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.
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•Water sample associated with mining tailings promoted histological alterations in gills and liver.•The environmental impact was demonstrated using biomarkers of Astyanax lacustris.•Mix of heavy metals contributed to morphological alterations related to ionic balance.•HSP70 and P-gp were modulated due to heavy metals impact.
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.
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•Exposure to abamectin alters the behavior of exposed zebrafish.•Abamectin alters HSP70 and P-gp expression in Danio rerio.•Prolonged exposure to abamectin cause liver damage in fish.
•Atrazine causes an increase of aneurysm in gills of Nile tilapia.•Atrazine promotes an immunomodulation in different concentrations.•The melanomacrophage centres are responsive to atrazine ...stimulation at 1 ppm.•Caspase 3 activity is increased in spleen after 15 days of atrazine exposure (2 ppm).
Atrazine is a herbicide that is banned in Europe but remains widely used on different types of crops in several countries in the American continent. Atrazine is known to be an endocrine disruptor and its effects on gonads have been extensively reported, but the toxic action on other organs is poorly documented. In this paper, we investigated the toxicity of atrazine on the gills and spleens of Nile tilapia (Oreochromis niloticus). The median lethal concentration (LC50), capable of killing one-half of the test animals was calculated, and sublethal concentrations of atrazine were used in a semistatic and subchronic assay. The following four experimental groups were formed: control not exposed to atrazine, a group exposed to 1 ppm atrazine for 15 days, a group exposed to 2 ppm for 7 days, and a group exposed to 2 ppm for 15 days. The concentrations were verified during the study by high performance liquid chromatography. The gills and spleens were stained with hematoxylin and eosin and histopathological findings were made. The Perls technique was used on the spleens to identify hemosiderin, lipofuscin, and melanin pigments in the cells from melanomacrophage centres (MMCs). The spleens were submitted to proliferating cell nuclear antigen (PCNA) and inducible nitric oxide synthase (iNOS) immunohistochemistry, and morphometry was used to assess splenocyte proliferation and melanomacrophage iNOS expression. Finally, a colorimetric assay for caspase-3 was performed on the spleens to identify apoptosis. Vascular and structural alterations, such as venous sinus congestion, aneurysm, hemorrhage, pillar cell hypertrophy, disarrangement of secondary lamellae, and epithelial lifting were observed in the gills. The frequency of individuals with aneurysms was higher in the groups treated with 2 ppm than in other groups. Atrazine had an immunomodulatory effect on the spleen, observed by the alteration in the percentage of red and white pulp, alteration of the MMC area, changes in the melanomacrophage pigment content, slight iNOS suppression, decrease in splenocyte proliferation under 1 ppm atrazine, and increased caspase 3 activity under 2 ppm atrazine after 7 and 15 d. Such effects could compromise oxygenation and the immune response and, ultimately, the survival and fitness of the fish.
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.
•The synthetic cannabinoid, WIN-55,212-2, shows strong and selective toxicity against glioblastoma cell lines.•WIN-55,212-2 reveals in vitro the inhibition of migration, invasion, and clonogenicity of GBM cell lines.•WIN-55,212-2 inhibited tumor growth in vivo and it can be safely administered parenterally.
Annonacea species have been reported to possess antitumor properties. However, the in vitro and in vivo antitumor activities of Xylopia aromatica (Annonacea) have not yet been elucidated. This study ...aimed to investigate the effects of Xylopia aromatica leaves hexane fraction (XaHF) on Ehrlich ascites carcinoma cells lines (EAC), both in vitro and in vivo. In vitro assays revealed a significant cytotoxic effect with the two lower XaHF concentrations (62.5 and 32.3mg/mL). EAC (2.5x10
cells) were inoculated in the right flank of Swiss mice, and the animals were treated intraperitoneally with 32.3mg kg
of XaHF daily, for 20 days. Our findings indicate that XaHF suppressed the growth of EAC in vivo, with a significant decrease (46%) in tumor volume. There was also a decrease in the necrosis area (71%), inflammatory infiltrate, and MMP-2 expression. High-Performance Liquid Chromatography with Diode Array Detector (HPLC-DAD) identified secondary metabolites possibly related to phenolic acids, flavonoids, and alkaloids. Thus, the results confirmed the antitumoral activity that may be related to the presence of the identified metabolites in XaHF extract.
