Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk / Purificação parcial da enzima isomerase do ácido linoleico da bactéria Lactobacillus paracasei isolada do leite

Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 µg.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 µg.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Is the use of Lactobacillus reuteri probiotic efficient as adjunctive therapy in the treatment of periodontitis? A systematic review / O uso do probiótico Lactobacillus reuteri como terapia adjuvante no tratamento da periodontite é eficaz? Uma revisão sistemática

Objective: This systematic review had the purpose to validate the probiotic Lactobacillus reuteri as adjuvant therapy in the periodontal treatment of periodontitis, by the analysis of randomized controlled trial, controlled clinical trial, and observational studies. Material and Methods: Search keys related to the subject were defined, and the following databases were used as search strategies: MEDLINE via PubMed, Scopus, Web of Science, Cochrane Central Controlled Trials Registry, and EMBASE. The data selection and study were performed by two independent evaluators: first, they selected the article by title and abstract and subsequently qualified according to the bias risk analysis. A narrative synthesis has been performed based on the data obtained from the best-quality articles. After data extraction, their heterogeneity was analyzed, and a meta-analysis was performed. Results: In general, the results of the meta-analysis were positive for the use of probiotics as an adjuvant treatment. Considering the limitations of the comparisons between the analyzed articles studied, the conclusion was that probiotics may provide supplementary benefits to the treatment of periodontitis, with improvement in bleeding on probing rates and probing depth (AU)

Objetivo: Esta revisão sistemática teve como objetivo validar o uso do probiótico Lactobacillus reuteri como terapia adjuvante no tratamento periodontal da periodontite, por meio da análise de ensaios clínicos randomizados, ensaios clínicos controlados e estudos observacionais. Material e Métodos: Chaves de busca relacionadas ao assunto foram definidas, e as seguintes bases de dados foram utilizadas como estratégias de busca: MEDLINE via PubMed, Scopus, Web of Science, Cochrane Central Controlled Trials Registry e EMBASE. A seleção dos dados e o estudo foram realizados por dois avaliadores independentes: primeiro, selecionaram o artigo pelo título e resumo e posteriormente qualificaram de acordo com a análise de risco de viés. Uma síntese narrativa foi realizada com base nos dados obtidos dos artigos de melhor qualidade. Após a extração dos dados, sua heterogeneidade foi analisada e uma meta-análise foi realizada. Resultados: Em geral, os resultados da meta-análise foram positivos para o uso de probióticos como tratamento adjuvante. Considerando as limitações das comparações entre os artigos analisados estudados, concluiu-se que os probióticos podem trazer benefícios complementares ao tratamento da periodontite, com melhora nas taxas de sangramento à sondagem e na profundidade de sondagem (AU)

Distribution and function of cariogenic bacteria in supragingival plaque in patients with type 2 diabetes mellitus / 口腔疾病防治

Objective@#To investigate the distribution characteristics and functional genes of cariogenic bacteria in oral microorganisms of patients with type 2 diabetes mellitus and to improve the understanding of the relationship between type 2 diabetes mellitus and dental caries.@*Methods@#The experimental group included 10 patients with type 2 diabetes treated in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University. The normal control group included healthy oral subjects without type 2 diabetes in the community population (10 cases). Samples of supragingival plaque from patients with type 2 diabetes mellitus and normal controls were collected and sequenced. Bioinformatics and statistical analysis of cariogenic bacteria such as Streptococcus mutans, Lactobacillus, Actinomyces viscosus and Candida albicans were carried out.@*Results@#There were slightly fewer cariogenic bacteria such as Streptococcus mutans, Lactobacillus, Actinomyces viscosus and Candida albicans in supragingival plaque samples of type 2 diabetic patients than in normal controls, but the difference was not statistically significant (P>0.05). The results of KEGG pathway functional metabolic differences showed that the metabolic pathways of D-arginine and D-ornithine metabolism, biofilm formation-Escherichia coli, carolactam degradation and arginine biosynthesis were more abundant in the T2DM group than in the normal control group, while metabolic pathways such as tyrosine metabolism, selenocompound metabolism and pyruvate metabolism showed the opposite trend. @*Conclusion @#There was no significant difference in the content of cariogenic microorganisms between type 2 diabetic patients and normal control group. The differential metabolic pathways of the functional genes indicated that an increase in the arginine metabolic pathway was beneficial to the maintenance of acid-base balance in the oral microecological environment.

Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Avaliação molecular de populações fecais de lactobacilos em crianças com prisão de ventre funcional / Molecular assessment of fecal lactobacilli populations in children with functional constipation

ABSTRACT Background: Investigation of the gut-specific bacterial strains including lactobacilli is essential for understanding the bacterial etiology of constipation. Objective: This study aimed to compare the prevalence and quantity of intestinal lactobacilli in constipated children and healthy controls. Methods: Forty children fulfilling Rome IV criteria for functional constipation and 40 healthy controls were recruited. Fecal samples were analyzed using species-specific polymerase chain reaction followed by random amplified polymorphic DNA-PCR and quantitative real-time PCR. Results: Totally, seven different species of lactobacilli were detected. Out of 80 volunteers, 65 (81.3%) were culture and species-specific PCR positive from which 25 (38.46%) constipated children and 40 (61.54%) healthy subjects. The most prevalent species were L. paracasei 21 (32.3%) followed by L. plantarum 18 (27.7%) among both healthy and patient groups. Analysis of the RAPD dendrograms displayed that strains isolated from constipated and non-constipated children have similarity coefficients of more than 90%. The qPCR assays demonstrated constipated children had a lower amount of total lactobacilli population (per gram of feces) than healthy controls. Conclusion: Our findings showed that the mere existence of various species of Lactobacillus in the gut does not enough to prevent some gastrointestinal disorders such as functional constipation, and their quantity plays a more important role.

RESUMO Contexto: A investigação das cepas bacterianas específicas do intestino, incluindo lactobacilos, é essencial para a compreensão da etiologia bacteriana da prisão de ventre. Objetivo: Este estudo teve como objetivo comparar a prevalência e a quantidade de lactobacilos intestinais em crianças constipadas e controles saudáveis. Métodos: Foram recrutadas quarenta crianças que preenchem os critérios de Roma IV para prisão de ventre funcional e 40 controles saudáveis. As amostras fecais foram analisadas utilizando-se uma reação da cadeia de polimerase específica da espécie, seguida por DNA polimórfico amplificado aleatório e PCR quantitativo em tempo real. Resultados: Foram detectadas sete espécies diferentes de lactobacilos. Dos 80 voluntários, 65 (81,3%) eram cultura em PCR específico de espécies, dos quais 25 (38,46%) crianças constipadas e 40 (61,54%) indivíduos saudáveis. As espécies mais prevalentes foram L. paracasei 21 (32,3%) seguidas por L. plantarum 18 (27,7%) entre grupos saudáveis e de pacientes. A análise dos dendrogramas do RAPD mostrou que cepas isoladas de crianças constipadas e não constipadas têm coeficientes de similaridade superiores a 90%. Os ensaios qPCR demonstraram que as crianças constipadas apresentavam uma quantidade menor de população total de lactobacilos (por grama de fezes) do que os controles saudáveis. Conclusão: Nossos achados mostraram que a mera existência de várias espécies de Lactobacillus no intestino não é suficiente para prevenir alguns distúrbios gastrointestinais, como a prisão de ventre funcional, e sua quantidade desempenha um papel mais importante.

Symbiotic pectin microparticles with native Jerusalem artichoke (Helianthus tuberosus L.) enhance Lactobadllus paracasei subsp. tolerans survival / La encapsulación de Lactobacillus paracasei subsp. toleraos en microgeles de pectina con inulina extraída de topinambur (Helianthus tuberosus L.) incrementa su supervivencia

Resumen El efecto sinérgico de la microencapsulación en geles de pectina e inulina extraída de cultivos nativos de topinambur se evaluó como una estrategia natural para aumentar la supervivencia de Lactobacillus paracasei subsp. toleraos F2, bacteria seleccionada por sus propiedades probióticas sobre la trucha arco iris (Oncorhynchus mykiss). La cepa fue capaz de crecer y fermentar la inulina de topinambur en caldo MRS modificado, lo que condujo al incremento del número de células (~+5 unidades logarítmicas) y la disminución neta del pH (6,2 ±0,2 a 4,0±0,5). La encapsulación de F2 en microgeles de pectina en presencia de inulina mejoró su supervivencia no solo a lo largo del almacenamiento sino también en condiciones donde se simuló el pasaje por el tracto gastrointestinal. Después de 56 días de almacenamiento a 4 °C, se registraron números significativamente mayores de células viables microencapsuladas en presencia del prebiótico (8,2-8,4 log UFC/g) que sin él (~7,00 log UFC/g). Estos resultados alientan la aplicación de estas técnicas para la formulación de alimentos funcionales utilizando fuentes alternativas naturales de inulina que incrementan la viabilidad del probiótico almacenado y su digestibilidad.

