Hydroxytakakiamide and Other Constituents from a Marine Sponge-Associated Fungus Aspergillus fischeri MMERU23, and Antinociceptive Activity of Ergosterol Acetate, Acetylaszonalenin and Helvolic Acid.

An unreported prenylated indole derivative hydroxytakakiamide (4) was isolated, together with the previously described ergosterol (1), ergosterol acetate (2), and (3R)-3-(1H-indol-3-ylmethyl)-3, 4-dihydro-1H-1,4-benzodiazepine-2,5-dione (3), from the column fractions of the crude ethyl acetate extract of the culture of a marine sponge-associated fungus, Aspergillus fischeri MMERU 23. The structure of 4 was elucidated by the interpretation of 1D and 2D NMR spectral data and high-resolution mass spectrum. The absolute configuration of the stereogenic carbon in 3 was proposed to be the same as those of the co-occurring congeners on the basis of their biogenetic consideration and was supported by the comparison of its sign of optical rotation with those of its steroisomers. The crude ethyl acetate extract and 2 were evaluated, together with acetylaszonalenin (5) and helvolic acid (6), which were previously isolated from the same extract, for the in vivo antinociceptive activity in the mice model. The crude ethyl acetate extract exhibited antinociceptive activity in the acetic acid-induced writhing and formalin tests, while 2, 5, and 6 displayed the effects in the late phase of the formalin test. On the other hand, neither the crude ethyl acetate extract nor 2, 5, and 6 affected the motor performance of mice in both open-field and rotarod tests. Additionally, docking studies of 2, 5, and 6 were performed with 5-lipoxygenase (5-LOX) and phosphodiesterase (PDE) enzymes, PDE4 and PDE7, which are directly related to pain and inflammatory processes. Molecular docking showed that 6 has low affinity energy to PDE4 and PDE7 targets while retaining high affinity to 5-LOX. On the other hand, while 2 did not display any hydrogen bond interactions in any of its complexes, it achieved overall better energy values than 6 on the three antinociceptive targets. On the other hand, 5 has the best energy profile of all the docked compounds and was able to reproduce the crystallographic interactions of the 5-LOX complex.

Antinociceptive and anti-inflammatory properties of α-D-mannan from the yeast Kluyveromyces marxianus: evidence for a role in interleukin-6 inhibition.

The management of inflammatory states typically involves non-steroidal anti-inflammatory drugs (NSAIDs) and opiates. Understanding the mechanisms underlying the processing of nociceptive information from potential alternatives such as some polysaccharides may enable new and meaningful therapeutic approaches. In this study, α-D-mannan isolated from the Kluyveromyces marxianus cell wall produced antinociceptive effects in models of inflammatory pain (formalin and complete Freund's adjuvant tests). Furthermore, α-D-mannan reduced paw edema and interleukin-6 (IL-6) production after carrageenan-induced inflammation. The polysaccharide α-D-mannan was characterized by gas chromatography-mass spectrometry, methylation analysis, and spectroscopic techniques. Moreover, the Doehlert experimental design was applied to find the optimal conditions for biomass production, with the best conditions being 10.8 g/L and 117 h for the glucose concentration and the fermentation time, respectively. These results indicate that α-D-mannan from K. marxianus exerts anti-inflammatory and antinociceptive effects in mice, possibly via a mechanism dependent on the inhibition of IL-6 production.

Unveiling Targets for Treating Postoperative Pain: The Role of the TNF-α/p38 MAPK/NF-κB/Nav1.8 and Nav1.9 Pathways in the Mouse Model of Incisional Pain.

