Echinodorus macrophyllus: Acute toxicological evaluation of hydroxycinnamoyl derivatives from SF1 subfractions.

ETHNOPHARMACOLOGICAL RELEVANCE: Echinodorus macrophyllus (Kunth.) Micheli (Alismataceae), known as chapéu-de-couro in Brazil, is popularly used to treat inflammatory diseases. We have previously demonstrated a significant reduction in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20) and described that hydroxycinnamoyl derivatives present in SF1 (Fr20 subfraction) showed higher anti-inflammatory properties by mechanisms that include a reduction of TNF-α, IL-1ß, CKCL1/KC, LTB4, and PGE2 levels in exudate. AIM OF THE STUDY: This work describes the acute toxicological effect of SF1 subfraction on SW mice treated orally for five days in the air pouch model by evaluating the hematological and biochemical determinations on the blood samples; the relative organ weight and its histopathological analysis; the liver genotoxicity assessment and the activity of liver enzymes from xenobiotic metabolism. MATERIALS AND METHODS: Fr20 was earlier fractionated on the Sephadex LH-20 column, yielding mainly four subfractions, including SF1. The SF1 toxicity was evaluated in mice challenged with carrageenan on the air pouch inflammation model and orally treated for five days. The body weight was monitored daily, and the organs were weighed after the euthanasia. Hematological and biochemical determinations were carried out using specific commercial kits and following the protocols provided by the manufacturers. The organs were fixed, sectioned, processed for hematoxylin and eosin staining, and analyzed by light microscopy. Genotoxicity assessment was performed by the alkaline single-cell gel electrophoresis. Livers were processed for ethoxyresorufin-O-deethylase (EROD) and Glutathione S-transferase (GST) assays. RESULTS: SF1 exhibited low toxicity, as no significant discrepancy was observed in the relative weight of the body organs of mice. Moreover, the daily treatment with SF1 did not alter the number and percentage of red blood cells or hemoglobin concentration in the blood. The treatment with SF1 did not affect the creatinine concentration, but the 25 mg/kg dose reduced the plasma urea level and uric acid, suggesting its use in treating acute renal failure. The parameters analyzed did not present biochemical alterations indicative of liver disease. Regarding serum triglyceride and cholesterol levels, a significant decrease was detected in both parameters in mice treated with SF1. In addition, the histopathological analysis showed that inflammatory focus in the livers seemed more relevant in the control groups than in those treated. There were no significant changes in the renal or splenic tissues of animals treated with SF1. Treatment with SF1 also does not have a genotoxic effect on liver cells. CONCLUSION: Treatment with SF1 showed no toxicity in mice at doses equivalent to those recommended for humans, which provides evidence of the safety of the therapeutic use of this subfraction.

LQB-118 Suppresses Migration and Invasion of Prostate Cancer Cells by Modulating the Akt/GSK3ß Pathway and MMP-9/Reck Gene Expression.

BACKGROUND/AIM: Prostate cancer (PCa) is one of the most common malignancies in adult men. LQB-118 is a pterocarpanquinone with antitumor activity toward prostate cancer cells. It inhibits cell proliferation by down-regulating cyclins D1 and B1 and up-regulating p21. However, the effects of LQB-118 on PCa cell migration are still unclear. Herein, the LQB-118 effects on PCa metastatic cell migration/invasion and its mechanism of action were evaluated. MATERIALS AND METHODS: PC3 cells were treated with LQB-118 or Paclitaxel (PTX), and cell migration (wound healing and Boyden chamber assays) and invasion (matrigel assay) were determined. The LQB-118 mechanisms were evaluated by αVßIII protein expression (flow cytometry), protein phosphorylation (Western blot), and mRNA expression (qPCR). RESULTS: LQB-118 impaired PCa cell migration and invasion, down-regulated Akt phosphorylation, and also reduced GSK3ß phosphorylation, through a FAK-independent pathway. Also, it was observed that LQB-118 controlled the invasiveness behavior by reducing matrix metalloproteinase-9 (MMP-9) and up-regulating reversion-inducing cysteine rich protein with Kazal motifs (Reck) mRNA levels. Interestingly, LQB-118 increased integrin αvßIII expression, but this effect was not related to its activation, since the cell adhesion ability was reduced after LQB-118 treatment. CONCLUSION: These data highlight novel LQB-118 mechanisms in prostate cancer cells. LQB-118 acts as a negative regulator of the Akt/GSK3 signaling pathway and can modulate PCa cell proliferation, death, and migration/invasion. The results also support the use of LQB-118 for the treatment of metastatic PCa, alone or combined with another chemotherapeutic agent, due to its demonstrated pleiotropic activities.