Abordaje de los probióticos en pediatría: el rol de Lactobacillus rhamnosus GG / Approach to probiotics in pediatrics: the role of Lactobacillus rhamnosus GG

El parto prematuro, las cesáreas, los antibióticos y la lactancia materna limitada contribuyen al aumento de enfermedades crónicas no transmisibles. El objetivo fue realizar una revisión descriptiva del uso de probióticos en pediatría, con foco en la cepa Lactobacillus rhamnosus GG. Ciertos probióticos han demostrado ser eficaces en la diarrea aguda y en la diarrea asociada a antibióticos. L. rhamnosus GG y Saccharomyces boulardii pueden acortar la duración y los síntomas. L. reuteri DSM 17938 y L. rhamnosus GG fueron efectivos para el abordaje del cólico del lactante. El uso de esta cepa en fórmulas infantiles para alergia a las proteínas de leche de vaca promovería la adquisición más temprana de tolerancia. En la prevención de dermatitis atópica, la administración de L. rhamnosus GG durante el embarazo redujo su manifestación en el bebé. El empleo de probióticos como coadyuvantes es una posibilidad para considerar en la práctica pediátrica actual.

Preterm birth, C-sections, antibiotics, and limited breastfeeding contribute to the increase in noncommunicable diseases. Our objective was to perform a descriptive review of probiotic use in pediatrics, focused on Lactobacillus rhamnosus GG. Certain probiotics have demonstrated to be effective in acute diarrhea and antibiotic-associated diarrhea. L. rhamnosus GG and Saccharomyces boulardii may shorten their duration and symptoms. L. reuteri DSM 17938 and L. rhamnosus GG were effective to manage infant colic. The use of this strain in infant formulas for cow's milk protein allergy may promote an earlier tolerance acquisition. In relation to the prevention of atopic dermatitis, the administration of L. rhamnosus GG during pregnancy reduced its development in the infant. The use of probiotics as adjuvants is a possibility to consider in current pediatric practice.

Bio prospecting of Riboflavin producing bacteria from different riboflavin enriched food sources

Riboflavin is an essential, water-soluble vitamin (B2) and a component of basic cellular metabolism. The aim of the present study is to isolate and characterize riboflavin producing bacteria from different food sources. Ten different riboflavin enriched food sources were collected from Vellore district. Totally 72 bacterial strains were isolated and cultured on nutrient agar plates. Out of these, 43 strains were identified as riboflavin producers. Isolated bacterial strains HDS27, HDS07, HDS14, HDS18, HDS38 and HDS54 isolated from milk, mushroom, spinach, lamb kidney, beef liver and mackerel fish were found to be potent riboflavin producers. Based on morphological, biochemical and molecular characterization, the potent strains were identified as Lactobacillus plantarum (HDS27), Bacillus cereus (HDS07), Delftia tsuruhatensis (HDS14), Citrobacter freundii (HDS18), Enterobacter cloacae (HDS38) and Bacillus cereus (HDS54). The selected potent isolates HDS27 from milk and HDS07 from mushroom showed a maximum riboflavin production of 3.69 mg/L and 2.9mg/L respectively. The present study explores the riboflavin producing novel bacteria from different food sources. This is the first report that the Enterobacter cloacae isolated from beef liver, Delftia tsuruhatensis from spinach and Citrobacter freundii from lamb kidney has the ability to produce riboflavin. These potent strains could be a better starter for substituting the conventional bacteria for large scale production of riboflavin in industry.

Lactobacilluscrispatus dominance in the vaginalmicrobiome reduces the occurrence of spontaneouspreterm birth in women with a short cervical length / Dominância de Lactobacillus crispatus no microbioma vaginal reduz a ocorrênciade parto prematuro espontâneo em mulheres com encurtamento do colo uterino

Introduction: The majority of pregnant women with a short cervix will deliver at term and, thus, may unnecessarily receive advanced monitoring and treatment. It is still necessary to define more accurately which sub-population of women with a short cervix is at elevated risk for early delivery. Objective: To determine if vaginal microbiome composition influenced the rate of spontaneous preterm birth in women with a short cervical length. Methods: In an exploratory, observational prospective study, vaginal secretions were obtained from 591 women at 21­24 week gestation. Vaginal microbiome composition was determined by analyzing the V1­V3 region of the bacterial 16S ribosomal RNA gene. Results: Lactobacillus crispatus was numerically dominant in the vagina in 41.7% of subjects, followed by L. iners in 32% and Gardnerella vaginalis in 12%. In women whose cervix was ≤25mm, the sensitivity to predict an spontaneous preterm birth was 11.8%. However, when L. crispatus was not the dominant vaginal bacterium, this sensitivity increased to 81.8%. Similarly, in women with a cervical length ≤30mm, the sensitivity to predict an spontaneous preterm birth increased from 21.7 to 78.3% when L. crispatus was not the dominant vaginal bacterium.In women with a prior spontaneous preterm birth and a cervix ≤25 or ≤30mm, L. crispatus dominance was also associated with a reduced rate of spontaneous preterm birth in the current pregnancy (p<0.001). Conclusion: In pregnant women with a cervix ≤25mm or ≤30mm, the risk for an spontaneous preterm birth is increased if L. crispatus is not dominant in the vagina.

