Toxicological effect of acetaminophen, metamizole, and nimesulide cocktail on early development of zebrafish.

BACKGROUND: Several countries' most incorrectly discarded medicines are acetaminophen (ACM), metamizole (MTZ), and nimesulide (NMS). These xenobiotics easily reach the aquatic environment; such contamination is very important for the health of humans and other species, yet little explored. OBJECTIVES: To evaluate the cocktail effect of ACM, MTZ, and NMS during zebrafish's initial development. METHODS: Zebrafish embryos 6-8 h post-fertilization (hpf) were exposed to different concentrations of ACM, MTZ, and NMS, separately, to obtain the 50% lethal concentrations (LC50). Next, the embryos were exposed to distinct concentrations of the cocktail (LC50/2, LC50/5, LC50/10, and LC50/20) in a semi-static system. Samples were analyzed 0, 24, 48, and 96 h after exposure, and the drugs' concentrations in E3 medium were assessed by high-performance liquid chromatography. For embryotoxicity evaluation, the mortality, hatching, and heart rates; total length; and pericardial and yolk sac areas were determined. In addition, body malformations, edemas, presence of pigmentation, and histopathological assessments were also recorded. RESULTS: The LC50 values obtained for MTZ, ACM, and NMS were 4.69 mgmL-1, 799.98 µgmL-1, and 0.92 µgmL-1, respectively. No difference was observed between the drugs' nominal and observed concentrations at each time point. The cocktail significantly induced mortality and decreased hatching in the LC50/10, LC50/5, and LC50/2 groups. Additionally, body malformations, pigmentation loss, and yolk sac and pericardial edemas were observed in the cocktail groups. The cocktail groups' larvae had decreased total length and slower heart rates compared to the controls (p < 0.05). The histopathological assessment showed that yolk sac edema promoted severe histological changes in the esophageal-intestine junction and intestine in larvae treated with cocktails. Moreover, PAS-positive structures decreased in the esophageal-intestine junction, intestine, and liver in larvae exposed to pharmaceutical cocktails. CONCLUSION: This study's findings suggest the cocktail of ACM, MTZ, and NMS may be hazardous to aquatic organisms in case of environmental contamination.

Tapirira guianensis is Selectively Cytotoxic, Induces Apoptosis to the Glioblastoma and Decreases Tumor Growth and Angiogenesis in vivo.

Glioblastoma is the most frequent primary malignant brain tumor without effective treatment, which makes this work extremely relevant. The study of the bioactive compounds from medicinal plants plays an important role in the discovery of new drugs.This research investigated the constituents of Tapirira guianensis and its antitumor potential (in vitro and in vivo) in glioblastoma. The T. guianensis extracts were characterized by mass spectrometry. The ethyl acetate partition (01ID) and its fractions 01ID-F2 and 01ID-F4 from T. guianensis showed potential antitumor treatment evidenced by selective cytotoxicity for GAMG with IC50 14.1 µg/mL, 83.07 µg/mL, 59.27 µg/mL and U251 with IC50 25.92 µg/mL, 37.3 µg/mL and 18.84 µg/mL. Fractions 01ID-F2 and 01ID-F4 were 10 times more selective when compared to TMZ and 01ID for the two evaluated cell lines. T. guianensis also reduced matrix metalloproteinases 2 - 01ID-F2 (21.84%), 01ID-F4 (29.6%) and 9 - 01ID-F4 (73.42%), ID-F4 (53.84%) activities, and induced apoptosis mainly through the extrinsic pathway. Furthermore, all treatments significantly reduced tumor size (01ID p < 0,01, 01ID-F2 p < 0,01 and 01ID-F4 p < 0,0001) and caused blood vessels to shrink in vivo. The present findings highlight that T. guianensis exhibits considerable antitumor potential in preclinical studies of glioblastoma. This ability may be related to the phenolic compounds and sesquiterpene derivatives identified in the extracts. This study deserves further in vivo research, followed by clinical investigation.

Toxicological effects of hydroxychloroquine sulfate and chloroquine diphosphate substances on the early-life stages of fish in the COVID-19 pandemic context.

