Identificação molecular de ostras Crassostrea spp (mollusca: bivalvia) dos dois maiores estuários do estado de Sergipe por PCR/RFLP

As espécies Crassostrea rhizophorae e C. brasiliana são ostras nativas do estado de Sergipe e desempenham importante papel ecológico no ecossistema manguezal, além de representarem um dos principais produtos de extrativismo. C. rhizophorae, conhecida popularmente como “ostra-do-mangue”, considerada de médio porte (10 cm) e a C. brasiliana, conhecida como “ostra-de-fundo”, considerada de grande porte (20 cm), apresentam-se similares em sua morfologia e coocorrem no ambiente em formas imaturas, contudo quando adultas pouco se sabe sobre esta coocorrência entre as espécies, podendo dificultar a identificação e diferenciação entre as espécies. Diante disso o objetivo do trabalho é a identificação molecular das espécies de ostras nativas adultas encontradas nos dois diferentes estuários do estado de Sergipe e verificar a coocorrência entre as mesmas. No presente estudo foram utilizadas ostras nativas adultas do gênero Crassostrea coletadas do ecossistema manguezal do litoral sergipano. As 28 amostras de ostras foram coletadas no mês de setembro de 2008, as quais foram retirados os músculos adutores de cada ostra para a extração de DNA total seguido por amplificações do segmento rDNA 16S e digerido com enzimas de restrição. O produto de amplificação das 28 amostras digerido com a enzima de restrição apresentou o mesmo perfil de restrição, correspondente à espécie Crassostrea brasiliana, resultado constatado por meio da comparação do produto da digestão com amostras padrão existente, referente às espécies C. brasiliana e C. rhizophorae

Alternativas Terapêuticas para o Cálculo Dentário: uma análise prospectiva científica e tecnológica

Dental calculus is the result of mutualistic interactions among microorganisms present in dental biofilm and is correlated with the development of periodontal disease. The present study aimed to present an overview of existing therapeutic alternatives for dental calculus, due to the need for innovative treatments. ScienceDirect and PubMed were used as databases for scientific prospecting, and the National Institute of Industrial Property and the World Intellectual Property Organizations were used for technological prospecting. Publications obtained were time-delimited between the years of 2012 and 2022, and, to encompass the maximum number of articles related to the study's objective, full-text documents were selected, using the descriptors "dental calculus," "periodontal debridement," and "treatment." Strategies such as the use of products based on medicinal plant extracts and essential oils were observed, along with the application of antiseptics and systemic antibiotics.O cálculo dentário é o resultado das interações mutualísticas dos microrganismos presentes no biofilme dental, e está correlacionado com o desenvolvimento da doença periodontal. O presente estudo objetivou apresentar um panorama de alternativas terapêuticas existentes frente ao cálculo dental devido à necessidade de tratamentos inovadores. Foram utilizados como banco de dados para prospecção científica o Science Direct e o PubMed e, para a prospecção tecnológica, o Instituto Nacional da Propriedade Industrial e o World Intellectual Property Organizations. Delimitou-se as publicações entre os anos de 2012-2022 e foram selecionados documentos disponibilizados na íntegra, a fim de abranger o máximo de artigos direcionados ao objetivo do estudo a partir de um refinamento com o uso dos descritores “dental calculus”, “periodontal debridement” e “treatment”. Estratégias como o uso de produtos à base de extratos de plantas medicinais e óleos essenciais foram observados, além da aplicação de antissépticos e antibióticos sistêmicos

Epítopos imunodominantes de proteína putativa Corynebacterium pseudotuberculosis apresentam interações com TLR-2 in silico / Immunodominant epitopes of putative protein Corynebacterium pseudotuberculosis interacts with TLR-2 in silico

