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Background: Luteolin, a flavonoid with well-documented antioxidant properties, has garnered significant attention for its potential therapeutic effects. Objectives: This study aims to investigate the antioxidant properties of luteolin under the influence of solvents, utilizing computational techniques to elucidate its interactions and its potential role as a modulator of enzymatic activities, particularly with Cytochrome 17A1. Methods: Density Functional Theory (DFT) calculations were employed to determine luteolin’s electronic and structural characteristics. Key aspects analyzed included electron density distribution and the energies of the frontier molecular orbitals (HOMO and LUMO). Free radical scavenging mechanisms were explored by comparing the dissociation enthalpy of the O–H bond in the absence and presence of water molecules. Additionally, molecular docking simulations were performed to assess the interactions of luteolin with Cytochrome 17A1, identifying preferred binding sites and interaction energies. Results: The findings indicate that luteolin possesses distinct structural and electronic features that contribute to its effectiveness in protecting against oxidative stress. However, hydrogen bonding interactions with water molecules were found to influence the dissociation enthalpy of the O–H bond. Docking simulations revealed significant interaction profiles between luteolin and Cytochrome 17A1, suggesting its potential role as a modulator of this protein. Conclusions: This study underscores the therapeutic potential of luteolin and highlights the importance of computational techniques in predicting and understanding the molecular interactions of bioactive compounds with biological targets. The results provide valuable insights that may aid in developing new therapeutic strategies for diseases associated with oxidative stress.

Background: Eugenia involucrata DC., a Cerrado native plant, is recognized for its medicinal properties. However, its bioactive compounds remain inadequately explored. Objectives: This study investigated bioactive compounds from a standardized liquid extract from E. involucrata leaves that can act with antioxidant, cytogenotoxic, cytoprotective, and genoprotective effects. Methods: The phenolic compounds in the standardized liquid extract from E. involucrata leaves were screened by HPLC-DAD. The capture of the free radicals DPPH, ABTS+, and the metal reduction power FRAP determined the antioxidant potential. Cytotoxicity was evaluated in RAW 264.7 macrophages (MTT assay), and (anti)cytotoxic and (anti)genotoxic effects were assessed in human lymphocytes using the Trypan blue exclusion method and comet assay, respectively. Results: The extracts present key phenolic compounds, such as ellagic acid, myricitrin, and epicatechin gallate. The standardized extract demonstrated antioxidant capacity, evidenced by its ability to reduce iron and scavenge free radicals. The liquid extract from E. involucrata leaves exhibited cytotoxic effects on RAW 264.7 macrophages at higher concentrations, while demonstrating (anti)cytotoxic activity on human lymphocytes from all tested concentrations. The highest concentration tested of the standardized liquid extract from E. involucrata leaves (250 µg/mL) showed genotoxicity against human lymphocytes compared to the negative control. In contrast, the lowest concentration (62.5 µg/mL) exhibited an antigenotoxic effect on human lymphocytes, reducing the genotoxicity of doxorubicin by approximately 27%. Conclusions: The bioactive compounds in the standardized liquid extract from E. involucrata leaves exhibited antioxidant and antigenotoxic properties, suggesting potential value for nutraceutical and pharmaceutical applications, particularly those related to oxidative stress associated withaging and disease progression.

