Uso da termografia e ultrassonografia power dopplerpara avaliação da resposta tecidual em falhas ósseas preenchidas com biomaterial em ovinos / Termography and power doppler sonography assessment for tissue response evaluation in sheep bone defects filled with biomaterial

Biomateriaistem diversas indicações como auxiliares no processo de reparação óssea, além de terem função de substituto ósseo em perdas extensas. Diversas são as vantagens de sua utilização, como por exemplo, auxílio na osteocondutividade, estímulo de neovascularização, potencial antimicrobiano, entre outros. Falhas ósseas foram realizadas nas tíbias de seis ovinos da raça Santa Inês e preenchidas com biomaterial à base de quitosana, colágeno e hidroxiapatita. Um membro foi considerado controle e outro membro tratado segundo estudo randomizado. Foram realizadas avaliações termográficas e por ultrassonografia Power Dopplerem todos os animais do estudo, semanalmente, nos dias D0, D7, D14, D21, D28, D35, D42 e D56. Não houve diferenças significativas com relação à temperatura mínima, máxima e média entre os grupos com biomaterial e controle nas imagens termográficas. Houveram variações com relação ao tempo dentro de ambos os grupos. Com relação à presença de vasos na ultrassonografia Power Dopplernão houve diferenças estatísticas entre os grupos, exceto no dia 21 (P=0,031). Dentro das possibilidades de avaliação que os exames de imagem fornecem, a termografia e a ultrassonografia Power Dopplermostraram-se ferramentas não invasivas de avaliação pós-operatória de processo inflamatório e neovascularização, sendo realizadas semanalmente, permitindo acompanhamento fidedigno e detalhado ao longo do experimento, sem gerar desconforto ou estresse aos animais. Não houve indícios de complicações relacionadas ao biomaterial.(AU)

Biomaterials have several indications supporting the bone repair process, besides having bone substitute function in extensive losses. There are several advantages of its use, such as contribution in osteoconductivity, stimulation of neovascularization, antimicrobial potential, among others.Tibial bone defects was performed in six Santa Inês breed ewes and implanted with chitosan, collagen and hydroxyapatite biomaterial. One limb was considered the control limb and the other one the biomateriallimb, chosenrandomly. Thermographic and Power Dopplersonography was performed in all animals, weekly for 56 days. The thermography showed no significant differences related to low, high and mean temperature between the control and the biomaterial groups. Statisticalvariations were found between time in both groups. The visualization of neovascularization with the Power Dopplersonography did not present statistical differences, except for day 21 (P=0,031). Within the possibilities provided by imaging exams, thermography and Power Doppler sonography demonstrate to be non-invasive methods for post-operative evaluation of inflammatory process and neovascularization. The weekly management allowed reliable and detailed monitoring throughout the experiment, without causingdiscomfort or stress to the animals. There was no evidence of biomaterial complications.(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.

A comparative study to evaluate the effects of antibiotics, plant extracts and fluoride-based toothpaste on the oral pathogens isolated from patients with gum diseases in Pakistan

Abstract Oral diseases caused by various microorganisms are common around the world. Scientific research has now been focusing on novel medicines to overcome bacterial resistance and antibiotics side effects; therefore, the current study was designed to assess the efficacy of certain antibiotics, toothpaste, and medicinal plant extracts (Ajuga bracteosa and Curcuma longa) versus the bacterial pathogens isolated from the human oral cavity. A total of 130 samples were collected from Khyber Teaching Hospital Peshawar, Pakistan, among those 27 species isolated, and eight bacterial species were identified from the samples. Among all the bacterial species, Staphylococcus aureus (29.62%) and Proteus mirabilis (22.2%) were found to be more prevalent oral pathogens. In comparison, the least pervasive microbes were Proteus vulgaris, Shigella sonnei, Escherichia coli and Aeromonas hydrophila. The study also suggested that dental problems were more prevalent in males (41-50 years of age) than females. Among the eight antibiotics used in the study, the most promising results were shown by Foxicillin against A. hydrophila. The survey of TP1 revealed that it showed more potent antagonist activity against Proteus vulgaris as compared TP2 and TP3 that might be due to the high content of fluoride. The Curcuma longa showed more significant activity than Ajuga bracteosa (Stem, leaves and root) extracts. The data obtained through this study revealed that antibiotics were more effective for oral bacterial pathogens than toothpaste and plant extracts which showed moderate and low activity, respectively. Therefore, it is suggested that the active compounds in individual medicinal plants like Curcuma longa and Ajuga bracteosa could replace the antibiotics when used in daily routine as tooth cleansers or mouth rinses.

Resumo As doenças bucais causadas por vários microrganismos são comuns em todo o mundo. A pesquisa científica agora tem se concentrado em novos medicamentos para superar a resistência bacteriana e os efeitos colaterais dos antibióticos; portanto, o presente estudo foi desenhado para avaliar a eficácia de certos antibióticos, pasta de dente e extratos de plantas medicinais (Ajuga bracteosa e Curcuma longa) contra os patógenos bacterianos isolados da cavidade oral humana. No total, 130 amostras foram coletadas do Khyber Teaching Hospital Peshawar, Paquistão, entre essas, 27 espécies foram isoladas e oito espécies bacterianas foram identificadas a partir das amostras. Entre todas as espécies bacterianas, Staphylococcus aureus (29.62%) e Proteus mirabilis (22.2%) foram os patógenos orais mais prevalentes. Em comparação, os micróbios menos difundidos foram Proteus vulgaris, Shigella sonnei, Escherichia coli e Aeromonas hydrophila. O estudo também sugeriu que os problemas dentários eram mais prevalentes em homens (41-50 anos de idade) do que em mulheres. Entre os oito antibióticos usados no estudo, os resultados mais promissores foram mostrados pelo Foxicillin contra A. hydrophila. A pesquisa de TP1 revelou que ele mostrou atividade antagonista mais potente contra Proteus vulgaris em comparação a TP2 e TP3, o que pode ser devido ao alto teor de flúor. A Curcuma longa apresentou atividade mais significativa em relação aos extratos de Ajuga bracteosa (caule, folhas e raiz). Os dados obtidos neste estudo revelaram que os antibióticos foram mais eficazes para os patógenos bacterianos orais do que os dentifrícios e os extratos vegetais que apresentaram atividade moderada e baixa, respectivamente. Portanto, sugere-se que os compostos ativos em plantas medicinais individuais como Curcuma longa e Ajuga bracteosa possam substituir os antibióticos quando usados na rotina diária como limpadores de dentes ou enxaguatórios bucais.

