Effect of Bacillus Subtilis on Immune Function of Hd11 Chicken Macrophages

Poultry is frequently contaminated by Salmonella, a pathogen leading to human health concern worldwide. This study aimed to evaluate the effect of Bacillus subtilis (BS)strain 048 (BS048) on the activation, phagocytosis, sterilization, cytokine secretion, and nitrogen oxide synthesis of HD11 chicken macrophages subjected to Salmonella enteritidis challenge, using lipopolysaccharide treatment as a negative control. The results showed: (1) BS048 had no significant effect on extracellular lactate dehydrogenase activity (p>0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p >0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p 0.05);(2)BS048 significantly upregulated the expression levels of pro-inflammatory cytokines (interleukin (IL)- 1â and IL-6), anti-inflammatory cytokines(IL-10 and transforming growth factor-â1), and anti-viral cytokine, interferon-â (p<0.01); ; (3) BS048 significantly upregulated the mRNA expression level of the inducible nitric oxide synthase and its activity as well as extracellular nitrogen oxide level (p <0.01). In conclusion, BS048 could improve antiinflammatory and immune functions of HD11 chicken macrophages, without cytotoxic effects on these cells.(AU)

Bisacurone gel ameliorated burn wounds in experimental rats via its anti-inflammatory, antioxidant, and angiogenic properties

Purpose: To investigate putative mechanism of wound healing for chitosan-based bisacurone gel against secondary burn wounds in rats. Methods: A second-degree burn wound with an open flame using mixed fuel (2 mL, 20 seconds) was induced in Sprague Dawley rats (male, 180-220 g, n = 15, each) followed by topical treatments with either vehicle control (white petroleum gel, 1%), silver sulfadiazine (1%) or bisacurone gel (2.5, 5, or 10%) for 20 days. Wound contraction rate and paw withdrawal threshold were monitored on various days. Oxidative stress (superoxide dismutase, glutathione, malondialdehyde, and nitric oxide), pro-inflammatory cytokines (tumour necrosis factor-alpha, interleukins by enzyme-linked immunosorbent assay), growth factors (transforming growth factor-ß, vascular endothelial growth factor C using real time polymerase chain reaction and Western blot assay) levels, and histology of wound skin were assessed at the end. Results: Bisacurone gel showed 98.72% drug release with a 420.90­442.70 cps viscosity. Bisacurone gel (5 and 10%) significantly (p < 0.05) improved wound contraction rate and paw withdrawal threshold. Bisacurone gel attenuated oxidative stress, pro-inflammatory cytokines, and water content. It also enhanced angiogenesis (hydroxyproline and growth factor) and granulation in wound tissue than vehicle control. Conclusions: These findings suggested that bisacurone gel can be a potential candidate to treat burn wounds via its anti-inflammatory, antioxidant, and angiogenic properties.

The in vitro effects of n-3 fatty acids on immune response regulation of bovine ex vivo endometrial explants / Os efeitos in vitro dos ácidos graxos n-3 na regulação da resposta imune de explantes endometriais bovinos ex vivo

Fatty acids are considered metabolic intermediaries, although new facts indicate they also work as signaling molecules with different roles in the immune response. Based on that, in this study, we investigated the anti-inflammatory effects of n-3 polyunsaturated fatty acids (PUFAs) as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and α-linolenic acid (LNA) in ex vivo bovine endometrial explants. For this, two groups were formed: (1) LPS-challenged and (2) control, both to evaluate the accumulation of proinflammatory cytokines as interleukin 1ß (IL1B) and interleukin 6 (IL6). To develop the study, bovine female reproductive tracts from non-pregnant Angus heifers without evidence of reproductive diseases were selected. Endometrial explants were processed and treated for 24 h with EPA, DHA, and LNA in five different concentrations (0µM, 50µM, 100 µM, 200µM and 400 µM) and then, challenged with LPS for 24 h. Supernatants were collected to evaluate the concentration of IL1B and IL6 by ELISA. Explants treated with EPA from control groups reduced the concentrations of ILB (200µM) and IL6 (400 µM), and IL6 (50 µM; 100 µM) from the LPS-challenged group. DHA decreased the accumulation of IL1B and IL6 at 200 µM on explants from the LPS-challenged group, and 200 µM reduced IL6 from the control group. In contrast, explants treated with LNA only reduced the accumulation of IL1B to 400µM (from both groups). In conclusion, the EPA acid is the best anti-inflammatory option to decrease the concentration of both pro-inflammatory cytokines (IL1B and IL6) from LPS-challenged and control groups in bovine endometrial explants; while LNA evidence to be the last option to promote an anti-inflammatory response.(AU)

