Do oral antiseptics used in the dental routine have anti-viral efficacy? a systematic review of in vitro studies / Os antissépticos orais utilizados na rotina odontológica têm eficácia anti-viral? Uma revisão sistemática de estudos in vitro / ¿Los antisépticos orales utilizados en la rutina odontológica tienen eficacia antiviral? Una revisión sistemática de estudios in vitro

Introduction: Mouthwashes play an important role in the dental clinic, but their role on viruses requires investigation. Objective:to review in vitro studies to identify the effect of different mouthwashes on the main viruses associated with routine dental care. Methodology:The following databases were searched in September 2023: PubMed, Embase, Scopus and Web of Science databases; the Cochrane Library and the Virtual Health Library (VHL); and grey literature. In vitro studies that used mouthwashes to reduce the viral load were selected. The PICOS strategy was considered to define eligibility criteria: the Population (viruses involved in the etiology of oral infection), the Intervention (oral antiseptics), the appropriate comparator (positive and negative controls), the Outcomes of interest (reduction of viral load) and the Study design (in vitro studies). Results:Considering the eligibility criteria, 19 articles were included in this review. The efficacy of povidone-iodine (PVP-I), chlorhexidine, Listerine®, essential oils, and cetylpyridinium chloride (CPC) rinses were investigated. PVP-I (0.23%) had its effects mainly associated with coronaviruses SARS(Severe Acute Respiratory Syndrome),demonstrating a significant reduction in viral load after 15 seconds of exposure. Chlorhexidine (0.05%; 0.1% and 0.5%) was ineffective against adenovirus, poliovirus, and rhinovirus respiratory viruses. Listerine® demonstrated superior efficacy against HSV-1 and 2 viruses and influenza A, and cetylpyridine chloride also demonstrated virucidal activity against influenza A. Conclusions:The type, concentration, and time of exposure to antiseptics varied between studies. PVP-I and chlorhexidine digluconate were the most studied substances, butin general, PVP-I was more effective in reducing viral titers, especially concerning coronaviruses. Other antiseptics such as CPC, H2O2 and Listerine® have also shown significant reduction in viral load, but this is a limited number of studies (AU).

Introdução: Os enxaguantes bucais desempenham um papel importante na clínica odontológico, porém seu papel sobre os vírus requer investigações. Objetivo: revisar estudos in vitro para identificar o efeito de diferentes colutórios sobre os principais vírus associados ao atendimento odontológico de rotina. Metodologia: As seguintes bases foram pesquisadas até setembro de 2023: PubMed, Embase, Scopus e Web of Science; a Biblioteca Cochrane e a Biblioteca Virtual em Saúde (BVS); e literatura cinzenta. Foram selecionados estudos in vitro que utilizaram bochechos com o objetivo de reduzir a carga viral. A estratégia PICOS foi considerada para a definição dos critérios de elegibilidade: População (vírus envolvidos na etiologia da infecção oral), Intervenção (antissépticos orais), Comparador (controles positivos e negativos), os Desfechos de interesse (redução da carga viral) e o desenho do estudo (estudos in vitro). Resultados: Considerando os critérios de elegibilidade, 19 artigos foram incluídos para esta revisão. A eficácia da povidona-iodo (PVP-I), clorexidina, Listerine®, óleos essenciais e lavagens com cloreto de cetilpiridínio foram investigadas. O PVP-I(0.23%)teve seus efeitos principalmente associados ao coronavírusSARS (Síndrome Respiratória Aguda Severa),demonstrando uma redução significativa da carga viral após 15 segundos de exposição. A clorexidina mostrou-se ineficaz contra vírus respiratórios de adenovírus, poliovírus e rinovírus. Listerine® demonstrou eficácia superior contra vírus HSV-1 e 2 e vírus influenza A, e cloreto de cetilpiridinio também demonstrou atividade virucida contra influenza A.Conclusões:O tipo, concentração e tempo de exposição aos antissépticos variaram entre os estudos. O PVP-I e o digluconato de clorexidina foram as substâncias mais estudadas, mas no geral, o PVP-I foi mais eficaz na redução dos títulos virais, principalmente no que diz respeito aos coronavírus. Outros antissépticos como CPC, H2O2 e Listerine® também mostraram redução significativa da carga viral, mas trata-se de um número limitado de estudos (AU).