INTRODUÇÃO: A Linfadenite Caseosa (LC) é uma doença infecciosa causada pela bactéria Corynebacterium pseudotuberculosis e vem ocasionando inúmeros prejuízos à indústria de carne ovinocaprina. Ela possui duas formas clínicas, a forma superficial, quando há a presença de abscessos em linfonodos periféricos ou forma visceral, quando esses abscessos encontram-se nos órgãos internos. A antibioticoterapia hoje existente se torna ineficaz por não conseguir cruzar essa barreira, como também, a vacina comercial hoje disponibilizada possui baixo poder de proteção, necessitando assim de uma nova dose a cada ano o que ocasiona um alto custo. OBJETIVO: realizar um desenho vacinal em busca de potenciais alvos proteicos de Corynebacterium pseudotuberculosis para formular uma vacina contra a Linfadenite Caseosa. METODOLOGIA: Para isso, análises in silico utilizando o Vaxign para busca de proteínas secretadas foram feitas e subsequentes análises relacionadas ao potencial antigênico, alergênico e caracterizações físico-químicas serviram para filtrar as proteínas de interesse. Das proteínas selecionadas foram obtidos os epítopos imunodominantes e sua interação com o TLR-2 foi determinada. RESULTADOS: Foram encontradas pelo Vaxign um total de 26 proteínas putativas secretadas, sendo realizada as suas caracterizações a fim de selecionar as proteínas aptas para possível formulação de uma vacina. Destas, nove delas se mostraram antigênicas e não alergênicas e após análise físico-quimicas apenas duas seguiram para análise dos epítopos imunodominantes de células B. As proteínas ADL20140.1 e ADL20298.1  apresentaram um total de 2 e 5 epítopos, respectivamente. Após determinação do potencial antigênico apenas os epítopos 4, 6 e 7 foram utilizados para o docking molecular com o TLR-2. Os epítopos 4 e 7 apresentaram possíveis interaõoes através dos domínios LRR enquanto o epítopo 6 apresentou possível interaçõo com o domínio TIR do TLR-2. CONCLUSÃO: Diante do uso de bioinformática é possivel verificar previsões in silico que possam diminuir etapas dispendiosas experimentais. Com isso, nesse trabalho foi possível selecionar uma proteina com características importantes para compor uma vacina contra LC e até epítopos imunodominantes que podem servir para formular uma vacina peptídica, esses epítopos apresentaram potenciais interações com o TLR-2 indicando uma possível ativação da imunidade inata, importante passo para um alvo vacinal. Portanto, esse trabalho fornece informções importantes que poderm servir para formular, de fato, uma vacina contra LC

Cymbopogon winterianus essential oil attenuates bleomycin-induced pulmonary fibrosis in a murine model

The essential oil of Cymbopogon winterianus (EOCW) is a natural product with antioxidant, anti-inflammatory, and antifibrotic properties. We studied the effect of EOCW in the progression of histological changes of pulmonary fibrosis (PF) in a rodent model. The oil was obtained by hydrodistillation and characterized using gas chromatographymass spectrometry. Intratracheal instillation of bleomycin was performed in 30 rats to induce PF, while Sham animals were subjected to instillation of saline solution. The treatment was performed using daily oral administration of distilled water, EOCW at 50, 100, and 200 mg/kg, and deflazacort (DFC). After 28 days, hemogram and bronchoalveolar lavage fluid (BALF), tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) were assayed. Histological grading of PF, immunohistochemical expression of -smooth muscle actin (-SMA), and transforming growth factor- (TGF-) were also analyzed. The EOCW major compounds were found to be citronellal, geraniol, and citronellol. EOCW significantly reduced inflammation in BALF, reduced MDA levels, and increased SOD activity. EOCW attenuated histological grading of PF and reduced immunohistochemical expression of -SMA and TGF- in a dose-dependent way, likely due to the reduction of oxidative stress, inflammation, and TGF--induced myofibroblast differentiation.This research was financed by the Coordenação Aperfeiçoamento de Pessoal de Nivel Superior (CAPES), Fundação de Amparo à Pesquisa do Estado de Sergipe (FAPITEC), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). EBS wishes to acknowledge the sponsorship of the project UIDB/04469/2020 (strategic fund) from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and was co-financed by FEDER, under the Partnership Agreement PT2020.info:eu-repo/semantics/publishedVersio

Immune-Informatic Analysis and Design of Peptide Vaccine From Multi-epitopes Against

Caseous lymphadenitis (CLA) is a disease caused by Corynebacterium pseudotuberculosis bacteria that affects sheep and goats. The absence of a serologic diagnose is a factor that contributes for the disease dissemination, and due to the formation of granuloma, the treatment is very expensive. Therefore, prophylaxis is the approach with best cost-benefit relation; however, it still lacks an effective vaccine. In this sense, this work seeks to apply bioinformatic tools to design an effective vaccine against CLA, using CP40 protein as standard for the design of immunodominant epitopes, from which a total of 6 sequences were obtained, varying from 10 to 16 amino acid residues. The evaluation of different properties of the vaccines showed that the vaccine is a potent and nonallergenic antigen remaining stable in a wide range of temperatures. The initial tertiary structure of the vaccine was then predicted and a model selected. Later, the process of CP40 protein and TLR2 receptor binding was performed, presenting interaction with this receptor, which plays an important role in the activation of the immune response

