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This study investigated Passiflora edulis peel flour (PEPF) intake and its effect against high-fat diet-induced obesity. PEPF is a source of fiber and phenolic compounds, which can decrease oxidative stress and inflammatory cytokines, both linked to chronic inflammatory response and fat deposition in obesity. Therefore, we hypothesized that PEPF intake could decrease inflammatory cytokines and oxidative stress observed in obesity, leading to decrease of fatness and chronic inflammatory response. The aims of the study were to evaluate the lipid peroxidation, the expression of antioxidants enzymes, and inflammatory parameters in obese rats. Male Sprague-Dawley rats were divided into 3 groups (n = 8 per group) according to the diets: control (based on AIN-93G), high-fat (HF, 35% fat w/w), and HF with PEPF (HFPF), and the experiment lasted for 10 weeks. PEPF showed high dietary fiber content and bioactive compounds, such as ferulic acid, and β-carotene. PEPF intake was effective in reducing body weight gain (13.31%) and total body fat (22.58%). The lipid peroxidation in the liver and adipose tissue decreased in the HFPF group compared to HF-fed animals, whereas hepatic glutathione peroxidase and glutathione reductase activity and their expressions in the liver were higher in HFPF than HF. In addition, the PEPF intake decreased inflammatory cytokines in serum. These results suggest that PEPF intake decreases oxidative stress, possibly by the increase of antioxidant enzymes expression. Furthermore, PEPF decreases inflammatory response and protects from adiposity. Then, PEPF could act as an adjuvant to control of early parameters in obesity dysfunction., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Cognitive flexibility, the ability to adjust behavior in response to new and unexpected conditions in the environment, is essential for adaptation to new challenges and survival. The cholinergic system is an important modulator of this complex behavior however, the exact cholinergic circuits involved in this modulation and the precise influence of acetylcholine (ACh) in the process is still not fully understood. Here we review the role of different cholinergic circuits in cognitive flexibility. Strong evidence indicates that cholinergic interneurons (CINs) from the dorsomedial striatum are essential for facilitating the establishment of a new selected strategy; an effect that seems to depend mainly on activation of muscarinic receptors. Cholinergic neurons from the nucleus basalis magnocellularis (nBM), which project to the prefrontal cortex, seem to modulate the initial inhibition of a previously learned strategy, however, this concept is still controversial. Additionally, some studies suggest that basal forebrain cholinergic neurons projecting to the hippocampus, basolateral amygdala, and posterior parietal cortex may also participate on the modulation of cognitive flexibility. We highlight the fact that when investigating effects of ACh on behavioral flexibility, or any other behavior, one has to keep in mind two important particularities of the cholinergic system: (1) Many cholinergic neurons in the brain co-release glutamate or GABA with ACh. Methodologies that rely on neuronal silencing or ablation lead to simultaneous elimination of both neurotransmitters, making interpretation of results complex. (2) The cholinergic gene locus has a unique organization, with the vesicular acetylcholine transporter (VAChT) gene present within the intron between the first and second exons of the choline acetyltransferase (ChAT) gene. Thus, behavioral studies using transgenic animals generated with ChAT bacterial artificial chromosome (BAC) clones should be considered carefully, taking into consideration that these mice may overexpress VAChT and therefore, present a hypercholinergic tone that can be a confounder in behavioral studies., (Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.)

Passiflora alata Curtis (P. alata) leaves have anti-inflammatory properties; the present study aimed to investigate the anti-diabetogenic properties of P. alata aqueous leaf extract. HPLC analysis identified the phenolic compounds catechin, epicatechin and rutin. The aqueous extract was administered for 30weeks to non-obese diabetic (NOD) mice presenting a decrease of 28.6% in diabetes incidence and the number of inflammatory cells in pancreatic islets, when compared with the control group (water). The P. alata group presented an antioxidant effect and decreased lipid peroxidation in the serum of NOD mice. Increased numbers of insulin-positive cells were also observed in the pancreatic islets of the treated group. The diabetic group exhibited higher levels in the glucose tolerance test and glycemic index, in comparison to the P. alata-treated group and non-diabetic control BALB/c mice. In addition, the P. alata extract reduced the percentage and the proliferation index of NOD mice lymphocytes submitted to in vitro dose/response mitogenic stimulation assays. These results suggest that the aqueous extract of P. alata has anti-inflammatory properties, contributing to the protection of beta cells in pancreatic islets in NOD mice, and presents potential for use a supporting approach to treat type 1 diabetes., (Copyright © 2016. Published by Elsevier B.V.)