Biocontrol potential of actinobacteria against Pantoea ananatis, the causal agent of maize white spot disease / Potencial de biocontrole de actinobactérias contra Pantoea ananatis, agente causal da doença mancha branca do milho

Pantoea ananatis is the causal agent of maize white spot, a foliar disease responsible for significant maize yield reduction worldwide, especially in Brazil. In general, the maize foliar diseases control involves the adoption of resistant genotypes and pesticides application. However, the use of agrochemicals can significantly cause increase production costs, damage to human health and negative environmental impacts. In this sense, the use of biological control agents has been considered among the most promising eco-friendly technologies for sustainable agriculture. Actinobacteria, particularly of Streptomyces genus, has been widely recognized as agroindustrially important microorganism due to its potential in producing diverse range of secondary metabolites, including antibiotics and enzymes. Thus, the aim of this work is to characterize and to evaluate the potential of soil actinobacteria for P. ananatis control. We observed that 59 actinobacteria strains (85%) exhibited proteolytic or chitinolytic activity. Only the strains Streptomyces pseudovenezuelae ACSL 470, that also exhibited high proteolytic activity, S. novaecaesareae ACSL 432 and S. laculatispora ACP 35 demonstrated high or moderate antagonist activity in vitro against P. ananatis. Temporal analysis of metabolites produced by these strains growth in different liquid media indicated greater antibacterial activity at 72 h. In this condition, chromatographic and mass spectrometry analysis revealed that S. pseudovenezuelae ACSL 470 strain produced neomycin, an aminoglycoside antibiotic that displayed high bactericidal activity in vitro against P. ananatis. This is the first report of actinobacteria acting as potential microbial antagonists for P. ananatis control. Further studies are needed to determine the control efficacy of maize white spot disease by Streptomyces strains or their metabolites in greenhouse and field conditions.

Pantoea ananatis é o agente causal da mancha branca do milho, doença foliar responsável pela redução significativa da produtividade do milho em todo o mundo, especialmente no Brasil. Em geral, o controle de doenças foliares do milho envolve a adoção de genótipos resistentes e a aplicação de agrotóxicos. No entanto, o uso de agroquímicos pode causar aumento significativo dos custos de produção, danos à saúde humana e impactos ambientais negativos. Nesse sentido, o uso de agentes de controle biológico tem sido considerado uma das tecnologias ecologicamente corretas mais promissoras para a agricultura sustentável. Actinobactérias, particularmente do gênero Streptomyces, têm sido amplamente reconhecidas como microrganismos de importância agroindustrial devido ao seu potencial de produção de diversos metabólitos secundários, incluindo antibióticos e enzimas. Assim, o objetivo deste trabalho é caracterizar e avaliar o potencial de actinobactérias do solo para o controle de P. ananatis. Observamos que 59 cepas de actinobactérias (85%) apresentaram atividade proteolítica ou quitinolítica. Apenas as cepas Streptomyces pseudovenezuelae ACSL 470, que também exibiu alta atividade proteolítica, S. novaecaesareae ACSL 432 e S. laculatispora ACP 35 demonstraram alta ou moderada atividade antagonista in vitro contra P. ananatis. A análise temporal do crescimento dos metabólitos produzidos por essas cepas em diferentes meios líquidos indicou maior atividade antibacteriana em 72 h. Nesta condição, análises cromatográficas e de espectrometria de massa revelaram que a cepa S. pseudovenezuelae ACSL 470 produziu neomicina, um antibiótico aminoglicosídeo que apresentou alta atividade bactericida in vitro contra P. ananatis. Este é o primeiro relato de actinobactérias atuando como potenciais antagonistas microbianos para o controle de P. ananatis. Mais estudos são necessários para determinar a eficácia do controle da doença da mancha branca do milho por cepas de Streptomyces ou seus metabólitos em condições de casa de vegetação e campo.

A threefold approach including quantum chemical, molecular docking and molecular dynamic studies to explore the natural compounds from Centaurea jacea as the potential inhibitors for COVID-19 / Uma abordagem tríplice incluindo química quântica, docking molecular e estudos dinâmicos moleculares para explorar os compostos naturais de Centaurea jacea como inibidores potenciais para Covid-19