Although the mouse model of incisional pain is broadly used, the mechanisms underlying plantar incision-induced nociception are not fully understood. This work investigates the role of Nav1.8 and Nav1.9 sodium channels in nociceptive sensitization following plantar incision in mice and the signaling pathway modulating these channels. A surgical incision was made in the plantar hind paw of male Swiss mice. Nociceptive thresholds were assessed by von Frey filaments. Gene expression of Nav1.8, Nav1.9, TNF-α, and COX-2 was evaluated by Real-Time PCR in dorsal root ganglia (DRG). Knockdown mice for Nav1.8 and Nav1.9 were produced by antisense oligodeoxynucleotides intrathecal treatments. Local levels of TNF-α and PGE2 were immunoenzymatically determined. Incised mice exhibited hypernociception and upregulated expression of Nav1.8 and Nav1.9 in DRG. Antisense oligodeoxynucleotides reduced hypernociception and downregulated Nav1.8 and Nav1.9. TNF-α and COX-2/PGE2 were upregulated in DRG and plantar skin. Inhibition of TNF-α and COX-2 reduced hypernociception, but only TNF-α inhibition downregulated Nav1.8 and Nav1.9. Antagonizing NF-κB and p38 mitogen-activated protein kinase (MAPK), but not ERK or JNK, reduced both hypernociception and hyperexpression of Nav1.8 and Nav1.9. This study proposes the contribution of the TNF-α/p38/NF-κB/Nav1.8 and Nav1.9 pathways to the pathophysiology of the mouse model of incisional pain.

Development of membranes based on carboxymethyl cellulose/acetylated arrowroot starch containing bromelain extract carried on nanoparticles and liposomes.

Polymeric membranes have been used in several applications, including their use as curatives in cutaneous wounds. Bromelain has long been used for anti-inflammatory purposes, so the objective of this work was to produce carboxymethylcellulose-acetylated blends, incorporate bromelain, characterize the systems, compare the blends with bromelain loaded in nanoparticles and liposomes and, finally, to evaluate their healing potential. Four membrane formulations were produced by solvent evaporation: the control, membranes containing free bromelain, bromelain-loaded nanoparticles (NPs) and bromelain-loaded liposomes (LIPs). The enzyme concentration was the same for all formulations. Transparent, flexible and intact films were obtained. The membranes containing free bromelain, bromelain-loaded NPs and bromelain-loaded LIPs had higher water content, lower water vapor permeability and maximum tensile strength, and greater elongation at rupture. The capacity to absorb simulated exudate was higher in samples containing free bromelain, and bioadhesion was reduced in the presence of free bromelain compared to the control. An in vivo assay was performed to verify the membranes' healing potential. Histological analysis revealed no edema on the 14th day in animals treated with membranes containing bromelain-loaded NPs and LIPs.

Antinociceptive and anti-inflammatory activity of α-d-mannan from Pseudozyma sp.

Pseudozyma sp. are yeasts that are commercially important due to their production of glycolipid biosurfactants, squalene, itaconic acid, and exopolysaccharide. The search for other analgesia inducing drugs, such as opiates and non-steroidal anti-inflammatory drugs (NSAIDs), as alternatives is beneficial. In this study, the antinociceptive and anti-inflammatory actions of α-d-mannan were studied using acetic acid-induced writhing, open field test, formalin test, and carrageenan-induced paw oedema tests in mice. The α-d-mannan obtained from Pseudozyma sp. was confirmed by methylation analysis, 1D and 2D NMR spectroscopic analysis, and GC-MS. The results show that α-d-mannan from Pseudozyma sp. has analgesic and anti-inflammatory activities. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02635-1.

Evaluation of Biological Activity of Polygala Boliviensis in Experimental Models.

