In silico prediction of ADMET parameters and in vitro evaluation of antioxidant and cytotoxic activities promoted by indole-thiosemicarbazone compounds.

Cancer is a complex and multifactorial disease characterized by uncontrolled cell growth and is one of the main causes of death in the world. This work aimed to evaluate a small series of 10 different indole-thiosemicarbazone compounds as potential antitumor agents. This is a pioneering study. For this, the antioxidant and cytotoxic capacity against normal and tumor cells was evaluated. The results showed that the compounds were able to promote moderate to low antioxidant activity for the ABTS radical scavenging assay. ADMET in silico assays showed that the compounds exhibited good oral bioavailability. As for toxicity, they were able to promote low cytotoxicity against normal cells, in addition to not being hemolytic. The compounds showed promising in vitro antitumor activity against the T47D, MCF-7, Jurkat and DU-145 strains, not being able to inhibit the growth of the Hepg2 strain. Through this in vitro study, it can be concluded that the compounds are potential candidates for antitumor agents.

Preliminary evaluation of the toxicological, antioxidant and antitumor activities promoted by the compounds 2,4-dihydroxy-benzylidene-thiosemicarbazones an in silico, in vitro and in vivo study.

Thiosemicarbazones are promising classes of compounds with antitumor activity. For this study, six 2,4-dihydroxy-benzylidene-thiosemicarbazones compounds were synthesized. These compounds were submitted to different assays in silico, in vitro and in vivo to evaluate the toxicological, antioxidant and antitumor effects. The in silico results were evaluated by the SwissADME and pkCSM platforms and showed that all compounds had good oral bioavailability profiles. The in vitro and in vivo toxicity assays showed that the compounds showed low cytotoxicity against different normal cells and did not promote hemolytic effects. The single dose acute toxicity test (2000 mg/kg) showed that none of the compounds were toxic to mice. In in vitro antioxidant activity assays, the compounds showed moderate to low activity, with PB17 standing out for the ABTS radical capture assay. The in vivo antioxidant activity highlighted the compounds 1, 6 and 8 that promoted a significant increase in the concentration of liver antioxidant enzymes. Finally, all compounds showed promising antitumor activity against different cell lines, especially MCF-7 and DU145 lines, in addition, they inhibited the growth of sarcoma 180 at concentrations lower than 50 mg/kg. These results showed that the evaluated compounds can be considered as potential antitumor agents.

Identification of runs of homozygosity (ROHs) in Curraleiro Pé-Duro and Pantaneiro cattle breeds.

This study aimed to identify and characterize runs of homozygosis (ROHs), genes involved in production characteristics and adaptation to tropical systems and to estimate the inbreeding coefficient of Curraleiro Pé-Duro (CPD) and Pantaneiro (PANT), two brazilian locally adapted cattle breeds. The results demonstrated that 79.25% and 54.29% of ROH segments were bigger than 8 Mb in CPD and PANT, respectively, indicating recent inbred matings in the studied population. Six homozygosis islands were identified simultaneously in both breeds, where 175 QTLs and 1072 genes previously described as associated with production traits are located. The inbreeding coefficient (FROH) estimated based on ROHs (FROH) showed that inbreeding is low (2 to 4%), which is different from expected for small populations such as locally adapted ones.

In silico ADMET prediction, evaluation of cytotoxicity in mouse splenocytes and preliminary evaluation of in vitro antimalarial activity of 4-(4-chlorophenyl)thiazole compounds.

In this work, an in silico study and evaluation of the cytotoxicity of 4-(4-chlorophenyl)thiazole compounds against mouse splenocytes and the chloroquine-sensitive Plasmodium falciparum 3D7 strain are reported. The in silico results showed that the compounds have important pharmacokinetic properties for compounds with potential drug candidates. Regarding cytotoxicity assays against splenocytes, the compounds have low cytotoxicity. In addition, they were able to promote activation of these cells by increasing nitric oxide production without promoting cell death. Finally, they were able to promote cell proliferation. Regarding the in vitro anti-P. falciparum activity assays, it was observed that the compounds were able to inhibit the parasite's growth, presenting IC50 values ​​ranging from 0.79 to greater than 10 µM. These results are promising when compared to chloroquine. Therefore, this study showed that 4-(4-chlorophenyl)thiazole compounds are promising candidates for antimalarials.