Os ácidos graxos são considerados intermediários metabólicos, embora novos fatos indiquem que eles também atuem como moléculas sinalizadoras com diferentes papéis na resposta imune. Dessa forma, este estudo investigou os efeitos anti-inflamatórios de ácidos graxos poliinsaturados n-3 (PUFAs) como ácido eicosapentaenóico (EPA), ácido docosahexaenóico (DHA) e ácido α-linolênico (LNA) em explantes endometriais ex vivo de bovinos. Para tal, o experimento foi divido em dois grupos: (1) Desafiado-LPS e (2) Controle, para que então pudesse avaliar o acúmulo de citocinas pró-inflamatórias como interleucina 1ß (IL1B) e interleucina 6 (IL6). Foram selecionados tratos reprodutivos de fêmeas bovinas de novilhas Angus não prenhes sem evidência de doenças reprodutivas. Explantes endometriais foram processados e tratados por 24h com EPA, DHA e LNA em cinco concentrações diferentes (0µM, 50µM, 100 µM, 200µM e 400 µM) e, em seguida, desafiados com LPS por mais 24h. Os sobrenadantes foram colhidos para avaliar a concentração de IL1B e IL6 pelo teste de ELISA. Os explantes tratados com EPA dos grupos de controle reduziram as concentrações de ILB (200 µM) e IL6 (400 µM) e no grupo desafiado com LPS houve redução das concentrações de IL6 (50 µM; 100 µM). Nos explantes do grupo desafiado com LPS, o DHA diminuiu o acúmulo de IL1B e IL6 nas concentrações de 200 µM, e no grupo controle reduziu IL6 nas concentrações de 200 µM, enquanto os explantes tratados com LNA reduziram apenas o acúmulo de IL1B a 400 µM (de ambos os grupos). Em conclusão, o ácido EPA provou ser a melhor opção anti-inflamatória para diminuir a concentração de ambas as citocinas pró-inflamatórias (IL1B e IL6) de grupos desafiados com LPS e controle em explantes endometriais bovinos; enquanto o LNA evidencia ser a opção menos viável para promover uma resposta anti-inflamatória.(AU)

Propolis anti-inflammatory effects on MAGE-1 and retinoic acid-treated dendritic cells and on Th1 and T regulatory cells

Background: Propolis exhibits huge potential in the pharmaceutical industry. In the present study, its effects were investigated on dendritic cells (DCs) stimulated with a tumor antigen (MAGE-1) and retinoic acid (RA) and on T lymphocytes to observe a possible differential activation of T lymphocytes, driving preferentially to Th1 or Treg cells. Methods: Cell viability, lymphocyte proliferation, gene expression (T-bet and FoxP3), and cytokine production by DCs (TNF-α, IL-10, IL-6 and IL-1ß) and lymphocytes (IFN-γ and TGF-ß) were analyzed. Results: MAGE-1 and RA alone or in combination with propolis inhibited TNF-α production and induced a higher lymphoproliferation compared to control, while MAGE1 + propolis induced IL-6 production. Propolis in combination with RA induced FoxP3 expression. MAGE-1 induced IFN-γ production while propolis inhibited it, returning to basal levels. RA inhibited TGF-ß production, what was counteracted by propolis. Conclusion: Propolis affected immunological parameters inhibiting pro-inflammatory cytokines and favoring the regulatory profile, opening perspectives for the control of inflammatory conditions.(AU)

Biochemical and immunohistochemical examination of the effects of ephedrine in rat ovary tissue

Purpose: It was aimed to investigate the biochemical and immunohistochemical effects of ephedrine (EPH) in bilateral ovariectomized rats. Methods: Twenty-four Sprague Dawley female rats were divided into three groups: control group: The abdomen was opened and closed without any treatment; ischemia-reperfusion (IR) group: 2 h of ischemia followed by 2 h of reperfusion were allowed to cause IR injury; IR+EPH group: oral EPH solution (5 mg/kg) was administered for 28 days. Results: Biochemical parameters were statistically significant in group comparisons. Increased interleukin-6 (IL-6) expression, degenerative preantral and antral follicle cells and inflammatory cells around blood vessels were seen in IR group. Negative IL-6 expression was observed in seminal epithelial cells, preantral and antral follicle cells in IR+EPH group. While caspase-3 activity increased in granulosa cells and stromal cells in IR group, caspase-3 expression was negative in preantral and antral follicle cells in the germinal epithelium and cortex in IR+EPH group. Conclusion: The effect of apoptosis, which occurs with the signaling that starts in the cell nucleus, caused the cessation of the stimulating effect at the nuclear level after EPH administration, and a decrease in the antioxidative effect in IR damage and inflammation in the apoptotic process.