Introdução: A maioria das mulheres grávidas com colo do útero curto dará à luz a termo e, portanto, pode receber desnecessariamente monitoramento e tratamento avançados. Permanece a necessidade de definir com mais precisão qual subpopulação de mulheres com colo do útero curto está em risco elevado de parto prematuro. Objetivo: Determinar se a composição do microbioma vaginal influenciou a taxa de parto prematuro espontâneo em mulheres com colo curto. Métodos: Em um estudo prospectivo exploratório observacional, os conteúdos vaginais foram obtidos de 591 mulheres com 21­24 semanas de gestação. A composição do microbioma vaginal foi determinada pela análise da região V1­V3 do gene de RNA ribossômico bacteriano 16S. Resultados: Lactobacilluscrispatus foi numericamente dominante na vagina em 41,7% dos indivíduos, seguido por L. iners em 32% e Gardnerella vaginalis em 12%. Em mulheres cujo colo do útero era <25 mm, a sensibilidade para prever uma taxa de parto prematuro espontâneo foi de 11,8%. No entanto, quando L. crispatus não era a bactéria vaginal dominante, essa sensibilidade aumentou para 81,8%. Da mesma forma, em mulheres com comprimento cervical <30 mm, a sensibilidade para prever uma taxa de parto prematuro espontâneo aumentou de 21,7 para 78,3% quando L. crispatus não era a bactéria vaginal dominante. Em mulheres com taxa de parto prematuro espontâneo anterior e colo do útero <25 ou <30 mm, a dominância de L. crispatus também foi associada a uma taxa reduzida de taxa de parto prematuro espontâneo na gravidez atual (p<0,001). Conclusão: Em mulheres grávidas com colo do útero <25 ou <30 mm, o risco de parto prematuro espontâneo é aumentado se L. crispatus não for dominante na vagina.

Aplicação do plasma de baixa temperatura sob pressão atmosférica no controle de biofilmes cariogênicos / Application of low temperature plasma under atmospheric pressure in the control cariogenic biofilms

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU) The general objective of this study was to evaluate the application of lowtemperature plasma under atmospheric pressure (PBTPA) of argon and helium flow, in the control of cariogenic biofilms. For this, the effective physical parameters of PBTPA-argon and helium in mono, dual and polymicrobial biofilms composed of combinations of the species Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans and Actinomyces naeslundii were established. The multi-species biofilms were treated by different PBTPA generating devices, one obtained commercially (kINPen09®) that used argon as working gas, and another prototype developed by FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) that used helium as working gas. Quantitative and microscopic analyzes (confocal, scanning electron microscopy) were performed. Negative control (no treatment), positive control (chlorhexidine 2%) and gas control (argon) were included. Besides that, cellular effects of PBTPA-argon and helium on dual and multi-species biofilms were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy. The results obtained for the treatments of mono, dual or multispecies biofilms with both PBTPA-argon and helium were all significant when compared to the negative control at all times analyzed. For PBTPA-argon, there was no recovery of S. gordonii and S. sanguinis at all analyzed times. For PBTPA-helium, the best results were obtained at 5 and 7 min of exposure of biofilms to PBTPA. All the tests were carried out in triplicate in three independent experiments. The results are tabulated and analyzed in terms of distribution. Next, the most suitable statistical tests were selected. The level of significance was 5%. The results obtained for the treatments of mono, dual or multi-species biofilms with PBTPA-argon and helium were all significant compared to the negative control at all analyzed times. Finally, both PBTPA generating could contribute to the development of new protocols for Minimal Intervention Dentistry (AU)

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU)

Advances in the study of intestinal flora for the treatment or prevention of spinal cord injury and complications / 中国医师杂志

Intestinal microorganisms are closely related to human health or disease status. Spinal cord injury is a serious traumatic disease of the nervous system. Recent studies have shown that intestinal flora can affect the progression of spinal cord injury. At the same time, intestinal flora is closely related to spinal cord injury complications such as anxiety, depression, aspiration pneumonia, malnutrition, constipation, neurocystitis and urinary system infection through intestinal-brain axis, intestinal-lung axis and oral-pulmonary axis. In this paper, the intestinal flora will be used as the starting point to explore the relationship between spinal cord injury, intestinal flora and spinal cord injury complications, providing new ideas for the treatment of spinal cord injury patients and the prevention of complications.