The plants of the genus Polygala (Polygalaceae) are employed in folk medicine for the treatment of several pathologies, including disorders of the bowel and kidney, as anesthetic, expectorant and anti-inflammatory. The present study was undertaken to investigate the antiedematogenic and antinociceptive activities of methanolic extract of Polygala boliviensis A. W. Benn (MEPB) in mice. The antinociceptive activity of MEPB was evaluated using the writhing, formalin, and tail immersion tests. The carrageenan-induced paw edema test was used to assess the antiedematogenic activity of MEPB. Mice motor performance was evaluated in the rota rod and open field tests and the acute toxicity were evaluated over 14 days. High-performance liquid chromatography was used to determine the fingerprint chromatogram of MEPB. Oral administration of MEPB (75- 600 mg/kg) reduced the number of writhing induced by acetic acid. In the formalin test, the oral pre-treatment with MEPB (75 - 600 mg/kg) produced a dose-related inhibition only of the late phase. MEPB (300 and 600 mg/kg) reduced the carrageenan-induced paw edema. In contrast, the treatment with MEPB (300 and 600 mg/kg) did not prevent the thermal nociception in the tail immersion test. MEPB (600 mg/kg)-treated mice did not show any motor performance alterations. Over the study duration of 14 days, there were no mortality or toxic signs recorded in the group mice given 6000 mg/kg of MEPB. The present study demonstrated, for the first time, the antinociceptive and antiedematogenic properties of Polygala boliviensis.

Antinociceptive effect of lupeol: evidence for a role of cytokines inhibition.

The present study investigates the antinociceptive properties of lupeol in models of inflammatory and post-operative pain, as well as its mechanisms of action. The effects of lupeol were tested against acetic acid-induced writhing, formalin test, carrageenan-induced hyperalgesia, and post-operative pain model. Cytokine levels were determined by ELISA. Mice motor performance was evaluated in the rota rod and open-field tests. Pre-treatment of mice with lupeol (5-100 mg/kg IP) produced a dose-related inhibition of writhing in mice. The maximal antinociception produced by lupeol (60 mg/kg) was unaffected in mice pre-treated with yohimbine (α2 adrenoceptor antagonist; 2 mg/kg IP), L-arginine (substrate for nitric oxide synthase; 600 mg/kg IP), glibenclamide (the KATP-channel blocker; 2 mg/kg IP), and methysergide maleate (serotoninergic receptors antagonist; 5 mg/kg IP). Furthermore, lupeol (25-100 mg/kg) inhibited the late phase of formalin test. Pre-treatment with lupeol (50 and 100 mg/kg) inhibited the hyperalgesia and the local increase in tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) levels induced by carrageenan. In contrast, lupeol did not inhibit the post-operative pain. Lupeol-treated mice did not show any motor performance alterations or apparent systemic toxicity. Our results demonstrate that lupeol has consistent antinociceptive properties during inflammatory pain, but not post-operative pain, acting through the inhibition of IL-1ß and TNF-α production.

Antinociceptive properties of bergenin.

Bergenin (1) is a C-glucoside of 4-O-methylgallic acid with known antiarthritic activity attributed to modulation of cytokine production. The present study was undertaken to evaluate whether 1 has antinociceptive properties in models of inflammatory pain and to investigate its possible mechanisms of action. Pretreatment with 1 (12.5-100 mg/kg, ip) produced a dose-related inhibition of acetic acid-induced writhing in mice. Furthermore, treatment with 1 (50 and 100 mg/kg) inhibited both the early and late phases in a formalin test. In addition, 1 (50 and 100 mg/kg, ip) inhibited mechanical hyperalgesia, edema, and paw production of hyperalgesic cytokines (TNF-α and IL-1ß) induced by complete Freund's adjuvant. However, the local production of IL-10, an anti-inflammatory cytokine, was not altered by 1 (100 mg/kg, ip). Treatment with 1 produced a similar profile of antinociception in wild-type and IL-10-deficient mice. Mice treated with 1 did not show any motor performance alterations or apparent systemic toxicity. The results presented herein demonstrate that bergenin has consistent antinociceptive and anti-inflammatory properties, acting by the inhibition of IL-1ß and TNF-α production, and suggest its potential for the control of inflammatory pain.

Evaluation of laser phototherapy in the inflammatory process of the rat's TMJ induced by carrageenan.

