N, S), ID/IG ratio obtained from RAMAN studies, and X-ray photoemission spectroscopy data of the C1s, N1s, and O1s regions. In addition, two more inputs were added: the cycle number and the applied C-rate. The ANN architecture consisted of a first hidden layer with a sigmoid transfer function and a second layer with a log-sigmoid transfer function. Finally, a sigmoid transfer function was used in the output layer. Each layer had 10 neurons. The training algorithm used was Bayesian regularization. The results show that the proposed ANN correctly predicts (R2 > 0.99) the performance of all materials. The proposed strategy provides critical insights into the variables that must be controlled during material synthesis to optimize the process and accelerate progress in developing tailored materials.

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The study of the adsorption of polycyclic aromatic hydrocarbons on microplastics (MPs) has attracted much attention as to how microplastics can act as carriers of these pollutants. Polyurethane (PU) is one of the MPs found in aquatic environments, containing different functional groups it can interact with polar and nonpolar molecules. PAH derivatives (dPAHs) present different properties and thus can be adsorbed by different interactions; thus, this study investigated the adsorption of fluorene (FLN), dibenzothiophene (DBT), dibenzofuran (DBF), and carbazole (CBZ) onto PU MP. The Langmuir, Freundlich, and BET isotherm models were examined, and the BET model best fitted. The adsorption was a nonspontaneous process, exothermic for mono- and multilayer formation for FLN, DBT, and CBZ, and endothermic for DBF monolayer formation. The adsorption monolayer was formed by van der Waals forces, H─bonding, and π─π interactions, while the formation of the multilayer can be explained by π─π and hydrophobic interactions. The pseudo-second-order model proved to be more consistent for the adsorption of dPAHs. The adsorption in artificial seawater shows no significant differences for the monolayer but favored the adsorption multilayer due to the salting-out effect. Due to the existence of several adsorption mechanisms, PU MP interacts with dPAHs in greater quantities when compared to a MP with a simpler structure.

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On an industrial scale, the residues accumulated in essential oil distilleries can be compared to the volume of residues produced in the textile industry. Although these residues are discarded, they possess molecules with diverse biological activities, including their application in phytopathogen control. In this study, the chemical profile of the residue from the hydrodistillation of Lantana camara L. leaves was determined using high-performance liquid chromatography (HPLC). Additionally, the effect of the residue on cells was assessed by determining plasma membrane integrity, levels of reactive oxygen species (ROS) production, and mitochondrial potential depolarization. The viability and cell density of Phytomonas serpens parasites significantly decreased after treatment with increasing concentrations of the lyophilized residue from accession LAC-038 (RL038). RL038 reduced cell viability by an average of 61.36%. ROS levels increased by approximately 2 × and 3 × at RL038 concentrations of 120 µg/mL and 180 µg/mL, respectively. It was observed that the same concentrations modified mitochondrial potential, reducing fluorescence by 44.6% and 46.8%, respectively. Analytical liquid chromatography of RL038 revealed the presence of 17 peaks subsequently classified as phenolic acids and flavonoids. RL038 from the hydrodistillation of Lantana camara L. leaves is a source of biologically active compounds with antiprotozoal potential.

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BACKGROUND: There is conflicting data regarding the optimal abductor mechanism (AM) repair technique after resection of proximal femur tumors. We sought to compare functional outcomes following tumor resection and reconstruction with proximal femoral replacement based on the AM repair technique utilized. METHODS: We conducted a systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. We established two study groups based on AM repair technique as follows: soft-tissue reattachment (STr) and greater trochanter preservation (GTp). In the STr group, the gluteus medius and minimus muscles were reattached to the endoprosthesis, whereas in the GTp group, the greater trochanter and gluteal tendons were preserved. The STr group was further subdivided into direct and indirect reattachments. Weighted means adjusting for sample size were calculated. RESULTS: A total of 658 patients from 12 articles were included. Patients who had STr displayed higher Musculoskeletal Tumor Society scores (75 versus 67.3%, P < 0.001), lower rates of Trendelenburg gait (33.9 versus 52.4%, P < 0.01), and ambulation with assistive devices (30.4 versus 54.9%, P < 0.001) compared to the GTp group. Within the STr group, indirect reattachment was associated with higher Musculoskeletal Tumor Society scores (87.2 versus 70.1%, P < 0.001) and lower rates of Trendelenburg gait (3.8 versus 36.3%, P < 0.001) and ambulation with assistive devices (0 versus 42.4%, P < 0.001) compared to direct reattachment. The reattachment hardware failure rate in GTp was 15%. CONCLUSION: A STr provided superior functional outcomes compared to GTp in tumor-related proximal femoral replacement. From a functional outcome perspective, the use of indirect STr was better compared to direct STr. LEVEL OF EVIDENCE: III.