Hydroxychloroquine sulfate (HCQ) and chloroquine diphosphate (CQ) have been used at increased rates to treat COVID-19 but can constitute a potential environmental risk. The objective was to evaluate the toxicity of sublethal concentrations of HCQ and CQ in zebrafish embryos/larvae. The 50% lethal concentrations (LC50) of HCQ and CQ at 96 h post-fertilization (hpf) were calculated by testing various concentrations on 2,160 embryos. The LC50 obtained were 560 and 800 µM for HCQ and CQ, respectively. Next, the embryotoxicity assay was performed, where 1,200 embryos were subjected to sublethal concentrations of HCQ and CQ. The hatching and heart rates were recorded. After euthanasia, photomicrographs of all larvae were taken to measure the total length, pericardial and yolk sac areas. The embryos exposed to sublethal concentrations of HCQ and CQ showed delayed hatching at 72 hpf, as well as an increase in the heart rate, larger pericardial and yolk sac areas, and body malformations at 96 hpf. The findings show that HCQ and CQ are toxic to fish in the early development phases. Understanding the mechanisms of toxicity will help extrapolate the effects of 4-aminoquinoline derivatives when they reach the aquatic environment in the context of the COVID-19 pandemic.

Exposição ao praguicida glifosato e a ocorrência do transtorno do espectro autista: existe associação? / Exposure to the pesticide glyphosate and the occurrence of autistic spectrum disorder: is there an association?

O presente estudo teve como objetivo estudar a associação entre a exposição ao glifosato e a ocorrência do Transtorno do Espectro Autista (TEA). Trata-se de um estudo documental com busca de dados acerca da prevalência do TEA (Centers of Disease Control and Prevention) e de um levantamento para verificar o princípio ativo do praguicida mais usado no Brasil (Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis) e no mundo (United States Evironmental Prevention Agency). Os resultados mostraram que a prevalência do TEA aumentou ao longo dos anos. Do mesmo modo, o uso e a compra de praguicidas também aumentaram, sendo o glifosato o princípio ativo mais usado. Ainda que os resultados apontem a existência de uma relação entre a exposição ao glifosato e a ocorrência de autismo, são necessários mais estudos para afirmar o potencial neurotóxico do glifosato e sua associação com o nascimento de crianças autistas.

The present study aimed to study the association between exposure to glyphosate and the occurrence of Autistic Spectrum Disorder (ASD). This is a documental study with search for data about the prevalence of ASD (Centers of Disease Control and Prevention) and a survey to verify the active principle of the most used pesticide in Brazil (Brazilian Institute of Environment and Renewable Natural Resources) and worldwide (United States Environmental Prevention Agency). The results showed that the prevalence of ASD increased over the years. Likewise, the use and purchase of pesticides have also increased, with glyphosate being the most widely used active ingredient in the world. Although the results point to the existence of a relationship between exposure to glyphosate and the occurrence of autism, further studies are needed to confirm the neurotoxic potential of glyphosate and its association with the birth of autistic children.

Abamectin promotes behavior changes and liver injury in zebrafish.

The indiscriminate use of pesticides is a worldwide concern due to the environment contamination since it can cause deleterious effects to non-target organisms including the fishes. The effects of abamectin, a pesticide from the avermectin family, were evaluated in adult zebrafish (Danio rerio) after exposure to a commercial formula commonly used in Brazil. The animals were submitted to acute (96 h) and to a short-term chronic exposure (15 days) of distinct concentrations of abamectin. LC50 was determined and a histological study followed by an immunohistochemistry analysis for P-gp and HSP70 identification were performed on livers of the animals submitted to the acute and chronic treatment, respectively. Moreover, behavior patterns were observed daily in both trials. A LC50 value of 105.68 µg/L was determined. The histological analysis revealed a morphological alteration of the hepatocytes, glycogen accumulation, degeneration, and disorganization of the cytoplasm, and a pyknotic, irregular, and laterally located nuclei. The immunohistochemistry for HSP70 and P-gp showed strong staining in the hepatocytes of the control groups and progressive decrease as the concentration of abamectin increased. Changes were observed in body posture, movement around the aquarium, opercular activity, body color and search for food in the groups treated with abamectin. The results presented suggest that abamectin can affect the behavioral pattern of the animals, promote morphological changes, and decrease the expression of HSP70 and P-gp in zebrafish liver.

Chemical composition, anti-inflammatory and antinociceptive effects of the butanolic fraction of Annona nutans (Annonaceae) leaves.

