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1.
Int J Mol Sci ; 24(18)2023 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-37762197

RESUMO

The dysregulation of complement system activation usually results in acute or chronic inflammation and can contribute to the development of various diseases. Although the activation of complement pathways is essential for innate defense, exacerbated activity of this system may be harmful to the host. Thus, drugs with the potential to inhibit the activation of the complement system may be important tools in therapy for diseases associated with complement system activation. The synthetic peptides Cp40 and PMX205 can be highlighted in this regard, given that they selectively inhibit the C3 and block the C5a receptor (C5aR1), respectively. The zebrafish (Danio rerio) is a robust model for studying the complement system. The aim of the present study was to use in silico computational modeling to investigate the hypothesis that these complement system inhibitor peptides interact with their target molecules in zebrafish, for subsequent in vivo validation. For this, we analyzed molecular docking interactions between peptides and target molecules. Our study demonstrated that Cp40 and the cyclic peptide PMX205 have positive interactions with their respective zebrafish targets, thus suggesting that zebrafish can be used as an animal model for therapeutic studies on these inhibitors.


Assuntos
Ativação do Complemento , Peixe-Zebra , Animais , Simulação de Acoplamento Molecular , Simulação por Computador , Inativadores do Complemento/farmacologia
3.
Sci Rep ; 13(1): 8060, 2023 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-37198208

RESUMO

Despite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-α and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans.


Assuntos
COVID-19 , SARS-CoV-2 , Animais , Humanos , Peixe-Zebra , Macrófagos , Peptídeos
4.
Int J Mol Sci, v. 24, n. 18, 13895, sep. 2023
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-5117

RESUMO

The dysregulation of complement system activation usually results in acute or chronic inflammation and can contribute to the development of various diseases. Although the activation of complement pathways is essential for innate defense, exacerbated activity of this system may be harmful to the host. Thus, drugs with the potential to inhibit the activation of the complement system may be important tools in therapy for diseases associated with complement system activation. The synthetic peptides Cp40 and PMX205 can be highlighted in this regard, given that they selectively inhibit the C3 and block the C5a receptor (C5aR1), respectively. The zebrafish (Danio rerio) is a robust model for studying the complement system. The aim of the present study was to use in silico computational modeling to investigate the hypothesis that these complement system inhibitor peptides interact with their target molecules in zebrafish, for subsequent in vivo validation. For this, we analyzed molecular docking interactions between peptides and target molecules. Our study demonstrated that Cp40 and the cyclic peptide PMX205 have positive interactions with their respective zebrafish targets, thus suggesting that zebrafish can be used as an animal model for therapeutic studies on these inhibitors.

5.
Int J Mol Sci, v. 24, 13895, set. 2023
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-5083

RESUMO

The dysregulation of complement system activation usually results in acute or chronic inflammation and can contribute to the development of various diseases. Although the activation of complement pathways is essential for innate defense, exacerbated activity of this system may be harmful to the host. Thus, drugs with the potential to inhibit the activation of the complement system may be important tools in therapy for diseases associated with complement system activation. The synthetic peptides Cp40 and PMX205 can be highlighted in this regard, given that they selectively inhibit the C3 and block the C5a receptor (C5aR1), respectively. The zebrafish (Danio rerio) is a robust model for studying the complement system. The aim of the present study was to use in silico computational modeling to investigate the hypothesis that these complement system inhibitor peptides interact with their target molecules in zebrafish, for subsequent in vivo validation. For this, we analyzed molecular docking interactions between peptides and target molecules. Our study demonstrated that Cp40 and the cyclic peptide PMX205 have positive interactions with their respective zebrafish targets, thus suggesting that zebrafish can be used as an animal model for therapeutic studies on these inhibitors.

6.
Sci Rep, v. 13, 8060, mai. 2023
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4913

RESUMO

Despite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-α and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans.