Efficacy of compound Lactobacillus acidophilus tablets combined with bismuth-containing quadruple therapy in the treatment of chronic non-atrophic gastritis and its effect on intestinal microflora / 中国基层医药

Objective:To investigate the efficacy of compound Lactobacillus acidophilus tablets combined with bismuth-containing quadruple therapy in the treatment of Helicobacter pylori-positive (Hp +) chronic non-atrophic gastritis and its effect on Lactobacillus and Escherichia coli.Methods:A total of 104 patients with Hp + chronic non-atrophic gastritis who received treatment in Affiliated Hospital of Shaoxing University from March 2019 to September 2020 were included in this study. They were randomly divided into control and observation groups, with 52 patients in each group. The control group was treated with bismuth-containing quadruple therapy. The observation group was treated with compound Lactobacillus acidophilus tablets (2 tablets once, three times per day) based on bismuth-containing quadruple therapy. Two groups were treated for 2 successive weeks. The endoscopic grading score of gastric mucosa, improvement in pathological grading of gastric mucosa, Hp eradication rate, and Lactobacillus and Escherichia coli levels were compared between the two groups. Results:After treatment, endoscopic grading score of gastric mucosa (gastric mucosa inflammation, inflammation activity, atrophy, and intestinal metaplasia) in the observation group was (1.31 ± 0.18) points, (1.24 ± 0.18) points, (1.15 ± 0.16) points, (1.04 ± 0.15) points, respectively, which was superior to (1.73 ± 0.25) points, (1.59 ± 0.21) points, (1.44 ± 0.19) points, (1.30 ± 0.18) points in the control group ( t = 3.67, 3.80, 4.17, 4.52, all P < 0.01). Improvement in pathological grading of gastric mucosa in the observation group was significantly higher than that in the control group [94.23% (49/52) vs. 78.85% (41/52), χ2 = 4.04, P < 0.05]. Hp eradication rate in the observation group was significantly higher than that in the control group [92.31% (48/52) vs. 75.00% (39/52), χ2 = 4.50, P < 0.05]. After treatment, the amount of Lactobacillus and Escherichia coli in the observation group were (8.30 ± 0.95) copies and (8.04 ± 0.95) copies, which were significantly greater than (7.51 ± 0.93) copies and (8.30 ± 0.95) copies in the control group ( t = 4.02, 3.75, both P < 0.01). Conclusion:Compound Lactobacillus acidophilus tablets combined with bismuth-containing quadruple therapy is highly effective on Hp + chronic non-atrophic gastritis and it can effectively improve intestinal microflora.

Progress of probiotics and allergic diseases in children / 国际儿科学杂志

Intestinal microbiota plays an important role in the establishment and maturation of the host immune system.In recent years, it has been found that the disorder of intestinal microecology is related to the occurrence and development of allergic diseases in children.Probiotics can improve intestinal microecological disorders, and its relationship with the prevention and treatment of allergic diseases is currently a research hotspot.In this review, we review the research progress on the relationship between intestinal microbiota and allergic diseases in children, the immunomodulatory mechanisms of probiotics rebalancing intestinal microecology, and the relationship between Lactobacillus paracei and allergic diseases such as atopic dermatitis, allergic rhinitis, and bronchial asthma.

Selective Cytotoxic effect of Probiotic, Paraprobiotic and Postbiotics of L. casei strains against Colorectal Cancer Cells: Invitro studies

Abstract This study highlights the cytotoxic effect of three L. casei strains on colorectal cell lines in invitro conditions. Different concentrations of live, heat killed (HK) and cell free supernatant (CFS) of three L.casei strains were subjected to CaCo2 and MRC5 cell lines. The viability of the treated and untreated cells was determined after 72 hrs by MTT assay, and IC50 estimated. Apoptosis was evaluated by Annexin V-propidium iodide method using flow cytometry. The live, HK and CFS of the L. casei strains showed cytotoxic effects on colorectal cell lines with significant differences. The cytotoxicity effects of live cells on CaCo2 cells were significantly higher (p˂0.01) than the HK cells. A dose dependent response was observed, as higher concentrations resulted in enhanced cytotoxicity effects. Live L.casei 1296-2cells inhibited 91% of CaCo2 cell growth, with IC50 of less than 108 cfu/ml. MRS medium and concentrations of CFS at above 20% v/v, were cytotoxic to the normal cell lines. Flow cytometry analyses of L. casei 1296-2 indicated that cytotoxicity effects on CaCo2 cells is related to apoptotic induction. Invitro studies indicate that Live and CFS of L. casei 1296-2 might be promising candidate for the control of colorectal cancers

Synthesis of silver nanoparticles using Lactobacillus bulgaricus and assessment of their antibacterial potential / Síntese de nanopartículas de prata usando Lactobacillus bulgaricus e avaliação de seu potencial antibacteriano