Pollutants found dispersed in water can cause irritations on the gills, challenge the immune system and prejudice the welfare of the fish. Here we investigated molecules linked to proliferation, ...survival, and cell death, as well as inflammatory and vascular control, in a model of fish gill remodeling, from injury to recovery. We assessed the gill histology and immunohistochemistry for PCNA, iNOS, HSP70, and Bax in Hypostomus francisci obtained from a river subjected to chronic anthropic influences and then after they were placed in water of good quality. A total of 30 H. francisci adult individuals were collected and distributed into two groups: euthanized on the day of capture (group 1) and maintained for 30 days in an aquarium (group 2). In all the fish from group 1, the primary and secondary lamellae showed hypertrophy of the respiratory epithelium, lamellar fusion, lifting of the epithelium, aneurysm, hyperemia, and vascular congestion. On the other hand, in all the fish from group 2, restoration of gill integrity was observed, and the primary and secondary lamellae showed a simple epithelium, absence of lamellar fusion, hypertrophy, and aneurysm. Gills of fish from group 1 had higher frequency of cells immunopositive for PCNA, iNOS, HSP70, and Bax than those of fish from group 2 (p < 0.05). The molecular and cellular mechanisms from injury to recovery were proposed, with a balance between survival and cell death signals being essential for determining the gill structure. In addition, the findings indicate that recovery of the structural organization of gills is possible if fishes are maintained in good-quality water, indicating the importance of the conservation of aquatic environments.
•Gill tissue changes can be used as biomarker of non-point source of pollution.•In gills injury, cell proliferation and vascular changes are the primary response.•Recovery gill tissue is possible in animals maintained in a good quality water.•A balance between survival or death, and inflammatory signs being essential for the gill recovery.
Sm16 is an immunomodulatory protein that seems to play a key role in the suppression of the cutaneous inflammatory response during Schistosoma mansoni penetration of the skin of definitive hosts. ...Therefore, Sm16 represents a potential target for protective immune responses induced by vaccination. In this work, we generated the recombinant protein rSm16 and produced polyclonal antibodies against this protein to evaluate its expression during different parasite life-cycle stages and its location on the surface of the parasite. In addition, we analyzed the immune responses elicited by immunization with rSm16 using two different vaccine formulations, as well as its ability to induce protection in Balb/c mice. In order to explore the biological function of Sm16 during the course of experimental infection, RNA interference was also employed. Our results demonstrated that Sm16 is expressed in cercaria and schistosomula and is located in the schistosomula surface. Despite humoral and cellular immune responses triggered by vaccination using rSm16 associated with either Freund’s or alum adjuvants, immunized mice presented no reduction in either parasite burden or parasite egg laying. Knockdown of Sm16 gene expression in schistosomula resulted in decreased parasite size in vitro but had no effect on parasite survival or egg production in vivo. Thus, our findings demonstrate that although the vaccine formulations used in this study succeeded in activating immune responses, these failed to promote parasite elimination. Finally, we have shown that Sm16 is not vital for parasite survival in the definitive host and hence may not represent a suitable target for vaccine development.
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.
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.
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.
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.
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.
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Objective
The aim of this study was to evaluate the immunohistochemical expressions of PD1, CD4
+
, and CD8
+
in premalignant lesions (OPML) that were transformed into oral squamous cell carcinoma ...OSCC (OPML-OSCC), in OSCC and also in premalignant lesions that were not transformed into OSCC (OPML-NOSSC).
Materials and methods
Retrospective analyses were performed in order to verify the demographic characteristics of the patients. CD4, CD8, and PD1 IMH studies were carried out on OPML and OSCC samples from 11 patients with OPML-OSCC and OPML, together with samples from 14 patients with OPML-NOSCC. The differences between OPML-OSCC and OPML-NOSCC were analyzed.
Results
Non-homogenous leukoplakia, together with the related oral subsite, and the lack of an exposure to tobacco, were all associated with malignant transformations. There were no statistical differences in the PD1 expression and the CD4
+
cells in OPML-OSCC and OPML-NOSCC. A significant increment in the CD8
+
cells was noted in the OPML that evolved into carcinomas when compared with OPML-NOSCC (
p
= 0.05), whereas there were higher CD8
+
cells levels in the carcinomas when compared with the OPML that evolved into carcinomas (
p
= 0.027).
Conclusions
CD8
+
cells infiltrate more in OPML-NOSCC than in OPML-OSCC. Carcinoma is more infiltrated by CD8
+
cells than its associated OPML.
Clinical relevance
Understanding immunological factors associated with malignant transformation of oral premalignant lesions can open a new way to treat this disease.
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