Purpose Infection of macrophages is a mandatory step for Lesihmania to promote mammalian infection and the evaluation of parasites proliferating inside macrophages reveals important information about parasites virulence and leishmanicidal drugs. Here we compare macrophage phagocytosis ability and amastigote proliferation of L. major or L. braziliensis by two different methods: fixation and staining of infected macrophages or recovery of promastigotes in culture. Methods Promastigote parasites were used to infect thioglycolate-elicited BALB/c mice peritoneal macrophages. Phagocytosis was evaluated at 3 h after infection and amastigote proliferation was evaluated directly or indirectly at 3, 6, and 9 days after infection. Results Phagocytosis of L. major and L. braziliensis by murine macrophages was respectively 54.38 ± 10.41% and 62.54 ± 19.01%, according to the fixation and staining method. The infection index (product of the percentage of infected cells X the number of parasites per cell) obtained by fixation and staining method showed proliferation of L. major inside macrophage from 108.42 ± 25.57 (3 h) to 510.09 ± 99.13 (9 days) and decrease of the number of L. braziliensis from 223.01 ± 58.46 (3 h) to 101.37 ± 20.06 (9 days). The parasites recovered/ mL in culture increased for L. major infected macrophage from 2,38 × 10 6  ± 2,31 × 10 6 (3 h) to 16,8 × 10 6  ± 8,3 × 10 6 (9 days) and decreased from 8.43 × 10 6  ± 6.8 × 10 6 (3 h) to 0.19 × 10 6  ± 0.21 × 10 6 (9 days) for L. braziliensis infected macrophage. The fixation and staining method allowed to observe that both parasite species proliferated inside of some macrophages, while other macrophages maintain the parasite number unaltered or even kill the parasite. Conclusion Both methods showed that L. major proliferates in vitro inside of murine macrophages while L. braziliensis are mostly eliminated by these cells. Only fixation and staining allowed to identify L. braziliensis susceptible macrophages in vitro.

Aim: The propouse of this study was to available the cytotoxicity, phagocytic activity, and leishmanicidal potential of extract standardized from the fruits of Pterodon emarginatus Vogel. Background: P. emarginatus is a Brazilian medicinal plant with several constituents shown to have anti-inflammatory, antinociceptive, antimicrobial, and antiproliferative activities. Materials and Methods: The standardized extract in geranylgeraniol obtained from the fruit of P. emarginatus was investigated for clastogenic and cytotoxic effects, as well as phagocytic activity against Leishmania guyanensis. Results: A 24-h exposure of Allium cepa roots to standardized geranylgeraniol extract (9.77 mg/mL) promoted clastogenic effects, with a mitotic index (MI) of 10.33 ± 0.61. Extending the exposure time to 48 or 72 h significantly reduced the MI (P < 0.05). The IC50for geranylgeraniol in 3T3 and RAW 264.7 cells was found to be 15.5 mg/mL and 12.2 mg/mL, respectively. The standardized extract in geranylgeraniol exhibited high phagocytic and microbicidal activities in Leishmania, promoting a >60% increase in macrophage infection, with greater internalization and destruction of the parasite. In silico study results suggest that geranylgeraniol acts in Leishmania by interfering with the biosynthesis of sterols through steric hindrance of the active site of the enzyme CYP51. Conclusion: The standardized extract in geranylgeraniol may be useful as an antiproliferative agent, macrophage immunomodulator, and as a cytotoxic treatment against infectious agents.

Background The interleukin 32 (IL-32) is a proinflammatory cytokine produced by immune and non-immune cells. It can be induced during bacterial and viral infections, but its production was never investigated in protozoan infections. American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan leading to cutaneous, nasal or oral lesions. The aim of this study was to evaluate the expression of IL-32 in cutaneous and mucosal lesions as well as in peripheral blood mononuclear cells (PBMC) exposed to Leishmania ( Viannia ) braziliensis . Methods IL-32, tumour necrosis factor (TNF) and IL-10 protein expression was evaluated by immunohistochemistry in cutaneous, mucosal lesions and compared to healthy specimens. The isoforms of IL-32α, β, δ, γ mRNA, TNF mRNA and IL-10 mRNA were assessed by qPCR in tissue biopsies of lesions and healthy skin and mucosa. In addition, PBMC from healthy donors were cultured with amastigotes of L. ( V. ) braziliensis . In lesions, the parasite subgenus was identified by PCR-RFLP. Results We showed that the mRNA expression of IL-32, in particular IL-32γ was similarly up-regulated in lesions of cutaneous (CL) or mucosal (ML) leishmaniasis patients. IL-32 protein was produced by epithelial, endothelial, mononuclear cells and giant cells. The IL-32 protein expression was associated with TNF in ML but not in CL. IL-32 was not associated with IL-10 in both CL and ML. Expression of TNF mRNA was higher in ML than in CL lesions, however levels of IL-10 mRNA were similar in both clinical forms. In all lesions in which the parasite was detected, L . ( Viannia ) subgenus was identified. Interestingly, L . ( V .) braziliensis induced only IL-32γ mRNA expression in PBMC from healthy individuals. Conclusions These data suggest that IL-32 plays a major role in the inflammatory process caused by L . ( Viannia ) sp or that IL-32 is crucial for controlling the L . ( Viannia ) sp infection.