Rodent models in placental research. Implications for fetal origins of adult disease

Rodent models in rats, mice, and guinea pigs have been extremely helpful to gain insight into pregnancy physiology and pathologies-related. Moreover, they have allowed understanding the mechanism that links an adverse intrauterine environment with the origin of adult disease. In this regard, the effects of diverse maternal conditions, such as undernutrition, obesity, hypoxia, and hyperandrogenism on placental function and its long-term consequences for the offspring, have been widely analyzed through rodents models involving dietary manipulations, modifications in environmental oxygen, surgical and pharmacological procedures that reduce uteroplacental blood flow and administrations of exogenous testosterone and dihydrotestosterone (DHT) mimicking maternal androgen excess. Both in human and in rodent models, these interventions induce modifications of placental morphology, transport of glucose, amino acid, and fatty acids, steroid synthesis, and signaling pathways control placental function. These changes are associated with the increase of pro-inflammatory and oxidative stress markers. For its part, offspring exhibit alterations in organs involved in metabolic control such as the hypothalamus, adipose tissue, liver, skeletal muscle, and pancreas altering the intake and preferences for certain foods, the metabolism of glucose and lipid, and hormonal function leading to fat accumulation, insulin resistance, fatty liver, dyslipidemia, and elevated glucose levels. Therefore, the present review discusses the evidence emerging from rodent models that relate maternal nutrition, hypoxia, and androgen exposure to the maternal mechanisms that lead to fetal programming and their metabolic consequences in postnatal life.(AU)

Treatment of colitis by oral negatively charged nanostructured curcumin in rats

Purpose: To examine the effects of a negatively charged nanostructured curcumin microemulsion in experimental ulcerative colitis (UC) in rats. Methods: Four percent acetic acid was used to induce UC. The animals were treated for seven days and randomly assigned to four groups: normal control (NC), colitis/normal saline (COL/NS), colitis/curcumin (COL/CUR), and colitis/mesalazine (COL/MES). The nanostructured curcumin was formulated with a negative zeta potential (-16.70 ± 1.66 mV). Dosage of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin 1-ß (IL-1ß), interleukin 6 (IL-6), and antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase), macro and microscopic evaluation of the colon tissue were analyzed. Results: The COL/CUR group had a higher level of antioxidant enzymes compared to the COL/MESgroup. The levels of TNF-α, IL-1ß and IL-6 were significantly lower in the colonic tissue of the COL/CUR group rats, when compared to the COL/NS and COL/MES groups (p < 0.001). The presence of ulcers in the colonic mucosa in rats of the COL/NSgroup was significantly higher than in the COL/MES group (p < 0.001). In the NC and COL/CUR groups, there were no ulcers in the colonic mucosa. Conclusions: The nanostructured microemulsion of curcumin, used orally, positively influenced the results of the treatment of UC in rats. The data also suggests that nanostructured curcumin with negative zeta potential is a promising phytopharmaceutical oral delivery system for UC therapy. Further research needs to be done to better understand the mechanisms of the negatively charged nanostructured curcumin microemulsion in UC therapy.

The ethanol extract of Periplaneta Americana L. improves ulcerative colitis induced by a combination of chronic stress and TNBS in rats

Purpose: To investigate the effects of Periplaneta americana L. on ulcerative colitis (UC) induced by a combination of chronic stress (CS) and 2,4,6-trinitrobenzene sulfonic acid enema (TNBS) in rats. Methods: The experiment UC model with CS was established in rats by a combination of chronic restraint stress, excess failure, improper, and TNBS. The body weight, disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS) and pro-inflammatory mediators were measured. The content of corticotropin-releasing hormone (CRH) in hypothalamus or adrenocorticotropic hormone (ACTH) and corticosteroids (CORT) in plasma were evaluated by enzyme-linked immunosorbent assay. The proportion of T lymphocyte subsets was detected by flow cytometry, and gut microbiota was detected by 16S rDNA amplicon sequencing. Results: Weight loss, DAI, CMDI, HS and proinflammatory mediators were reversed in rats by P. americana L. treatment after UC with CS. Increased epidermal growth factor (EGF) was observed in P. americana L. groups. In addition, P. americana L. could reduce the content of CRH and ACTH and regulate the ratio of CD3+, CD3+CD8+ and CD3+CD4+CD25+/CD4+ in spleen. Comparably, P. americana L. changes composition of gut microbiota. Conclusions: The ethanol extract of Periplaneta Americana L. improves UC induced by a combination of CS and TNBS in rats.