Abstract Many pathogenic strains have acquired multidrug-resistant patterns in recent a year, which poses a major public health concern. The growing need for effective antimicrobial agents as novel therapies against multidrug-resistant pathogens has drawn scientist attention toward nanotechnology. Silver nanoparticles are considered capable of killing multidrug-resistant isolates due to their oligo-dynamic effect on microorganisms. In this research study NPs were synthesized using the gram-positive bacteria Lactobacillus bulgaricus and its activity against selected pathogenic strains. Lactobacillus bulgaricus pure cultures were isolated from raw milk and grown in "De Man, Rogasa, and Sharp" broth for synthesis of nanoparticles. Lactobacillus bulgaricus culture was centrifuged and Cell- free supernatant of it was employed with aqueous silvery ions and evaluated their antibacterial activities against bacterial strains i.e. Staphylococcus aureus, Staphylococcus epidermidis and Salmonella typhi using agar well diffusion assay. Antibiotic profiling against selected pathogenic strains were also conducted using disc diffusion method. The synthesis and characterization of silver nanoparticles were monitored primarily by the conversion of the pale-yellow color of the mixture into a dark-brown color and via ultraviolet-visible absorption spectroscopy and Scanning electron microscopy respectively. The result showed that that AgNPs with size (30.65-100 nm) obtained from Lactobacillus bulgaricus were found to exhibit antibacterial activities against selected bacterial strains. Taken together, these findings suggest that Lactobacillus bulgaricus has great potential for the production of AgNPs with antibacterial activities and highly effective in comparison to tested antibiotics.

Resumo Muitas cepas patogênicas adquiriram padrões multirresistentes nos últimos anos, o que representa um grande problema de saúde pública. A crescente necessidade de agentes antimicrobianos eficazes como novas terapias contra patógenos multirresistentes atraiu a atenção dos cientistas para a nanotecnologia. As nanopartículas de prata são consideradas capazes de matar isolados multirresistentes por causa de seu efeito oligodinâmico em microrganismos. Neste estudo de pesquisa, as NPs foram sintetizadas usando a bactéria Gram-positiva Lactobacillus bulgaricus e sua atividade contra cepas patogênicas selecionadas. Culturas puras de Lactobacillus bulgaricus foram isoladas do leite cru e cultivadas em caldo "De Man, Rogasa e Sharp" para síntese de nanopartículas. A cultura de Lactobacillus bulgaricus foi centrifugada, e o sobrenadante livre de células foi empregado com íons prateados aquosos, avaliando-se suas atividades antibacterianas contra cepas bacterianas, isto é, Staphylococcus aureus, Staphylococcus epidermidis e Salmonella typhi usando ensaio de difusão em poço de ágar. O perfil de antibióticos contra cepas patogênicas selecionadas também foi conduzido usando o método de difusão em disco. A síntese e a caracterização das nanopartículas de prata foram monitoradas principalmente pela conversão da cor amarelo-pálida da mistura em uma cor marrom-escura e por espectroscopia de absorção visível e ultravioleta e por microscopia eletrônica de varredura, respectivamente. O resultado mostrou que AgNPs com tamanho de 30,65-100 nm, obtidas de Lactobacillus bulgaricus, exibiram atividades antibacterianas contra cepas bacterianas selecionadas. Tomados em conjunto, esses achados sugerem que o Lactobacillus bulgaricus tem um grande potencial para a produção de AgNPs com atividades antibacterianas e altamente eficazes em comparação aos antibióticos testados.

Desenvolvimento de formulações probióticas de isolados clínicos de Lactobacillus spp. para controle de Candida albicans / Development of probiotic formulations of clinical isolates of Lactobacillus spp. for the control of Candida albicans

O uso de probióticos tem sido considerado uma alternativa para o controle das candidoses bucais, como a estomatite protética. Recentemente, nosso grupo de pesquisa isolou cepas clínicas bucais de Lactobacillus rhamnosus (5.2) e Lactobacillus fermentum (20.4) com atividade antifúngica contra Candida albicans. Entretanto, é necessário o desenvolvimento de formulações para uso clínico dessas cepas probióticas na cavidade bucal. Assim, o objetivo desse projeto foi desenvolver formulações probióticas em hidrogel gellan gum com atividade antifúngica que possam ser usadas no controle da estomatite protética. Para o preparo das formulações probióticas, as cepas L. rhamnosus 5.2 ou L. fermentum 20.4 foram incorporadas em formulações com diferentes concentrações de gellan gum (0,6 a 1%). A viabilidade das células de Lactobacillus nas formulações e o sistema de liberação foram testados em diferentes condições de armazenamento, incluindo tempo e temperaturas de estocagem. A seguir, os efeitos das formulações probióticas foram avaliados sobre C. albicans pelo método de difusão em ágar e sobre biofilmes microcosmos por contagem de células viáveis e análise de microscopia eletrônica de varredura. Os resultados demonstraram que todas as concentrações do gellan foram capazes de manter a viabilidade das células de L. rhamnosus e L. fermentum em torno de 5 a 7 log, durante 14 semanas, independentemente da temperatura de armazenamento (4°C ou temperatura ambiente). No teste de difusão em ágar, verificou-se que as formulações probióticas levaram à inibição de cepas padrão e clínicas de C. albicans, demonstrando que as cepas de Lactobacillus incorporadas no gellan mantiveram sua atividade antifúngica. As formulações probióticas também conseguiram inibir os biofilmes microcosmos provenientes de pacientes com estomatite protética, reduzindo o número de células viáveis de Candida e desestruturando a arquitetura dos biofilmes. Concluiu-se que o gellan gum mostrou ser um biomaterial promissor como carreador das cepas probióticas de L. rhamnosus 5.2 e L. fermentum 20.4. Além disso, as formulações probióticas desenvolvidas apresentaram ação antifúngica sobre células de C. albicans e biofilmes de estomatite protética, podendo desta forma atuar na prevenção da candidose bucal. (AU)