The interleukin 32 (IL-32) is a proinflammatory cytokine produced by immune and non-immune cells. It can be induced during bacterial and viral infections, but its production was never investigated in protozoan infections. American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan leading to cutaneous, nasal or oral lesions. The aim of this study was to evaluate the expression of IL-32 in cutaneous and mucosal lesions as well as in peripheral blood mononuclear cells (PBMC) exposed to Leishmania (Viannia) braziliensis. IL-32, tumour necrosis factor (TNF) and IL-10 protein expression was evaluated by immunohistochemistry in cutaneous, mucosal lesions and compared to healthy specimens. The isoforms of IL-32[alpha], [beta], [delta], [gamma] mRNA, TNF mRNA and IL-10 mRNA were assessed by qPCR in tissue biopsies of lesions and healthy skin and mucosa. In addition, PBMC from healthy donors were cultured with amastigotes of L. (V.) braziliensis. In lesions, the parasite subgenus was identified by PCR-RFLP. We showed that the mRNA expression of IL-32, in particular IL-32[gamma] was similarly up-regulated in lesions of cutaneous (CL) or mucosal (ML) leishmaniasis patients. IL-32 protein was produced by epithelial, endothelial, mononuclear cells and giant cells. The IL-32 protein expression was associated with TNF in ML but not in CL. IL-32 was not associated with IL-10 in both CL and ML. Expression of TNF mRNA was higher in ML than in CL lesions, however levels of IL-10 mRNA were similar in both clinical forms. In all lesions in which the parasite was detected, L. (Viannia) subgenus was identified. Interestingly, L. (V.) braziliensis induced only IL-32[gamma] mRNA expression in PBMC from healthy individuals. These data suggest that IL-32 plays a major role in the inflammatory process caused by L. (Viannia) sp or that IL-32 is crucial for controlling the L. (Viannia) sp infection.

Doc number: 249 Abstract Background: The interleukin 32 (IL-32) is a proinflammatory cytokine produced by immune and non-immune cells. It can be induced during bacterial and viral infections, but its production was never investigated in protozoan infections. American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan leading to cutaneous, nasal or oral lesions. The aim of this study was to evaluate the expression of IL-32 in cutaneous and mucosal lesions as well as in peripheral blood mononuclear cells (PBMC) exposed to Leishmania (Viannia ) braziliensis . Methods: IL-32, tumour necrosis factor (TNF) and IL-10 protein expression was evaluated by immunohistochemistry in cutaneous, mucosal lesions and compared to healthy specimens. The isoforms of IL-32α, β, δ, γ mRNA, TNF mRNA and IL-10 mRNA were assessed by qPCR in tissue biopsies of lesions and healthy skin and mucosa. In addition, PBMC from healthy donors were cultured with amastigotes of L. (V. ) braziliensis . In lesions, the parasite subgenus was identified by PCR-RFLP. Results: We showed that the mRNA expression of IL-32, in particular IL-32γ was similarly up-regulated in lesions of cutaneous (CL) or mucosal (ML) leishmaniasis patients. IL-32 protein was produced by epithelial, endothelial, mononuclear cells and giant cells. The IL-32 protein expression was associated with TNF in ML but not in CL. IL-32 was not associated with IL-10 in both CL and ML. Expression of TNF mRNA was higher in ML than in CL lesions, however levels of IL-10 mRNA were similar in both clinical forms. In all lesions in which the parasite was detected, L . (Viannia ) subgenus was identified. Interestingly, L . (V .) braziliensis induced only IL-32γ mRNA expression in PBMC from healthy individuals. Conclusions: These data suggest that IL-32 plays a major role in the inflammatory process caused by L . (Viannia ) sp or that IL-32 is crucial for controlling the L . (Viannia ) sp infection.