Toxicogenetic assessment of a pre-workout supplement: In vitro mutagenicity, cytotoxicity, genotoxicity and glutathione determination in liver cell lines and in silico ADMET approaches.

The benefits of practicing physical activity, such as weight loss and control, are commonly associated with caloric restriction diets and may be improved by the ingestion of thermogenic and ergogenic supplements. However, there is a lack of safety data on commonly marketed nutritional supplements. Therefore, this investigation aims to evaluate a pre-workout supplement for mutagenicity using the Ames test, hepatocytoxicity in HepG2 and F C3H cells after 24 h, 48 h and 72 h, genotoxicity using the CBMN assay, determination of gluthatione activity and computational prediction of the three major isolated compounds present in the supplement. The mutagenicity test showed a mutagenic response in TA98 His+ revertants of 5 mg/plate in the presence of metabolic activation, cytotoxicity in TA98 of 5 mg/plate in the absence of metabolic conditions, and in TA102 of 0.5 mg/plate both in the presence and absence of metabolic activation. In our in vitro eukaryotic cell viability, WST-1, LDH and alkaline phosphatase assays, the supplement showed hepatocytotoxicity both dose-dependently and time-dependently. In the cytokinesis blocking micronuclei assay, the supplement induced micronuclei, nuclear buds, nucleoplasmatic, bridge formation, and a decreased in nuclear division. In addition, the supplement decreased intra and extracellular GSH. Computational analysis showed that the three isolated compounds most present in the supplement have the potential to cause hepatotoxicity. In the present investigation, the pre-workout supplement induced mutagenic, genotoxic, and cytotoxic responses and GSH decrease. Thus, considering food safety and public health sanitary vigilance, the consumption of this pre-workout supplement may harm the health of its consumers.

Echinodorus macrophyllus fraction with a high level of flavonoid inhibits peripheral and central mechanisms of nociception.

BACKGROUND AND AIM: Echinodorus macrophyllus (Kunth.) Micheli is popularly used for acute and chronic inflammatory conditions. The anti-inflammatory activity was previously demonstrated for its flavonoid-enriched fractions. The aim of this work assessed the antinociceptive properties of both aqueous extract and its fractions. EXPERIMENTAL PROCEDURE: The antinociceptive activity was determined by acetic acid-induced writhing, formalin test, tail immersion test, hot-plate test, xylene-induced ear edema methods, and the evaluation of its mechanism was performed in the writhing model. The aqueous extract of Echinodorus macrophyllus (AEEm) was fractionated, yielding Fr20, and Fr40. Fr40 composition was determined by HPLC-DAD-ESI-MS. RESULTS AND CONCLUSION: Fr20 (all doses) and Fr40 (100 mg/kg) reduced the nociception in the tail-flick model. Both fractions increased the percentage of maximum possible effect with 25 mg/kg, in the hot-plate assay, at 60 min, while AEEm reduced pain only with 50 and 100 mg/kg. There was a reduction in xylene-edema index, with Fr40 (25 mg/kg), AEEm (50 mg/kg) and Fr20 (50 mg/kg). All doses of AEEm, Fr20, and Fr40 reduced both phases of the formalin model. In the abdominal contortion model, Fr40 presented the highest activity, reducing 96% of contortions and its antinociceptive mechanism was evaluated. The results indicated the involvement of NO and adrenergic activation pathways. The main components of Fr40 are swertisin, swertiajaponin, isoorientin 7,3'-dimethyl ether, swertisin-O-rhamnoside, isoorientin, isovitexin, isovitexin-Orhamnoside, and isovitexin-7-O-glucoside. The aqueous extract of E. macrophyllus leaves and its fractions exhibited significant analgesic effect, mediated through both peripheral and central mechanisms being considered a potentially antinociceptive drug.

Echinodorus macrophyllus: Hydroxycinnamoyl derivatives reduces neutrophil migration through modulation of cytokines, chemokines, and prostaglandin in the air-pouch model.