The species Annona nutans (R. E. Fries) is a plant found in Bolivia, Paraguay, Argentina and the Brazilian cerrado. Considering the anti-inflammatory and antinociceptive activities of the hydrometanolic fraction (FHMeOH) of A. nutans leaves previously reported, the present study aimed to evaluate in vivo anti-inflammatory and antinociceptive activities of a subfraction obtained from FHMeOH, the butanolic fraction (FBuOHf). Intraperitoneal (i.p.) treatment with FBuOHf (50 and 100 mg · kg-1) inhibited paw edema induced by carrageenan. Moreover, FBuOHf (100 mg · kg-1, i.p.) also suppressed polymorphonuclear (PMN) leukocyte migration in the footpad. Regarding the antinociceptive activity, FBuOHf (50, 100, and 200 mg · kg-1, i.p.) inhibited acetic acid-induced abdominal writhing. In the formalin test, this fraction (200 mg · kg-1, i.p.) reduced licking time only in the inflammatory phase. The FBuOHf contents flavonoids and cinnamic acid derivatives, such as quercetin-3-O-galactoside, quercetin-3-O-glucoside, isorhamnetin-3-O-galactoside, quercetin-3-O-ß-D-apio-furanosyl-(1→2)-galactopyranoside and chlorogenic acid, identified and quantified by LC-MS. The FBuOHf possesses anti-inflammatory and peripheral antinociceptive activities.

Anti-inflammatory and antinociceptive activities of a phenylpropanoid-enriched fraction of Duguetia furfuracea.

A previous study reported the in vivo anti-inflammatory and antinociceptive activities of essential oil of the underground stem bark of Duguetia furfuracea, termed EODf. This study aimed to obtain a phenylpropanoid-enriched fraction from the D. furfuracea (EFDf) essential oil and to investigate its anti-inflammatory and antinociceptive effects. The chemical composition of the EFDf was determined by gas chromatography-mass spectrometry (GC-MS). The in vivo anti-inflammatory activity was evaluated with a lipopolysaccharide (LPS)-induced paw oedema model. The effects of the EFDf on the polymorphonuclear leukocyte recruitment and the inducible nitric oxide synthase (iNOS) expression were evaluated in mice footpads. Moreover, the in vivo antinociceptive effect was assayed using the formalin test and the LPS-induced thermal hyperalgesia model. In the EFDf, 8 major compounds were identified, with α-asarone (36.4%) and 2,4,5-trimethoxystyrene (27.8%) the main constituents. A higher concentration of phenylpropanoid derivatives was found in the EFDf, 64.2% compared to the EODf (38%). The oral (p.o.) treatment with the EFDf at a dose of 3 mg/kg significantly attenuated the paw oedema, polymorphonuclear leukocyte migration, iNOS expression, and tumour necrosis factor alpha (TNF-α) production. The EFDf (10 and 30 mg/kg) also inhibited both phases of the formalin test and caused a significant increase in the reaction time in the LPS-induced thermal hyperalgesia model. Finally, EFDf-treated animals did not show any alteration of motor coordination. The results suggest that the enrichment of 2,4,5-trimethoxystyrene and α-asarone enhances the anti-inflammatory activity of the EFDf compared to the EODf. In contrast, the antinociception promoted by the EFDf was similar to the EODf and was mediated via activation of adenosinergic and opioidergic receptors.

Assessment of the Toxicological Effects of Pesticides and Detergent Mixtures on Zebrafish Gills: a Histological Study

Abstract Pesticides, as Temephos and Roundup, and surfactants, like Sodium Dodecyl Sulfate (SDS), when discharged into the environment, are harmful to non-target wildlife, especially fish. To evaluate the effects of those chemicals alone and their binary combinations, we assessed the histological alterations in Danio rerio gills after 15 days of exposure to Temephos (1ppm), Roundup (10ppm), SDS (2ppm), SDS plus Temephos (2ppm+1ppm), SDS plus Roundup (2ppm+10ppm) and Temephos plus Roundup (1ppm+10ppm). Hyperplasia, a regressive change, was significant in all exposed groups. The progressive change, complete fusion of lamellae, was significantly intensive in Roundup plus Temephos group. Regarding circulatory disturbances, vascular congestion was significant intensive in SDS group, while severe aneurysm was observed in Roundup group. Although xenobiotics in the blend are biologically complex, mainly for long periods, combinatorial exposure incremented only complete fusion of lamellae. Our results reinforce the rationalization of indiscriminate use of those compounds alone or in combination.