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Metacaspases are a distinct class of cysteine proteases predominantly found in plants, fungi, and protozoa, crucial for regulating programmed cell death (PCD). They possess unique structural features and differ markedly from caspases in their activation mechanisms and substrate specificities, with a notable preference for binding basic residues in substrates. In this study, we introduced vanillin-derived oximic compounds to explore their pharmaceutical potential. We evaluated these compounds for their inhibitory effects on TbMCA2, a metacaspase in Trypanosoma brucei, identifying AO-7, AO-12, and EO-20 as promising inhibitors. AO-12 showed significant potential as a non-competitive inhibitor with notable IC50 values. Molecular docking studies were also conducted to evaluate the binding affinity of these compounds for TbMCA2. This research is particularly relevant given the urgent need for more effective and less toxic treatments for trypanosomiasis, a parasitic disease caused by trypanosomes. The absence of available vaccines and the limitations imposed by drug toxicity underscore the importance of these findings. Our study represents a significant advancement in developing therapeutic agents targeting metacaspases in trypanosomatids and highlights the necessity of understanding metacaspase regulation across various species. It provides valuable insights into inhibitor sensitivity and potential species-specific therapeutic strategies. In conclusion, this research opens promising avenues for novel therapeutic agents targeting metacaspases in trypanosomatids, addressing a critical gap in combating neglected diseases associated with these pathogens. Further research is essential to refine the efficacy and safety profiles of these compounds, aiming to deliver more accessible and effective therapeutic solutions to populations afflicted by these debilitating diseases.

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The passive stay apparatus works by blocking flexion of the knee and tarsus joints in the horse, preventing muscle fatigue. During ambulation, if this mechanism fails to release, the animal will present upward fixation of the patella, which in severe cases, can only be treated by medial patellar desmotomy (MPD). The objective of this study was to investigate the possible electromyographic and behavioral changes, after the impairment of the passive stay apparatus, in horses who have undergone MPD surgery. Five horses presenting dorsal fixation of the patella underwent electromyographic measurements at preoperatively and immediately postoperatively, 24 h, seven, 14 and 21 days after surgery. The samples were collected on a computerized surface electromyograph designed in accordance with the standards of international society of electrophysiology and kinesiology (ISEK). To capture the signals, electrodes positioned on the tensor fasciae lata, vastus lateralis, biceps femoris and semitendinosus muscles were used, bilaterally. These muscles were chosen due to their participation in the passive stay apparatus. The electromyographic signals were collected and processed using a software calibrated to collect data with a sampling frequency of 1000 Hz per channel. The collections took place over 60 min periods, and the results were obtained from the root means square (RMS) analysis. It was noted that the preoperative average of rest in tripedal support was 62%. On the other hand, change in weight bearing member and rest in three limbs in the postoperative period did not occur. This inability to maintain tripedal support in the postoperative period prevents the horse from resting while standing. Furthermore, there was a gradual increase in the need for post-surgical muscle recruitment, resulting in long periods of decubitus due to muscle fatigue, which is less than ideal for such large animals. Therefore, it was concluded that the animals submitted to medial patellar desmotomy, in this study, presented an inability to maintain tripedal support, in addition to a greater need for recruitment of muscle fibers, evidenced by the gradual increase in the electromyographic tracing and total RMS value of the muscles evaluated.

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Staphylococcus aureus is a bacterium responsible for resistance to multiple drugs and the efflux system is widely studied among the resistance mechanisms developed by this species. The present study evaluates the inhibition of the MepA efflux pump by thiadiazine-derived compounds. For this purpose, thiadiazine-derived compounds (IJ-14 to IJ-20) were tested against S. aureus K2068 strains. Microdilution tests were initially conducted to assess the Minimum Inhibitory Concentration (MIC) of the compounds and their efflux pump inhibition activity. In addition, fluorimetry tests were performed using BrEt emission and tests were conducted to inhibit the expression of the mepA gene. This involved comparing the bacterial gene expression with the antibiotic alone to the gene expression after combining compounds (IJ-17 and IJ-20) with the antibiotic. Furthermore, membrane permeability assessment tests and in silico molecular docking tests were performed. It was observed that the IJ17 and IJ20 compounds exhibited direct activity against the tested strain. The IJ17 compound produced significant results in the gene inhibition tests, which was also evidenced through the membrane permeability alteration test. These findings suggest that thiadiazine-derived compounds have promising effects against one of the main resistance mechanisms, with the IJ17 compound presenting observable mechanisms of action.