The use of probiotics has been considered an alternative for the control of oral candidiasis such as denture stomatitis. Recently, our research group isolated oral clinical strains of Lactobacillus rhamnosus (5.2) and Lactobacillus fermentum (20.4) with antifungal activity against Candida albicans. However, it is necessary to develop formulations for clinical use of these probiotic strains in the oral cavity. Thus, the purpose of this project was to develop probiotic formulations in gellan gum hydrogel with antifungal activity that could be used in the control of denture stomatitis. For the preparation of probiotic formulations, the strains L. rhamnosus 5.2 or L. fermentum 20.4 were incorporated into formulations with different concentrations of gellan gum (0.6 to 1%). The viability of Lactobacillus cells in the formulations and the delivery system were tested under different storage conditions, including storage time and temperatures. Next, the effects of the probiotic formulations were evaluated on C. albicans by the agar diffusion method and on microcosm biofilms by viable cell count and scanning electron microscopy analysis. The results showed that all concentrations of gellan were able to maintain the viability of L. rhamnosus and L. fermentum cells around 5 to 7 log for 14 weeks, regardless of storage temperature (4°C or room temperature). In the agar diffusion test, it was found that the probiotic formulations led to the inhibition of standard and clinical strains of C. albicans, demonstrating that the Lactobacillus strains incorporated into gellan maintained their antifungal activity. Probiotic formulations were also able to inhibit microcosm biofilms from patients with denture stomatitis, reducing the number of viable Candida cells and disrupting the architecture of biofilms. As a conclusion, the gellan gum proved to be a promising biomaterial as a carrier of the probiotic strains of L. rhamnosus 5.2 and L. fermentum 20.4. In addition, the probiotic formulations developed showed antifungal action on C. albicans cells and denture stomatitis biofilms, thus being able to act in the prevention of oral candidiasis. (AU)

Alendronate combined with Lactobacillus rhamnosus to combat bone loss in ovariectomized mice / 中华创伤骨科杂志

Objective:To study the protective effect of alendronate combined with Lactobacillus rhamnosus on bone loss in ovariectomized mice.Methods:Fifty female C57BL/6 mice were divided into 5 equal groups ( n=10). Ovariotomy was performed in groups A, B, C and D while a sham operation was performed in group E. Group A was subjected to combined administration of alendronate and Lactobacillus rhamnosus, group B to administration of alendronate, group C to administration of Lactobacillus rhamnosus, and groups D and E to administration of physiological saline only. At 3 months after operation, all the mice were sacrificed to harvest their femurs. Micro CT scanning was performed to detect the bone mineral density (BMD), trabecular relative volume, bone surface area/bone volume, and trabecular thickness and number of trabecular bone. Three-point bending test was used to detect the maximum load, stiffness, ultimate load, Young＇s modulus, and fracture energy. Osteocalcin and alkaline phosphatase levels were measured using blood samples from the mice eyeballs. The 2 groups were compared in terms of all the above indexes. Results:The BMD [(669.87±67.87) mg/cm 3], maximum load [(14.35±0.75) N] and fracture energy [(1,497.43±38.29) J/m 2] in group A were significantly higher than those in group B [(520.07±9.01) mg/cm 3, (11.94±0.82) N and(1,277.61±35.12) J/m 2] and group C [(388.15±25.61) mg/cm 3, (11.10±0.93) N and (1,115.27±63.24) J/m 2] (all P<0.05). The osteocalcin level in group A [(22.25±1.78) ng/mL] was significantly higher than that in group B [(19.08±1.45) ng/mL] and group D [(19.33±1.66) ng/mL] (both P<0.05). The alkaline phosphatase level in group A [(83.21±9.69) ng/mL] was significantly lower than that in group C [(113.16±14.44) ng/mL] and group D [(137.96±14.01) g/mL] (both P<0.05). Conclusion:Alendronate combined with Lactobacillus rhamnosus may play a synergistic role in prevention of bone loss in ovariectomized mice, because combined administration of the two is more effective than administration of either of the two.

