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In Brief: The natriuretic peptide system, particularly the C-type natriuretic peptide (CNP) and natriuretic peptide receptor 2 (NPR2), plays a critical role in regulating mammalian ovarian functions, including oocyte-cumulus cell communication and meiotic maturation. However, the contribution of natriuretic peptide receptor 3 (NPR3) to these processes has not been thoroughly investigated. Data from this study provide compelling evidence that NPR3 is involved in modulating gonadotropin signaling and regulating cumulus cell expansion in cattle., Abstract: The significant role of CNP and its receptor 2 (NPR2) in regulating oocyte meiotic maturation and facilitating communication between oocytes and surrounding cumulus cells has been well documented in various mammalian species including mice, cattle and swine. However, further investigation is needed to ascertain whether natriuretic peptide receptors (NPRs) are involved in regulating other essential ovarian functions. Hence, this study aimed to explore the potential involvement of NPRs in the regulation of cumulus expansion and oocyte meiotic maturation in bovine cumulus-oocyte complexes (COCs). The findings revealed that NPR3 mRNA abundance was downregulated by follicle-stimulating hormone and luteinizing hormone in cumulus cells of bovine COCs during in vitro maturation (IVM), while NPR2 mRNA levels were not affected by gonadotropins. Inhibition of the epidermal growth factor receptor (EGFR) during IVM of COCs prevented the NPR3 mRNA downregulation induced by gonadotropins in cumulus cells. Additionally, treatment of COCs during IVM with an NPR3 agonist (cANP4-23) inhibited cumulus expansion induced by gonadotropins. This inhibitory effect was further intensified when COCs were cotreated with cANP4-23 and CNP. These findings provide robust evidence indicating that normal cumulus expansion in bovine COCs involves an inhibitory effect of gonadotropins on NPR3 mRNA expression, which is mediated via EGFR signaling. The study also provides evidence that CNP and NPR3 interact synergistically to regulate cumulus expansion in response to gonadotropins.

Leishmaniases are among the neglected tropical diseases that still cause devastating health, social, and economic consequences to more than 350 million people worldwide. Despite efforts to combat these vector-borne diseases, their incidence does not decrease. Meanwhile, current antileishmanial drugs are old and highly toxic, and safer presentations are unaffordable to the most severely affected human populations. In a previous study by our research group, we synthesized 17 flavonoid derivatives that demonstrated impressive inhibition capacity against rCPB2.8, rCPB3, and rH84Y. These cysteine proteases are highly expressed in the amastigote stage, the target form of the parasite. However, although these compounds have been already described in the literature, until now, the amastigote effect of any of these molecules has not been proven. In this work, we aimed to deeply analyze the antileishmanial action of this set of synthetic flavonoid derivatives by correlating their ability to inhibit cysteine proteases with the action against the parasite. Among all the synthesized flavonoid derivatives, 11 of them showed high activity against amastigotes of Leishmania amazonensis , also providing safety to mammalian host cells. Furthermore, the high production of nitric oxide by infected cells treated with the most active cysteine protease B (CPB) inhibitors confirms a potential immunomodulatory response of macrophages. Besides, considering flavonoids as multitarget drugs, we also investigated other potential antileishmanial mechanisms. The most active compounds were selected to investigate another potential biological pathway behind their antileishmanial action using flow cytometry analysis. The results confirmed an oxidative stress after 48 h of treatment. These data represent an important step toward the validation of CPB as an antileishmanial target, as well as aiding in new drug discovery studies based on this protease.