7.
Fish Shellfish Immunol ; 130: 323-331, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36122633

RESUMO

To identify activation pathways and effector mechanisms of innate immunity in fish has become relevant for the sanitary management of intensive fish farming. However, little is known about the blocking of cysteinyl leukotrienes receptors (CysLTRs) and their effects in teleost fish. Our study evaluated the anti-inflammatory effect of 250 and 500 µg zafirlukast (antagonist of CysLTRs)/kg b.w., administered orally in the diet, during acute inflammatory reaction induced by Aeromonas hydrophila bacterins in Oreochromis niloticus. 80 tilapia were distributed in 10 aquariums (100L of water each, n = 8) to constitute three treatments: Control (inoculated with A. hydrophila bacterin and untreated); Treated with 250 µg or 500 µg of zafirlukast/kg b.w. and inoculated. To be evaluated in three periods: 6, 24 and 48 h post-inoculation (HPI), totaling nine aquariums. A tenth group was sampled without any stimulus to constitute reference values (Physiological standards). Tilapia treated with zafirlukast demonstrated dose-response effect in the decrease of accumulated inflammatory cells, strongly influenced by granulocytes and macrophages. Zafirlukast treated-tilapia showed decrease in blood leukocyte counts (mainly neutrophils, and monocytes) and reactive oxygen species production. Treatment with zafirlukast resulted in down-regulation of ceruloplasmin, complement 3, alpha2-macroglobulin, transferrin and apolipoprotein A1, as well as up-regulation of haptoglobin. Our study provided convincing results in the pathophysiology of tilapia inflammatory reaction, considering that treatment with zafirlukast, antagonist of cysteinyl leukotriene receptors, resulted in a dose-response effect by suppressing the dynamics between leukocytes in the bloodstream and cell accumulation in the inflamed focus, as well as modulated the leukocyte oxidative burst and the acute phase protein response.


Assuntos
Ciclídeos , Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , alfa 2-Macroglobulinas Associadas à Gravidez , Tilápia , Aeromonas hydrophila/fisiologia , Animais , Anti-Inflamatórios , Apolipoproteína A-I , Vacinas Bacterianas , Ceruloplasmina , Complemento C3 , Feminino , Haptoglobinas , Indóis , Fenilcarbamatos , Gravidez , Espécies Reativas de Oxigênio , Receptores de Leucotrienos/genética , Sulfonamidas , Transferrinas , Água
8.
Sci Rep ; 12(1): 3890, 2022 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-35273234

RESUMO

The new outbreak of coronavirus disease 2019 (COVID-19) has infected and caused the death of millions of people worldwide. Intensive efforts are underway around the world to establish effective treatments. Immunoglobulin from immunized animals or plasma from convalescent patients might constitute a specific treatment to guarantee the neutralization of the virus in the early stages of infection, especially in patients with risk factors and a high probability of progressing to severe disease. Worldwide, a few clinical trials using anti-SARS-CoV-2 immunoglobulins from horses immunized with the entire spike protein or fragments of it in the treatment of patients with COVID-19 are underway. Here, we describe the development of an anti-SARS-CoV-2 equine F(ab')2 immunoglobulin using a newly developed SARS-CoV-2 viral antigen that was purified and inactivated by radiation. Cell-based and preclinical assays showed that the F(ab')2 immunoglobulin successfully neutralizes the virus, is safe in animal models, and reduces the severity of the disease in a hamster model of SARS-CoV-2 infection and disease.


Assuntos
COVID-19/terapia , Imunoglobulinas/uso terapêutico , Receptores Imunológicos/uso terapêutico , SARS-CoV-2/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Feminino , Cavalos/imunologia , Humanos , Imunoglobulinas/imunologia , Imunoglobulinas/isolamento & purificação , Masculino , Mesocricetus/imunologia , Plasmaferese/veterinária , Receptores Imunológicos/imunologia
9.
J Sci Food Agric ; 102(10): 4287-4295, 2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-35038166