AIM: The aim of this study was to evaluate, by light microscopy, the effects of laser phototherapy (LPT) at 780 nm or a combination of 660 and 790 nm, on the inflammatory process of the rat temporomandibular joint (TMJ) induced by carrageen. BACKGROUND: Temporomandibular disorders (TMDs) are frequent in the population and generally present an inflammatory component. Previous studies have evidenced positive effects of laser phototherapy on TMDs. However, its mechanism of action on the inflammation of the TMJ is not known yet. MATERIALS AND METHODS: Eighty-five Wistar rats were divided into 9 groups: G1, Saline; G2, Saline + LPT IR; G3, Saline + LPT IR + R; G4, Carrageenan; G5, Carrageenan + LPT IR; G6, Carrageenan + LPT IR + R; G7, previous LPT + Carrageenan; G8, previous LPT + carrageenan + LPT IR; and G9, previous LPT + carrageenan + LPT IR + R, and then subdivided in subgroups of 3 and 7 days. After animal death, specimens were taken, routinely cut and stained with HE, Sirius Red, and Toluidine Blue. Descriptive analysis of components of the TMJ was done. The synovial cell layers were counted. RESULTS: Injection of saline did not produced inflammatory reaction and the irradiated groups did not present differences compared to nonirradiated ones. After carrageenan injection, intense inflammatory infiltration and synovial cell layers proliferation were observed. The infrared irradiated group presented less inflammation and less synovial cell layers number compared to other groups. Previous laser irradiation did not improve the results. CONCLUSION: It was concluded that the LPT presented positive effects on inflammatory infiltration reduction and accelerated the inflammation process, mainly with IR laser irradiation. The number of synovial cell layers was reduced on irradiated group.

Mechanisms involved in the antinociceptive effects of 7-hydroxycoumarin.

7-Hydroxycoumarin (umbelliferone, 1), the main metabolite of coumarin, has been reported to produce potent antinociceptive effects in animal models of pain. However, the biochemical events involved in antinociception mediated by 1 are currently not well understood. In the present study, the mechanisms by which 1 exerts its pharmacological actions were investigated. Acute pretreatment of mice with 1 produced a long-lasting antinociceptive effect against complete Freund's adjuvant (CFA)-induced hyperalgesia. The subchronic administration of 1 inhibited CFA-induced hyperalgesia and paw edema, while it did not cause any apparent toxicity. Another set of experiments showed that 1 inhibited carrageenan-induced mechanical hyperalgesia, but not mechanical hyperalgesia induced by prostaglandin E(2) (PGE(2)), suggesting that it acts upstream of PGE(2.) Treatment with 1 was able to prevent the plantar tissue neutrophil influx induced by local inflammatory stimuli. In addition, 1 exhibited inhibitory effects on the release of hyperalgesic cytokines (TNF-α and IL-1ß) and the production of PGE(2), a directly acting hyperalgesic mediator. The present results suggest that the antinociceptive effect of 1 is correlated with the inhibition of neutrophil migration, cytokine release, and PGE(2) production and are supportive of the further investigation of the therapeutic potential of 1 to control inflammatory pain.

Direct blockade of inflammatory hypernociception by peripheral A1 adenosine receptors: involvement of the NO/cGMP/PKG/KATP signaling pathway.

Through activation of the A1 adenosine receptors (A1Rs) at both the central and peripheral level, adenosine produces antinociception in a wide range of tests. However, the mechanisms involved in the peripheral effect are still not fully understood. Therefore, the mechanisms by which peripheral activation of A1Rs reduces inflammatory hypernociception (a decrease in the nociceptive threshold) were addressed in the present study. Immunofluorescence of rat dorsal root ganglion revealed significant expression of A1Rs in primary sensory neurons associated with nociceptive pathways. Functionally, peripheral activation of A1Rs reduced inflammatory hypernociception because intraplantar (i.pl.) administration of an A1R antagonist (DPCPX) enhanced carrageenan-induced hypernociception. On the other hand, local (paw) administration of CPA (a selective A1R agonist) reversed mechanical hypernociception induced by carrageenan or by the directly acting hypernociceptive mediator prostaglandin E(2) (PGE(2)). Down-regulation of A1Rs expression in primary nociceptive neurons by intrathecal treatment with antisense oligodeoxinucleotides significantly reduced peripheral antinociceptive action of CPA. Direct blockade of PGE(2) inflammatory hypernociception by the activation of A1Rs depends on the nitric oxide/cGMP/Protein Kinase G/KATP signaling pathway because the peripheral antinociceptive effect of CPA was prevented by pretreatment with inhibitors of neuronal nitric oxide synthase (N-propyl-l-arginine), guanylyl cyclase (ODQ), and Protein Kinase G (KT5823) as well as with a KATP blocker (glibenclamide). However, this effect of CPA was not reduced by naloxone, excluding the participation of endogenous opioids. These results suggest that the peripheral activation of A1R plays a role in the regulation of inflammatory hypernociception by a mechanism that involves the NO/cGMP/PKG/KATP intracellular signaling pathway.