Efeitos tóxicos à saúde humana e ao ambiente causados pelo derramamento de rejeitos de minério da barragem de Fundão / Toxic effects to human health and the environment caused by the spillage of ore tailings from the Fundão dam

Objetivo: analisar as evidências científicas existentes até o presente momento que descrevem os efeitos tóxicos causados à saúde humana e ao ambiente, devido à liberação dos rejeitos de minério, decorrentes do rompimento da barragem de Fundão em Mariana - MG. Métodos: a revisão narrativa da literatura baseou-se na busca de artigos em quatro bases de dados: PubMed, SciELO, Science Direct e LILACS, utilizando os descritores "Dam", "Fundão" e "Mariana" junto com o operador booleano "AND". Resultados: trinta e quatro das oitenta e cinco publicações encontradas atenderam aos critérios de inclusão e foram utilizadas para compor a presente revisão. Foram observados os seguintes efeitos tóxicos: contaminação da água e do solo, principalmente, por metais pesados, potencial genotóxico, citotóxico e mutagênico da água, diminuição da diversidade microbiana, contaminação de peixes, perda da vegetação e prejuízos ao metabolismo de crescimento de plantas. Entre esses metais, foi relatado aumento na concentração de ferro, manganês, cobre, cádmio, chumbo, arsênio, cromo, magnésio, alumínio, mercúrio, cobalto, bário, níquel e selênio. Além disso, o desastre promoveu o aumento no número de casos de doenças humanas, como dengue, doenças respiratórias, dermatológicas, intestinais, psicológicas e comportamentais. Conclusão: todas as evidências científicas expostas na presente revisão sugerem que houve a contaminação ambiental, bem como da população local, por agentes tóxicos presentes nos rejeitos de minério, em especial metais pesados, liberados após o rompimento da barragem de Fundão. Assim, torna-se essencial a realização de mais estudos que visem esclarecer os efeitos tóxicos crônicos decorrentes dos rejeitos de minério liberados ao ambiente.

Objective: to analyze the scientific evidence existing to date that describes the toxic effects caused to human and the environmental health, due to the release of ore tailings, resulting from the rupture of the Fundão dam in Mariana-MG. Methods: the narrative literature review was based on a search for articles in four databases: PubMed, SciELO, Science Direct and LILACS, using the descriptors "Dam", "Fundão" and "Mariana" together with the Boolean operator "AND". Results: thirty-four, of the eighty-five publications found, met the inclusion criteria and were used to compose the present review. The following toxic effects were observed: contamination of water and soil mainly by heavy metals, genotoxic, cytotoxic and mutagenic potential of water, decrease in microbial diversity, contamination of fish, loss of vegetation, and impairment of plant growth metabolism. Among these metals, increased concentrations of iron, manganese, copper, cadmium, lead, arsenic, chromium, magnesium, aluminum, mercury, cobalt, barium, nickel and selenium were reported. In addition, the disaster promoted an increase in the number of cases of human diseases, such as dengue, respiratory, dermatological, intestinal, psychological and behavioral diseases. Conclusion: all the scientific evidence exposed in the present review that there was environmental contamination, as well as of the local population, by toxic agents present in the ore tailings, especially heavy metals, released after the collapse of the Fundão dam. Thus, further studies aimed at clarifying the chronic toxic effects resulting from the ore tailings released into the environment are essential.

Primary culture macrophages from fish as potential experimental model in toxicity studies with multiwalled carbon nanotubes

Multiwalled carbon nanotube (MWCNT) has been broadly used in several sectors of society. This material when exposed to the environment might reach the aquatic animals and cause toxic effects. Here, it was evaluated the MWCNTs toxicity in melanomacrophages primary culture that was submitted to 1 µ gm L-1 MWCNTs for 24 hours. After exposition to MWCNT, 48 and 59% liver and spleen melanomacrophages were healthy, respectively. The control group presented 85% viability. Phagocytosis activity of melanomacrophages was observed by presence of black inclusions in cytoplasm. The findings indicate MWCNT was cytotoxic to melanomacrophages, where its release and effect into aquatic environment must be more studied. Finally, the melanomacrophages present large potential as experimental model for evaluation of carbon-based nanomaterial toxicity.

Histological and molecular changes in gill and liver of fish (Astyanax lacustris Lütken, 1875) exposed to water from the Doce basin after the rupture of a mining tailings dam in Mariana, MG, Brazil.