Resposta metabólica ao trauma cirúrgico / Metabolic response to surgical trauma

Os efeitos do trauma cirúrgico no organismo, os quais são amplamente estudados pela medicina humana e veterinária, resultam em uma complexa resposta neuroendócrina e imunológica, desencadeadas para restabelecer a homeostase e preservar a vida. A resposta metabólica ao trauma é caracterizada pelo aumento da secreção de hormônios hipofisários, ativação do sistema nervoso simpático e secreção de citocinas pró-inflamatórias. Essa resposta tem por objetivo manter a perfusão tecidual, controlar a proliferação de microrganismos e mobilizar substratos para garantir a integridade das funções celulares e promover reparação tecidual. Diferentes fatores podem influenciar a intensidade da resposta do organismo ao trauma, incluindo magnitude e duração dos estímulos, enfermidades ou eventos traumáticos pré-existentes, protocolos anestésicos e técnicas cirúrgicas utilizadas. Respostas metabólicas exacerbadas ocasionam complicações pós-operatórias, tempo prolongado de recuperação dos pacientes e óbito. Através da identificação dos principais eventos relacionados à resposta metabólica ao trauma cirúrgico, é possível atuar de forma a reduzir os danos associados, melhorando a qualidade e minimizando os riscos do período pós-operatório. Sendo assim, este trabalho tem como objetivo apresentar uma revisão descrevendo a fisiopatologia e as consequências da resposta metabólica ao trauma cirúrgico, em suas diferentes formas e origens, no período perioperatório.

The effects of surgical trauma on the organism, which are widely studied by human and veterinary medicine, result in a complex neuroendocrine and immune responses triggered to restore homeostasis and preserve life. The metabolic response to trauma is characterized by increased secretion of hypophyseal hormones, activation of the sympathetic nervous system, and secretion of pro-inflammatory cytokines. This response aims to maintain tissue perfusion, control the proliferation of microorganisms, and mobilize substrates to ensure the integrity of cellular functions and promote tissue repair. Different factors can influence the intensity of the organism's response to trauma, including magnitude and duration of stimuli, pre-existing infirmities or traumas, anaesthetic protocols, and surgical techniques applied. Exacerbated metabolic responses can cause post-operative complications, prolonged recovery time, and death. By identifying the main events related to the metabolic response to surgical trauma, it is possible to act to reduce the associated damages, improving quality and minimizing the risks of the postoperative period. Thus, the present paper aims to present a review describing the physiopathology and the consequences of the metabolic response to surgical trauma, in its different forms and origins, in the perioperative period.

Tarantula cubensis extract (Theranekron®) inhibits inflammation in carrageenan-induced acute paw edema in rats / [Extrato de tarântula cubensis (Theranekron®) inibe a inflamação no edema induzido por algas vermelhas na pata de ratos]

The aim of this study was to investigate the anti-inflammatory effect of alcoholic extract of Tarantula cubensis alcoholic extract (TCAE) in experimentally induced inflammation in rats. Fifty-four adult Sprague-Dawley male rats were randomly divided into nine groups. Paw edema was induced by 0.2mL subplantar (s.p.) injection of 1% carrageenan (CAR) into the right hind paw. Rats were treated with the nonsteroidal anti-inflammatory drug (NSAID) indomethacin (INDO) (10mg/kg, p.o.) or TCAE at different doses (1, 10 or 100µg/kg) injected s.c. for systemic or s.p. for local anti-inflammatory effect. Saline was used as control. Changes in paw thickness, volume, and weight were calculated as percentages. Formalin-fixed paws were used for histopathological examination. We detected that TCAE applied s.c. at 10µg/kg and 100µg/kg doses resulted in thinner paw thickness, lower paw volume, and lower paw weights four hours after the induction of inflammation when compared with the INDO group (p<0.05). The paw edema inhibitory effect of TCAE applied at a dose of 10µg/kg, s.c. was 68% when compared with the INDO which had an inhibitory effect of 56%. These results were verified with similar histopathological findings. The anti-inflammatory feature of 10µg/kg of TCAE given systematically was similar to the effects of INDO. Our results suggest that TCAE has anti-inflammatory effects by reducing edema and decreasing inflammatory reaction. These results may be attributed to the inhibition of the production of proinflammatory mediators. Thus, TCAE may be considered as a potential anti-inflammatory agent for treating acute inflammatory conditions.

O objetivo deste estudo foi investigar o efeito anti-inflamatório do extrato alcoólico de Tarantula cubensis (TCAE) na inflamação induzida experimentalmente em ratos. Cinqüenta e quatro ratos Sprague-Dawley adultos machos foram divididos aleatoriamente em nove grupos. O edema da pata foi induzido pela injeção de 0,2mL de subplantar (s.p.) de 1% de carragena (CAR) na pata traseira direita. Ratos foram tratados com o medicamento antiinflamatório não esteróide (NSAID) indometacina (INDO) (10mg/kg, p.o.) ou TCAE em doses diferentes (1, 10 ou 100µg/kg) injetado s.c. para efeito sistêmico ou s.p. para efeito antiinflamatório local. A soro fisiológico foi usado como controle. As mudanças na espessura da pata, volume e peso foram calculadas como porcentagens. As patas fixadas com fórmalina foram usadas para exame histopatológico. Detectamos que o TCAE aplicado s.c. em doses de 10µg/kg e 100µg/kg resultou em menor espessura da pata, menor volume da pata e menor peso da pata quatro horas após a indução da inflamação quando comparado com o grupo INDO (p<0,05). O efeito inibidor do edema da pata de TCAE aplicado na dose de 10µg/kg, s.c. foi de 68% quando comparado com o INDO que teve um efeito inibidor de 56%. Estes resultados foram verificados com resultados histopatológicos semelhantes. A característica anti-inflamatória de 10µg/kg de TCAE dada sistematicamente foi semelhante aos efeitos do INDO. Nossos resultados sugerem que o TCAE tem efeitos anti-inflamatórios reduzindo o edema e diminuindo a reação inflamatória. Estes resultados podem ser atribuídos à inibição da produção de mediadores pró-inflamatórios. Assim, o TCAE pode ser considerado como um agente antiinflamatório potencial para o tratamento de condições inflamatórias agudas.