Abstract In the current report, we studied the possible inhibitors of COVID-19 from bioactive constituents of Centaurea jacea using a threefold approach consisting of quantum chemical, molecular docking and molecular dynamic techniques. Centaurea jacea is a perennial herb often used in folk medicines of dermatological complaints and fever. Moreover, anticancer, antioxidant, antibacterial and antiviral properties of its bioactive compounds are also reported. The Mpro (Main proteases) was docked with different compounds of Centaurea jacea through molecular docking. All the studied compounds including apigenin, axillarin, Centaureidin, Cirsiliol, Eupatorin and Isokaempferide, show suitable binding affinities to the binding site of SARS-CoV-2 main protease with their binding energies -6.7 kcal/mol, -7.4 kcal/mol, -7.0 kcal/mol, -5.8 kcal/mol, -6.2 kcal/mol and -6.8 kcal/mol, respectively. Among all studied compounds, axillarin was found to have maximum inhibitor efficiency followed by Centaureidin, Isokaempferide, Apigenin, Eupatorin and Cirsiliol. Our results suggested that axillarin binds with the most crucial catalytic residues CYS145 and HIS41 of the Mpro, moreover axillarin shows 5 hydrogen bond interactions and 5 hydrophobic interactions with various residues of Mpro. Furthermore, the molecular dynamic calculations over 60 ns (6×106 femtosecond) time scale also shown significant insights into the binding effects of axillarin with Mpro of SARS-CoV-2 by imitating protein like aqueous environment. From molecular dynamic calculations, the RMSD and RMSF computations indicate the stability and dynamics of the best docked complex in aqueous environment. The ADME properties and toxicity prediction analysis of axillarin also recommended it as safe drug candidate. Further, in vivo and in vitro investigations are essential to ensure the anti SARS-CoV-2 activity of all bioactive compounds particularly axillarin to encourage preventive use of Centaurea jacea against COVID-19 infections.

Resumo No presente relatório, estudamos os possíveis inibidores de Covid-19 de constituintes bioativos de Centaurea jacea usando uma abordagem tripla que consiste em técnicas de química quântica, docking molecular e dinâmica molecular. Centaurea jacea é uma erva perene frequentemente usada em remédios populares de doenças dermatológicas e febre. Além disso, as propriedades anticâncer, antioxidante, antibacteriana e antiviral de seus compostos bioativos também são relatadas. A Mpro (proteases principais) foi acoplada a diferentes compostos de Centaurea jacea por meio de docking molecular. Todos os compostos estudados, incluindo apigenina, axilarina, Centaureidina, Cirsiliol, Eupatorina e Isokaempferide, mostram afinidades de ligação adequadas ao sítio de ligação da protease principal SARS-CoV-2 com suas energias de ligação -6,7 kcal / mol, -7,4 kcal / mol, - 7,0 kcal / mol, -5,8 kcal / mol, -6,2 kcal / mol e -6,8 kcal / mol, respectivamente. Dentre todos os compostos estudados, a axilarina apresentou eficiência máxima de inibidor, seguida pela Centaureidina, Isokaempferida, Apigenina, Eupatorina e Cirsiliol. Nossos resultados sugeriram que a axilarina se liga aos resíduos catalíticos mais cruciais CYS145 e HIS41 do Mpro, além disso a axilarina mostra 5 interações de ligações de hidrogênio e 5 interações hidrofóbicas com vários resíduos de Mpro. Além disso, os cálculos de dinâmica molecular em uma escala de tempo de 60 ns (6 × 106 femtossegundos) também mostraram percepções significativas sobre os efeitos de ligação da axilarina com Mpro de SARS-CoV-2 por imitação de proteínas como o ambiente aquoso. A partir de cálculos de dinâmica molecular, os cálculos RMSD e RMSF indicam a estabilidade e dinâmica do melhor complexo ancorado em ambiente aquoso. As propriedades ADME e a análise de previsão de toxicidade da axilarina também a recomendaram como um candidato a medicamento seguro. Além disso, as investigações in vivo e in vitro são essenciais para garantir a atividade anti-SARS-CoV-2 de todos os compostos bioativos, particularmente a axilarina, para encorajar o uso preventivo de Centaurea jacea contra infecções por Covid-19.

Evaluation of the antinociceptive effect generated by citronellal monoterpene isomers / Avaliação do efeito antinociceptivo gerado por isômeros do monoterpeno citronelal

Due to the complex nature of pain and the participation of physical, cognitive, psychological and behavioral aspects, pain management has several approaches. The use of medicinal plants in developing countries is quite expressive. Seeking new options for the treatment of emerging or debilitating diseases. Therefore, the present study seeks to elucidate the effects of the monoterpene, citronellal, differentiating its activity by isomers (R)-(+) and (S)-(-) citronellal. The study used several methods to evaluate the effects of citronellal isomers on motor coordination, nociceptive response, and the involvement of opioid, glutamatergic, and transient receptor pathways. The methods included rota-rod, hot-plate, and formalin tests, as well as the use of specific inhibitors and agonists. Data were analyzed using inferential statistics with a 95% confidence level. Both isomers did not significantly affect the motor coordination of the studied animals. The isomer (S)-(-) citronellal showed better results in relation to its structural counterpart, managing to have an antinociceptive effect in the formalin and hot plate tests with a lower concentration (100 mg/kg) and presenting fewer side effects, however, the this study was not able to elucidate the mechanism of action of this isomer despite having activity in studies with substances that act on specific targets such as glutamate and capsaicin, its activity was not reversed with the use of antagonists for pathways related to nociception. While the (R)-(+) citronellal isomer, despite showing total activity only at a concentration of 150 mg/kg, was able to determine its mechanism of action related to the opioid pathway by reversing its activity by the antagonist naloxone, being this is a pathway already correlated with nociception control treatments, however, it is also related to some unwanted side effects. In this way, new studies are sought to elucidate the mechanism related to the isomer (S)-(-) citronellal and a possibility of use in other areas related to the treatment of pain or inflammation.