In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay

Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new antigens to compose diagnosis assays or vaccines are fundamental. Therefore, this study aimed to use bioinformatics software to evaluate the critical chemical characteristics of unknown proteins of C. pseudotuberculosis by selecting them for heterologous expression in Escherichia coli. For this purpose, six protein sequences of ascorbate transporter subunit, UPF protein, MMPL family transporter, Ribonuclease, Iron ABC transporter domain-containing permease, and fimbrial subunit were obtained. In silico analyses were performed using amino acid sequences to access immunodominant epitopes and their antigenic and allergenic potential and physicochemical characterization. The expressed proteins were used as an antigen for serological diagnosis by ELISA. All proteins showed distinct immunodominant epitopes and potential antigenic characteristics. The only proteins expressed were PTS and Ribonuclease. In parallel, we expressed CP40 and all were used with ELISA antigen in 49 CLA positive sera and 26 CLA negative sera. The proteins alone showed 100% sensitivity and 96.2%, 92.3%, and 88.5% specificity for rPTS, rRibonuclease, and rCP40, respectively. When proteins were combined, they showed 100% sensitivity and 84.6%, 92.3%, 88.5%, and 92.3% specificity for rPTS/rCp40, rRibonuclease/rCP40, rPTS/rRibonuclease, and rPTS/rRibonuclease/rCP40, respectively. The results of this study show an excellent correlation of sensitivity and specificity with all proteins. None of the specificity values preclude the potential of rPTS, rRibonuclease, or rCP40 for use in ELISA diagnostic assays since the results of this work are superior to those of other studies on CLA diagnosis described in the literature

Hancornia speciosa extract presents antimicrobial action against Pseudomonas aeruginosa and Staphylococcus aureus Extract shows antimicrobial activity / O extracto de Hancornia speciosa apresenta acção antimicrobiana contra Pseudomonas aeruginosa e Staphylococcus aureus O extracto mostra actividade antimicrobiana

Aims: To evaluate the preliminary phytochemical profile of the leaf and fruit ethanolic extracts of Hancornia speciosa Gomes, as well as its antimicrobial and anti-biofilm activities.Methods and Results:  Samples of the leaves and fruits of H. speciosa Gomes, collected in the city of Estância, Sergipe state, Brazil, were used to obtain the extract. Each material was macerated in 96% alcohol and submitted to the ultrasonic bath. The extracts showed effectiveness against gram positive and Gram-negative planktonic bacteria, where the leaves presented better results, with a minimum inhibitory concentration (MIC) of 0.625 mg.ml-1 against hospital Staphylococcus aureus, and 1.25 mg.ml-1 against the standard strain of this species. Regarding the percentage of inhibition of biofilms, leaf extract remained more efficient, although the effects were quite varied, strongly dependent on the individual features of each lineage. Conclusions:  The provided data demonstrate that crude extracts were able to inhibit some multi-resistant microorganisms.Significance and Impact of Study: This study evaluated another potential pharmacological property of the extract of Hancornia speciosa Gomes, in a context of dissemination bacterial multiresistance, which demands the discovery of new alternative methods for the treatment of infections

Multiplex PCR for molecular characterization of isolated strains of granuloma in sheep from state of Sergipe and Antimicrobial Susceptibility Test (AST) / PCR Multiplex para caracterizaçao molecular de linhagem isolada de granuloma em ovelha no estado de Sergipe e Teste de suceptibilidade antimicrobiana

Corynebacterium pseudotuberculosis is the etiologic agent responsible for Caseous Lymphadenitis (CLA), an infectious disease that affects cattle and horses, but mostly small ruminants such as sheep and goats. This pathology, also known as the "bad seed," causes abscesses formation in the contaminated animal tissue, which can be superficial or visceral. As it is present in several Brazilian states, it is necessary to find out effective ways of diagnostic, prophylactic and effective treatment for CLA in attempt to eliminate this pathogen in rural areas. In Sergipe, the goat farming has progressively gained recognition in recent years and has surpassed over 50% of sheep flocks in the last decade. Therefore, Sergipe has presented a considerable development in sheep farming as well as it has expanded its potential commercialization regarding these animals and their derivatives. However, with the lack of epidemiological analysis, it becomes difficult to know both the presence and prevalence of the disease in the state. Thus, this study aimed to identify the microorganism isolated from a sheep granuloma in Sergipe not only by biochemical tests, considered the gold standard for C. pseudotuberculosis but also by molecular analysis through multiplex PCR, as well as to evaluate the antimicrobial susceptibility test

Eficácia do chip de gluconato de clorexidina na terapia da peri-implantite

 A Peri-implantite é um processo inflamatório que afeta os tecidos ao redor do implante dentário. O tratamento consiste  no procedimento de raspagem e alisamento radicular, todavia a recorrência e a dificuldade de tratar a doença levaram ao desenvolvimento de meios associativos, entre eles o chip de clorexidina. Nesse sentido, o  objetivo deste artigo foi nortear uma investigação sobre a eficácia do chip de gluconato de clorexidina como terapia da peri-implantite. Foi realizada uma revisão de literatura a partir das bases de dados PubMed, ScienceDirect e Lilacs dos últimos 5 anos. As buscas foram conduzidas com os  seguintes   descritores:  “Chip”, “Chlorhexidine”, “Peri-implantitis”, “Chip AND Chlorhexidine” e “Chlorexidine AND Peri-implantitis”, “Chip” AND “Chlorhexidine” AND “Peri-implantitis” combinadas entre si por meio do operador booleano AND. A entrega local de clorexidina através do chip mostrou-se eficaz na redução de sinais clínicos de doença peri-implantar