Introducción: Los enjuagues bucales son importantesen la clínica dental, sin embargo, su efecto sobre los virus requiere investigaciones. Objetivo: Revisar estudios in vitro para identificar el efecto de enjuagues bucales sobre los principales virus asociados con larutinaodontológica. Metodología: Las siguientes bases de datos fueron investigadas hasta septiembrede 2023: PubMed, Embase, Scopus y Web of Science; Biblioteca Cochrane y Biblioteca Virtual en Salud (BVS); yliteratura gris. Se seleccionaron estudios in vitro que utilizaron enjuagues bucales con el objetivo de reducir la carga viral. Se consideró la estrategia PICOS para definir los criterios de elegibilidad: Población (virus implicados en la etiología de la infección oral), Intervención (antisépticos bucales), Comparador (controles positivos y negativos), Resultados de interés (reducción de la carga viral) y diseño del estudio (in vitro). Resultados: Considerando los criterios de elegibilidad, se incluyeron 19 artículos.Se investigó la eficacia de povidona yodada (PVP-I), clorhexidina, Listerine®,aceites esenciales y enjuagues de cloruro de cetilpiridinio (CPC). PVP-I(0.23%)mostró sus efectos principalmente asociados al coronavirus SARS(Síndrome Respiratorio Agudo Severo), demostrando una reducción significativa de la carga viral después de 15 segundos. Se ha demostrado que la clorhexidina es ineficaz contra losvirus respiratorios adenovirus, poliovirus y rinovirus. Listerine® demostró una eficacia superior contra los virus HSV-1 y 2 y el virus de la influenza A, y el CPCtambién mostró actividad virucida contra la influenza A.Conclusiones: El tipo, la concentración y el tiempo de exposiciónvariaron entre los estudios. PVP-I y digluconato de clorhexidina fueron las sustancias más estudiadas, pero,PVP-I fue más efectiva en la reducción de los títulos virales, especialmente en lo que respecta a los coronavirus. Otros antisépticos como CPC, H2O2 y Listerine® también mostraron una reducción significativa de la carga viral, pero se trata de un número limitado de estudios (AU).

In vitro production of cat-restricted Toxoplasma pre-sexual stages.

Sexual reproduction of Toxoplasma gondii, confined to the felid gut, remains largely uncharted owing to ethical concerns regarding the use of cats as model organisms. Chromatin modifiers dictate the developmental fate of the parasite during its multistage life cycle, but their targeting to stage-specific cistromes is poorly described1,2. Here we found that the transcription factors AP2XII-1 and AP2XI-2 operate during the tachyzoite stage, a hallmark of acute toxoplasmosis, to silence genes necessary for merozoites, a developmental stage critical for subsequent sexual commitment and transmission to the next host, including humans. Their conditional and simultaneous depletion leads to a marked change in the transcriptional program, promoting a full transition from tachyzoites to merozoites. These in vitro-cultured pre-gametes have unique protein markers and undergo typical asexual endopolygenic division cycles. In tachyzoites, AP2XII-1 and AP2XI-2 bind DNA as heterodimers at merozoite promoters and recruit MORC and HDAC3 (ref. 1), thereby limiting chromatin accessibility and transcription. Consequently, the commitment to merogony stems from a profound epigenetic rewiring orchestrated by AP2XII-1 and AP2XI-2. Successful production of merozoites in vitro paves the way for future studies on Toxoplasma sexual development without the need for cat infections and holds promise for the development of therapies to prevent parasite transmission.

Effect of Water and Chlorhexidine with Different Pressures of Oral Irrigation Device on the Surface Roughness and Topography of Giomer

Abstract Objective: To evaluate the effect of different pressures of an oral irrigation device (OID) and the irrigation solution type on the surface roughness of the giomer restorative material. Material and Methods: In this in vitro study, disk-shaped giomer samples were fabricated and assigned to 5 groups (n=23): Group 1, storage in distilled water (control); Group 2, OID #7 pressure/ water; Group 3, OID #10 pressure/ water; Group 4, OID #7 pressure/ 0.05% CHX; Group 5, OID #10 pressure/ 0.05% CHX. The samples' treatment simulated a one-year application of OID. Surface roughness (Ra) and topography of the giomer were evaluated using profilometry and scanning electron microscopy. The data were analyzed with Paired t-test, Tukey, and ANOVA tests (α=0.05). Results: The Ra of the samples increased significantly after treatment with OID (p<0.001). The roughness increase in groups with a pressure of 10 was higher than those with a pressure of 7 (p<0.001). The effect of pressure on surface changes was significant (p<0.001). However, the solution type and the cumulative effect of these two factors were insignificant (p=0.08 and p=0.43, respectively). Conclusion: Oral irrigation device with both solutions significantly increased the surface roughness and topographic changes of the giomer. The severity of these changes was related to the device's pressure.