Devido à natureza complexa da dor e a sua participação de aspectos físicos, cognitivos, psicológicos e comportamentais, o manejo da dor possui diversas abordagens. O uso de plantas medicinais em países em desenvolvimento é bastante expressivo. Buscando novas opções para o tratamento de denças emergentes ou debilitantes. Portanto, o presente estudo busca elucidar os efeitos do monoterpeno, citronelal, diferenciando sua atividade pelos isômeros (R)-(+) e (S)-(-) citronelal. O estudo utilizou diversos métodos para avaliar os efeitos dos isômeros de citronelal na coordenação motora, resposta nociceptiva e o envolvimento de vias opioides, glutamatérgicas e de receptores transitórios. Os métodos incluíram testes de rota-rod, placa quente e formalina, além do uso de inibidores e agonistas específicos. Os dados foram analisados estatisticamente com um intervalo de confiança de 95%. Ambos os isomeros não afetaram significativamente a coordenação motora dos animais em estudo. O isômero (S)-(-) citronelal apresentou melhores resultados em relação ao seu homólogo estrutural, conseguindo ter um efeito antinociceptivo nos testes de formalina e placa quente com menor concentração (100 mg/kg) e apresentando menos efeitos colaterais, entretanto, o presente estudo não foi capaz de elucidar o mecanismo de ação deste isomero apesar de ter atividadade em estudos com substancias que agem em alvos específicos como glutamato e capsaicina, sua atividade não foi revertida com a utilização de antagonistas para as vias relacionadas à nocicepção. Enquanto o isômero (R)-(+) citronelal, apesar de apresentar de apresentar total atividade somente na concentração de 150 mg/kg, foi capaz de determinar seu mecanismo de ação relacionado à via opióide pela reversão da sua atividade pelo antagonista naloxona, sendo esta uma via já correlacionada com os tratamentos de controle da a nocicepção, no entanto, também está relacionada a alguns efeitos colaterais indesejados. Desta forma busca-se novos estudos para elucidação do mecanismo relacionados ao isomero (S)-(-) citronellal e uma possibilidade de utilização em outras areas relacionadas ao tratamento da dor ou inflamação.

A comparative study to evaluate the effects of antibiotics, plant extracts and fluoride-based toothpaste on the oral pathogens isolated from patients with gum diseases in Pakistan / Um estudo comparativo para avaliar os efeitos de antibióticos, extratos vegetais e creme dental à base de flúor sobre os patógenos orais isolados de pacientes com doenças gengivais no Paquistão

Oral diseases caused by various microorganisms are common around the world. Scientific research has now been focusing on novel medicines to overcome bacterial resistance and antibiotics side effects; therefore, the current study was designed to assess the efficacy of certain antibiotics, toothpaste, and medicinal plant extracts (Ajuga bracteosa and Curcuma longa) versus the bacterial pathogens isolated from the human oral cavity. A total of 130 samples were collected from Khyber Teaching Hospital Peshawar, Pakistan, among those 27 species isolated, and eight bacterial species were identified from the samples. Among all the bacterial species, Staphylococcus aureus (29.62%) and Proteus mirabilis (22.2%) were found to be more prevalent oral pathogens. In comparison, the least pervasive microbes were Proteus vulgaris, Shigella sonnei, Escherichia coli and Aeromonas hydrophila. The study also suggested that dental problems were more prevalent in males (41-50 years of age) than females. Among the eight antibiotics used in the study, the most promising results were shown by Foxicillin against A. hydrophila. The survey of TP1 revealed that it showed more potent antagonist activity against Proteus vulgaris as compared TP2 and TP3 that might be due to the high content of fluoride. The Curcuma longa showed more significant activity than Ajuga bracteosa (Stem, leaves and root) extracts. The data obtained through this study revealed that antibiotics were more effective for oral bacterial pathogens than toothpaste and plant extracts which showed moderate and low activity, respectively. Therefore, it is suggested that the active compounds in individual medicinal plants like Curcuma longa and Ajuga bracteosa could replace the antibiotics when used in daily routine as tooth cleansers or mouth rinses.

As doenças bucais causadas por vários microrganismos são comuns em todo o mundo. A pesquisa científica agora tem se concentrado em novos medicamentos para superar a resistência bacteriana e os efeitos colaterais dos antibióticos; portanto, o presente estudo foi desenhado para avaliar a eficácia de certos antibióticos, pasta de dente e extratos de plantas medicinais (Ajuga bracteosa e Curcuma longa) contra os patógenos bacterianos isolados da cavidade oral humana. No total, 130 amostras foram coletadas do Khyber Teaching Hospital Peshawar, Paquistão, entre essas, 27 espécies foram isoladas e oito espécies bacterianas foram identificadas a partir das amostras. Entre todas as espécies bacterianas, Staphylococcus aureus (29.62%) e Proteus mirabilis (22.2%) foram os patógenos orais mais prevalentes. Em comparação, os micróbios menos difundidos foram Proteus vulgaris, Shigella sonnei, Escherichia coli e Aeromonas hydrophila. O estudo também sugeriu que os problemas dentários eram mais prevalentes em homens (41-50 anos de idade) do que em mulheres. Entre os oito antibióticos usados no estudo, os resultados mais promissores foram mostrados pelo Foxicillin contra A. hydrophila. A pesquisa de TP1 revelou que ele mostrou atividade antagonista mais potente contra Proteus vulgaris em comparação a TP2 e TP3, o que pode ser devido ao alto teor de flúor. A Curcuma longa apresentou atividade mais significativa em relação aos extratos de Ajuga bracteosa (caule, folhas e raiz). Os dados obtidos neste estudo revelaram que os antibióticos foram mais eficazes para os patógenos bacterianos orais do que os dentifrícios e os extratos vegetais que apresentaram atividade moderada e baixa, respectivamente. Portanto, sugere-se que os compostos ativos em plantas medicinais individuais como Curcuma longa e Ajuga bracteosa possam substituir os antibióticos quando usados na rotina diária como limpadores de dentes ou enxaguatórios bucais.