Optimization of the extraction methodology for antioxidant phenolic compounds in red grape jam was performed with an ultrasound-assisted system. The antioxidant phenolic compounds were extracted and analyzed by determining the total phenolic content (Folin Ciocalteu), as well as by employing free radical DPPH() and the beta-carotene/linoleic acid system. To optimize the parameters of solvent concentration, time and extraction temperature, the experiments were carried out using the central composite rotatable design (CCRD) method. Using response surface methodology (RSM), the best combinations achieved were with 60% ethanol and water for 20min at 50°C. The optimized parameters for this method were compared to an extraction method that has been commonly noted in the literature, which used to be the standard method, and the results were expressed in the milligram equivalent of quercetin per gram of jam (mg E.Q/g Jam). With the new method, the antioxidant potential measured by DPPH(ⁱ) was 70% higher than that obtained with the standard extraction method, and the antioxidant potential measured using the beta-carotene/linoleic acid system was 65% higher. In addition, a significant decrease in the total analysis time was achieved (from 10h to 30min), when compared to the standard method., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The key neural substrates involved in memory and cognitive tasks have been reported to receive important modulation from ovarian hormones. In fact, neurochemical systems associated with cognitive functions, such as the cholinergic system, are, at least in part, under modulation of estrogens. Here we show that vesicular acetylcholine transporter (VAChT) mutant mice, which express lower levels of the VAChT (VAChT KD) and reduced acetylcholine release, present sexual dimorphism on memory. We evaluate short- and long-term object recognition memories (STM and LTM) in both sexes. We have showed previously, and confirm here, that VAChT KDHET male mice present deficits in both STM and LTM object recognition memories in comparison with WT. In contrast, VAChT KDHET female mice present deficit in LTM, but not in STM. To test if the female hormones levels could be a determinant factor on sexual dimorphism observed, we submitted female mice to ovariectomy (OVX) or sham-surgery. After 1 week (1 w), we evaluate STM. Female hormone deprivation promotes STM impairment in VAChT KDHET, but not in WT female mice. Our results strongly suggest that the sexual dimorphism observed in VAChT KDHET mice on STM is due to modulation of cholinergic system by ovarian hormones., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Two cDNAs, isolated from a Trypanosoma cruzi amastigote library immunoscreened with sera from patients with Chagas disease, encode proteins with sequence homology to eukaryotic components of the cellular sorting and recycling machinery. These proteins, denominated TcAGL, present an N-terminal lectin domain and a C-terminal region containing repetitive amino acids and a poly-glutamine tract. They are products of polymorphic alleles of a single copy gene constitutively expressed during the parasite life cycle. Polyclonal antibodies obtained from mice immunized with the recombinant antigen recognize proteins with apparent molecular weight ranging from 95 to 120 kDa in cell lysates from all three life stages and in various strains of the parasite. Sera from Chagas disease patients recognize the recombinant antigen in ELISA and immunoprecipitation assays but not in Western blot assays under denaturing conditions. Consistent with its proposed role in the glycoprotein secreting pathway, immunofluorescence analyses and expression of a green fluorescent protein-tagged TcAGL protein indicate a sub-cellular localization in the vicinity of the flagellar pocket membrane and the Golgi complex of the parasite.