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Candida species are among the priority pathogens in the area of research and development. Due to the problems associated with resistance to antifungals, new therapeutic alternatives are necessary. In this regard, drug repositioning has gained prominence. The objective of this study was to evaluate the activity of three tricyclic antidepressants (TCAs) - amitriptyline (AMT), nortriptyline (NOR) and clomipramine (CLO) - isolated or associated with antifungals against strains of Candida spp., as well as to analyze the possible mechanism of action. Among the methods used were broth microdilution tests, tolerance level assessment, checkerboard assays, flow cytometry and fluorescence microscopy. Furthermore, Candida cells were visualized after treatments by scanning electron microscopy (SEM). AMT presented MIC 50% in the range of 16 to 128 µg/mL, NOR from 8 to 128 µg/mL, and CLO from 8 to 64 µg/mL, with all three TCAs having a fungicidal inhibitory action profile. For these TCAs, there was synergism with amphotericin B (AMB) in 100% of the isolates. In association with fluconazole (FLC) and itraconazole (ITR), there were mostly indifferent interactions. TCAs isolated and associated with AMB reduced cell viability, promoted DNA fragmentation and damage, caused mitochondrial depolarization, externalization of phosphatidylserine, produced reactive oxygen species (ROS), decreased reduced glutathione (GSH) and increased carbonyl protein levels, causing morphological changes. The results suggest the antifungal mechanism of the TCAs works via the apoptotic pathway.

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In this research work, we examined the decomposition mechanisms of N-substituted diacetamides. We focused on the substituent effect on the nitrogen lone-pair electron delocalization, with electron-withdrawing and electron donor groups. DFT functionals used the following: B1LYP, B3PW91, CAMB3LYP, LC-BLYP, and X3LYP. Dispersion corrections (d3bj) with Becke-Johnson damping were applied when necessary to improve non-covalent interactions in the transition state. Pople basis sets with higher angular moments and def2-TZVP basis sets were also applied and were crucial for obtaining consistent thermodynamic parameters. The proposed mechanism involves a six-membered transition state with the extraction of an α hydrogen. Several conformers of N-diacetamides were used to account for the decrease in entropy in the transition state in the rate-determining state. All calculations, including natural bond orbital (NBO) analyses, were performed using the Gaussian16 computational package and its GaussView 6.0 visualizer, along with VMD and GNUPLOT software. The isosurfaces and IBSIs were calculated using MultiWFN and IGMPlot, respectively.

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Nanoparticles play a crucial role in the field of nanotechnology, offering different properties due to their surface area attributed to their small size. Among them, silver nanoparticles (AgNPs) have attracted significant attention due to their antimicrobial properties, with applications that date back from ancient medicinal practices to contemporary commercial products containing ions or silver nanoparticles. AgNPs possess broad-spectrum biocidal potential against bacteria, fungi, viruses, and Mycobacterium, in addition to exhibiting synergistic effects when combined with certain antibiotics. The mechanisms underlying its antimicrobial action include the generation of oxygen-reactive species, damage to DNA, rupture of bacterial cell membranes and inhibition of protein synthesis. Recent studies have highlighted the effectiveness of AgNPs against various clinically relevant bacterial strains through their potential to combat antibiotic-resistant pathogens. This review investigates the proteomic mechanisms by which AgNPs exert their antimicrobial effects, with a special focus on their activity against planktonic bacteria and in biofilms. Furthermore, it discusses the biomedical applications of AgNPs and their potential non-preparation of antibiotic formulations, also addressing the issue of resistance to antibiotics.

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The increase in fungal resistance is a major public health concern. In this context, Candida spp. is an important genus related to invasive diseases, especially in immunosuppressed patients. The relevance of alternative approaches to increasing fungal resistance stands out, in which products of natural origin demonstrate potential antifungal activity in vitro against Candida spp. In this sense, this work aimed to evaluate the in vitro activity of tannic acid against Candida spp. Minimum inhibitory concentration (MIC) was determined for tannic acid and the antifungals, and the checkerboard assay was performed to analyze the interactions between them. Furthermore, we evaluated the tannic acid antibiofilm activity and its possible mechanism of action. Tannic acid showed MIC ranging to 0.06 to 0.5 µg/ml and showed no loss of effectiveness when combined with antifungals. Also, is safe at the concentrations it exerts its antifungal activity in pre-formed biofilms, as demonstrated by IC50 in murine fibroblasts cells and the hemolytic assay. Additionally, its mechanisms of action can be related with induction of signals that lead to apoptosis in fungal cells.