The effects of the rupture of a mining tailings dam were investigated using the gills and liver of Astyanax lacustris as a proxy for environmental quality. The fish were exposed for seven days to water sampled forming four groups: upstream of the dam rupture (P1), and 22 km (P2); 48 km (P3); and 70 km (P4) downstream from the dam rupture in the Doce River basin. The control group received dechlorinated tap water. The dissolved concentrations of metals were determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). We evaluated the histology of the gills and liver, as well as, immunohistochemistry for HSP70 and Na+/K+ ATPase (NKA) in the gills, and for P-gp in liver. In all sites we observed a mix of metals, with higher concentrations of Mn, Cd, As, and Cu/Cr in P1, P2, P3, and P4, respectively. All treatments groups showed histological changes in gills and liver, with the highest amount of these alterations found in the P2 group. Disorganization of the secondary lamellae, epithelial lifting, and mitochondria-rich cells (MRC) were observed in the gills. The parenchyma of the liver was rather disorganized, and hepatocytes and nuclei showed hypertrophy, vacuolization and cytoplasmic degeneration. A higher immunoreaction of HSP70 in P2 when compared with the other groups and lower labeling of HSP70 in the P4 was registered. In P2 and P3, NKA-positive cells were observed with hypertrophy and disorganization. Morphometric analyses of the liver revealed that all treatment groups presented a lower immunolabeling of P-gp when compared with the control group. Thus, the experimental approach revealed that the water from Doce basin can promote histological alterations in fish's liver and gills, as well as modulation of disruption of ionic balance, cellular responses to stress, and cell detoxification pathways.

Using the plants of Brazilian Cerrado for wound healing: From traditional use to scientific approach.

ETHNOPHARMACOLOGICAL RELEVANCE: The Brazilian Cerrado is a biome with a remarkable diversity of plant species, many of which are used mainly by local communities as a source of treatment to several pathologic processes, especially for the treatment of wounds. However, no systematic review exists focusing on the plants used in this respect and on the appropriate pharmacological investigations that substantiate the actions that are reported. This study revisits the traditional use of medicinal plants from the Brazilian Cerrado in the treatment of wounds and the pharmacological characteristics of the reported plant species. METHOD: ology: For the present article, previous studies on plants of the Brazilian Cerrado used for wound healing carried out between 1996 and 2018 were researched on various academic databases (PubMed, Elsevier, Springer, Lilacs, Google Escolar, and Scielo). RESULTS: A total of 33 studies were carried out on 29 plant species distributed into 18 families, mainly Fabaceae or Leguminosae (9), Bignoniaceae (2), Asteraceae (2), Euphorbiaceae (2). Considering the great diversity of Cerrado plants, only a small number of wound healing studies were carried out between 1996 and 2018. It was observed that there is a large gap between experimentation assay and traditional use. There are only few connections between the form of use by the population and the experiments conducted in the laboratory. We found that only about 12% of these studies considered to use the methodologies, or at least in parts, to obtain extracts such as those used in folk medicine. Approximately 37% of the experiments were performed using the bark as well as the same ratio for leaves, 6% using the fruits, and 9% using the seeds, roots or flowers. In several studies, there are reports of chemical constituents such as flavonoids and tannins, followed by steroid terpenes, saponins, and fatty acids, and alkaloids. However, approximately 35% of the studies did not supply information about compounds present in the preparation or the effect which could be attributed to these agents in respect to wound healing. Regarding treatment, most of the studies employed a topical treatment, though intraperitoneal and oral treatment were also described using either topical, oil-based formulations, but also gel- or saline-based formulations. CONCLUSIONS: Although, there has been an increase in knowledge about the biological actions of plants from Cerrado biome, the scientific basis for the traditional use of these local medicinal plants in wound healing does not provide sufficient information on the efficacy of the treatment, the molecular mechanisms, or, in particular, the effective doses used and the drug interactions. Thus, focused research investigating these hypotheses from traditional knowledge is necessary to prove the evidence of the potential pharmacological action.

Anti-inflammatory and central and peripheral anti-nociceptive activities of α-asarone through the inhibition of TNF-α production, leukocyte recruitment and iNOS expression, and participation of the adenosinergic and opioidergic systems.