ETHNOPHARMACOLOGICAL RELEVANCE: In Brazil, Echinodorus macrophyllus (Alismataceae), popularly known as chapéu-de-couro, is used to treat inflammatory diseases. Previous studies have shown a significant decrease in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20). AIM OF THE STUDY: This work fractionated Fr20, identified the fraction and substances responsible for the in vivo anti-inflammatory property, and demonstrated important immunomodulatory mechanisms of action. MATERIALS AND METHODS: Fr20 was fractionated using Sephadex LH-20, and the most active fraction was chromatographically analyzed (HPLC-DAD and UPLC-ESI-TOF-MS). Leukotriene B4, Prostaglandin E2, and cytokines were determined by the enzyme-linked immunosorbent assay and in vivo acute inflammation by the air pouch model. RESULTS: The subfractions SF1, SF3, and mainly the SF4 decreased NO levels (p < 0.05). SF3 and SF4 showed high DPPH scavenger activity. SF1 was more effective than SF4 in reducing vasodilation, redness, and leukocyte migration into the 4-h air pouch. SF1 inhibited 90.5% (100 mg/kg) and SF4 54.0% (50 mg/kg), mainly affecting the number of neutrophils. SF1 and SF4 reduced the protein level in the exudate. SF1 was also more effective in inhibiting neutrophil migration in a transwell assay (46.3%) and reduced (86.1%) the Leukotriene B4 level in the exudate. After five days of treatment, some SF1 anti-inflammatory mechanisms were evaluated in the air pouch's 24 h exudate and tissue. Despite the high level of inflammation of the control group in this condition, SF1 confirmed the decrease in the protein level and neutrophils migration into the pouch. It decreased the number of bone marrow cells, indicating a systemic effect of SF1. SF1 also decreased TNF-α (87%), IL-1ß (77%), CKCL1/KC (71.3%), and PGE2 (97.8%) and increased IL-10 (74.1%) levels in the air pouch exudate. Phytochemical analysis of SF1 indicates mainly hydroxycinnamoyl derivatives. CONCLUSION: Hydroxycinnamoyl derivatives present in SF1 are related to the crucial anti-inflammatory mechanisms of E. macrophyllus, decreasing the levels of TNF-α, IL-1ß, CKCL1/KC, LTB4, and PGE2 on the exudate. These results explain the reduction of vasodilatation, erythema, and neutrophil migration into the air pouch model, confirming this plant's anti-inflammatory potential.

Anti-inflammatory potential of invasive sun corals (Scleractinia: Tubastraea spp.) from Brazil: alternative use for management?

OBJECTIVES: The objective was to analyse the anti-inflammatory potential of the invasive coral species Tubastraea coccinea and Tubastraea tagusensis. METHODS: Methanolic extracts, fractions and synthesized compounds were evaluated for their anti-inflammatory ability, and their composition was elucidated through chemical analysis. KEY FINDINGS: The genus Tubastraea (Order Scleractinia, Family Dendrophylliidae) (known as sun corals) presents compounds with pharmacological value. The introduction of these azooxanthellate hard corals into Brazil, initially in Rio de Janeiro state, occurred through their fouling of oil and gas platforms from the Campos oil Basin. The two invasive species have successfully expanded along the Brazilian coast and threaten endemic species and biodiversity. The HPLC-MS and GC-MS data suggest the presence of aplysinopsin analogues (alkaloids). Anti-inflammatory activity was observed in all samples tested in in-vivo assays, especially in T. coccinea. The ethyl acetate fraction from this sample was more effective in in-vitro assays for anti-inflammatory activity. Depending on the concentration, this fraction showed cytotoxic responses. CONCLUSIONS: These species have potential pharmacological use, and considering their invasive nature, this study presents a potential alternative use, which may enhance the management of this biological invasion.

Anti-inflammatory effect of diterpenes-enriched fractions from Pterodon polygalaeflorus through inhibition of macrophage migration and cytokine production.