A comparative study to evaluate the effects of antibiotics, plant extracts and fluoride-based toothpaste on the oral pathogens isolated from patients with gum diseases in Pakistan / Um estudo comparativo para avaliar os efeitos de antibióticos, extratos vegetais e creme dental à base de flúor sobre os patógenos orais isolados de pacientes com doenças gengivais no Paquistão

Abstract Oral diseases caused by various microorganisms are common around the world. Scientific research has now been focusing on novel medicines to overcome bacterial resistance and antibiotics side effects; therefore, the current study was designed to assess the efficacy of certain antibiotics, toothpaste, and medicinal plant extracts (Ajuga bracteosa and Curcuma longa) versus the bacterial pathogens isolated from the human oral cavity. A total of 130 samples were collected from Khyber Teaching Hospital Peshawar, Pakistan, among those 27 species isolated, and eight bacterial species were identified from the samples. Among all the bacterial species, Staphylococcus aureus (29.62%) and Proteus mirabilis (22.2%) were found to be more prevalent oral pathogens. In comparison, the least pervasive microbes were Proteus vulgaris, Shigella sonnei, Escherichia coli and Aeromonas hydrophila. The study also suggested that dental problems were more prevalent in males (41-50 years of age) than females. Among the eight antibiotics used in the study, the most promising results were shown by Foxicillin against A. hydrophila. The survey of TP1 revealed that it showed more potent antagonist activity against Proteus vulgaris as compared TP2 and TP3 that might be due to the high content of fluoride. The Curcuma longa showed more significant activity than Ajuga bracteosa (Stem, leaves and root) extracts. The data obtained through this study revealed that antibiotics were more effective for oral bacterial pathogens than toothpaste and plant extracts which showed moderate and low activity, respectively. Therefore, it is suggested that the active compounds in individual medicinal plants like Curcuma longa and Ajuga bracteosa could replace the antibiotics when used in daily routine as tooth cleansers or mouth rinses.

Resumo As doenças bucais causadas por vários microrganismos são comuns em todo o mundo. A pesquisa científica agora tem se concentrado em novos medicamentos para superar a resistência bacteriana e os efeitos colaterais dos antibióticos; portanto, o presente estudo foi desenhado para avaliar a eficácia de certos antibióticos, pasta de dente e extratos de plantas medicinais (Ajuga bracteosa e Curcuma longa) contra os patógenos bacterianos isolados da cavidade oral humana. No total, 130 amostras foram coletadas do Khyber Teaching Hospital Peshawar, Paquistão, entre essas, 27 espécies foram isoladas e oito espécies bacterianas foram identificadas a partir das amostras. Entre todas as espécies bacterianas, Staphylococcus aureus (29.62%) e Proteus mirabilis (22.2%) foram os patógenos orais mais prevalentes. Em comparação, os micróbios menos difundidos foram Proteus vulgaris, Shigella sonnei, Escherichia coli e Aeromonas hydrophila. O estudo também sugeriu que os problemas dentários eram mais prevalentes em homens (41-50 anos de idade) do que em mulheres. Entre os oito antibióticos usados ​​no estudo, os resultados mais promissores foram mostrados pelo Foxicillin contra A. hydrophila. A pesquisa de TP1 revelou que ele mostrou atividade antagonista mais potente contra Proteus vulgaris em comparação a TP2 e TP3, o que pode ser devido ao alto teor de flúor. A Curcuma longa apresentou atividade mais significativa em relação aos extratos de Ajuga bracteosa (caule, folhas e raiz). Os dados obtidos neste estudo revelaram que os antibióticos foram mais eficazes para os patógenos bacterianos orais do que os dentifrícios e os extratos vegetais que apresentaram atividade moderada e baixa, respectivamente. Portanto, sugere-se que os compostos ativos em plantas medicinais individuais como Curcuma longa e Ajuga bracteosa possam substituir os antibióticos quando usados ​​na rotina diária como limpadores de dentes ou enxaguatórios bucais.

A comparative study to evaluate the effects of antibiotics, plant extracts and fluoride-based toothpaste on the oral pathogens isolated from patients with gum diseases in Pakistan / Um estudo comparativo para avaliar os efeitos de antibióticos, extratos vegetais e creme dental à base de flúor sobre os patógenos orais isolados de pacientes com doenças gengivais no Paquistão

Oral diseases caused by various microorganisms are common around the world. Scientific research has now been focusing on novel medicines to overcome bacterial resistance and antibiotics side effects; therefore, the current study was designed to assess the efficacy of certain antibiotics, toothpaste, and medicinal plant extracts (Ajuga bracteosa and Curcuma longa) versus the bacterial pathogens isolated from the human oral cavity. A total of 130 samples were collected from Khyber Teaching Hospital Peshawar, Pakistan, among those 27 species isolated, and eight bacterial species were identified from the samples. Among all the bacterial species, Staphylococcus aureus (29.62%) and Proteus mirabilis (22.2%) were found to be more prevalent oral pathogens. In comparison, the least pervasive microbes were Proteus vulgaris, Shigella sonnei, Escherichia coli and Aeromonas hydrophila. The study also suggested that dental problems were more prevalent in males (41-50 years of age) than females. Among the eight antibiotics used in the study, the most promising results were shown by Foxicillin against A. hydrophila. The survey of TP1 revealed that it showed more potent antagonist activity against Proteus vulgaris as compared TP2 and TP3 that might be due to the high content of fluoride. The Curcuma longa showed more significant activity than Ajuga bracteosa (Stem, leaves and root) extracts. The data obtained through this study revealed that antibiotics were more effective for oral bacterial pathogens than toothpaste and plant extracts which showed moderate and low activity, respectively. Therefore, it is suggested that the active compounds in individual medicinal plants like Curcuma longa and Ajuga bracteosa could replace the antibiotics when used in daily routine as tooth cleansers or mouth rinses.(AU)