Alpha-asarone has been found to possess many pharmacological activities, which can improve cognitive function and exert anti-oxidant, anxiolytic, anti-epileptic and protective effects against endothelial cell injury. The anti-inflammatory activity of α-asarone was evaluated using lipopolysaccharide (LPS)-induced paw oedema. Moreover, leukocyte migration, inducible nitric oxide synthase (iNOS) expression and tumour necrosis factor-alpha (TNF-α) levels were quantified in footpads. Formalin and LPS-induced thermal hyperalgesia models were generated using adenosinergic, opioidergic, serotonergic and muscarinic receptor antagonists. The effects on motor coordination were evaluated by means of the rota-rod test. Oral treatment (p.o.) with α-asarone (3 mg/kg) significantly inhibited paw oedema by 62.12 and 72.22%, 2 and 4 h post LPS injection, respectively. Alpha-asarone (3 mg/kg, p.o.) attenuated the inflammatory infiltrate 1, 3 and 6 h after LPS injection. Furthermore, α-asarone (3 mg/kg, p.o.) suppressed iNOS expression and TNF-α production, 6 and 1 h after inflammatory stimulus, respectively. Alpha-asarone (3, 10 and 30 mg/kg, p.o.) inhibited both phases of formalin-induced licking. In the hot-plate test, α-asarone (10 and 30 mg/kg, p.o.) increased the latency to response 3 and 5 h post LPS stimulus. Caffeine and naloxone abolished the central anti-nociceptive effect of α-asarone (neurogenic phase of formalin and hot plate tests), suggesting the participation of the adenosinergic and opioidergic systems. Furthermore, naloxone reversed the peripheral activity of α-asarone (inflammatory phase of formalin test), indicating the possible involvement of the opioidergic pathway. In the rota-rod test, α-asarone did not change motor coordination. These findings suggest that α-asarone has anti-inflammatory, peripheral and central anti-nociceptive effects and could represent a promising agent for future research.

Matteucinol, isolated from Miconia chamissois, induces apoptosis in human glioblastoma lines via the intrinsic pathway and inhibits angiogenesis and tumor growth in vivo.

Gliomas account for nearly 70% of the central nervous system tumors and present a median survival of approximately 12-17 months. Studies have shown that administration of novel natural antineoplastic agents is been highly effective for treating gliomas. This study was conducted to investigate the antitumor potential (in vitro and in vivo) of Miconia chamissois Naudin for treating glioblastomas. We investigated the cytotoxicity of the chloroform partition and its sub-fraction in glioblastoma cell lines (GAMG and U251MG) and one normal cell line of astrocytes. The fraction showed cytotoxicity and was selective for tumor cells. Characterization of this fraction revealed a single compound, Matteucinol, which was first identified in the species M. chamissois. Matteucinol promoted cell death via intrinsic apoptosis in the adult glioblastoma lines. In addition, Matteucinol significantly reduced the migration, invasion, and clonogenicity of the tumor cells. Notably, it also reduced tumor growth and angiogenesis in vivo. Moreover, this agent showed synergistic effects with temozolomide, a chemotherapeutic agent commonly used in clinical practice. Our study demonstrates that Matteucinol from M chamissois is a promising compound for the treatment of glioblastomas and may be used along with the existing chemotherapeutic agents for more effective treatment.

Immunohistochemical Analysis of P-gp, LC3-II, and Cathepsin-D Associated with Histological Changes in Fish Liver: from the Impacted Environment to Clean Water

Abstract Herein we evaluated the histopathological alterations and expression patterns of multixenobiotic resistence (MXR) and autophagic proteins in liver samples of fish chronically exposed to anthropogenic contaminants in a highly polluted river, and then again after they had been transferred to good quality water. Two groups were established: euthanized on the day of capture (0 h), and maintained for 30 days in a tank (30 d). The fish of 0 h presented liver with vacuolated and hypertrophic hepatocytes. Also, it was observed strong immunostaining of cathepsin-D, LC3-II and P-gp. Necrosis and apoptosis were also observed throughout the liver. Conversely, the second group (30 d) showed recovery of the liver normal histology and weak immunoreaction of the studied proteins. So, our results indicated that there was a hepatic recovery in the fish kept in good quality water, as showed by the decreased expression of cathepsin-D, LC3-II, and the MXR (P-gp). Therefore, the alterations here observed could be proposed as potential biomarkers to be tested for following the impacts of remediation or mitigation measures to environmental impacts.

Estuarine fish assemblages present a species-specific difference in the multixenobiotics resistance activity.