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BACKGROUND AND PURPOSE: Gastrointestinal tumours overexpress voltage-gated calcium (CaV3) channels (CaV3.1, 3.2 and 3.3). CaV3 channels regulate cell growth and apoptosis colorectal cancer. Gossypol, a polyphenolic aldehyde found in the cotton plant, has anti-tumour properties and inhibits CaV3 currents. A systematic study was performed on gossypol blocking mechanism on CaV3 channels and its potential anticancer effects in colon cancer cells, which express CaV3 isoforms. EXPERIMENTAL APPROACH: Transcripts for CaV3 proteins were analysed in gastrointestinal cancers using public repositories and in human colorectal cancer cell lines HCT116, SW480 and SW620. The gossypol blocking mechanism on CaV3 channels was investigated by combining heterologous expression systems and patch-clamp experiments. The anti-tumoural properties of gossypol were estimated by cell proliferation, viability and cell cycle assays. Ca2+ dynamics were evaluated with cytosolic and endoplasmic reticulum (ER) Ca2+ indicators. KEY RESULTS: High levels of CaV3 transcripts correlate with poor prognosis in gastrointestinal cancers. Gossypol blockade of CaV3 isoforms is concentration- and use-dependent interacting with the closed, activated and inactivated conformations of CaV3 channels. Gossypol and CaV3 channels down-regulation inhibit colorectal cancer cell proliferation by arresting cell cycles at the G0/G1 and G2/M phases, respectively. CaV3 channels underlie the vectorial Ca2+ uptake by endoplasmic reticulum in colorectal cancer cells. CONCLUSION AND IMPLICATIONS: Gossypol differentially blocked CaV3 channel and its anticancer activity was correlated with high levels of CaV3.1 and CaV3.2 in colorectal cancer cells. The CaV3 regulates cell proliferation and Ca2+ dynamics in colorectal cancer cells. Understanding this blocking mechanism maybe improve cancer therapies.

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Metallocompounds are a class of anticancer compounds largely used in the treatment of several types of solid tumors, including bone cancer. Osteosarcoma (OS) is a primary malignant bone tumor that frequently affects children, adolescents and young adults. It is a very invasive type of tumor, so ∼40% of patients develop distant metastases, showing elevated mortality rates. In this review, we present an outline of the chemistry and antitumor properties of metal-based compounds in preclinical (in vitro and in vivo) and clinical OS models, focusing on the relationship between structure-activity, molecular targets and the study of the mechanism of action involved in metallocompound anticancer activity.

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Introduction: Salicylic acid has shown promise in alleviating water stress in cultivated plants. However, there is a lack of studies confirming its effectiveness in cowpea plants grown in field conditions. Therefore, this research aimed to evaluate the use of salicylic acid as a water stress mitigator in cowpea cultivars under different irrigation depths in field conditions. Methods: Four cowpea cultivars (BRS Novaera, BRS Tapaihum, BRS Pujante, and BRS Pajeú) were subjected to different treatments: control (W100: 100% replacement of crop evapotranspiration - ETc), W50 (50% of ETc), W50+SA2 (50% of ETc + 276 mg L-1 of SA), and W50+SA4 (50% of ETc + 552 mg L-1 of SA). The treatments were combined in a 4×4 factorial scheme with three replications, arranged in a randomized block design. Results: Water restriction had a negative impact on the water status, growth, gas exchange, and production of the cultivars while also leading to changes in the antioxidant metabolism and osmolyte concentration. The application of SA enhanced antioxidant activity and the synthesis of osmotic adjusters under stress conditions. The most effective concentration was 276 mg L-1 in stage R2 and 552 mg L-1 in stage V7, respectively. The BRS Pujante cultivar showed increased productivity under water restriction with SA application, while the BRS Tapaihum was the most tolerant among the cultivars studied. Discussion: In summary, our findings underscore the importance of using SA to mitigate the effects of water restriction on cowpea cultivation. These discoveries are crucial for the sustainability of cowpea production in regions susceptible to drought, which can contribute to food security. We further add that the adoption of new agricultural practices can enhance the resilience and productivity of cowpea as an essential and sustainable food source for vulnerable populations in various parts of the world.