Synthesis of 4-(4-chlorophenyl)thiazole compounds: in silico and in vitro evaluations as leishmanicidal and trypanocidal agents.

Neglected tropical diseases are a diverse group of communicable pathologies that mainly prevail in tropical and subtropical regions. Thus, the objective of this work was to evaluate the biological potential of eight 4-(4-chlorophenyl)thiazole compounds. Tests were carried out in silico to evaluate the pharmacokinetic properties, the antioxidant, cytotoxic activities in animal cells and antiparasitic activities were evaluated against the different forms of Leishmania amazonensis and Trypanosoma cruzi in vitro. The in silico study showed that the evaluated compounds showed good oral availability. In a preliminary in vitro study, the compounds showed moderate to low antioxidant activity. Cytotoxicity assays show that the compounds showed moderate to low toxicity. In relation to leishmanicidal activity, the compounds presented IC50 values that ranged from 19.86 to 200 µM for the promastigote form, while for the amastigote forms, IC50 ranged from 101 to more than 200 µM. The compounds showed better results against the forms of T. cruzi with IC50 ranging from 1.67 to 100 µM for the trypomastigote form and 1.96 to values greater than 200 µM for the amastigote form. This study showed that thiazole compounds can be used as future antiparasitic agents.

Estimating genetic parameters of reproductive, carcass, and meat quality traits in Polled Nellore cattle.

Considering the economic and commercial efficiency of the beef production chain, the yield and quality of the meat produced must also be included in breeding programs. For the Nellore breed, including the polled herd, these aspects have not been much studied. The aim of this study was to estimate genetic parameters for scrotal circumference adjusted to 365 (SC365) and 450 (SC450) days of age, age at first calving (AFC), accumulated productivity (AP), stayability (STAY), longissimus muscle area (LMA), thickness of subcutaneous fat over the 12th-13th ribs (BF), thickness of subcutaneous fat over the rump (RF), and shear force measured by Warner-Bratzler shear force (WBSF) of polled Nellore cattle. Bayesian analyses were performed by adopting a linear animal model, whereas STAY analyses used the linear threshold model. Heritability estimates were 0.31 (SC365), 0.37 (SC450), 0.16 (AFC), 0.25 (AP), 0.16 (STAY), 0.30 (LMA), 0.13 (BF), 0.24 (RF), and 0.15 (WBSF), indicating moderate response to selection. Genetic and residual correlations between SC365 and SC450 were high (0.91 and 0.74, respectively), as well as the genetic correlations of AP with SC365, SC450, AFC, and STAY (0.61, 0.62, - 0.69, and 0.83, respectively). Genetic and residual correlations of WBSF with reproductive and carcass characteristics exhibited high standard deviations, however favorable. Based on the results, it is expected that in the medium term, animals with greater sexual precocity will also have greater accumulated productivity and longer permanence of females in the herd, along with superior carcass traits. However, due to the low heritabilities and small genetic associations with reproductive traits, fat thickness characteristics (BF and RF) will still require direct selection.

Thiazolidine derivatives: In vitro toxicity assessment against promastigote and amastigote forms of Leishmania infantum and ultrastructural study.

Neglected diseases, such as Leishmaniasis, constitute a group of communicable diseases that occur mainly in tropical countries. Considered a public health problem with limited treatment. Therefore, there is a need for new therapies. In this sense, our proposal was to evaluate in vitro two series of thiazolidine compounds (7a-7e and 8a-8e) against Leishmania infantum. We performed in vitro evaluations through macrophage cytotoxicity assays (J774) and nitric oxide production, activity against promastigotes and amastigotes, as well as ultrastructural analyzes in promastigotes. In the evaluation of cytotoxicity, the thiazolidine compounds presented CC50 values between 8.52 and 126.83 µM. Regarding the evaluation against the promastigote forms, the IC50 values ranged between 0.42 and 142.43 µM. Compound 7a was the most promising, as it had the lowest IC50. The parasites treated with compound 7a showed several changes, such as cell body shrinkage, shortening and loss of the flagellum, intense mitochondrial edema and cytoplasmic vacuolization, leading the parasite to cell inviability. In assays against the amastigote forms, the compound showed a low IC50 (0.65 µM). These results indicate that compound 7a was efficient for both evolutionary forms of the parasite. In silico studies suggest that the compound has good oral bioavailability. These results show that compound 7a is a potential drug candidate for the treatment of Leishmaniasis.