We investigated the activity of the multixenobiotic resistance (MXR) phenotype, a biological defense system in aquatic organisms, in the fish assemblages of two tropical estuaries with different degrees of environmental impacts, the Paraiba River and Mamanguape River Estuaries. The aim of this work was to compare the activity of the MXR phenotype of different fishes to test the hypothesis that each species has an inherent activity level and to use this activity as a bioindicator of aquatic contamination. We assessed the MXR activity of the gills, using rhodamine B (RB) accumulation assay. The results demonstrated a species-specific difference in the MXR activity of fishes caught in the same estuarine zone. Also, the pelagic species Eucinostomus melanopterus, Eucinostomus argenteus, and Lutjanus jocu had higher RB accumulation, while the demersal species Sphoeroides testudineus and Sphoeroides greeleyi had the lowest RB accumulation, suggesting that the ecological characteristic of fish in the water column exerts an influence on MXR activity. Besides, we demonstrated the potential of using the gill MXR activity of the key estuarine species, the Brazilian silversides Atherinella brasiliensis, as a tool for biomonitoring estuaries.

WIN55,212-2 induces caspase-independent apoptosis on human glioblastoma cells by regulating HSP70, p53 and Cathepsin D.

Despite the standard approaches to treat the highly aggressive and invasive glioblastoma (GBM), it remains incurable. In this sense, cannabinoids highlight as a promising tool, because this tumor overexpresses CB1 and/or CB2 receptors and being, therefore, can be susceptible to cannabinoids treatment. Thus, this work investigated the action of the cannabinoid agonist WIN55-212-2 on GBM cell lines and non-malignant cell lines, in vitro and in vivo. WIN was selectively cytotoxic to GBM cells. These presented blebbing and nuclear alterations in addition to cell shrinkage and chromatin condensation. WIN also significantly inhibited the migration of GAMG and U251 cells. Finally, the data also showed that the antitumor effects of WIN are exerted, at least to some extent, by the expression of p53 and increased cathepsin D in addition to the decreased expression of HSP70.This data can indicate caspase-independent cell death mechanism. In addition, WIN decreased tumoral perimeter as well as caused a reduction the blood vessels in this area, without causing lysis, hemorrhage or blood clotting. So, the findings herein presented reinforce the usefulness of cannabinoids as a candidate for further evaluation in treatment in glioblastoma treatment.

Chemical composition and evaluation of the anti-inflammatory and antinociceptive activities of Duguetia furfuracea essential oil: Effect on edema, leukocyte recruitment, tumor necrosis factor alpha production, iNOS expression, and adenosinergic and opioidergic systems.

ETHNOPHARMACOLOGICAL RELEVANCE: Duguetia furfuracea (A. St. -Hil.) Saff. (Annonaceae) is commonly known in Brazil as "araticum-seco," and its root is used in folk medicine to treat inflammatory and painful disorders. However, no studies have been performed to evaluate these therapeutic activities. AIM OF THE STUDY: Investigate the chemical composition, anti-inflammatory and antinociceptive effects, and elucidate the possible antinociceptive mechanisms of action from the essential oil of D. furfuracea (EODf) underground stem bark. MATERIALS AND METHODS: Chemical composition was determined by gas chromatography and mass spectrometry (GC/MS). The paw edema induced by LPS, formalin-induced nociception, LPS-induced thermal hyperalgesia and rota-rod tests in vivo were used to evaluate the anti-inflammatory and antinociceptive effects in addition to the alteration on motor coordination. Histological analyses and an immunohistochemistry assay for iNOS were performed on mouse footpads of naive, control, 10 mg/kg EODf, and 10 mg/kg indomethacin (Ind) groups. The samples were removed at 1, 3, and 6 h after subplantar injection of LPS. In addition, the involvement of the adenosinergic, opioidergic, serotonergic, and cholinergic systems were investigated, in order to elucidate possible antinociceptive mechanisms. RESULTS: Twenty-four volatile constituents were detected and identified. (E)-asarone (21.9%), bicyclogermacrene (16.7%), 2,4,5-trimethoxystyrene (16.1%), α-gurjunene (15%), cyperene (7.8%), and (E)-caryophyllene (4.6%) were major compounds found in EODf. Oral treatment (p.o.) with EODf (1, 3, and 10 mg/kg) significantly inhibited the paw edema induced by LPS. At 10 mg/kg EODf promoted inhibition of tumor necrosis factor alpha (TNF-α) production, recruitment of polymorphonuclear (PMN) leukocytes and inducible nitric oxide synthase (iNOS) expression in paw tissue. EODf (10 and 30 mg/kg, p.o.) also reduced licking time in both phases of the formalin test and it had a significant effect on the LPS-induced thermal hyperalgesia model. The administration of caffeine (Caf) and naloxone (Nal) reversed the antinociceptive activity of EODf, in the first phase of the formalin test and in the LPS-induced thermal hyperalgesia model. Moreover, Nal was also able to abolish the antinociception caused by EODf, in the second phase of formalin test. In the rota-rod test, EODf-treated animals did not show any alteration of motor coordination. CONCLUSIONS: Our findings indicate that EODf underground stem bark produces anti-inflammatory and both central and peripheral antinociceptive effects. Furthermore, the antinociceptive activity of EODf underground stem bark is possibly mediated by adenosinergic and opioidergic pathways, and its properties do not induce effects on motor coordination. These results support the use of the folk medicine, D. furfuracea root, to treat inflammation and painful conditions.