RESUMO

BACKGROUND: Astaxanthin, classified as a xanthophyll, has antioxidant properties about 500 times greater than α-tocopherols and ten times greater than ß-carotenes. Based on the antioxidant activity of this carotenoid, this study aimed to evaluate the shelf-life of tilapia fillets (Oreochromis niloticus) fed with astaxanthin, by determining the microbiological quality (colimetry, counts of mesophilic and psychrotrophic microorganisms), physicochemical analyses (colorimetry, pH, thiobarbituric acid reactive substances (TBARS)) and sensory analysis. RESULTS: Tilapia supplemented with astaxanthin presented a reduction in the counts of microorganisms (mesophiles and psychrotrophics) and lower lipid oxidation index (TBARS), when compared to fillets of control fish. Colorimetric changes of fillet degradation were observed, associated with increased pH during storage, as well as loss of brightness and texture in addition to worsening of appearance and odor. These deteriorating changes were minimized using astaxanthin. CONCLUSION: Our results demonstrate the beneficial performance of astaxanthin in the shelf-life of tilapia fillets stored under refrigeration. Therefore, dietary supplementation with astaxanthin (100 and 200 mg kg-1 of feed) improves the microbiological and physicochemical quality of tilapia fillets during 50 days of shelf-life. © 2022 Society of Chemical Industry.


Assuntos
Tilápia , Animais , Conservação de Alimentos/métodos , Refrigeração , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Xantofilas/análise
10.
Sci Total Environ ; 813: 152345, 2022 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-34942250

RESUMO

Despite the significant increase in the generation of SARS-CoV-2 contaminated domestic and hospital wastewater, little is known about the ecotoxicological effects of the virus or its structural components in freshwater vertebrates. In this context, this study evaluated the deleterious effects caused by SARS-CoV-2 Spike protein on the health of Danio rerio, zebrafish. We demonstrated, for the first time, that zebrafish injected with fragment 16 to 165 (rSpike), which corresponds to the N-terminal portion of the protein, presented mortalities and adverse effects on liver, kidney, ovary and brain tissues. The conserved genetic homology between zebrafish and humans might be one of the reasons for the intense toxic effects followed inflammatory reaction from the immune system of zebrafish to rSpike which provoked damage to organs in a similar pattern as happen in severe cases of COVID-19 in humans, and, resulted in 78,6% of survival rate in female adults during the first seven days. The application of spike protein in zebrafish was highly toxic that is suitable for future studies to gather valuable information about ecotoxicological impacts, as well as vaccine responses and therapeutic approaches in human medicine. Therefore, besides representing an important tool to assess the harmful effects of SARS-CoV-2 in the aquatic environment, we present the zebrafish as an animal model for translational COVID-19 research.


Assuntos
COVID-19 , Glicoproteína da Espícula de Coronavírus , Animais , Feminino , Humanos , SARS-CoV-2 , Peixe-Zebra
11.
Aquac Res, v. 7, n. 53, p.2930-2936, mar. 2022
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4265

RESUMO

Aeromonas hydrophila is an opportunistic pathogen frequently isolated from outbreaks with higher mortality rates in tropical farms, causing disease in pacus Piaractus mesopotamicus. The objective of this study was to obtain a purified polyclonal immunoglobulin Y (IgY) by immunizing laying White Leghorn hens with A. hydrophila. Antibodies anti-A. hydrophila derived from the yolks and serum of hens showed antibody titres increased at 60 days and remained detectable until 90 days after immunization. Pacus were challenged intraperitoneally with A. hydrophila and the capacity of antibodies to detect A. hydrophila was tested by immunohistochemistry using tissues from pacus. The results observed in this study demonstrated the utility of IgY for the diagnosis of A. hydrophila within the phagosomes of phagocytes and extracellular bacteria colonizing host tissues in pacus.

12.
Sci Rep, v. 12, 3890, mar. 2022
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4257

RESUMO

The new outbreak of coronavirus disease 2019 (COVID-19) has infected and caused the death of millions of people worldwide. Intensive efforts are underway around the world to establish effective treatments. Immunoglobulin from immunized animals or plasma from convalescent patients might constitute a specific treatment to guarantee the neutralization of the virus in the early stages of infection, especially in patients with risk factors and a high probability of progressing to severe disease. Worldwide, a few clinical trials using anti-SARS-CoV-2 immunoglobulins from horses immunized with the entire spike protein or fragments of it in the treatment of patients with COVID-19 are underway. Here, we describe the development of an anti-SARS-CoV-2 equine F(ab′)2 immunoglobulin using a newly developed SARS-CoV-2 viral antigen that was purified and inactivated by radiation. Cell-based and preclinical assays showed that the F(ab′)2 immunoglobulin successfully neutralizes the virus, is safe in animal models, and reduces the severity of the disease in a hamster model of SARS-CoV-2 infection and disease.