Aspartame (ASP) is a common sweetener, but studies show it can harm the nervous system, causing learning and memory deficits. β-caryophyllene (BCP), a natural compound found in foods, including bread, coffee, alcoholic beverages, and spices, has already described as a neuroprotector agent. Remarkably, ASP and BCP are commonly consumed, including in the same meal. Therefore, considering that (a) the BCP displays plenty of beneficial effects; (b) the ASP toxicity; and (c) that they can be consumed in the same meal, this study sought to investigate if the BCP would mitigate the memory impairment induced by ASP in rats and investigate the involvement of the brain-derived neurotrophic factor (BDNF)/ tropomyosin receptor kinase B (TrKB) signaling pathway and acetylcholinesterase (AChE) activity. Young male Wistar rats received ASP (75 mg/kg; i.g.) and/or BCP (100 mg/kg; i.p.) once daily, for 14 days. At the end of the treatment, the animals were evaluated in the open field and object recognition tests. The cerebral cortex and hippocampus samples were collected for biochemical and molecular analyses. Results showed that the BCP effectively protected against the cognitive damage caused by ASP in short and long-term memories. In addition, BCP mitigated the increase in AChE activity caused by ASP. Molecular insights revealed augmented BDNF and TrKB levels in the hippocampus of rats treated with BCP, indicating greater activation of this pathway. In conclusion, BCP protected against ASP-induced memory impairment. AChE activity and the BDNF/TrkB signaling pathway seem to be potential targets of BCP modulatory role in this study., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Aflatoxin B 1 (AFB 1 ) is a mycotoxin highly toxic and carcinogenic to humans due to its potential to induce oxidative stress. The Beta-caryophyllene (BCP) have been highlighted for its broad spectrum of pharmacological effects. The present study aimed to investigate the beneficial effects of BCP against the susceptibility of hepatic and renal tissues to AFB 1 toxicity, in biochemical parameters to assess organ function, tissue oxidation, and the immunocontent of oxidative and inflammatory proteins. Male Wistar rats was exposed to AFB 1 (250 μg/kg, i.g.) and/or BCP (100 mg/kg, i.p.) for 14 successive days. It was found that exposure to AFB 1 did not change the measured renal toxicity parameters. Also, AFB 1 increased liver injury biomarkers (gamma glutamyl transferase and alkaline phosphatase) and reduced levels of non-enzymatic antioxidant defenses (ascorbic acid and non-protein thiol), however did not cause changes in the lipid peroxidation levels. Moreover, AFB 1 interfered in oxidative pathway regulated by Kelch-like ECH-associated protein (Keap1)/nuclear factor (erythroid-derived 2)-like 2 (Nrf2), overacting Glutathione-S-Transferase (GST) activity. Lastly, a main effect of AFB 1 on the total interleukin 1 beta (IL-1β) was observed. Remarkably, the associated treatment of AFB 1  + BCP improved altered liver parameters. In addition, BCP and AFB 1  + BCP groups showed an increase in the levels of inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ). Thus, these results indicated that BCP has potential protective effect against AFB 1 induced hepatotoxicity., (Copyright © 2021. Published by Elsevier B.V.)

Nanotechnology has been an important tool for the production of nanoparticles with controlled release of drugs for therapeutic applications. Here, we produced solid lipid nanoparticles (SLN) loaded with curcumin and capsaicin (NCC) following the overarching goals of green chemistry. Currently, besides evaluating the composition, and size of these, it is necessary to understand the interactions between nanoparticles and the biomolecules present in the biological medium. For this, assays were conducted in order to evaluate the potential formation of the protein 'corona', and to better understand the results obtained in vitro, we also performed an interaction study, in silico, between the NCC components and the main serum protein, albumin. In the first hour of contact between the NCC and the culture medium showed fluctuation in the diameter of the NCC. However, after 24 and 48 h of the incubation period, all NCC concentrations showed an increase in size, which can be attributed to plasma protein adsorption. Since, hard corona takes a few seconds, while the soft corona can be formed in minutes up to a few hours. On the other hand, best docking binding-poses of interaction for the formed docking complexes evaluated suggest interactions