Ageing characteristics related to cation exchange and interlayer spacing of some Brazilian bentonites.

This study investigated the influence of cation type on the sodium deactivation of Brazilian bentonite varieties. Four bentonite clays were studied, including three mixed-cationic and one magnesium. Swelling and the main exchangeable cations (Na+, K+, Mg2+ and Ca2+) were the main evaluation parameters in this study. Periodic washes of the sodium-activated bentonites were performed based on the hypothesis that a possible desorption of the cations (primarily Na+) could cause the deactivation. Sodium activation was monitored using XRD measurements and an increase in swelling. Sodium deactivation was observed and monitored via a decrease in swelling. Positive and negative effects, caused by the proportions of the cations and the dominance of Mg2+, were emphasized by the set of cationically different samples applied in this study, which helped to answer the influence of main exchangeable cations, specially Mg2+, on the deactivation process.

As dificuldades da participação: desencontro de interesses na recuperação do rio doce / The difficulties of participation: interests mismatchs in rio doce recovery / Las dificultades de la participación: desencuentro de intereses en la recuperación del rio doce / Les difficultés de la participation: intérêts introuvable dans la récupération de rio doce

O exame de experiência de implementação de uma tecnologia participativa (CRCMOP) nas comunidades atingidas pelo desastre do Rio Doce, no Espírito Santo, ao longo de dois anos, permite atentar para as dificuldades da participação da população atingida na governança da gestão do desastre, especificamente na definição das ações de reparação, compensação e indenização a serem realizadas pela Fundação Renova. Argumentamos que a formulação de demandas e sua projeção às instituições competentes para acolhe-las e lhes dar resposta depende de pressupostos relativos a duas dimensões: cognitivo simbólica e sócio- institucional. Ambas, em boa medida, ausentes no caso das comunidades atingidas, particularmente daquelas situadas na foz do rio. Por isso, argumentamos, a gestão do desastre socioambiental ocasionado pelo rompimento da barragem de Fundão acusa uma forma peculiar de desencontro de interesses entre comunidades atingidas e instituições incumbidas dessa gestão. As dificuldades diagnosticadas podem ser comuns a outros contextos de grandes desastres.

Examining the experience of implementing a participatory technology (CRCMOP) in the communities affected by the Rio Doce disaster in Espírito Santo over two years reveals the difficulties of affected population participation in disaster management governance, specifically in taking part in the definition of reparation, compensation and indemnity measures to be carried out by the Renova Foundation. We argue that the formulation of claims and their statement vis-à-vis the competent institutions that are supposed to receive and respond to them depends on assumptions concerning two dimensions: symbolic-cognitive and socio-institutional. Both are largely absent in affected communities, particularly those at the mouth of the river. Therefore, we argue, the management of the socio-environmental disaster caused by the Fundão dam burst rupture presents a peculiar form difficulty: a mismatch of interests between affected communities and institutions responsible for managing it. Diagnosed difficulties may be common to other major disaster contexts. These difficulties, here diagnosed, may also be common to other major disaster contexts.

El examen de una experiencia de implantación de una tecnología participativa (CRCMOP) en las comunidades afectadas por el desastre del Rio Doce (Espírito Santo), durante dos años, permite iluminar las dificultades de la participación de la población afectada en la gobernanza de la gestión del desastre, específicamente, en las acciones de reparación, compensación e indemnización que debe realizar la Fundación Renova. Argumentamos que la formulación de demandas y su proyección hacia las instituciones competentes para acogerlas y darles respuesta depende de presupuestos relativos a dos dimensiones: cognitivo-simbólica e socio-institucional. Ambas, en buena medida, ausentes en el caso de las comunidades afectadas, particularmente de aquellas situadas en la boca del rio. Argumentamos que la gestión del desastre socioambiental causado por la ruptura de la presa de Fundão acusa una forma peculiar de desencuentro de intereses entre comunidades afectadas e instituciones responsables por esa gestión. Las dificultades diagnosticadas pueden ser comunes a otros contextos de grandes desastres.