13.
J Sci Food Agric, v. 102, n. 10, p. 4287-4295, ago. 2022
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4099

RESUMO

Background: Astaxanthin, classified as a xanthophyll, has antioxidant properties about 500 times greater than α-tocopherols and 10 times greater than β-carotenes. Based on the antioxidant activity of this carotenoid, this study aimed to evaluate the shelf-life of tilapia fillets (Oreochromis niloticus) fed with astaxanthin, by determining the microbiological quality (colimetry, counts of mesophilic and psychrotrophic microorganisms), physicochemical analyzes (colorimetry, pH, TBARs) and sensory analysis. Results: Tilapia supplemented with astaxanthin presented reduction in the counts of microorganisms (mesophiles and psychrotrophics) and lower lipid oxidation index (TBARs), when compared to fillets of control fish. Colorimetric changes of fillet degradation were observed, associated with increased pH during storage, as well as loss of brightness and texture in addition to worsening in appearance and odor. These deteriorating changes were minimized using astaxanthin. Conclusion: Our results demonstrate the beneficial performance of astaxanthin in the shelf-life of tilapia fillets stored under refrigeration. Therefore, dietary supplementation with astaxanthin (100 and 200mg kg-1 of feed) improves the microbiological and physicochemical quality of tilapia fillets during 50 days of shelf-life.

14.
Sci Total Environ ; 813: 152345, 2022.
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4093

RESUMO

Despite the significant increase in the generation of SARS-CoV-2 contaminated domestic and hospital wastewater, little is known about the ecotoxicological effects of the virus or its structural components in freshwater vertebrates. In this context, this study evaluated the deleterious effects caused by SARS-CoV-2 Spike protein on the health of Danio rerio, zebrafish. We demonstrated, for the first time, that zebrafish injected with fragment 16 to 165 (rSpike), which corresponds to the N-terminal portion of the protein, presented mortalities and adverse effects on liver, kidney, ovary and brain tissues. The conserved genetic homology between zebrafish and humans might be one of the reasons for the intense toxic effects followed inflammatory reaction from the immune system of zebrafish to rSpike which provoked damage to organs in a similar pattern as happen in severe cases of COVID-19 in humans, and, resulted in 78,6% of survival rate in female adults during the first seven days. The application of spike protein in zebrafish was highly toxic that is suitable for future studies to gather valuable information about ecotoxicological impacts, as well as vaccine responses and therapeutic approaches in human medicine. Therefore, besides representing an important tool to assess the harmful effects of SARS-CoV-2 in the aquatic environment, we present the zebrafish as an animal model for translational COVID-19 research.

15.
Fish Shellfish Immunol ; 118: 34-50, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34464686

RESUMO

Group B Streptococcus (GBS) causes meningitis in neonates and Nile tilapia (Oreochromis niloticus). The molecular mechanisms regulating the intracellular survival of this pathogen in the host cell are complex and crucial for the progression of infection. Thus, we propose the use of GBS-infected Nile tilapia microglia as an in vitro model system simulating infection caused by homologous bacteria in humans. We used this model to evaluate the phagocytic activity, as well as the functional aspects of the capsular proteins A, B, C, and D and the major redox enzymes, and the synergistic role of mechanisms/proteins involved in blocking phagocytic process. We observed that in the intracellular phase, GBS showed enhanced synthesis of the polysaccharide capsule and used superoxide dismutase, thioredoxin, NADH oxidase, and alkyl hydroperoxide reductase to scavenge reactive oxygen species and reactive nitrogen species produced by the host cell. Furthermore, although these virulence mechanisms were effective during the initial hours of infection, they were not able to subvert microglial responses, which partially neutralized the infection. Altogether, our findings provided important information regarding the intracellular survival mechanisms of GBS and perspectives for the production of new drugs and vaccines, through the druggability analysis of specific proteins. In conclusion, tilapia microglia serve as a potent in vitro experimental model for the study of meningitis.