Advances in Lactobacillus rhamnosus GG and pediatric nonalcoholic fatty liver disease / 中华实用儿科临床杂志

Nonalcoholic fatty liver disease (NAFLD)has become one of the most common chronic liver diseases in children, which has a wide spectrum including the nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis and hepatocellular carcinoma (HCC)with its clinical consequences.At least 50% of obese children present NAFLD.Increased levels of free fatty acids and insulin resistance play major roles in NAFLD progression, resulting in excessive accumulation of triglycerides in hepatocytes.Lactobacillus rhamnoides is one of the most thoroughly studied intestinal probiotics, and its role in NAFLD progress has been growingly concerned.Lactobacillus rhamnosus GG alleviates NAFLD in children by improving intestinal microbiota dysbiosis, increasing intestinal barrier integrity, promoting fatty acid β-oxidation, and reducing hepatic triglyceride accumulation via regulating bile acid metabolism.Recent studies have shown that microRNAs are involved in multiple pathways to alleviate NAFLD-associated lipid metabolism disorders, which are functional as noninvasive diagnostic marker of fatty liver disease in children.This review mainly summarizes that Lactobacillus rhamnosus GG improves NAFLD in children from multiple metabolic pathways, aiming to provide a new strategy and target for the treatment and early diagnosis of NAFLD in children.

Mechanism of intestinal flora imbalance in the development and progression of liver cancer and the role of probiotics in preventing liver cancer / 临床肝胆病杂志

Intestinal flora imbalance plays a certain role in the development and progression of liver cancer, while probiotics have a certain impact on liver cancer, both of which are the focus of clinical research. This article introduces the mechanism of action of intestinal flora imbalance in the pathogenesis of liver cancer and the preventive effect of probiotics against liver cancer. Intestinal flora imbalance can participate in the pathological process of liver cancer by activating Toll-like receptor 4, regulating the level of metabolites, producing endotoxin, and inducing bacterial translocation and intestinal bacterial overgrowth, while probiotics can effectively prevent liver cancer by maintaining enterohepatic circulation, enhancing immune function, promoting the reproduction of intestinal probiotics, and reducing the toxicity of carcinogens, which can be further studied as the focus of subsequent liver cancer prevention in clinical practice.

Uso de probióticos en la prevención de enterocolitis necrotizante / Use of probiotics in the prevention of necrotizing enterocolitis

INTRODUCCIÓN: La enterocolitis necrotizante (ECN) es una patología inflamatoria del sistema digestivo, potencialmente mortal, que afecta a los recién nacidos. Ocurre con mayor frecuencia en recién nacidos prematuros y en especial en aquellos con muy bajo peso de nacimiento (RNMBPN) y extremo bajo peso de nacimiento (RNEBPN). Su fisiopatología es multifactorial, sin embargo, se postula disbiosis intestinal es un factor patogénico importante en su desarrollo. Por consiguiente, se ha propuesto que la administración de probióticos podría tener un rol en la prevención de la ECN en pacientes con alto riesgo. OBJETIVO: Realizar una búsqueda bibliográfica con el objetivo de conocer la evidencia existente en relación al efecto de los probióticos en recién nacidos. RESULTADOS: Los resultados de las revisiones sistemáticas y metanálisis revisados indican que la administración de probióticos reduce el riesgo de ECN, muerte y sepsis tardía en los RNMBPN, no obstante, en el grupo de RNEBPN aun hacen falta estudios. Los estudios que comparan diferentes estrategias de administración de probióticos señalan que los esquemas que incluyen la administración de una mezcla de 2 o más probióticos obtienen los mejores resultados en la prevención de ECN y muerte. Adicionalmente, algunos artículos sugieren un mejor efecto si los esquemas probióticos a utilizar, incluyen especies de los géneros Bifidobacterium y Lactobacillus. CONCLUSIÓN: Existe evidencia en la literatura especializada que apoya el uso de los probióticos en la prevención de ECN y otros desenlaces adversos en los recién nacido prematuros.

Necrotizing enterocolitis (NEC) is a life-threatening inflammatory disease of the digestive system that affects newborns. It occurs most frequently in preterm infants and especially in very low birth weight (VLBW) and extremely low birth weight (ELBW) infants. Its pathophysiology is multifactorial; however, intestinal dysbiosis is postulated to be an important pathogenic factor in its development. Therefore, it has been proposed that the administration of probiotics may have a role in the prevention of NEC in high-risk patients. This update presents a brief overview of the evidence regarding the effect of probiotics in neonates. The results of the systematic reviews and meta-analyses reviewed indicate that probiotic administration reduces the risk of NEC, death and late sepsis in VLBW infants, however, studies are still lacking in the ELBW group. Studies comparing different probiotic administration strategies indicate that schemes that include the administration of a mixture of 2 or more probiotics obtain the best results in the prevention of NEC and death. Additionally, there is some evidence to favor schemes that include species of the genera Bifidobacterium and Lactobacillus. CONCLUSION: There is evidence in the specialized literature that supports probiotic administration may reduces the risk of NEC and other adverse outcomes in preterm infants.