OBJECTIVES: To evaluate the anti-inflammatory potential of Pterodon polygalaeflorus hexane extract (HE) and its fractions on macrophage migration in vitro and in vivo. METHODS: Hexane extract from P. polygalaeflorus fruits was fractionated and yielded four fractions. RAW 264.7 cells were treated with samples to evaluate cell viability (MTT assay), cell migration (wound healing and transwell assays), CD14 expression (flow cytometry), iNOS and cytokine mRNA expression (RT-qPCR), NO (Griess reaction) and cytokine (ELISA) production. In vivo migration was evaluated on the thioglycollate-induced peritonitis model. Qualitative analysis was performed by GC-MS. KEY FINDINGS: All fractions inhibited the NO production by LPS-stimulated RAW 264.7 cells. Fr3 and Fr4 presented the lowest IC50 values. The expressions of iNOS and IL-1ß, TNF-α and IL-10 cytokines were inhibited by Fr3 and Fr4, whereas the CD14 expression was only inhibited by Fr3. All the samples inhibited RAW 264.7 migration in the wound healing and transwell assays. Fr3 and Fr4 reduced the migration of Mac-1+ Gr-1- cells to the peritoneum and presented in their compositions: 6α-hydroxy-7ß-acetoxyvouacapan-17ß-oate, methyl 6α,7ß-dihydroxyvouacapan-17ß-oate, methyl 6α-acetoxy-7ß-hydroxyvouacapan-17ß-oate, geranylgeraniol and 14,15-epoxy-geranylgeraniol. CONCLUSIONS: The anti-inflammatory effects of Fr3 and Fr4 involve inhibition of cell migration, iNOS expression and NO production, cytokine expression (mRNA and proteins) and CD14 expression (Fr3).

Flavonoid-enriched fraction from Echinodorus macrophyllus aqueous extract exhibits high in-vitro and in-vivo anti-inflammatory activity.

OBJECTIVES: Echinodorus macrophyllus (Kunth) Micheli (Alismataceae) is popularly used as an infusion to treat inflammatory diseases. This work fractionated the aqueous extract of E. macrophyllus (AEEm) to improve its anti-inflammatory effects. METHODS: Aqueous extract of E. macrophyllus was fractionated by Sephadex LH-20 and analysed by HPLC-DAD. Anti-inflammatory action was evaluated, in vivo, by air pouch model (total leucocyte, protein and leukotriene B4 (LTB4 )), and, in vitro, by neutrophil migration (transwell assay) and its Mac1 expression (flow cytometry), and RAW 264.7 nitric oxide (NO) production (Griess reaction). KEY FINDINGS: Fr20 reduced total leucocyte at 2.5 mg/kg (29.7%) while ethanolic extract of E. macrophyllus (EAEm) increased it (94.0%). Fr20 showed higher (P < 0.05) inhibition (89.8%) of LTB4 in exudate than EAEm (75.0%). Fr20 and EAEm decreased exudate protein and inflammatory infiltrate in pouch tissues, in-vitro neutrophil migration, and NO production. Otherwise, Fr40 did not reduce leucocytes and exudate protein (until 50 mg/kg) nor tissue inflammation, and increased in-vitro NO production. The inhibition of neutrophil migration by EAEm, but not Fr20, was dependent on reduced Mac-1 expression. CONCLUSIONS: The fractionation of AEEm provided a more potent anti-inflammatory fraction containing flavonoids (Fr20) that reduces the migration of neutrophils and LTB4 release, probably contributing to its mechanism of action.

Preclinical Studies Evaluating Subacute Toxicity and Therapeutic Efficacy of LQB-118 in Experimental Visceral Leishmaniasis.

Visceral leishmaniasis (VL) is the most severe form of leishmaniasis and is the second major cause of death by parasites, after malaria. The arsenal of drugs against leishmaniasis is small, and each has a disadvantage in terms of toxicity, efficacy, price, or treatment regimen. Our group has focused on studying new drug candidates as alternatives to current treatments. The pterocarpanquinone LQB-118 was designed and synthesized based on molecular hybridization, and it exhibited antiprotozoal and anti-leukemic cell line activities. Our previous work demonstrated that LQB-118 was an effective treatment for experimental cutaneous leishmaniasis. In this study, we observed that treatment with 10 mg/kg of body weight/day LQB-118 orally inhibited the development of hepatosplenomegaly with a 99% reduction in parasite load. An in vivo toxicological analysis showed no change in the clinical, biochemical, or hematological parameters. Histologically, all of the analyzed organs were normal, with the exception of the liver, where focal points of necrosis with leukocytic infiltration were observed at treatment doses 5 times higher than the therapeutic dose; however, these changes were not accompanied by an increase in transaminases. Our findings indicate that LQB-118 is effective at treating different clinical forms of leishmaniasis and presents no relevant signs of toxicity at therapeutic doses; thus, this framework is demonstrated suitable for developing promising drug candidates for the oral treatment of leishmaniasis.

Terpenic fraction of Pterodon pubescens inhibits nuclear factor kappa B and extracellular signal-regulated protein kinase 1/2 activation and deregulates gene expression in leukemia cells.