Assuntos
Ciclídeos , Doenças dos Peixes , Infecções Estreptocócicas , Animais , Doenças dos Peixes/microbiologia , Microglia , Oxirredução , Proteômica , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae
16.
Fish Shellfish Immunol ; 115: 134-141, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34098067

RESUMO

Streptococcosis causes great economic losses in intensive culture of tilapia. Vaccination is the most effective and safest way to tackle infectious diseases. Thus, this study sought the more effective and safer antigenic fraction after sonication of Streptococcus agalactiae to elaborate a vaccine against streptococcosis in Nile tilapia. For this, twenty-one days after vaccination with different fractions (soluble and insoluble) of S. agalactiae, the fish were challenged with the homologous strain (LD50). Then, samples were taken at zero, 14, 28, 60 and 90 days post-vaccination (DPV, n = 7). Blood and organs (cranial kidney, spleen and liver) were collected from vaccinated and unvaccinated fish. Finally, insoluble fraction vaccine presented the best effect, resulting in a 100% relative percent of survival (RPS) and without clinical manifestations. In view of the results, it was to evaluate the role of the insoluble fraction of the antigen in the protective immunity against streptococcosis. The results indicate that the spleen might be the main organ in the vaccine response in Nile tilapia due to the great morphological and immunological differences in vaccinated fish, evidenced by the greater of melanomacrophage centers (MMC) and IgM + lymphocytes in relation to the non-vaccinated fish. At 60 DPV, it was observed the peak of the protective immunity related to the maximum concentration of proteins, circulating leukocytes, antibody titers in the serum and tissue changes with greater expression of IgM + and MMC number in the spleen and kidney of Oreochromis niloticus. Vaccination with insoluble fraction of S. agalactiae was safe and provided effective protection against streptococcosis with maximum protective response at 60 DPV.


Assuntos
Antígenos de Bactérias/administração & dosagem , Ciclídeos/imunologia , Doenças dos Peixes/imunologia , Imunogenicidade da Vacina , Vacinas Estreptocócicas/administração & dosagem , Streptococcus agalactiae/imunologia , Vacinação/veterinária , Animais , Sonicação/veterinária , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/veterinária
17.
ARS Vet, v. 37, n. 2, 067-073, jun. 2021
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4879

RESUMO

Little is known about the toxicity of immune modulators in fish. Zafirlukast is an anti-inflammatory that antagonizes cysteine leukotriene receptors (CysLTR1). Aiming to study immunomodulatory treatments on fish health, this study evaluated the clinical safety of oral zafirlukast treatment, through biochemical and hematological analyzes during acute inflammatory reaction in Nile tilapia (Oreochromis niloticus), induced by Aeromonas hydrophilabacterins. 72 young tilapias were randomly divided in 9 aquariums (100 L each, n=8) to compose the following treatments: T0 (control), T1 (Treatment with 250 μg zafirlukast) and T2 (Treatment with 500 μg zafirlukast). Eight animals were evaluated per treatment in three periods: six, 24 and 48 hours post-inoculation (HPI), blood collection was performed for hematological and serum biochemical evaluation. The study of hepatic and renal functionality revealed that treatment with both doses of zafirlukast did not result in changes in the circulating values of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, creatinine, triglycerides, cholesterol, and total protein, suggesting that the drug has not presented hepatotoxicity, as well as compromised liver and kidney functions. Tilapia submitted to treatment with 500 μg showed adverse hematological effects characterizedby polycythemia associated with microcytosis. Therefore, oral treatment with zafirlukast has demonstrated clinical safety at a therapeutic dose of 250 μg in tilapia during acute aerocystitis, although hematological changes were observed in tilapia treatedwith overdose of this leukotriene blocker.