Are the Statins promising antifungal agents against invasive candidiasis?

The medical importance of intra-abdominal candidiasis (IAC) contrasts with the limited number of pharmacological treatment options available and the increasing rate of resistance to antifungal drugs. Thus, the repositioning of compounds in clinical use can contribute to the broadening of treatment possibilities for this infection. Statins, a class of drugs used to reduce cardiovascular event risk, have shown antiparasitic, antibacterial, and antiviral activities; however, their antifungal effects remain poorly studied. In this context, the present study aimed to elucidate the antifungal potential of six statins in vitro, as well as to evaluate the therapeutic use of fluvastatin in a mouse model of IAC. The biological effects of statins were evaluated against Candida spp., through the determination of the minimum inhibitory concentration (MIC). For the statins that showed activity, the fungicidal concentration, toxicity/selectivity, synergism with azoles and polyenes, phenotypic effects, and activity against virulence factors were also determined. Atorvastatin, rosuvastatin and fluvastatin were highly active, especially against C. albicans (MIC < 1-128 µg.mL-1) and C. glabrata (MIC 32-64 µg.mL-1). Fluvastatin and atorvastatin were selective for C. albicans in baby hamster kidney (BHK) cells. Moreover, all active statins in the antifungal assay showed high selectivity for fungal cells over bacteria. The combination of atorvastatin, rosuvastatin, and fluvastatin with azoles was associated with a synergistic effect. Active statins do not act on the fungal membrane or wall, but instead stimulate farnesol-dependent pathogenicity factors such as yeast-to-hyphal transition and biofilm generation. Fluvastatin treatment was evaluated in a mouse model of IAC, showing stimulation of the extra-hepatic dissemination of C. albicans but improvements in renal, splenic, and hepatic histological aspects. In conclusion, statins have potent antifungal activity in vitro, but the therapeutic effect in vivo is restricted to their anti-inflammatory activity.

21­Benzylidene digoxin, a novel digoxin hemi-synthetic derivative, presents an anti-inflammatory activity through inhibition of edema, tumour necrosis factor alpha production, inducible nitric oxide synthase expression and leucocyte migration.

Recent findings have demonstrated new therapeutic functions of cardiotonic steroids, a process that is termed drug repositioning. Despite the confirmed anti-inflammatory effects of cardiotonic steroids, their clinical use has been discouraged due to toxicity related to inhibition of the Na+/K+ ATPase. A novel synthetic compound derived from digoxin, 21­benzylidene digoxin (21­BD), does not inhibit this enzyme. Herein, we evaluated the anti-inflammatory and antinociceptive effects and acute toxicity of 21­BD. Murine (Swiss mice) models of paw oedema induced by carrageenan, acetic acid-induced abdominal writhing, and formalin and acute toxicity tests were used. Oral administration of 21­BD (0.3 mg/kg) showed a significant and prolonged inhibition of paw oedema. Histological analysis demonstrated a reduction in inflammatory cells and expression of inducible nitric oxide synthase (iNOS) in footpads 6 h after administration of carrageenan. 21­BD (0.3 mg/kg) also reduced the levels of tumour necrosis factor (TNF)-α 2 and 4 h after carrageenan. 21­BD demonstrated antinociceptive activity, inhibiting abdominal writhes at all tested doses. However, in the formalin test, 21­BD did not present antinociceptive activity. In the acute toxicity test, 21­BD did not cause symptoms of toxicity or mortality. The present study demonstrated, for the first time, that 21­BD is safe and exhibits a marked anti-inflammatory activity in acute local inflammation. This effect might be a consequence of its ability to inhibit the release of the PMN leucocyte-derived mediators, including TNF-α, and iNOS expression as well as its inhibitory effect on oedema and PMN leucocyte infiltration.