The effects of valproic acid on skin healing: experimental study in rats

Purpose: To recognize the effects of valproic acid (VPA), an epigenetic drug, on the skin healing process. Methods: Sixty male Wistar rats were divided into two groups: the experiment treated with VPA (100 mg/kg/day); and the control, with 0.9% sodium chloride by gavage. Skin healing was studied in three moments (the third, the seventh, and the 14th day), evaluating the parameters: inflammatory reaction and its intensity (anti-LCA), angiogenesis (anti-CD34), collagen I and III (anti-collagen I, anti-collagen III and Picrosirius-red F3BA) and myofibroblasts (anti-alpha-AMS). Results: The inflammatory reaction was acute or sub-acute in both groups on the third day. On the seventh and the 14th day, chronic predominated in the control (p=0.006), and sub-acute in the experiment (p=0.020). There was a greater number of leukocytes in the group treated only on the third day (p=0.036). The number of vessels was lower in the treated group at the three times (p3=0.002, p7<0.001, and p14=0.027). Myofibroblasts were rare in the third day and moderate quantity in the remaining periods. Collagen I density was higher in the control at the three times (p<0.001) and collagen III in the treated group (p<0.001). Conclusions: VPA led to a more intense inflammatory reaction, decreased angiogenesis and collagen deposition, especially type I collagen.

Protective effects of melatonin in cisplatin-induced cardiac toxicity: possible role of BDNF-TNF-α signaling pathway

Purpose: The present study explored the role of melatonin in cisplatin-induced cardiac injury along with the possible role of brain-derived neurotrophic factor (BDNF) in melatonin-mediated effects. Methods: Wistar rats were administered cisplatin (10 mg/kg), and cardiac injury was assessed by measuring the levels of cardiac troponin (cTnT) and lactate dehydrogenase (LDH-1).The extent of apoptosis was measured by measuring caspase-3 (pro-apoptotic) and Bcl-2 (anti-apoptotic) in hearts. The levels of BDNF, tumour necrosis factor α (TNF-α) and reduced glutathione were measured in heart. Melatonin (5 and 10 mg/kg) was administered for 15 days, and the role of BDNF was identified by co-administering BDNF inhibitor, ANA-12 (0.25 and 0.5 mg/kg). Results: Melatonin attenuated cTnT and LDH-1 levels along with reduction in caspase-3 and increase in Bcl-2. It also increased cisplatin-induced decrease in BDNF, increase in TNF-α and decrease in reduced glutathione levels. Moreover, ANA-12 abolished the cardioprotective effects, anti-inflammatory and antioxidant effects of melatonin suggesting the role of BDNF in melatonin-mediated effects in cisplatin-induced cardiac injury. Conclusions: Melatonin is useful in cisplatin-induced cardiac injury, which may be due to an increase in BDNF, decrease in inflammation and increase in antioxidant activities.

Pharmacological screening of mucus produced by the pacamã fish, Lophiosilurus alexandri / Triagem farmacológica de muco produzido pelo pacamã, Lophiosilurus alexandri

Recently, fish byproducts have been appraised as a source of structurally diverse bioactive compounds. In this context, fish mucus has revealed a myriad of pharmacological activities. This study describes a new bioactive molecule release system composed of protein nanoparticles from pacamã (Lophiosilurus alexandri) mucus, an endemic fish to the São Francisco River (Brazil). The process of obtaining and applying these nanoparticles, composed of proteins present in the pacamã mucus, as biocompatible carriers of bioactive molecules is also addressed herein. Therefore, the external mucus of the fish was collected, lyophilized and hydrated in deionized water containing the template molecule. After pH adjustment to 7.5, followed by sonication and centrifugation, the formed nanoparticles were collected in the supernatant. Additionally, the mucus and the particles were characterized, and pharmacological effects were evaluated regarding their antifungal, antibacterial, anticancer, anti or pro-inflammatory and antinociceptive proprieties. NP-Mucus did not exhibit antibacterial activity against Pseudomonas aeruginosa or Staphylococcus aureus. However, it showed potential effects against Candida albicans with a minimum inhibitory concentration (MIC) of 1 µg/mL, and significant pro-inflammatory role, reflected by cellular recruitment activity and healing effects. Therefore, future studies are now needed in order to identify specific compounds in NP-mucus responsible for the observed effects in order to provide new pharmacological and therapeutic strategies.