As doenças bucais causadas por vários microrganismos são comuns em todo o mundo. A pesquisa científica agora tem se concentrado em novos medicamentos para superar a resistência bacteriana e os efeitos colaterais dos antibióticos; portanto, o presente estudo foi desenhado para avaliar a eficácia de certos antibióticos, pasta de dente e extratos de plantas medicinais (Ajuga bracteosa e Curcuma longa) contra os patógenos bacterianos isolados da cavidade oral humana. No total, 130 amostras foram coletadas do Khyber Teaching Hospital Peshawar, Paquistão, entre essas, 27 espécies foram isoladas e oito espécies bacterianas foram identificadas a partir das amostras. Entre todas as espécies bacterianas, Staphylococcus aureus (29.62%) e Proteus mirabilis (22.2%) foram os patógenos orais mais prevalentes. Em comparação, os micróbios menos difundidos foram Proteus vulgaris, Shigella sonnei, Escherichia coli e Aeromonas hydrophila. O estudo também sugeriu que os problemas dentários eram mais prevalentes em homens (41-50 anos de idade) do que em mulheres. Entre os oito antibióticos usados no estudo, os resultados mais promissores foram mostrados pelo Foxicillin contra A. hydrophila. A pesquisa de TP1 revelou que ele mostrou atividade antagonista mais potente contra Proteus vulgaris em comparação a TP2 e TP3, o que pode ser devido ao alto teor de flúor. A Curcuma longa apresentou atividade mais significativa em relação aos extratos de Ajuga bracteosa (caule, folhas e raiz). Os dados obtidos neste estudo revelaram que os antibióticos foram mais eficazes para os patógenos bacterianos orais do que os dentifrícios e os extratos vegetais que apresentaram atividade moderada e baixa, respectivamente. Portanto, sugere-se que os compostos ativos em plantas medicinais individuais como Curcuma longa e Ajuga bracteosa possam substituir os antibióticos quando usados na rotina diária como limpadores de dentes ou enxaguatórios bucais.(AU)

Ulinastatin alleviates early brain injury after traumatic brain injury by inhibiting oxidative stress and apoptosis

Purpose: Traumatic brain injury (TBI) remains a major public health problem and cause of death. Ulinastatin (UTI), a serine protease inhibitor, has been reported to have an anti-inflammatory effect and play a role in immunoregulation and organ protection by reducing reactive oxygen species (ROS) production, oxidative stress and inflammation. However, the neuroprotective of UTI in TBI has not been confirmed. Therefore, this study aimed to investigate the neuroprotection and potential molecular mechanisms of UTI in TBI-induced EBI in a C57BL/6 mouse model. Methods: The neurological score and brain water content were evaluated. Enzyme-linked immunosorbent assay was used to detect neuroinflammatory cytokine levels, ROS and malondialdehyde detection to evaluate oxidative stress levels, and TUNEL staining and western blotting to examine neuronal damages and their related mechanisms. Results: Treatment with UTI markedly increased the neurological score; alleviated brain oedema; decreased the inflammatory cytokine tumour necrosis factor a, interleukin-1ß (IL-1ß), IL-6 and nuclear factor kappa B (NF-kB) levels; inhibited oxidative stress; decreased caspase-3 and Bax protein expressions; and increased the Bcl-2 levels, indicating that UTI-mediated inhibition of neuroinflammation, oxidative stress and apoptosis ameliorated neuronal death after TBI. The neuroprotective capacity of UTI is partly dependent on the TLR4/NF-kB/p65 signalling pathway. Conclusions: Therefore, this study reveals that UTI improves neurological outcomes in mice and reduces neuronal death by protecting against neural neuroinflammation, oxidative stress and apoptosis.

Effects of systemic inflammation due to hepatic ischemia-reperfusion injury upon lean or obese visceral adipose tissue

Purpose: To evaluate how the induction of liver damage by ischemia and reperfusion affects the adipose tissue of lean and obese mice. Methods: Lean and diet-induced obese mice were subjected to liver ischemia (30 min) followed by 6 h of reperfusion. The vascular stromal fraction of visceral adipose tissue was analyzed by cytometry, and gene expression was evaluated by an Array assay and by RT-qPCR. Intestinal permeability was assessed by oral administration of fluorescein isothiocyanate (FITC)-dextran and endotoxemia by serum endotoxin measurements using a limulus amebocyte lysate assay. Results: It was found that, after liver ischemia and reperfusion, there is an infiltration of neutrophils, monocytes, and lymphocytes, as well as an increase in the gene expression that encode cytokines, chemokines and their receptors in the visceral adipose tissue of lean mice. This inflammatory response was associated with the presence of endotoxemia in lean mice. However, these changes were not observed in the visceral adipose tissue of obese mice. Conclusions: Liver ischemia and reperfusion induce an acute inflammatory response in adipose tissue of lean mice characterized by an intense chemokine induction and leukocyte infiltration; however, inflammatory alterations are already present at baseline in the obese adipose tissue and liver ischemia and reperfusion do not injure further.

Microorganisms from corn stigma with biocontrol potential of Fusarium verticillioides / Microrganismos do estigma do milho com potencial de biocontrole de Fusarium verticillioides

The mycotoxigenic fungus Fusarium verticillioides is the primary maize pathogen and causes the maize stalk and ear rot diseases with significant economic losses. Furthermore, the excessive use of fungicides to control F. verticillioides constitutes threats to the environment and human health. Thus, sustainable alternatives such as biological control are needed to minimize the hazards associated with the current method. Although much is known about the vulnerability of the maize silks as a gateway for several fungal pathogens invading the developing grains, studies on the chemical properties of silk extracts and their resident microbiota are scarce. This study isolated and characterized bacteria and fungi that colonize the maize stigma to assess new potential biocontrol agents. The samples were collected from maize fields in the Brazilian localities of Sete Lagoas-MG, Sidrolândia-MS, Sertaneja-PR, and Goiânia-GO. One hundred sixty-seven microorganisms were isolated, 46% endophytic and 54% epiphytic. First, the antagonist activity was evaluated by the agar disc diffusion method performed in triplicate, and 83% of the isolates showed antagonist activity against F. verticillioides. Then, the 42 most efficient isolates were identified based on the partial sequencing of the bacterial 16S rRNA gene and fungi ITS region. The bacteria belong to the genera Bacillus (57.1%), Burkholderia (23.8%), Achromobacter (7.1%), Pseudomonas (2.4%), and Serratia (2.4%), while the fungi are Penicillium (2.4%), Candida (2.4), and Aspergillus (2.4%). The results showed that microorganisms from maize stigma might represent new promising agents for F. verticillioides control.