BACKGROUND: Plant derived compounds have been shown to be important sources of several anti-cancer agents. As cell cycle deregulation and tumor growth are intimately linked, the discovery of new substances targeting events in this biochemical pathway would be of great value. The anti-leukemic effect of an ethanolic extract of Pterodon pubescens seeds (EEPp) has been previously demonstrated and now we show that a terpenic subfraction (SF5) of EEPp containing farnesol, geranylgeraniol and vouacapan derivatives induces apoptosis in the human chronic myelogenous leukemia cell line K562. This work addresses SF5's antiproliferative mechanisms in these cells since they are still unclear. METHODS: DNA synthesis in K562 cells was assessed by [3H]-methyl-thymidine incorporation and cell cycle status by flow cytometry. The expression of cyclins D1 and E2, of the cell cycle inhibitor p21 and of the proto-oncogene c-myc was evaluated by semi-quantitative RT-PCR. Extracellular-signal-regulated kinases (ERK) 1/2 and nuclear factor kappa B (NF-κB) activation was evaluated by western blotting. RESULTS: In K562 cells, SF5 treatment induced a higher inhibition of DNA synthesis and cell growth than the original EEPp hexanic fraction from which SF5 originated, and also arrested the cell cycle in G1. Exposure of these cells to SF5 led to a decrease in cyclin E2 and c-myc expression while p21 mRNA levels were increased. Furthermore, SF5 inhibited the activation of mitogen-activated protein kinase (MAPK) ERK 1/2 and NF-κB. CONCLUSIONS: This work suggests that the anti-leukemic action of SF5 is linked to the inhibition of ERKs, NF-κB and c-myc signaling pathways resulting in reduced cyclin E2 mRNA expression and cell cycle arrest in the G1 phase.

Heme-induced ROS in Trypanosoma cruzi activates CaMKII-like that triggers epimastigote proliferation. One helpful effect of ROS.

Heme is a ubiquitous molecule that has a number of physiological roles. The toxic effects of this molecule have been demonstrated in various models, based on both its pro-oxidant nature and through a detergent mechanism. It is estimated that about 10 mM of heme is released during blood digestion in the blood-sucking bug's midgut. The parasite Trypanosoma cruzi, the agent of Chagas' disease, proliferates in the midgut of the insect vector; however, heme metabolism in trypanosomatids remains to be elucidated. Here we provide a mechanistic explanation for the proliferative effects of heme on trypanosomatids. Heme, but not other porphyrins, induced T. cruzi proliferation, and this phenomenon was accompanied by a marked increase in reactive oxygen species (ROS) formation in epimastigotes when monitored by ROS-sensitive fluorescent probes. Heme-induced ROS production was time- and concentration-dependent. In addition, lipid peroxidation and the formation of 4-hydroxy-2-nonenal (4-HNE) adducts with parasite proteins were increased in epimastigotes in the presence of heme. Conversely, the antioxidants urate and GSH reversed the heme-induced ROS. Urate also decreased parasite proliferation. Among several protein kinase inhibitors tested only specific inhibitors of CaMKII, KN93 and Myr-AIP, were able to abolish heme-induced ROS formation in epimastigotes leading to parasite growth impairment. Taken together, these data provide new insight into T. cruzi- insect vector interactions: heme, a molecule from the blood digestion, triggers epimastigote proliferation through a redox-sensitive signalling mechanism.

The heme uptake process in Trypanosoma cruzi epimastigotes is inhibited by heme analogues and by inhibitors of ABC transporters.

Heme (iron protoporphyrin IX) is an important molecule involved in many biological reactions, including oxygen transport, respiration, photosynthesis and drug detoxification. Trypanosoma cruzi parasites, the etiological agent of Chagas' disease, take up heme from the environment to supply their nutritional needs because they do not synthesize this cofactor. However, the mechanisms involved in heme transport across biological membranes are poorly understood. Indeed, in T. cruzi, no heme transporter has yet been characterized. In the present work, we evaluate the heme uptake processes by T. cruzi epimastigotes using fluorescent heme-analogues. Heme uptake decreased significantly when cells were pretreated with different concentrations of SnPPIX, PdMPIX or ZnMPIX, this observed competition suggests that they are taken up by the same transport system. We studied the growth behavior of epimastigotes using the same heme-analogues and the treatments with SnPPIX or PdMPIX impaired cell growth but when heme was added to the culture medium the observed inhibition was partially reversed. In addition, we tested how the heme uptake processes are affected by the presence of different transporter inhibitors. When the cells were treated with inhibitors and then incubated with heme, heme uptake decreased significantly for all treatments. These results constitute a strong indication for the existence of a protein associated with porphyrin transport in T. cruzi, possibly ATP-binding cassette transporters (ABC-transporter).