Effect of Toothpaste Fluoride Concentration on Color Stability of Resin Composites: An In Vitro Study

ABSTRACT Objective: To investigate the effects of two different dentifrice fluoride concentrations on the color stability of the composite. Material and Methods: Twenty-seven specimens (2×4×5 mm) each of microfilled (Gradia, GC, Japan) and nanohybrid (Grandio, VOCO, Germany) composites were prepared. The specimens were randomly divided into six groups (control, Fluoflor caries protection toothpaste with 1450ppm Fluoride (EXW, France), and Fluoflor kids toothpaste with 500ppm Fluoride (EXW, France) (n = 9). The specimens were immersed in a mixture of artificial saliva and toothpaste in a ratio of 1:3 and applied for 60 seconds every 12 hours for 42 days. The control samples were incubated in artificial saliva at 37°C. Primary and secondary color measurements were performed using color parameters (L∗a∗b) with a spectrophotoshade (MHT Optic Research AG, Niederhasli, Switzerland). Data were analyzed using a two-way analysis of variance at a significance level of 0.05. Results: According to the two-way ANOVA analysis, there was no significant difference in color change between the composites and no difference in the level of discoloration between different fluoride concentrations(p>0.05). Also, None of the dentifrices caused clinically significant color changes(∆E˂3.3). Conclusion: No clinically unacceptable color changes were observed in the microfilled and nanofilled composites with different concentrations of fluoride toothpaste.

In vitro activity of ceftobiprole and dalbavancin against a collection of coagulase-negative staphylococci isolates from clinical samples with reduced susceptibility to daptomycin and/or resistant to linezolid or glycopeptides / Evaluación de la actividad in vitro de ceftobiprol y dalbavancina frente a estafilococos coagulasa negativos con sensibilidad disminuida a daptomicina y/o resistentes a linezolid o glucopéptidos aislados de muestras clínicas

Introduction:The aim was to investigate the in vitro activity of ceftobiprole and dalbavancin against a collection of coagulase-negative staphylococci (CoNS) isolates with reduced susceptibility to daptomycin or resistant to linezolid and/or glycopeptides. Methods: A total of 228 CoNS were tested using the Vitek-2 AST-626 cards (bioMérieux) and MIC of daptomycin, linezolid, vancomycin and teicoplanin were confirmed by Etest Strips (bioMérieux). Susceptibility testing for ceftobiprole and dalbavancin were performed by CLSI broth microdilution methodology. Results were interpreted according to 2021 EUCAST clinical breakpoints. Results: Ceftobiprole and dalbavancin were active against 96.0% and 93.0% of CoNS, respectively, MIC90 were 2 and 0.125mg/L. MICs of ceptobiprole were higher against S. hominis and S. haemolyticus (MIC90 4mg/L). Dalbavancin exhibited higher MICs against S. haemolyticus and CoNS with reduced susceptibility to daptomycin and resistant to teicoplanin. Conclusion: Ceftobiprole and dalbavancin demonstrated a high in vitro activity against our collection of CoNS isolates.(AU)

Introducción: El objetivo fue evaluar la actividad in vitro de dalbavancina y ceftobiprol frente a estafilococos coagulasa negativos (ECN) con sensibilidad disminuida a daptomicina y/o resistentes a linezolid o glucopéptidos. Métodos: Se testó la sensibilidad de 228 ECN con tarjetas VITEK®2 AST-626 (bioMérieux) y las CMI de daptomicina, linezolid, vancomicina y teicoplanina fueron confirmadas con tiras Etest® (bioMérieux). El ensayo de sensibilidad frente a ceftobiprol y dalbavancina se realizó mediante microdilución en caldo (metodología CLSI). Los resultados se interpretaron siguiendo los puntos de corte de EUCAST 2021. Resultados: Ceftobiprol y dalbavancina fueron activos en el 96,0 y 93% de ECN, las CMI90 fueron 2 y 0,125mg/L, respectivamente. Las CMI de ceftobiprol fueron superiores en Staphylococcus hominis y Staphylococcus haemolyticus (CMI90 4mg/L). Dalbavancina exhibió mayores CMI en S. haemolyticus y en ECN con sensibilidad disminuida a daptomicina o resistentes a teicoplanina. Conclusión: Ceftobiprol y dalbavancina han demostrado una potente actividad in vitro frente a esta colección de ECN.(AU)