O fungo micotoxigênico Fusarium verticillioides é o principal patógeno do milho e causa doenças do colmo e da podridão da espiga com perdas econômicas significativas. Além disso, o uso excessivo de fungicidas no controle de F. verticillioides constitui uma ameaça ao meio ambiente e à saúde humana. Assim, alternativas sustentáveis, como o controle biológico, são necessárias para minimizar os riscos associados ao método atual. Este estudo isolou e caracterizou bactérias e fungos que colonizam o estigma do milho para avaliar novos agentes de biocontrole em potencial. As amostras foram coletadas em campos de milho nas localidades brasileiras de Sete Lagoas-MG, Sidrolândia-MS, Sertaneja-PR e Goiânia-GO. Cento e sessenta e sete microrganismos foram isolados, 46% endofíticos e 54% epifíticos. O teste de antagonismo empregando a técnica de disco de difusão em meio sólido, mostrou que 83% dos isolados apresentaram atividade antagonista contra F. verticillioides. Em seguida, 42 isolados mais eficientes foram identificados a partir do sequenciamento parcial do gene 16S rRNA bacteriano e da região ITS de fungos. Os isolados bacterianos pertencem ao gênero Bacillus (57,1%), Burkholderia (23,8%), Achromobacter (7,1%), Pseudomonas (2,4%) e Serratia (2,4%), enquanto os fungos são Penicillium (2,4%), Candida (2.4), e Aspergillus (2,4%). Os resultados mostraram que microrganismos do estigma do milho podem representar novos agentes promissores para o controle de F. verticillioides.

Efficiency of different strains of Trichoderma on the control of Sclerotinia sclerotiorum, Sclerotium rolfsii and Sclerotium cepivorum

The aim of this work was to verify the efficiency of different isolates of Trichoderma spp.on the control of Sclerotiniasclerotiorum, Sclerotiumrolfsii and Sclerotiumcepivorum, and the influence they pose on the conidia production of Trichoderma spp.For mycelial growth, discs with inoculum of phytopathogens were placed on the center of the Petri dishes followed by the addition of two Trichodermasp. discs on the opposite sides of the plate after 24hours. Every 12hoursdata were collected from colonies diameters and used for the analyses of Mycelial Growth Index (MGI) and Area Under the Curve of Mycelial Growth (AUCMG). The analyses were performed by a completely randomized design with two controls, a negative one without Trichodermasp. and one with acommercial strain of Trichodermaharzianum. Spore solution for evaluation of conidia production were made by adding 10 mL of distilled water and scratching the surface of the colonies. For S. cepivorum, all Trichoderma spp. strains reduced both indexes tested. However, while for MGI S. sclerotiorum also presented some reduction on the growth rate, the total area of this fungus was not affected. Sclerotiumrolfsii strains of Trichodermasp. from Lages and Curitibanosshowed an effect on the reduction of AUCMG of this fungus, although none of the Trichodermaaffected the growth rate of this phytopathogen. On the presence of S. sclerotiorumand S. cepivorum, none of the Trichodermaspp. showed any difference on conidia production when compared among themselves, nonetheless we did notice that on the presence of S. cepivorum, the strain from Rio do Sul retained its reproductive ability compared to control. Results obtained from this research can demonstrate the importance of biocontrol agents against different plant pathogens since it might have a specific antagonist-pathogen relation.(AU)

Ulinastatin alleviates early brain injury after intracerebral hemorrhage by inhibiting oxidative stress and neuroinflammation via ROS/MAPK/Nrf2 signaling pathway

Purpose: Spontaneous intracerebral hemorrhage (ICH) is still a major public health problem, with high mortality and disability. Ulinastatin (UTI) was purified from human urine and has been reported to be anti-inflammatory, organ protective, and antioxidative stress. However, the neuroprotection of UTI in ICH has not been confirmed, and the potential mechanism is unclear. In the present study, we aimed to investigate the neuroprotection and potential molecular mechanisms of UTI in ICH-induced early brain injury in a C57BL/6 mouse model. Methods: The neurological score, brain water content, neuroinflammatory cytokine levels, oxidative stress levels, and neuronal damage were evaluated. Results: UTI treatment markedly increased the neurological score, alleviated brain edema, decreased the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, and NF-κB, decreased the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and upregulated the levels of glutathione (GSH), superoxide dismutase (SOD), and Nrf2. This finding indicated that UTI-mediated inhibition of neuroinflammation and oxidative stress alleviated neuronal damage after ICH. The neuroprotective capacity of UTI is partly dependent on the ROS/MAPK/Nrf2 signaling pathway. Conclusions: UTI improves neurological outcomes in mice and reduces neuronal death by protecting against neural neuroinflammation and oxidative stress.