Recentemente, os subprodutos de peixes foram avaliados como uma fonte de compostos bioativos estruturalmente diversos. Nesse contexto, o muco dos peixes revelou uma miríade de atividades farmacológicas. Este estudo descreve um novo sistema de liberação de moléculas bioativas compostas por nanopartículas de proteínas do muco do pacamã (Lophiosilurus alexandri), um peixe do Rio São Francisco (Brasil). O processo de obtenção e aplicação dessas nanopartículas, compostas por proteínas presentes no muco do pacamã, como carreadores biocompatíveis de moléculas bioativas também são abordados neste artigo. Para tanto, o muco externo dos peixes foi coletado, liofilizado e hidratado em água deionizada contendo a molécula template. Após ajuste do pH para 7,5, seguido de sonicação e centrifugação, as nanopartículas formadas foram coletadas no sobrenadante. Adicionalmente, foram caracterizados o muco e as partículas, sendo avaliados os efeitos farmacológicos quanto às propriedades antifúngicas, antibacterianas, anticancerígenas, anti ou pró-inflamatórias e antinociceptivas. NP-Mucus não exibiu atividade antibacteriana contra Pseudomonas aeruginosa ou Staphylococcus aureus. No entanto, apresentou efeitos potenciais contra Candida albicans com concentração inibitória mínima (MIC) de 1 µg/mL, além de importante papel próinflamatório, refletido pela atividade de recrutamento celular e efeitos cicatrizantes. Portanto, estudos são necessários para identificar compostos no NP-muco responsáveis pelos efeitos observados, a fim de fornecer novas estratégias farmacológicas e terapêuticas.

Pharmacological screening of mucus produced by the pacamã fish, Lophiosilurus alexandri / Triagem farmacológica de muco produzido pelo pacamã, Lophiosilurus alexandri

Recently, fish byproducts have been appraised as a source of structurally diverse bioactive compounds. In this context, fish mucus has revealed a myriad of pharmacological activities. This study describes a new bioactive molecule release system composed of protein nanoparticles from pacamã (Lophiosilurus alexandri) mucus, an endemic fish to the São Francisco River (Brazil). The process of obtaining and applying these nanoparticles, composed of proteins present in the pacamã mucus, as biocompatible carriers of bioactive molecules is also addressed herein. Therefore, the external mucus of the fish was collected, lyophilized and hydrated in deionized water containing the template molecule. After pH adjustment to 7.5, followed by sonication and centrifugation, the formed nanoparticles were collected in the supernatant. Additionally, the mucus and the particles were characterized, and pharmacological effects were evaluated regarding their antifungal, antibacterial, anticancer, anti or pro-inflammatory and antinociceptive proprieties. NP-Mucus did not exhibit antibacterial activity against Pseudomonas aeruginosa or Staphylococcus aureus. However, it showed potential effects against Candida albicans with a minimum inhibitory concentration (MIC) of 1 µg/mL, and significant pro-inflammatory role, reflected by cellular recruitment activity and healing effects. Therefore, future studies are now needed in order to identify specific compounds in NP-mucus responsible for the observed effects in order to provide new pharmacological and therapeutic strategies.(AU)

Recentemente, os subprodutos de peixes foram avaliados como uma fonte de compostos bioativos estruturalmente diversos. Nesse contexto, o muco dos peixes revelou uma miríade de atividades farmacológicas. Este estudo descreve um novo sistema de liberação de moléculas bioativas compostas por nanopartículas de proteínas do muco do pacamã (Lophiosilurus alexandri), um peixe do Rio São Francisco (Brasil). O processo de obtenção e aplicação dessas nanopartículas, compostas por proteínas presentes no muco do pacamã, como carreadores biocompatíveis de moléculas bioativas também são abordados neste artigo. Para tanto, o muco externo dos peixes foi coletado, liofilizado e hidratado em água deionizada contendo a molécula template. Após ajuste do pH para 7,5, seguido de sonicação e centrifugação, as nanopartículas formadas foram coletadas no sobrenadante. Adicionalmente, foram caracterizados o muco e as partículas, sendo avaliados os efeitos farmacológicos quanto às propriedades antifúngicas, antibacterianas, anticancerígenas, anti ou pró-inflamatórias e antinociceptivas. NP-Mucus não exibiu atividade antibacteriana contra Pseudomonas aeruginosa ou Staphylococcus aureus. No entanto, apresentou efeitos potenciais contra Candida albicans com concentração inibitória mínima (MIC) de 1 µg/mL, além de importante papel próinflamatório, refletido pela atividade de recrutamento celular e efeitos cicatrizantes. Portanto, estudos são necessários para identificar compostos no NP-muco responsáveis pelos efeitos observados, a fim de fornecer novas estratégias farmacológicas e terapêuticas.(AU)