Avaliação antimicrobiana de uma resina Bulk Fill Fluida com inserção do xilitol / Antimicrobial evaluation of a Fluid Bulk Fill resin with xylitol insertion

Introdução: As resinas Bulk Fill apresentam uma boa procura pelos profissionais, pois o seu uso diminui o tempo clínico, como também a melhora qualidade das restaurações, porém não possuem atividade antibacteriana, sendo um dos fatores que ainda causam impacto negativo na vida das pessoas. A biomodificação com o xilitol tem o sentido de produzir ação microbiana e com isso aperfeiçoar as suas características clínicas. Objetivo: Avaliar a ação antimicrobiana de uma resina Bulk Fill flow após a inserção do xilitol. Metodologia: A resina Tetric® N-Flow Bulk Fill foi misturada às concentrações de xilitol (0% [Controle], 2,5% e 5% p/p). Amostras cilíndricas (n=5 do grupo controle e n=3 dos demais grupos experimentais) foram confeccionadas em moldes acrílicos de diâmetro de 2mm espessura, fotoativadas por 10s e armazenadas a 37ºC por 24h. Os espécimes foram esterilizados por luz ultravioleta por 20 minutos antes de serem acomodadas em uma placa de 48 poços estéril, sendo adicionado em cada poço 0,5mL de caldo Mueller Hinton. Então, adicionou-se 50µL do inóculo de S. mutans nos poços correspondentes. A placa foi incubada a 37 ± 1 ˚C durante 48 horas. Após o período de incubação, os espécimes foram gentilmente removidos e o crescimento microbiano foi indicado pela adição de 100µL da solução aquosa de resazurina (SigmaAldrich) a 0,01% com a posterior incubação a 37 ± 1 ˚C por duas horas. Micro-organismos viáveis reduzem o corante mudando sua coloração azul para rosa e a CIM foi definida como a menor concentração da substância que inibiu a mudança de coloração da resazurina. Em um poço contendo o grupo controle foi acrescentado clorexidina a 0,12% com o intuito de comparar o resultado gerado dos grupos testes. Resultado: Não houve inibição do crescimento bacteriano nos poços com inóculos que continham S. mutans e corpo de prova de resina acrescida de xilitol. Conclusão: Esse estudo mostrou que o acréscimo de 2,5% e 5% de Xilitol à resina Tetric® N-Flow Bulk Fill não apresentou inibição do crescimento bacteriano (AU).

Introduction: Bulk Fill resins are in good demand among professionals, as their use reduces clinical time and improves the quality of restorations, but they do not have antibacterial activity, which is one of the factors that still hurt people's lives. Biomodification with xylitol aims to improve its clinical characteristics. Objective: To evaluate the antimicrobial action of a Bulk Fill flow resin after inserting xylitol. Methodology: Tetric® N-Flow Bulk Fill resin was mixed with xylitol concentrations (0% [Control], 2.5% and 5% w/w). Cylindrical samples (n=5 from the control group and n=3 from the other groups) were made in acrylic molds with a diameter of 2 mm thick, light-cured for 10 s and stored at 37ºC for 24h. The specimens were sterilized by ultraviolet light for 20 minutes before being placed in a sterile 48-well plate, with 0.5 mL of Mueller Hinton broth added to each well. The plate was incubated at 37 ± 1 ˚C for 48 hours. After the incubation period, the specimens were gently removed, and microbial growth was indicated by adding 100 µL of 0.01% resazurin aqueous solution with subsequent incubation at 37 ± 1 ˚C for two hours. Viable microorganisms reduce the dye, changing its color from blue to pink. The MIC was defined as the lowest concentration of the substance that inhibited the color change of resazurin. In 0.12%, chlorhexidine was added to a well containing the control group to compare the results generated from the test groups. Result: There was no inhibition of bacterial growth in the wells with inocula containing S. mutans and the resin specimen with xylitol added. Conclusion: This study showed that adding 2.5% and 5% Xylitol to the Tetric® N-Flow Bulk Fill resin did not inhibit bacterial growth (AU).