Pouco se sabe sobre a toxicidade de imunomoduladores em peixes. Zafirlukast é um anti-inflamatório que antagoniza os receptores de leucotrienos cisteínicos (CysLTR1). Com o objetivo de estudar o efeito de tratamentos imunomoduladores sobre a saúde dos peixes, este estudo avaliou a segurança clínica do tratamento com zafirlucaste oral, por meio de análises bioquímicas e hematológicas durante reação inflamatória aguda em tilápia do Nilo (Oreochromis niloticus), induzida por bacterinas de Aeromonas hydrophila.Para tal, 72 tilápias jovens foram divididas aleatoriamente em 9 aquários (100 L cada, n=8) para compor os seguintes tratamentos: T0 (controle), T1 (Tratamento com 250 μg de zafirlucaste) e T2 (Tratamento com 500 μg de zafirlucaste). Oito animais foram avaliados por tratamento em três períodos: seis, 24 e 48 horas pós-inoculação (HPI), foi realizada coleta de sangue para avaliação hematológica e bioquímica sérica. O estudo da funcionalidade hepática e renal revelou que o tratamento com ambas as doses de zafirlucaste não resultou em alterações nos valores circulantes de aspartato aminotransferase (AST), alanina aminotransferase (ALT), fosfatase alcalina, creatinina, triglicerídeos, colesterol e proteína total, sugerindo que a droga não comprometeu as funçõeshepáticas e renais. As tilápias tratadas com 500 μg apresentaram efeitos hematológicos adversos caracterizados por policitemia associada a microcitose. Portanto, o tratamento oral com zafirlucaste demonstrou segurança clínica nadose de 250 μg em tilápiasdurante aerocistite aguda, embora alterações hematológicas tenham sido observadas em tilápias tratadas com sobredosagem deste bloqueador de leucotrieno.

18.
Biomed Pharmacother, v. 144, 112310, dez. 2021
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-3975

RESUMO

This study aims to demonstrate the applicability and importance of zebrafish (Danio rerio) model to study acute and chronic inflammatory responses induced by different stimuli: carrageenan phlogogen (nonimmune); acute infection by bacteria (immune); foreign body reaction (chronic inflammation by round glass coverslip implantation); reaction induced by xenotransplantation. In addition to the advantages of presenting low breeding cost, high prolificity, transparent embryos, high number of individuals belonging to the same spawning and high genetic similarity that favor translational responses to vertebrate organisms like humans, zebrafish proved to be an excellent tool, allowing the evaluation of edema formation, accumulation of inflammatory cells in the exudate, mediators, signaling pathways, gene expression and production of specific proteins. Our studies demonstrated the versatility of fish models to investigate the inflammatory response and its pathophysiology, essential for the successful development of studies to discover innovative pharmacological strategies.

19.
Fish Shellfish Immunol, v. 118, p. 34-50, nov. 2021
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-3934

RESUMO

Group B Streptococcus (GBS) causes meningitis in neonates and Nile tilapia (Oreochromis niloticus). The molecular mechanisms regulating the intracellular survival of this pathogen in the host cell are complex and crucial for the progression of infection. Thus, we propose the use of GBS-infected Nile tilapia microglia as an in vitro model system simulating infection caused by homologous bacteria in humans. We used this model to evaluate the phagocytic activity, as well as the functional aspects of the capsular proteins A, B, C, and D and the major redox enzymes, and the synergistic role of mechanisms/proteins involved in blocking phagocytic process. We observed that in the intracellular phase, GBS showed enhanced synthesis of the polysaccharide capsule and used superoxide dismutase, thioredoxin, NADH oxidase, and alkyl hydroperoxide reductase to scavenge reactive oxygen species and reactive nitrogen species produced by the host cell. Furthermore, although these virulence mechanisms were effective during the initial hours of infection, they were not able to subvert microglial responses, which partially neutralized the infection. Altogether, our findings provided important information regarding the intracellular survival mechanisms of GBS and perspectives for the production of new drugs and vaccines, through the druggability analysis of specific proteins. In conclusion, tilapia microglia serve as a potent in vitro experimental model for the study of meningitis.

20.
Fish Shellfish Immunol, v. 148-153, p. 148-153, abr. 2021
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-3664

RESUMO

The mechanism of extracellular traps (ETs) is important in the cellular response against bacteria. Thus, in the present study, we describe for the first time the capacity of the Nile tilapia (Oreochromis niloticus) microglia in the formation of ETs in Weissella cibaria in vitro infection. Thus, we evaluated the ultrastructure of the microglia culture and observed the formation of ETs 6 hours after stimulation with lipopolysaccharide (LPS) and during the course of infection. Our results shed light on the mechanism of formation of ETs in the microglia of teleost fish and the ability of W. cibaria to infect these cells.

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