Several pharmacopoeias recommend the membrane filtration method for the sterility test of ophthalmic ointments. Isopropyl myristate, a fatty acid ester that exhibits high toxicity mainly against Gram-negative microorganisms, is indicated as a solvent for ointments. In this study, six fatty acid esters (diethyl adipate, diisopropyl adipate, ethyl laurate, ethyl myristate, methyl caprylate and isopropyl palmitate) were evaluated as solvents to replace isopropyl myristate in the sterility test for ophthalmic ointments. The logarithm of the partition coefficient (logP) of the fatty acid esters was calculated from the sum of the substituent hydrophobicity constants (pi) of the functional groups present in their molecules. The ability of the solvents to dissolve an ophthalmic ointment base was investigated. The D-value method was used to assess the antimicrobial activity of isopropyl palmitate, ethyl myristate, ethyl laurate and isopropyl myristate against Pseudomonas aeruginosa. Isopropyl palmitate was the least toxic solvent to this microorganism, since it had the highest D-value (171.1 min). No significant difference was observed between the D-values of ethyl myristate (89.4 min) and isopropyl myristate (92.5 min). Ethyl laurate exhibited the lowest D-value (27.2 min). Using gas chromatography coupled to mass spectrometry, other fatty acid esters were detected as the predominant impurities in the solvents, as well as acid contaminants in low or insignificant amounts.

The mechanism of action of volatile anesthetics is not completely understood. Calcium release from internal stores may alter signaling pathways that influence neurotransmission. Abnormalities of the regulation of intracellular calcium concentration ([Ca2+]i) from patients with malignant hyperthermia is a hallmark of this syndrome indicating the potential of these agents to interact with proteins involved in Ca2+ signaling. In the present study, a cholinergic cell line (SN56) was used to examine whether the release of calcium from intracellular stores occurs in the presence of sevoflurane. Changes in [Ca2+]i were measured using fluo-4, a fluorescent calcium sensitive dye and laser scanning confocal microscopy. Sevoflurane induced an increase on [Ca2+]i from SN56 cells. The sevoflurane-induced increase on [Ca2+]i remained even when the cells were perfused with medium lacking extracellular calcium. However, this effect was abolished by BAPTA-AM, a chelator of intracellular calcium, suggesting the involvement of intracellular Ca2+ stores. Using cyclopiazonic acid, an inhibitor of sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase, we investigated whether the depletion of intracellular Ca2+ stores interfered with the effect of sevoflurane. In the presence of this agent, sevoflurane caused a small but not significant rise on [Ca2+]i of the SN56 cells. Dantrolene, an inhibitor of ryanodine-sensitive calcium stores did not modify the sevoflurane increase on [Ca2+]i. Carbachol, a drug that releases Ca2+ from the IP3 pool, abolished the effect of sevoflurane. In addition, xestospongin D, a cell-permeant IP3 receptor antagonist, decreased significantly the sevoflurane increase on [Ca2+]i. Our data suggest that the sevoflurane-induced increase on [Ca2+]i from SN56 cells occurs through the release of calcium from IP3-sensitive calcium stores.

Protein kinase C (PKC) is a signal transducing enzyme that is an important regulator of multiple physiologic processes and a potential molecular target for volatile anaesthetic actions. However, the effects of these agents on PKC activity are not yet fully understood. Volatile anaesthetics increase intracellular calcium concentration ([Ca(2+)](i)) in a variety of cells, thus their effects on PKC activity may be indirect due to [Ca(2+)](i) increase. Alternatively, the anaesthetics could directly stimulate PKC activity. In order to distinguish these two possibilities in intact cells, we used a fully functional green fluorescent protein conjugated PKCbetaII (GFP-PKCbetaII) and confocal microscopy to evaluate the dynamic redistribution of PKC in living SN56 cells, a cholinergic cell line, in response to halothane. Halothane induced PKC translocation in SN56 cells transfected with GFP-PKCbetaII. This effect was not suppressed by dantrolene, a drug that blocks halothane-induced Ca(2+) release from intracellular stores in these cells. These findings indicate that halothane induces PKC translocation in SN56 cells independently of its ability to release calcium from internal stores., (Copyright 2002 Elsevier Science Inc.)