following the docking affinity like free energy FEB (Tween 80-bovine serum albumin) ≈ FEB (Span 80-bovine serum albumin) showing a pharmacodynamic pattern based in non-covalent hydrophobic interactions with the bovine serum albumin binding-site. Our in silico results clarify and reinforce our in vitro findings of corona formation, which represents the real interaction with cell membranes in vivo., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Although widely known for their involvement in the control of carbohydrate and lipid metabolism of vertebrates, the participation of catecholamines (CAs) in the metabolism of invertebrates is less understood. This study was designed to identify the physiological role of Epinephrine (E) in the intermediary metabolism of the burrowing crab Neohelice granulata and how E regulates the metabolism in crabs fed with a high-carbohydrate (HC) or a high-protein (HP) diet. To answer these questions, we evaluated in vivo the effects of E injections on glucose and triglycerides in the hemolymph and tissue glycogen levels of crabs fed with HC or HP diet. An in vitro investigation was carried out to assess the direct effects of E on glycogenolysis, lipolysis and glycolysis pathways in the hepatopancreas, mandibular muscle and anterior and posterior gills of this crab. E injections increased glucose and did not affect triglycerides levels in the hemolymph of either group of crabs, and E decreased glycogen in the hepatopancreas and mandibular muscle only in HP crabs, suggesting that these effects may be mediated by the crustacean hyperglycemic hormone (CHH). When the tissues were incubated with different concentrations of E, the concentration of glucose released to the medium decreased in the hepatopancreas and posterior gills, while glucose oxidation increased in the posterior gills of HP crabs. Incubation with E did not alter any parameter in tissues of HC crabs. These effects suggest that E may be involved in the metabolic response to osmotic stress., (Copyright © 2019 Elsevier Inc. All rights reserved.)

During folliculogenesis, the luteinizing hormone (LH) surge triggers dynamic events in granulosa cells that culminate with ovulation. The aim of this study was to evaluate if the epidermal growth factor receptor (EGFR) is required for ovulation in cattle, and if it regulates the expression of the natriuretic peptide (NP) system in granulosa cells after gonadotropin-releasing hormone (GnRH)/LH stimulation. It was observed that GnRH induces amphiregulin (AREG) and epiregulin (EREG) mRNA at 3 and 6 h after in vivo treatment, but the expression of these genes was not regulated by atrial (ANP) and C-type (CNP) NPs in granulosa cells cultured in vitro. The abundance of mRNA encoding the NP receptors (NPR1, 2 and 3) was not altered by LH supplementation and/or EGFR inhibition (AG1478; AG) in granulosa cells after 6 h of in vitro culture. However, in the same conditions, mRNA encoding the natriuretic peptide precursor C (NPPC) was upregulated by LH, whereas AG (0.5 and 5 μM) inhibited the LH effect. In order to confirm those results, 5 μM AG or saline were intrafollicularly injected in preovulatory follicles and cows were simultaneously treated with GnRH intramuscularly. Granulosa cells harvested at 6 h after GnRH injection revealed higher NPR3 and lower NPPC mRNA levels in AG-treated, compared to control cows. However, intrafollicular injection of AG did not inhibit GnRH-induced ovulation. In granulosa cells cultured in vitro, ANP associated with LH increased prostaglandin-endoperoxide synthase 2 (PTGS2) mRNA abundance. In conclusion, we inferred that LH modulated NPPC and NPR3 mRNA abundance through EGFR in bovine granulosa cells, but ovulation in cattle did not seem to depend on EGFR activation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The relationship between metabolic disturbances and clinical events related to diabetes is well known. Yerba mate has presented a potential use as preventive and therapeutic agent on diabetes. The aim of this study was to evaluate the effect of yerba mate on different tissues of diabetic rats, focusing on energetic metabolism. Diabetes was induced by streptozotocin, followed by daily yerba mate treatment. After 30 days, the animals were euthanized to evaluate metabolic parameters on liver, adipose tissue, muscle and serum. The results showed mate treatment promoted a decrease in retroperitoneal adipose tissue in healthy animals. Muscle weight returned to control levels in diabetic rats treated with mate. There was improvement on serum