L'examen d'une expérience de mise en oeuvre d'une technologie participative (CRCMOP) dans les communautés affectées par la catastrophe de Rio Doce (Espirito Santo) au cours des deux années, nous permet d'éclairer les difficultés dela participation despersonnes affectées dans la gestion de la catastrophe, en particulier, dans les actions de réparation et d'indemnisation qui doivent être menées par la Fondation Renova. Nous soutenons que la formulation des demandes a les institutions responsable de recevoir les demandes et d'y répondre dépend a deux conditions concernant a les dimensions cognitive-symbolique et socio-institutionnelle. Les deux sont, dans une large mesure, absents dans le cas des communautés affectées, en particulier celles situées à l'embouchure de la rivière du Doce. Nous soutenons ausssi que la gestion de la catastrophe socio-environnementale causée par la rupture du barrage de Fundão montre une forme particulière de décalage des intérêts entre les communautés affectées et les institutions responsables de cette gestion. Les difficultés diagnostiquées peuvent être communes à d'autres contextes de catastrophes majeures.

Chronic invasive rhinosinusitis by Conidiobolus coronatus, an emerging microorganism.

Fungi from the Conidiobolus genus have been implicated in the development of chronic invasive fungal rhinosinusitis, mainly in tropical countries. The mycosis associated to these fungi may cause irreversible facial deformities and is potentially fatal. The authors present the first case of a chronic invasive fungal rhinosinusitis due to Conidiobolus coronatus diagnosed in a 66-year-old Caucasian male patient, living in Portugal without any travels abroad and complaining of progressive refractory nasal obstruction, facial pain and anosmia. Upon the culture of samples collected during sinus endoscopic surgery, colonies that presented a macroscopic aspect remembering wax were detected. The microscopic evaluation allowed the observation of simple conidiophores forming spherical conidia, and of conidiophores that presented hair-like appendages. Together, these characteristics allowed the identification of the fungi as a Conidiobolus, which was confirmed upon the DNA sequencing. The authors emphasised the role of this fungi as an emergent microorganism as well as the difficulties associated to the diagnosis and treatment.

Effect of Heavy Metals Contamination from Cigarette Smoke on Sound and Caries-Like Enamel.

In this study, we sought to evaluate the influence of cigarette smoke and pH cycling on the chemical composition and surface/cross-sectional enamel microhardness. A total of 40 dental blocks obtained from bovine incisors were divided into four groups (n=10): no treatment (control); exposure to cigarette smoke (CS); exposure to pH cycling (PC); and exposure to cigarette smoke and pH cycling (CS-PC). The samples were analyzed by synchrotron radiation micro X-ray fluorescence, bench mode X-ray fluorescence, as well as surface microhardness (SMH) and cross-sectional microhardness (CSMH) testing. The SMH results were submitted to analysis of variance (ANOVA) and Tukey's test. The CSMH results were evaluated using split-plot ANOVA and Tukey's test. A high amount of Cd and Pb and traces of Ni and As were observed in enamel and dentin after exposure to cigarette smoke (CS and CS-PC). The SMH and CSMH of CS were statistically higher when compared with the control. The PC and CS-PC showed lower SMH and CSMH. We conclude that exposure to cigarette smoke promoted heavy metal deposition in enamel/dentin. In addition, it increased the enamel microhardness but did not promote a protective effect on the in vitro development of caries. The clinical significance of this work is that there is significant bioaccumulation of heavy metals from cigarette smoke on the surface and in the enamel and dentin.

Correlation between DNA/HSA-interactions and antimalarial activity of acridine derivatives: Proposing a possible mechanism of action.