Efeitos do D-limoneno no metabolismo energético de camundongos C57/Bl6 com obesidade induzida pela dieta / Effects of D-limonene on the energy metabolism of C57/Bl6 mice with diet-induced obesity

A obesidade está associada ao desenvolvimento de doenças crônicas não transmissíveis como hipertensão, resistência insulínica, dislipidemia e esteatose hepática. O consumo de compostos bioativos impacta na manutenção da saúde e na prevenção de risco de desenvolvimento dessas doenças. Entre os compostos bioativos, os monoterpenos são pouco investigados, apesar da literatura demonstrar efeitos promissores desses compostos sobre o metabolismo. O D-limoneno, o principal monoterpeno encontrado na laranja, é caracterizado por possuir efeitos hipolipemiantes, anti-inflamatórios e anti-obesogênicos. Estudos in vitro e in vivo descrevem sua capacidade de promover a ß-oxidação de ácidos graxos em adipócitos e redução da inflamação. Este estudo teve como objetivo investigar o efeito do D-limoneno no metabolismo e inflamação em um modelo de obesidade induzida por dieta. Para isso, quarenta camundongos machos (C57/Bl6) de 11 semanas de idade, foram distribuídos em 4 grupos, sendo que um dos grupos recebeu ração normolipídica e os demais, ração hiperlipídica. O D-limoneno foi suplementado na ração de dois grupos que receberam dieta hiperlipídica nas concentrações de 0,1%, e 0,8%. Considerando-se a ingestão alimentar dos animais, a ração suplementada com 0,1% D-limoneno correspondeu à ingestão de 0,15 g/kg/dia e ração com 0,8% de D-limoneno correspondeu a 1,3 g/kg/dia. Os animais tiveram o peso e a ingestão alimentar monitorados ao longo da intervenção com duração de 7 semanas. Os camundongos que receberam D-limoneno a 0,1% apresentaram menor ganho de peso e de acúmulo de tecido adiposo, comparado com os animais sem suplementação alimentados com a dieta hiperlipídica. Além disso, o D-limoneno promoveu a diminuição da concentração plasmática de marcadores inflamatórios incluindo TNF-α, INF-γ e IL-6 nos animais dos grupos que foram suplementados com D-limoneno. Entretanto, não houve diferença nos marcadores bioquímicos e metabólicos. Uma limitação do estudo foi o fato das complicações metabólicas associadas ao modelo de obesidade não terem sido plenamente estabelecidas, dados o alojamento individual, à curta duração da exposição à ração hiperlipídica e idade dos animais no início da suplementação. Esse fato pode ter dificultado a observação dos efeitos do D-limoneno na reversão dos parâmetros que seriam normalmente deteriorados pelo desenvolvimento da obesidade. Concluímos que o D-limoneno pode interferir no metabolismo energético, com possível efeito anti-obesogênico e anti-inflamatório. Devido às limitações do modelo, são necessários mais estudos para confirmar esses resultados

Obesity is associated with the development of chronic non-communicable diseases such as hypertension, insulin resistance, dyslipidemia, and hepatic steatosis. The intake of dietary bioactive compounds is associated with the maintenance of health and the prevention of chronic diseases. Among the group of bioactive compounds, monoterpenes are poorly investigated, in spite of several reports of their promising effects on metabolism. D-limonene is the main monoterpene found in oranges, known for its hypolipemic, anti-inflammatory, and anti-obesogenic effects. in vitro and in vivo studies associate D-limonene to increased ß-oxidation of fatty acids in adipocytes and reduced inflammation. This study aimed at investigating the effects of D-limonene on metabolism and inflammation in a diet-induced obesity model. For this purpose, forty male mice (C57/Bl6) were distributed in 4 groups, with one group receiving a normolipidic diet and the others, a high-fat diet. D-limonene was supplemented in the diets of two groups that received high-fat diet at the concentrations of 0.1% and 0.8%. Considering the feed intake, mice receiving D-limonene supplementation at 0.1% ingested in average 0.15 g/kg/day, while the mice receiving the supplemmentation at 0.8%, ingested approximately 1.3 g of D-limonene /kg/day. The animals had their weight and food intake monitored throughout the intervention. Mice that received Dlimonene supplementation at 0.1% showed reduced weight gain and accumulation of adipose tissue compared to the non-supplemented mice fed the high-fat diet. In addition, D-limonene promoted a decrease in hepatic inflammatory markers including TNF-α, INF-γ, and IL-6. However, there was no difference in biochemical and metabolic markers. A limitation of the study was that the metabolic complications associated with the obesity model were not fully established, probably due to the age at the start of the protocol (11 weeks), individual housing and short duration of the exposure to the high-fat feed. This fact may have prevented the observation of the positive effects of D-limonene in reversing parameters that would normally be impaired by the development of obesity. We conclude that D-limonene may interfere in energy metabolism, with a possible anti-obesogenic and anti-inflammatory effect. Due to the limitations of the model, further studies are needed to confirm these findings