The modulation of neurotransmitter release by calcium channels is well established, yet, sodium channels were regarded mainly as charge carriers. Many lines of evidence suggest a more fine-tuning role played by sodium channels. Using rat cerebrocortical isolated nerve endings (synaptosomes) and two toxins that have separate sites of action on sodium channels and provoke distinct changes in channel kinetics, we were able to show that depending on the rate of increase in channel conductance, the outcome in terms of neurotransmitter release and calcium channel types coupled to that event are different. Mainly, our study focused on veratridine, an alkaloid from lilaceous plants that binds to sodium channel toxin site 2, and tityustoxin, a toxin purified from the venom of the Brazilian yellow scorpion Tityus serrulatus that binds to site 3. Veratridine induces a slower increase in intrasynaptosomal sodium and calcium concentrations, slower depolarization, delayed exocytosis and a slower and predominantly calcium-independent glutamate release, when compared to tityustoxin.Thus, we have used these two toxins to investigate the events that start with sodium entry and culminate with the release of glutamate in isolated nerve endings (synaptosomes) from rat cerebral cortex. With that in mind we measured intrasynaptosomal free sodium concentration [Na(+)](i), intrasynaptosomal free calcium concentration [Ca(2+)](i), membrane potential, exocytosis and glutamate release using fluorescent probes.

In this paper we describe the effects of the beta scorpion toxin Tityus gamma (TiTX gamma) and spider neurotoxins Tx3-3 and Tx3-4 in the (45)Ca(2+) uptake in synaptosomes. The TiTX gamma-stimulatory effect on (45)Ca(2+) uptake in synaptosomes was inhibited omega-Conotoxin MVIIC (omega-CgTX MVIIC) (0.1 microM) and omega-Agatoxin IVA (0.1 microM) by 70% and 41%, respectively. omega-CgTX MVIIC (1.0 microM) almost completely blocked the TiTX gamma-induced (45)Ca(2+) uptake in synaptosomes. Verapamil (1.0 microM) and omega-Conotoxin GVIA (0.1 microM) had no effect in the scorpion toxin-induced (45)Ca(2+) influx. The spider neurotoxins Tx3-3 and Tx3-4 inhibited the TiTX gamma-induced calcium uptake with an IC(50) of 10.0 and 30.0 nM, respectively. It is suggested that spider neurotoxins Tx3-3 and Tx3-4 blocking effect in the TiTX gamma-induced calcium uptake involves P/Q-type calcium channels.

Volatile general anaesthetics are believed to affect synaptic transmission, but their actions in the central nervous system (CNS) remains unclear. Acetylcholine (ACh) is one of the most important neurotransmitter in the CNS and thus, it is possible that its release could be one of the targets for volatile anaesthetic action. However, the effects of these agents on the release of ACh are not yet fully understood. Rat brain cortical slices were loaded with [(3)H]-choline in order to study the effect of isoflurane on the release of [(3)H]-ACh from this preparation. Isoflurane (28, 43, 54, 95 and 182 nM) significantly increased the basal release of [(3)H]-ACh. This effect was independent of the extracellular sodium and calcium concentration but was decreased by tetracaine and dantrolene, inhibitors of Ca(2+-)release from intracellular stores. These findings indicate that isoflurane may cause a Ca(2+-)release from internal stores that increases [(3)H]-ACh release in rat brain cortical slices.

Glutamate is the major excitatory neurotransmitter in the CNS. The recent characterization of glutamate as a neurotransmitter in the enteric nervous system opened a new line of investigation concerning the role of glutamate in that system. The present study aimed to further characterize the enteric glutamate release and the calcium channels coupled to it. For this study the myenteric plexus-longitudinal muscle of guinea-pig ileum was stimulated with potassium chloride or with electrical pulses. The released glutamate was detected by spectrofluorimetry. Laser scanning confocal microscopy was used for analysis of immunolabeled enteric tissue for co-localization studies of calcium channels (N- and P/Q-type) and glutamate transporters (EAAC1). Here we report the effects of known Ca(2+)-channel blockers on glutamate release evoked by KCl-depolarization or electrical stimulation in the myenteric plexus. We find that N-type Ca(2+) channels control a major portion of evoked glutamate release from this system, with a very small contribution from L-type Ca(2+) channels. Moreover, alpha(1A)-like (P-type Ca(2+) channel) and alpha(1B)-like (N-type Ca(2+ )channel) immunoreactivity co-localized with glutamate transporters in the myenteric plexus. In addition, KCl-evoked or electrically stimulated glutamate release was sensitive to omega-agatoxin IVA, in a frequency-dependent manner, suggesting that P-type channels are also coupled to the release of glutamate. We, thus, conclude that both N-type and P-type Ca(2+) channels control most of the evoked glutamate release from the enteric nervous system, as also occurs in some parts of the CNS.