Effects of venoms on neutrophil respiratory burst: a major inflammatory function

Neutrophils play a pivotal role in innate immunity and in the inflammatory response. Neutrophils are very motile cells that are rapidly recruited to the inflammatory site as the body first line of defense. Their bactericidal activity is due to the release into the phagocytic vacuole, called phagosome, of several toxic molecules directed against microbes. Neutrophil stimulation induces release of this arsenal into the phagosome and induces the assembly at the membrane of subunits of the NAPDH oxidase, the enzyme responsible for the production of superoxide anion that gives rise to other reactive oxygen species (ROS), a process called respiratory burst. Altogether, they are responsible for the bactericidal activity of the neutrophils. Excessive activation of neutrophils can lead to extensive release of these toxic agents, inducing tissue injury and the inflammatory reaction. Envenomation, caused by different animal species (bees, wasps, scorpions, snakes etc.), is well known to induce a local and acute inflammatory reaction, characterized by recruitment and activation of leukocytes and the release of several inflammatory mediators, including prostaglandins and cytokines. Venoms contain several molecules such as enzymes (phospholipase A2, L-amino acid oxidase and proteases, among others) and peptides (disintegrins, mastoporan, parabutoporin etc.). These molecules are able to stimulate or inhibit ROS production by neutrophils. The present review article gives a general overview of the main neutrophil functions focusing on ROS production and summarizes how venoms and venom molecules can affect this function.(AU)

Effects of venoms on neutrophil respiratory burst: a major inflammatory function

Neutrophils play a pivotal role in innate immunity and in the inflammatory response. Neutrophils are very motile cells that are rapidly recruited to the inflammatory site as the body first line of defense. Their bactericidal activity is due to the release into the phagocytic vacuole, called phagosome, of several toxic molecules directed against microbes. Neutrophil stimulation induces release of this arsenal into the phagosome and induces the assembly at the membrane of subunits of the NAPDH oxidase, the enzyme responsible for the production of superoxide anion that gives rise to other reactive oxygen species (ROS), a process called respiratory burst. Altogether, they are responsible for the bactericidal activity of the neutrophils. Excessive activation of neutrophils can lead to extensive release of these toxic agents, inducing tissue injury and the inflammatory reaction. Envenomation, caused by different animal species (bees, wasps, scorpions, snakes etc.), is well known to induce a local and acute inflammatory reaction, characterized by recruitment and activation of leukocytes and the release of several inflammatory mediators, including prostaglandins and cytokines. Venoms contain several molecules such as enzymes (phospholipase A2, L-amino acid oxidase and proteases, among others) and peptides (disintegrins, mastoporan, parabutoporin etc.). These molecules are able to stimulate or inhibit ROS production by neutrophils. The present review article gives a general overview of the main neutrophil functions focusing on ROS production and summarizes how venoms and venom molecules can affect this function.(AU)

Cordycepin prevents the esophageal stricture formation in the alkali-burn rat model by exerting anti-inflammatory and antifibrotic effects

Purpose To investigate the efficacy of cordycepin, an adenosine analogue, on prevention of esophageal damage and stricture formation due to esophageal caustic burns in rat model comparing with prednisolone. Methods Caustic esophageal burn was introduced by 37.5% of NaOH to distal esophagus. Thirty-two Wistar albino rats were divided in four groups: sham rats undergone laparotomy, treated with 0.9% NaCl; control rats injured with NaOH without cordycepin treatment; cordycepin group injured with NaOH, treated with 20 mg/kg cordycepin; prednisolone group injured with NaOH, treated with 1 mg/kg prednisolone for 28 days. Efficacy was assessed by histopathological and immunohistochemical analysis of esophageal tissues. Results Cordycepin treatment significantly decreased inflammation, granulation tissue and fibrous tissue formation and prevented formation of esophageal strictures shown by histopathological damage score and stenosis indexes compared to control group (p 0.01). These effects are relatively more substantial than prednisolone, probably based on attenuation of elevation of proinflammatory cytokines hypoxia-inducible factor 1-alpha (HIF-1?), tumor necrosis factor alpha (TNF-?), proliferative and fibrotic factor fibroblast growth factor 2 (FGF2) and angiogenic factor vascular endothelial growth factor A (VEGFA) (p 0.05). Conclusions The findings suggest that cordycepin has a complex multifactorial healing process in alkali-burned tissue, more successful than prednisolone in preventing the formation of esophageal strictures and may be used as a therapeutic agent in the acute phase of esophageal alkali-burn.(AU)

Role of crotoxin in coagulation: novel insights into anticoagulant mechanisms and impairment of inflammation-induced coagulation