Acridines are considered an important class of compounds due to their wide variety of biological activities. In this work, we synthesized four acridine derivatives (1-4) and evaluated their biological activity against the Plasmodium falciparum W2 line, as well as studied the interaction with ctDNA and HSA using spectroscopic techniques and molecular docking. The acridine derivative 2 (IC50 = 0.90 ±â€¯0.08 µM) was more effective against P. falciparum than primaquine (IC50 = 1.70 ±â€¯0.10 µM) and similar to amsacrine (IC50 = 0.80 ±â€¯0.10 µM). In the fluorescence and UV-vis assays, it was verified that the acridine derivatives interact with ctDNA and HSA leading to a non-fluorescent supramolecular complex formation. The non-covalent binding constants ranged from 2.09 to 7.76 × 103 M-1, indicating moderate interaction with ctDNA. Through experiments with KI, fluorescence contact energy transfer and competition assays were possible to characterize the main non-covalent binding mode of the acridines evaluated with ctDNA as intercalation. The binding constants obtained showed a high linear correlation with the IC50 values against the antimalarial activity, suggesting that DNA may be the main biological target of these molecules. Finally, HSA interaction studies were performed and all evaluated compounds bind to the site II of the protein. The less active compounds (1 and 3) presented the highest affinity to HSA, indicating that the interaction with carrier protein can affect the (bio)availability of these compounds to the biological target.

Solid dispersions to enhance the delivery of a potential drug candidate LPSF/FZ4 for the treatment of schistosomiasis.

Drug candidate LPSF/FZ4 with promising schistosomicidal properties in vitro was previously synthesized. However, LPSF/FZ4 has limited aqueous solubility (<1 µg/mL), leading to ineffective dissolution and, therefore, no meaningful in vivo comparative studies could be pursued. This study was aimed to develop a proper amorphous solid dispersion (SD) to enhance the solubility and dissolution rate of LPSF/FZ4 such that its biological activity could be investigated. To better understand its physiological behavior, the pKa of LPSF/FZ4, a monoprotic weak acid with NH group at the imidazolidine ring, was first determined to be 8.13 using an automated SiriusT3. The development of SD systems for LPSF/FZ4 involved the evaluation of various water-soluble polymer carriers such as PVP K-29/32, PVP K-90, HPMC K4M, PVPVA 64 and SOLUPLUS®. The most promising SD systems were selected through in vitro dissolution studies under nonsink conditions, together with physicochemical characterization as well as accelerated stability study. It was shown that SD of 10% LPSF/FZ4 in SOLUPLUS® and PVP K-90 could significantly increase the area-under-the-curve value of the nonsink dissolution profile (AUC values of the SD in SOLUPLUS® and PVP K-90 were 1381.03 and 1342.34 µL/mL·min, respectively, and that of the pure crystalline drug was 0.02 µL/mL·min), a useful surrogate for the in vivo bioavailability. Cmax values for the SD in SOLUPLUS® (12.50 µL/mL) and PVP K-90 (25.86 µL/mL) were also higher than the one of the crystalline drug (0.02 µL/mL). The SD system of LPSF/FZ4 in SOLUPLUS® showed a significant increase in schistosomicidal activity in an animal model as compared with the conventional treatment using crystalline drug, consistent with the AUC trend from the nonsink dissolution. Thus this SD system of LPSF/FZ4 could be useful as a potential formulation for treating schistosomiasis.

Mismatch novelty exploration training enhances hippocampal synaptic plasticity: A tool for cognitive stimulation?

Memory formation relies on experience-dependent changes in synaptic strength such as long-term potentiation (LTP) or long-term depression (LTD) of synaptic activity, that in turn depend on previous learning experiences through metaplasticity. Novelty detection is a particularly important cognitive stimulus in this respect, and mismatch novelty has been associated with the activation of the hippocampal CA1 area in human studies. A single exposure to a new location of known objects in a familiar environment, a behavioural mismatch novelty paradigm, is known to favour the expression of LTD in hippocampal CA3 to CA1 synaptic transmission in vivo, through short-term metaplasticity. Aiming to shape hippocampal responsiveness to synaptic plasticity phenomena we developed a training program based on exploration of a known environment containing familiar objects, everyday presented in a new location. Repeated exposure to this new location of objects for two weeks caused a mild long-lasting decrease in synaptic efficacy. Furthermore, it enhanced both LTP evoked by theta-burst stimulation and depotentiation evoked by low-frequency stimulation of CA3 to CA1hippocampal synaptic transmission in juvenile rats. This suggests that training programs using these behavioural tasks involving mismatch novelty can be used to reshape brain circuits and promote cognitive recovery in pathologies where LTP/LTD imbalance occurs, such as epilepsy, aging or Down's syndrome, an approach that requires further investigation at the behavioural level.