O efeito do tratamento com rapamicina em órgãos linfoides de camundongos propensos à senescência acelerada / The effect of rapamycin treatment on lymphoid organs of senescence accelerated mice prone

Envelhecer compreende um fenômeno complexo, natural e irreversível, que submete o organismo a inúmeras alterações nos processos biológicos, fisiológicos, ambientais, psicológicos, comportamentais e sociais. Esse processo é caracterizado por um declínio gradual dos mecanismos homeostáticos do organismo, intimamente relacionados com o estado senescente. A senescência, quando diz respeito ao sistema imunológico, é denominada de imunossenescência, que pode ser definida como uma parada estável do ciclo celular associada a mudanças, com uma resposta que limita a proliferação de células envelhecidas ou danificadas. A autofagia está diretamente relacionada com a manutenção do fenótipo senescente, em que a atividade autofágica exerce um papel essencial e ativo na influência da biossíntese de proteínas e organelas. Essa via é regulada naturalmente pela proteína mTOR e quimicamente pelo fármaco rapamicina. Assim, pretendemos investigar: (1) as alterações no perfil corporal e hematimêtrico dos animais ao longo do tratamento com rapamicina; (2) avaliar o perfil de citocinas; (3) observar as modificações histológicas em órgãos linfoides primários e secundário; (4) analisar as populações de células linfoides e mieloides; e (5) avaliar a capacidade proliferativa de linfócitos in vitro. Camundongos SAMP-8 e SAMR-1 foram tratados com rapamicina durante dois meses. A mensuração da massa corporal e análises hematológicas foram realizadas antes e durante o tratamento. Amostras de soro, medula óssea, timo e baço foram analisados em ensaios de ELISA, histologia, população e subpopulações de células. Alterações na massa corporal, parâmetros hematológicos e celularidade de células foram nítidas entre os dois modelos utilizados. Diferenças também foram percebidas na detecção de citocinas IL-1ß. IL-6 e TNF-α, com resultados significantes nas amostras de baço, timo e medula óssea. As citocinas IL-7 e IL-15 apresentaram diferenças de secreção entre os grupos, sendo a primeira maior detectada em camundongos com senescência acelerada tratados com rapamicina. Em nossa análise histológica observamos que os camundongos SAM-P8 apresentaram involução tímica. E nas subpopulações de linfócitos T do baço, células TCD4+ e TCD8+ estavam, respectivamente, em maior e menor quantidade nos camundongos SAM-P8 tratados com rapamicina. Dessa forma, o camundongo da linhagem SAM-P8 é um excelente modelo para se estudar as alterações da senescência, em que o mesmo apresenta características fisiológicas distintas dos camundongos utilizados como controle (SAM-R1). Além disso, verificamos que a dose de rapamicina empregada não desencadeou alterações que pudessem comprometer a resposta imunológica desses camundongos, bem como na possibilidade de atuar na resposta contra os efeitos complexos do envelhecimento