Snake venom phospholipases A2 (svPLA2) are biologically active toxins, capable of triggering and modulating a wide range of biological functions. Among the svPLA2s, crotoxin (CTX) has been in the spotlight of bioprospecting research due to its role in modulating immune response and hemostasis. In the present study, novel anticoagulant mechanisms of CTX, and the modulation of inflammation-induced coagulation were investigated. Methods: CTX anticoagulant activity was evaluated using platelet poor plasma (PPP) and whole blood (WB), and also using isolated coagulation factors and complexes. The toxin modulation of procoagulant and pro-inflammatory effects was evaluated using the expression of tissue factor (TF) and cytokines in lipopolysaccharide (LPS)-treated peripheral blood mononuclear cells (PBMC) and in WB. Results: The results showed that CTX impaired clot formation in both PPP and WB, and was responsible for the inhibition of both intrinsic (TF/factor VIIa) and extrinsic (factor IXa/factor VIIIa) tenase complexes, but not for factor Xa and thrombin alone. In addition, the PLA2 mitigated the prothrombinase complex by modulating the coagulation phospholipid role in the complex. In regards to the inflammation-coagulation cross talk, the toxin was capable of reducing the production of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α, and was followed by decreased levels of TF and procoagulant activity from LPS-treated PBMC either isolated or in WB. Conclusion: The results obtained in the present study recognize the toxin as a novel medicinal candidate to be applied in inflammatory diseases with coagulation disorders.(AU)

Role of crotoxin in coagulation: novel insights into anticoagulant mechanisms and impairment of inflammation-induced coagulation

Snake venom phospholipases A2 (svPLA2) are biologically active toxins, capable of triggering and modulating a wide range of biological functions. Among the svPLA2s, crotoxin (CTX) has been in the spotlight of bioprospecting research due to its role in modulating immune response and hemostasis. In the present study, novel anticoagulant mechanisms of CTX, and the modulation of inflammation-induced coagulation were investigated. Methods: CTX anticoagulant activity was evaluated using platelet poor plasma (PPP) and whole blood (WB), and also using isolated coagulation factors and complexes. The toxin modulation of procoagulant and pro-inflammatory effects was evaluated using the expression of tissue factor (TF) and cytokines in lipopolysaccharide (LPS)-treated peripheral blood mononuclear cells (PBMC) and in WB. Results: The results showed that CTX impaired clot formation in both PPP and WB, and was responsible for the inhibition of both intrinsic (TF/factor VIIa) and extrinsic (factor IXa/factor VIIIa) tenase complexes, but not for factor Xa and thrombin alone. In addition, the PLA2 mitigated the prothrombinase complex by modulating the coagulation phospholipid role in the complex. In regards to the inflammation-coagulation cross talk, the toxin was capable of reducing the production of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α, and was followed by decreased levels of TF and procoagulant activity from LPS-treated PBMC either isolated or in WB. Conclusion: The results obtained in the present study recognize the toxin as a novel medicinal candidate to be applied in inflammatory diseases with coagulation disorders.(AU)

Neuroprotection and immunomodulation by dimethyl fumarate and a heterologous fibrin biopolymer after ventral root avulsion and reimplantation

Ventral root avulsion (VRA) is an experimental approach in which there is an abrupt separation of the motor roots from the surface of the spinal cord. As a result, most of the axotomized motoneurons degenerate by the second week after injury, and the significant loss of synapses and increased glial reaction triggers a chronic inflammatory state. Pharmacological treatment associated with root reimplantation is thought to overcome the degenerative effects of VRA. Therefore, treatment with dimethyl fumarate (DMF), a drug with neuroprotective and immunomodulatory effects, in combination with a heterologous fibrin sealant/biopolymer (FS), a biological glue, may improve the regenerative response. Methods: Adult female Lewis rats were subjected to VRA of L4-L6 roots followed by reimplantation and daily treatment with DMF for four weeks. Survival times were evaluated 1, 4 or 12 weeks after surgery. Neuronal survival assessed by Nissl staining, glial reactivity (anti-GFAP for astrocytes and anti-Iba-1 for microglia) and synapse preservation (anti-VGLUT1 for glutamatergic inputs and anti-GAD65 for GABAergic inputs) evaluated by immunofluorescence, gene expression (pro- and anti-inflammatory molecules) and motor function recovery were measured. Results: Treatment with DMF at a dose of 15 mg/kg was found to be neuroprotective and immunomodulatory because it preserved motoneurons and synapses and decreased astrogliosis and microglial reactions, as well as downregulated the expression of pro-inflammatory gene transcripts. Conclusion: The pharmacological benefit was further enhanced when associated with root reimplantation with FS, in which animals recovered at least 50% of motor function, showing the efficacy of employing multiple regenerative approaches following spinal cord root injury.(AU)