glucose, creatinine, urea and total protein levels associated with mate treatment. Muscle parameters, such as glucose uptake and carbon dioxide production, were improved by mate treatment to control levels. The results evidenced the beneficial actions mate can have on metabolic disturbances of diabetes., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Chagas disease (CD) is caused by Trypanosoma cruzi, an intracellular protozoan which is a potent stimulator of cell-mediated immunity. In the indeterminate form of CD (IFCD) a modulation between pro- and anti-inflammatory responses establishes a host-parasite adaptation. It was previously demonstrated that purinergic ecto-enzymes regulates extracellular ATP and adenosine levels, influencing immune and inflammatory processes during IFCD. In inflammatory sites ATP, as well as its degradation product, adenosine, function as signaling molecules and immunoregulators through the activation of purinergic receptors. In this work, it was analyzed the gene and protein expression of P2X7 purinergic receptor in peripheral lymphocytes and serum immunoregulatory cytokines from IFCD patients. Gene and protein expression of P2X7 receptor (P2X7R), and serum cytokines (IL-2, IL-10, IL-17 and IFN-γ) were unaltered. However, IFCD group showed significantly higher IL-4 and IL-6 levels while TNF-α was significantly decreased. These results indicate that imune profile of IFCD patients displays anti-inflammatory characteristics, consistent with the establishment of an immunomodulatory response. Further study about the molecular knowledge of P2X7R in IFCD is useful to clarify the participation of purinergic system in the regulatory mechanism which avoid the progression of CD., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This study evaluated (1) the effects of in vivo GnRH treatment on mRNA expression of TNF-α system (TNF-α, TNFR1 and TNFR2) in granulosa cells of bovine preovulatory follicles, (2) the in vitro influence of gonadotropins on mRNA expression of TNF-α system in cultured cumulus cells, (3) the protein expression of the TNF-α system in late antral follicles and, (4) the influence of TNF-α on cumulus cells expansion, ultrastructure and on expression of HAS2, CASP3 and CASP6 in follicular cells cultured for 24 h. An increased expression of TNF-α and TNFR1 was observed after 3, 6 and 12 h of GnRH treatment when compared to 0 and 24h. Higher TNFR2 mRNA levels were observed 3, 6 and 12 h after GnRH, when compared to 0 and 24 h. Proteins of TNF-α system were also expressed in late antral follicles. In vitro, TNF-α did not affect cumulus cells expansion, but reduced the HAS2, CASP3 and CASP6 mRNA levels in cumulus cells after 12 h. After 24 h of culture, TNF-α increased the mRNA levels for CASP6 in mural granulosa cells, while the TNF-α, TNFR1 and TNFR2 mRNA levels were increased in cumulus-oocyte complexes (COCs) cultured for 12 h with gonadotropins, but not after 24 h. Ultrastructural analysis confirmed the integrity of COCs cultured in presence of TNF-α. In conclusion, TNF-α system members are present in bovine antral follicles and expression of TNF-α is influenced by gonadotropins in vivo and in vitro. In vitro, TNF-α maintained cumulus cells ultrastructure during COC culture., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The present study evaluated whether the gonadotrophin surge modulates components of the renin-angiotensin system and whether angiotensin II (Ang II) plays a role in the production of hormones by follicular cells during the ovulatory process. In Experiment 1, cows were ovariectomised at various times (0, 3, 6, 12 and 24 h) after GnRH injection to obtain preovulatory follicles. The concentration of Ang II in follicular fluid increased after GnRH and reached a peak at 24 h, concomitant with the peak of angiotensinogen (AGT) mRNA expression in granulosa cells. AGT mRNA was not expressed in theca cells. Ang II receptor type 2 and angiotensin-converting enzyme mRNA levels were transiently upregulated in theca cells. In Experiment 2, an in vitro culture was used to determine whether Ang II could modulate hormone production by healthy dominant follicles. In the absence of LH, Ang II did not alter hormonal production by either theca or granulosa cells. Ang II plus LH increased progesterone and prostaglandin secretion by granulosa cells. In summary, the renin-angiotensin system is actively controlled during the preovulatory period and Ang II amplifies the stimulatory effects of LH on the secretion of progesterone and prostaglandins by granulosa cells.