Aging comprises a complex, natural, and irreversible phenomenon, which subjects the organism to countless alterations in biological, physiological, environmental, psychological, behavioral, and social processes. This process is characterized by a gradual decline in the organism's homeostatic mechanisms, closely related to senescence effects. Senescence, when it concerns the immune system, is called immunosenescence, which can be defined as a stable cell cycle arrest associated with changes and is a response that limits the proliferation of aged or damaged cells. Autophagy is a genetically regulated, conserved cellular process and a metabolic pathway essential for maintaining cellular homeostasis, which plays a constitutive and active role in controlling the biosynthesis of proteins and organelles. This pathway is regulated naturally by mTOR or chemically by the drug rapamycin, having a direct relationship with cellular homeostasis and maintenance of the senescent phenotype. Thus, we intend to investigate: (1) the changes in the body and hematimetic profile of the animals throughout the rapamycin treatment; (2) evaluate the cytokine profile; (3) observe histological changes in primary and secondary lymphoid organs; (4) analyze lymphoid and myeloid cell populations; and (5) evaluate the proliferative capacity of lymphocytes in vitro. SAMP-8 and SAMR-1 mice were treated with rapamycin for two months. Body mass measurement and hematological analyses were performed before and during treatment. Serum, bone marrow, thymus and spleen samples were analyzed in ELISA assays, histology, cell population and subpopulations. Changes in body mass, hematological parameters, and cellularity were clear between the two models used. Differences were also noticed in the detection of cytokines IL-1ß. IL-6 and TNF-α, with significant results in the spleen, thymus and bone marrow samples. The cytokines IL-7 and IL-15 showed differences in secretion between groups, the former being higher detected in mice with accelerated senescence treated with rapamycin. In our histological analysis we observed that SAM-P8 mice showed thymic involution. And in the spleen T-lymphocyte subpopulations, TCD4+ and TCD8+ cells were, respectively, in higher and lower quantities in SAM-P8 mice treated with rapamycin. Thus, the SAM-P8 mouse is an excellent model to study the changes of senescence, since it presents physiological characteristics different from the control mice (SAM-R1). Furthermore, we verified that the dose of rapamycin used did not trigger changes that could compromise the immune response of these mice, as well as the possibility of acting in the modulatory response against the complex effects of aging

Moderate Toxicity of Potential Boron-containing Therapeutic, Dipotassium-trioxohydroxytetrafl uorotriborate - K2 (B3 O3 F4 OH) in Rats and Mice

Abstract Biological activity of boron-containing compounds (BCCs) has been well-known. Growing interest and numerous applications for BCCs have been reported. Boron and boron-containing acids show low acute toxicity in mammals but data on halogenated boroxine (HB) - dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH) acute toxicity have not been reported before. This compound, characterized as a potential therapeutic for skin changes, exhibits no observable genotoxicity in doses lower that 0.1 mg/ml in vitro and 55 mg/kg in vivo. It has also been confirmed as an antitumour agent both in vitro and in vivo as well as an inhibitor of enzymes involved in antioxidant mechanisms. The aim of this study was to assess the acute toxicity of HB and to determine the maximum tolerated dose as well as a dose free of any signs of toxicity in different test organisms. Acute toxicity of HB was tested in Sprague-Dawley and Wistar rats and BALB/c mice after single parenteral application of different doses. We determined doses free of any sign of toxicity and LD50 after single dose administration. LD50 of HB ranges from 63 to 75 mg/kg in different test models, meaning that HB shows moderate toxicity

Investigation on formulation parameters of donepezil HCl loaded solid lipid nanoparticles

Abstract Donepezil-HCl is a member of the acetylcholinesterase inhibitors that is indicated for the symptomatic treatment of Alzheimer's disease (AD) and has many side effects. In this study, to reduce the side effects of Donepezil-HCl and increase the penetration of the drug through the blood-brain barrier, we aimed to design a solid lipid nanoparticle (SLN) formulation. The effects of the different formulation parameters, such as homogenization speed, sonication time, lipid and drug concentration, surfactant type and concentration, and volume of the aqueous phase, were assessed for optimization. The particle size and PDI increased with increasing lipid concentration but decreased with increasing amounts of surfactant (Tween 80) and co-surfactant (lecithin). When the homogenization rate and sonication time increased, the particle size decreased and the encapsulation efficiency increased. The optimized formulation exhibited particle size, PDI, encapsulation efficiency, and zeta potential of 87.2±0.11 nm; 0.22±0.02; 93.84±0.01 %; -17.0±0.12 mV respectively. The in vitro release investigation revealed that approximately 70% of Donepezil-HCl was cumulatively released after 24 hours. TEM analysis proved that spherical and smooth particles were obtained and formulations had no toxic effect on cells. The final optimized formulation could be a candidate for Donepezil-HCl application in Alzheimer's treatment with reduced side effects and doses for patients

Effect of polycaprolactone nanocapsules loaded with essential oils on biofilm formation by Staphylococcus aureus strains isolated from bovine mastitis cases