Synthesis and Anticancer Evaluation of Thiazacridine Derivatives Reveals New Selective Molecules to Hematopoietic Neoplastic Cells.

AIM AND OBJECTIVE: Cancer has become one of the leading causes of morbidity and mortality worldwide. Limitations associated with existing agents increase the need to develop more effective anticancer drugs to improve the therapeutic arsenal available. The aim of this study was to synthesize and evaluate the antiproliferative effects of three new thiazacridine derivatives. MATERIAL AND METHODS: Using a three steps synthesis reaction, three novel thiazacridine derivatives were obtained and characterized: (Z)-5-acridin-9-ylmethylene-3-(4-methyl-benzyl)-4-thioxo-thiazolidin- 2-one (LPSF/AC-99), (Z)-5-acridin-9-ylmethylene-3-(4-chloro-benzyl)-4-thioxo-thiazolidin-2- one (LPSF/AC-119) and (Z)-5-acridin-9-ylmethylene-3-(3-chloro-benzyl)-4-thioxo-thiazolidin-2- one (LPSF/AC-129). Toxicity and selectivity assays were performed by colorimetric assay. Then, changes in cell cycle and cell death induction mechanisms were assessed by flow cytometry. RESULTS: All compounds exhibited cytotoxicity to Raji (Burkitt's lymphoma) and Jurkat (acute T cell leukemia) cells, where LPSF/AC-119 showed best IC50 values (0.6 and 1.53 µ M, respectively). LPSF/AC-129 was the only cytotoxic compound in glioblastoma cell line NG97 (IC50 = 55.77 µ M). None of the compounds were toxic to normal human cells and induced neoplastic cell death primarily by apoptosis. CONCLUSION: All derivatives were more cytotoxic to hematopoietic neoplastic cells when compared to solid tumor derived cells. All three compounds are promising for in vivo and combination therapy studies against cancer.

Accuracy of genomic predictions in Gyr (Bos indicus) dairy cattle.

Genomic selection may accelerate genetic progress in breeding programs of indicine breeds when compared with traditional selection methods. We present results of genomic predictions in Gyr (Bos indicus) dairy cattle of Brazil for milk yield (MY), fat yield (FY), protein yield (PY), and age at first calving using information from bulls and cows. Four different single nucleotide polymorphism (SNP) chips were studied. Additionally, the effect of the use of imputed data on genomic prediction accuracy was studied. A total of 474 bulls and 1,688 cows were genotyped with the Illumina BovineHD (HD; San Diego, CA) and BovineSNP50 (50K) chip, respectively. Genotypes of cows were imputed to HD using FImpute v2.2. After quality check of data, 496,606 markers remained. The HD markers present on the GeneSeek SGGP-20Ki (15,727; Lincoln, NE), 50K (22,152), and GeneSeek GGP-75Ki (65,018) were subset and used to assess the effect of lower SNP density on accuracy of prediction. Deregressed breeding values were used as pseudophenotypes for model training. Data were split into reference and validation to mimic a forward prediction scheme. The reference population consisted of animals whose birth year was ≤2004 and consisted of either only bulls (TR1) or a combination of bulls and dams (TR2), whereas the validation set consisted of younger bulls (born after 2004). Genomic BLUP was used to estimate genomic breeding values (GEBV) and reliability of GEBV (R2PEV) was based on the prediction error variance approach. Reliability of GEBV ranged from ∼0.46 (FY and PY) to 0.56 (MY) with TR1 and from 0.51 (PY) to 0.65 (MY) with TR2. When averaged across all traits, R2PEV were substantially higher (R2PEV of TR1 = 0.50 and TR2 = 0.57) compared with reliabilities of parent averages (0.35) computed from pedigree data and based on diagonals of the coefficient matrix (prediction error variance approach). Reliability was similar for all the 4 marker panels using either TR1 or TR2, except that imputed HD cow data set led to an inflation of reliability. Reliability of GEBV could be increased by enlarging the limited bull reference population with cow information. A reduced panel of ∼15K markers resulted in reliabilities similar to using HD markers. Reliability of GEBV could be increased by enlarging the limited bull reference population with cow information.

Analysis of tryptase-positive mast cells and immunoexpression of MMP-9 and MMP-13 in periapical lesions.