Terpenic subfraction of Pterodon pubescens induces apoptosis of K562 leukemic cells by modulating gene expression.

Deregulation of cell proliferation and apoptosis is linked to malignant cell development. Leukemia is the most frequent cancer in children, and plants are important sources for new potential anti-cancer agents. Although anti-tumoral effects have been shown for Pterodon pubescens extracts, the mechanisms are still obscure. This study describes in Pterodon pubescens a furane diterpene only reported in Pterodon polygalaeflorus, the methyl-6α-acetoxy-7ß-hydroxyvouacapan-17ß-oate, indicated by HRMS and 13C-NMR analysis, and demonstrates some mechanisms of the anti-leukemia action of its terpene subfraction SF5. SF5 induced cytotoxic and anti-proliferative effects on K562 cells. Increased sub-G1 nuclei and Annexin V+-FITC cells confirmed apoptosis of leukemic cells by treatment of these cells with SF5. Down-regulation of DNMT1 gene transcription and over-expression of Apaf-1 mRNA suggested that SF5 may be inducing apoptosis of K562 cells by epigenetic up-regulation of pro-apoptotic proteins involved in the mitochondrial intrinsic pathway.

Pterodon pubescens seed extract induces the cell cycle arrest of leukemic cells by deregulating cyclin D1 and E2 mRNA levels.

Plant-derived compounds are important sources of effective anti-cancer agents. Pterodon pubescens is a native Brazilian plant popularly known for its anti-inflammatory and anti-arthritic effects. The ethanolic extract of its seeds (EEPp) is a viscous, brown and fragrant oil containing geranylgeraniol, farnesol, naphthalene, dimethyldodecatrienol and vouacapan diterpene derivatives, in addition to other compounds. This study investigated the in vitro anti-leukemic properties of EEPp using the resistant human leukemia cell line K562. The EEPp anti-proliferative effect was demonstrated by the inhibition of DNA synthesis and cell growth, and the induction of cell cycle arrest in the G(1) phase. Furthermore, cyclin E2 mRNA levels were down-regulated, while those of cyclin D1 were up-regulated. An EEPp anti-leukemic effect may have also triggered apoptosis, as it increased the number of shrunken cells and phosphatidylserine cell membrane exposure. These observations suggest that EEPp deregulates cyclin D1 and E2 expression, inducing cell cycle arrest and apoptosis of leukemic cells.

Antiplatelet activity of geranylgeraniol isolated from Pterodon pubescens fruit oil is mediated by inhibition of cyclooxygenase-1.

Geranylgeraniol is a natural isoprenoid with anti-inflammatory properties extracted from the Pterodon pubescens Benth. fruit oil (PpO). In this work, the antiplatelet effect of both PpO and geranylgeraniol is investigated. ADP-, thrombin- and arachidonic acid (AA)-induced aggregation in human and rabbit platelets showed a prime involvement of PpO and geranylgeraniol in the arachidonic acid cascade. The lack of any significant inhibition of platelet aggregation induced by U-46 619 and thrombin, associated with PpO and geranylgeraniol suppression of prostaglandin E(2) and thromboxane A(2) formation demonstrate, for the first time, the involvement of geranylgeraniol in the AA metabolisation by inhibiting the cyclooxygenase enzyme.

Immunosuppressive effects of Echinodorus macrophyllus aqueous extract.

Echinodorus macrophyllus is a medicinal plant, popularly known in Brazil as "chapéu de couro", used to treat rheumatic diseases, which are usually characterized by exacerbated T and B lymphocyte response. We have evaluated the effects of the aqueous extract of Echinodorus macrophyllus (AEEm) on these cell functions, proliferation, and nitric oxide production. Mice treated orally for 7 days with AEEm had inhibited B cell antibody production (0.5mg/kg b.w.) and delayed type hypersensitivity (0.5 and 5mg/kg b.w.) mediated by T cells, reducing subcutaneous tissue leukocyte infiltration. AEEm inhibited, in vitro, NO production by stimulating J774 cells in a dose-dependent manner, with no cytotoxicity. We have demonstrated, for the first time, its immunosuppressive effect. This immunosuppressive effect supports a potential therapeutic use of AEEm to control exacerbated humoral and/or cellular immune response, as in autoimmune rheumatic diseases. However, it is important to be cautious about its indiscriminate popular use to avoid side effects, mainly in immunodeficiency diseases.