Assessment of melatonin-alpha adrenergic receptor complexes by molecular docking analysis / Avaliação dos complexos melatonina-receptores alfa-adrenérgicos por análises de docagem molecular

The pineal melatonin (N-acetyl-5-methoxytryptamine) is a molecule associated in a way or another with probably all physiological systems, aiming to fulfil its functional integrative roles in central nervous system activity, sleep and wakefulness cycles, energy metabolism and thermoregulation, immune, reproductive, endocrine, cardiovascular, respiratory and excretory systems. Within this context, the present study aimed to assess in silico the formation of complexes between ligand melatonin and other potential receptor proteins by molecular docking analyses. The main steps established in this experimental procedure were: a) search and selection of the 3D structure of the melatonin from DrugBank; b) search and selection of 3D structures of other target receptor proteins using STRING, protein BLAST and database PDB; and c) formation of the complexes between melatonin and receptors selected using AutoDock4.0 server by molecular docking analyses. High reliability score and significant similarity were only identified between type 1B melatonin and alpha-2A adrenergic receptor. Thus, molecular docking assays were carried out using ligand melatonin and crystallographic structures of the alpha-2A adrenergic receptor coupled to an antagonist (ID PDB 6kux) and a partial agonist (ID PDB 6kuy) available in the database PDB. Binding energy values of -6.79 and -6.98 kcal/mol and structural stability by non-covalent intermolecular interactions were predicted during the formation of complexes between melatonin and alpha-2A adrenergic receptor 6kux and 6kuy, respectively. In this way, the findings described in current study may indicate strong interactions between melatonin and adrenoceptors, suggesting its possible partial agonist effect on the activation of the alfa-2A adrenergic receptor.(AU)

A melatonina pineal (N-acetil-5-metoxitriptamina) é uma molécula associada de um modo ou outro com provavelmente todos os sistemas fisiológicos, visando cumprir seus papéis funcionais integradores na atividade do sistema nervoso central, ciclos de sono e vigília, metabolismo energético e termorregulação, sistemas imunológico, reprodutivo, endócrino, cardiovascular, respiratório e excretor. Assim, o presente estudo objetivou avaliar in silico a formação de complexos entre o ligante melatonina e outras proteínas potenciais receptoras por meio de análises de docagem molecular. As principais etapas estabelecidas neste procedimento experimental foram: a) busca e seleção da estrutura 3D da melatonina a partir do banco de dados DrugBank; b) busca e seleção de estruturas 3D de outras proteínas receptoras-alvo utilizando STRING, proteína BLAST e o banco de dados PDB; e c) avaliação da formação dos complexos entre melatonina e receptores selecionados a partir do servidor AutoDock4.0 para análises de docagem molecular. Alto escore de confiabilidade e similaridade significativa foram identificados apenas entre a melatonina do tipo 1B e o receptor alfa-2A adrenérgico. Valores de energia de ligação de -6,79 e -6,98 kcal/mol e estabilidade estrutural pela presença de interações intermoleculares não covalentes foram preditos durante a formação de complexos entre o ligante melatonina e os receptores adrenérgico alfa-2A 6kux e 6kuy, respectivamente. Dessa forma, os achados descritos no presente estudo podem indicar fortes interações entre melatonina e adrenoceptores, sugerindo seu possível efeito agonista parcial na ativação do receptor alfa-2A adrenérgico.(AU)

In vitro and in silico evaluation of the schistosomicidal activity of eugenol derivatives using biochemical, molecular, and morphological tools

Background Eugenol shows both antibacterial and antiparasitic activities, suggesting that it might be evaluated as an option for the treatment of praziquantel-resistant schistosome. Methods The in vitro activities of three eugenol derivatives (FB1, FB4 and FB9) on adult worms from Schistosoma mansoni were examined by fluorescence and scanning electron microscopy to analyze effects on the excretory system and integument damage, respectively. Biochemical tests with verapamil (a calcium channel antagonist) and ouabain (a Na+/K+-ATPase pump inhibitor) were used to characterize eugenol derivative interactions with calcium channels and the Na+/K+-ATPase, while in silico analysis identified potential Na+/K+-ATPase binding sites. Results The compounds showed effective doses (ED50) of 0.324 mM (FB1), 0.167 mM (FB4), and 0.340 mM (FB9). In addition, FB4 (0.322 mM), which showed the lowest ED50, ED90 and ED100 (p < 0.05), caused the most damage to the excretory system and integument, according to both fluorescence and scanning electron microscopy analysis. The death of adult worms was delayed by ouabain treatment plus FB1 (192 versus 72 hours) and FB9 (192 versus 168 hours), but the response to FB4 was the same in the presence or absence of ouabain. Besides, no changes were noted when all of the eugenol derivatives were combined with verapamil. Moreover, FB1 and FB9 inhibited Na+/K+-ATPase activity according to in silico analysis but FB4 did not show a time-dependent relationship and may act on targets other than the parasite Na+/K+-ATPase. Conclusion Eugenol derivatives, mainly FB4 when compared to FB1 and FB9, seem to act more effectively on the integument of adult S. mansoni worms.(AU)

Clinical safety of zafirlukast treatment during acute inflammatory reaction in nile tilapia (Oreochromis niloticus) / Segurança clínica do tratamento com zafirlucaste durante a reação inflamatória aguda em tilápia do nilo (Oreochromis niloticus)

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 hydrophila bacterins. 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 characterized by 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 treated with 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 funções hepáticas 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 na dose de 250 µg em tilápias durante aerocistite aguda, embora alterações hematológicas tenham sido observadas em tilápias tratadas com sobredosagem deste bloqueador de leucotrieno.

Clinical safety of zafirlukast treatment during acute inflammatory reaction in nile tilapia (Oreochromis niloticus) / Segurança clínica do tratamento com zafirlucaste durante a reação inflamatória aguda em tilápia do nilo (Oreochromis niloticus)

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 hydrophila bacterins. 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 characterized by 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 treated with overdose of this leukotriene blocker(AU)

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 funções hepáticas 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 na dose de 250 µg em tilápias durante aerocistite aguda, embora alterações hematológicas tenham sido observadas em tilápias tratadas com sobredosagem deste bloqueador de leucotrieno.(AU)

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae) / Identificação molecular, caracterização e análise de expressão de um gene inibidor de serina protease da lagarta Helicoverpa armigera (Lepidoptera: noctuidae) na cultura de algodão

Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341N350), the serpin signature, (F367F375) and a predicted P1P1 cleavage site (L357S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.(AU)

Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.(AU)

P-I metalloproteinases and L-amino acid oxidases from Bothrops species inhibit angiogenesis

Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)