Adenomatoid odontogenic tumor: an unusual case exhibiting cribriform aspect.

The occurrence of areas exhibiting cribriform morphology in odontogenic tumors is not a very frequent finding. This report describes the case of a 22-year-old man who presented with swelling in the left anterior maxilla. An excisional biopsy was performed under local anesthesia. Histopathologically, the lesion consisted of a cystic wall exhibiting an odontogenic epithelium lining composed of oval, angular, and elongated cells forming a cribriform pattern. No recurrence has been detected after 2 years.

Hipotireoidismo altera secreção e composição salivar em ratos sépticos / Hypothyroidism alters salivary secretion and composition in septic rats

Objetivo: Este estudo investigou os efeitos do hipotireoidismo na secreção salivar total, nas concentrações de proteína e de eletrólitos (Na+, K+ e Ca+2) da saliva estimulada de ratos sépticos. Metodologia: Foram utilizados ratos Wistar (200-280g) separados em dois grupos: tireoidectomizados (TX) e controle (N). Os ratos receberam injeção intraperitoneal de lipopolissacarídeo (LPS), 250 μg/100g de peso corporal (p.c.), ou salina (SL), 90 min antes da estimulação salivar com pilocarpina (5mg/kg de p.c.) para coleta de fluxo salivar total por 15 min (μL/min/100g de p.c.). A concentração de proteínas totais foi analisada pelo método de Lorwy e as concentrações de eletrólitos através de reagente padrão (Doles) para fotômetro de chama (Na+, K+) e teste colorimétrico (Ca+2). Resultados: O hipotireoidismo e a endotoxemia induzida pelo LPS diminuíram o fluxo salivar (Teste de Mann-Whitney, P < 0,05). A sepse reduziu significativamente a concentração de proteínas totais salivar de ratos TX e a concentração de íon sódio na saliva de ratos N. Não houve alteração nas concentrações de íon potássio em nenhum dos grupos, mas o hipotireoidismo provocou aumento da concentração de íon cálcio na saliva. Conclusão: A secreção salivar e a composição da saliva são alteradas pelo hipotireoidismo em ratos sépticos.

Purpose: We investigated the effects of hypothyroidism on salivary flow rate, total protein and electrolytes concentrations (Na+, K+ and Ca+2) in septic rats. Methods: Male Wistar rats (200-280g) were thyroidectomized (TX) or sham operated (N) and treated with lypopolysaccharide (LPS) (250 μg/100g b.w.; i.p.) or saline (SL), 90 min before salivary stimulation with pilocarpine (5mg/kg; i.p.). Saliva was collected over 15 min (μL/min/100g of b.w.). Total proteins were analyzed by Lorwy method and electrolytes concentrations were determined by flame fotometry (Na+, K+) and colorimetric test (Ca+2).Results: Hypothyroidism and sepsis by LPS injection caused a decrease in salivary flow rates (Mann-Whitney Test, P < 0.05). LPS injection decreased salivary total protein concentration in TX rats and sodium concentration in N rats. Sepsis and thyroid hormones do not participate in the molecular mechanism of potassium transport, but hypothyroidism increased calcium concentration in saliva. Conclusion: Salivary flow rate and composition are modified by hypothyroidism in rats with sepsis.