AIM: To evaluate the immunoexpression of tryptase, MMP-9 and MMP-13 in periapical lesions, correlating them with the type of lesion, intensity of the inflammatory infiltrate and thickness of the epithelial lining. METHODOLOGY: Twenty periapical granulomas (PGs), twenty radicular cysts (RCs) and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using antitryptase, anti-MMP-9 and anti-MMP-13 antibodies. Immunoexpression of MMP-9 and MMP-13 was quantitatively evaluated both in the connective tissue of all lesions and in the epithelial lining of RCs and RRCs. Tryptase-positive mast cells were counted only in the connective tissue. The data were analysed using Kruskal-Wallis and Fisher's exact tests, as well as Spearman's correlation test (P < 0.05). RESULTS: In comparison with RCs and RRCs, PGs exhibited higher immunoexpression of tryptase, MMP-9 and MMP-13 (P = 0.002, P = < 0.001 and P < 0.001, respectively). In comparison with lesions with inflammatory infiltrates grades I and II, lesions with inflammatory infiltrate grade III had higher median percentages of MMP-13-positive cells (P = 0.003) and a tendency for higher expression of MMP-9 (P = 0.059). No significant difference was observed between the expression of the studied markers and epithelial thickness (P > 0.05). There were positive correlations between the number of tryptase-positive mast cells and the immunoexpression of MMP-9, as well as between the immunoexpression of MMP-9 and MMP-13. CONCLUSION: A larger number of tryptase-positive mast cells and greater enzymatic activity of MMP-9 and MMP-13 were found in PGs compared to RCs and RRCs. These findings are a characteristic of the dynamics of periapical diseases.

Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil).

Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of Paullinia cupana. The Actinobacteria was identified as Streptomyces hygroscopicus ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with n-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C54H88O18) based on 1H and 13C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC50 of 1 µg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC50 of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and <3.9 µg/mL, respectively). In conclusion, the EB1 fraction and elaiophylin of S. hygroscopicus have potent antimicrobial, antifungal, and anticancer activities.

Description of Loxtox protein family and identification of a new group of Phospholipases D from Loxosceles similis venom gland.

Envenoming resulting from Loxosceles spider bites (loxoscelism) is a recognized public health problem in Brazil. However, the pathophysiology of loxoscelism caused by L. similis bites, which is widespread in Brazil, remains poorly understood. In the present work, the RNA sequencing (RNA-Seq - Next Generation sequencing - NGS) of the L. similis venom gland was performed to identify and analyze the sequences of the key component phospholipase D. The sequences were aligned based on their classical domains, and a phylogenetic tree was constructed. In the bioinformatics analysis, 23 complete sequences of phospholipase D proteins were found and classified as Loxtox proteins, as they contained the characteristic domains of phospholipase D: the active site, the Mg(2+)-binding domain, and the catalytic loop. Three phospholipase D sequences with non-canonical domains were also found in this work. They were analyzed separately and named PLDs from L. similis (PLD-Ls). This study is the first to characterize phospholipase D sequences from Loxosceles spiders by RNA-Seq. These results contribute new knowledge about the composition of L. similis venom, revealing novel tools that could be used for pharmacological, immunological, and biotechnological applications.

Mineral requirements of hair sheep in tropical climates.

In this study, we evaluated the requirements of calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na) and potassium (K) for sheep hair growth. Experimental diets contained different levels of metabolizable energy [ME; 0.96, 1.28, 1.72, 2.18 and 2.62 Mcal/kg of dry matter, (DM), corresponding to 4.23, 5.64, 7.58, 9.61 and 11.55 MJ/kg DM]. The lambs' hair (n = 48) at 2 months of age presented an average body weight (BW) of 12.05 ± 1.81 kg. At the beginning of the experiment, eight animals were slaughtered as a reference group to estimate the initial empty body weight and body composition. Net mineral requirements (g/day) ranged from 0.73 to 0.71 g of Ca, 0.51 to 0.49 g of P, 0.026 to 0.026 g of Mg, 0.16 to 0.19 g of Na and 0.15 to 0.13 g of K for animals with a BW ranging from 15 to 30 kg and a daily gain of 100 g. The results of this study indicate that the net macromineral requirements for weight gain in Morada Nova lambs are different from the values commonly recommended by the Agricultural and Food Research Council.