The kallikrein-kinin system (KKS) has been described as an important mediator of physiologic processes. Kallikreins use kininogen (KNG) as substrate to generate bradykinin, the main active peptide of the KKS that acts through two types of receptors, the B(1)R and the B(2)R. The goal of this study was to characterize some components of the KKS in different compartments of the ovary during the bovine ovulation process. The KNG, B(1)R and B(2)R mRNA expression patterns were assessed in theca and granulosa cells, as well as the bradykinin concentration and kallikrein-like activity in follicular fluid of bovine periovulatory follicles. To obtain a periovulatory follicle (≥12 mm), twenty-seven cows were submitted to estrus synchronization protocol and ovariectomized by colpotomy at 0, 3, 6, 12 or 24h after a GnRH-analog injection (gonadorelin; 100 μg, IM). Follicular fluid was aspirated for enzymatic assays while granulosa and theca cells were harvested for mRNA analysis. The mRNA expressions in follicular cells were evaluated by real-time RT-PCR and data representation related to the cyclophilin housekeeping gene. The bradykinin concentration and kallikrein-like activity were measured in follicular fluid by enzymatic immunoassay and selective substrate cleavage, respectively. The B(2)R expression in theca cells and B(1)R expression in theca and granulosa cells showed different profiles during the periovulatory period (P<0.05). The bradykinin concentration and kallikrein-like activity in the follicular fluid were different (P<0.05) due to the time during the ovulation process. KNG mRNA expression was similar for both follicular cell types (P>0.05). Taken together, these results provide an important characterization of the presence and possible KKS regulation during the bovine ovulation., (Copyright © 2011. Published by Elsevier Inc.)

A total of 117 species of freshwater decapod crustaceans are known from Brazil. Knowledge regarding the fauna of Decapoda from inland waters in the state of Bahia, northeast Brazil, is incipient. In spite of its wide territory and rich hydrographic net, only 13 species of limnetic decapods have been reported from that state. The objective of this contribution was to survey decapod crustaceans of some hydrographic basins in southeastern Bahia. The material described herein was obtained in samplings conducted between 1997 and 2005. Voucher specimens were deposited in the carcinological collections of the Museu de Zoologia, Universidade Estadual de Santa Cruz, Ilhéus, Brazil, and Departamento de Oceanografia, Universidade Federal de Pernambuco, Recife, Brazil. A total of 13 species was collected. The carideans were represented by the atyids Atya scabra (Leach, 1815) and Potimirim potimirim (Müller, 1881) and the palaemonids Macrobrachium acanthurus (Wiegmann, 1836), M. amazonicum (Heller, 1862), M. carcinus (Linnaeus, 1758), M. heterochirus (Wiegmann, 1836), M. jelskii (Miers, 1877), M. olfersi (Wiegmann, 1836), and Palaemon (Palaemon) pandaliformis (Stimpson, 1871). The brachyurans were represented by the portunids Callinectes bocourti A. Milne-Edwards, 1879 and C. sapidus Rathbun, 1895, the trichodactylid Trichodactylus fluviatilis Latreille, 1828 and the panopeid Panopeus rugosus A. Milne-Edwards, 1881. Macrobrachium heterochirus represents a new record from Bahia, and M. amazonicum is reported for the first time in southeast Bahia. The occurrence of two extreme different forms of T. fluviatilis was observed. Form A is characterized by the frontal margin of carapace bordered by conspicuous granules, the anterolateral margin provided with developed teeth plus granules, and the posterolateral margin provided with granulation similar to that found on the front. In form B the frontal margin is smooth or has an inconspicuous granulation; the anterolateral margin is usually provided with 1-3 notches, and teeth (1-2), if present, are small; and the posterolateral margin is smooth or slightly granulated.