Marcadores séricos de cartilagem na artrite reumatóide avançada / Serum cartilage biomarkers in late rheumatoid arthritis

O objetivo deste trabalho foi avaliar o valor clínico de dois novos marcadores séricos de cartilagem em pacientes com artrite reumatóide crônica. Cento e quarenta pacientes foram seguidos por um período médio de 12 anos e avaliados para a presença de glicoproteína-39 de cartilagem humana e para matrix de proteína oligomérica sérica e seu respectivo coeficiente de correlação entre os marcadores e um escore de atividade da doença. Os valores médios da matrix de proteína oligomérica sérica e glicoproteína-39 de cartilagem humana foram, respectivamente, 9 ± 6 ug/ml e 36 ± 16 ug/ml (p > 0,001) quando comparados aos controles. Uma correlação positiva foi observada entre os escores de atividade da doença e os biomarcadores (r = 0,67 para a glicoproteína-39 de cartilagem humana e r = 0,83 para a matrix de proteína oligomérica sérica). Conclusão: O presente estudo mostrade maneira inequívoca que pacientes com artrite reumatóide de longa duração mostram valores séricos mais elevados de marcadores bioquímicos do metabolismo da cartilagem e umacorrelação positiva com o escore de atividade da doença.

Antinociceptive properties of ethanolic extract and fractions of Pterodon pubescens Benth. seeds.

We have previously demonstrated that the hydroalcoholic extract from Pterodon pubescens Benth. seeds (sucupira branca, Leguminosae) exhibits anti-arthritic activity and that its oleaginous extract (OEP) and PF1 fraction exhibit acute and topic anti-edematogenic activities. In this work, we studied the antinociceptive activity of OEP and its fractions on the acetic acid-induced abdominal constriction and formalin assays in SW male mice. OEP was obtained by ethanol extraction and its four fractions by sequential liquid-liquid extraction. PF2 GC/MS profile indicated it contains furane diterpenes derivatives of vouacapan and non-vouacapan compounds. The antinociceptive properties were demonstrated to OEP and predominantly to PF1 and PF2 by the writhing test. In the formalin assay, PF1 inhibited both phases and PF2 inhibited mainly the late one. Then, PF1 and PF2 seemed to present antinociceptive effects by different mechanisms, peripheral and/or central inhibitory ones, and showed maximum antinociceptive properties with very low doses, providing a rationale for its popular use in pain disorders.

Avaliação da proteína amilóide A sérica na atividade clínica da artrite reumatóide / Evaluation of the serum amyloid A protein in the rheumatoid arthritis clinical activity

A artrite reumatóide (AR) é uma doença auto-imune, crônica, caracterizada pelo comprometimento inflamatório das articulações sinoviais periféricas. A proteína amilóide A sérica (SAA) é uma das principais proteínas de fase aguda (PFA), porém seu uso na rotina do laboratório clínico ainda é pouco difundido. Objetivo: O objetivo deste trabalho foi analisar a utilidade da SAA na avaliação da atividade clínica da AR. Métodos: Foram estudados 113 pacientes com AR, diagnosticados segundo os critérios do Colégio Americano de Reumatologia. Para a caracterização da atividade de doença, foi utilizado o Índice de Atividade de Doença (IAD), proposto pela Liga Européia Contra o Reumatismo. Resultados: A SAA apresentou correlação positiva, estatisticamente significativa, com a proteína C-Reativa (PCR), tanto como a alfa-1-glicoproteína ácida (AGP), quanto com o IAD. Nossos resultados demonstraram que a SAA apresentou, particularmente, uma maior sensibilidade na determinação da atividade inflamatória da AR, em comparação às outras PFA. Apresentou, também, uma boa capacidade de discriminar os grupos de atividade moderada e alta do IAD. Como o IAD não mede unicamente o componente inflamatório do AR, a dosagem de uma PFA é de grande utilidade para a caracterização da atividade dessa enfermidade. Conclusões: Os resultados deste estudo sugerem que a SAA pode ser de grande valor na determinação da atividade inflamatória da AR