Abstract Bovine infectious mastitis is largely resistant to antibacterial treatment, mainly due to mechanisms of bacterial resistance in the biofilms formed by Staphylococcus aureus. Melaleuca (MEO) and citronella essential oils (CEO) are promising agents for reducing or eliminating biofilms. Free melaleuca oil presented a medium Minimum Inhibitory Concentration (MIC) of 0.625% and a Minimum Bactericidal Concentration (MBC) of 1.250%, while free citronella oil showed medium MIC and MBC of 0.313%. Thus, free CEO and MEO demonstrate bacteriostatic and bactericidal potential. We generated polymeric nanocapsules containing MEO or CEO and evaluated their efficacy at reducing biofilms formed by S. aureus. Glass and polypropylene spheres were used as test surfaces. To compare the responses of free and encapsulated oils, strains were submitted to 10 different procedures, using free and nanoencapsulated essential oils (EOs) in vitro. We observed no biofilm reduction by MEO, free or nanoencapsulated. However, CEO nanocapsules reduced biofilm formation on glass (p=0.03) and showed a tendency to diminish biofilms on polypropylene (p=0.051). Despite nanoencapsulated CEO reducing biofilms in vitro, the formulation could be improved to modify the CEO component polarity and, including MEO, to obtain more interactions with surfaces and the biofilm matrix

Development and characterization of photoprotective nanoemulsions containing Babassu (Orbignya phalerata Mart. ) lipophilic extract

Abstract Oil-in-water photoprotective nanoemulsions (NEs) were developed using Babassu (BBS) lipophilic extract, nonionic surfactants, and low concentrations of organic sunscreens by ultrasonic processing. BBS extract was chosen due to its suitable physicochemical properties (acidity index, peroxide index, refraction index, and relative density) and predominance of saturated fatty acids, identified by gas chromatography-mass spectrometry (GC-MS), which promote biological activities and high oxidative stability. NEs were characterized by mean droplet size, morphology, polydispersity index (PdI), pH, and organoleptic properties, and the physical stability of the NEs was evaluated for 120 days at room temperature. The sun protection factor (SPF) was determined, and the photostability and in vitro cytotoxicity assays were performed for NEs. All NEs remained stable for 120 days, with a droplet size <150 nm and a monomodal distribution profile. The pH values were compatible with the skin's pH. NE3 showed a spherical morphology, with a mean droplet size of 125.15 ± 0.16 nm and PdI of 0.145 ± 0.032. NE3 containing BBS extract and sunscreens presented an SPF of 35.5 ± 3.0, was photostable after 6 h of radiation and was non-cytotoxic to fibroblast cells. Thus, NE3 could be considered a promising formulation for developing synergic plant-extract sunscreen photoprotective products for the market

Clinical Evaluation of Composite Restorations with and without Polyethylene Fiber in First Permanent Molars: A 24-Month Randomized Clinical Trial

ABSTRACT Objective: To evaluate the 24-month clinical performance of composite resin restorations with and without polyethylene fiber in the first permanent molars of pediatric patients with extensive caries. Material and Methods: In total, 75 Class II restorations were placed in the permanent molar teeth of 75 children (mean age 11.3 years) with (FC; n=38) or without (C; n=37) fiber. One operator placed all the restorations. The restorations were evaluated using the modified USPHS criteria in terms of retention, color match, marginal discoloration, anatomic form, marginal adaptation, secondary caries, and postoperative sensitivity. Statistical data were analyzed using chi-square and Cochran tests (p<0.05). Results: At the end of two years, 65 restorations (FC:31; C:34) were followed up. No changes were observed during the first six months. After 24 months of follow-up, there were minor changes in marginal adaptation and marginal color in both groups; however, no statistically significant difference was observed between the clinical performances for all criteria (p>0.05). Conclusion: Extensive composite restorations with or without fibers displayed good clinical performance in high load-bearing areas after 24 months.

Comparison Between Radiographs, White and Fluorescent Images in the Diagnosis and Treatment Decisions for Occlusal Caries: An Ex Vivo Study

ABSTRACT Objective: To compare the agreement of images in white light (WL), fluorescence (FL), and digital radiographs (DR), on the diagnosis and treatment decisions for occlusal caries lesions against a micro-CT gold standard. Material and Methods: Ten extracted third molars, with enamel and/or dentin caries (ICDAS 2-4), were included. Occlusal surface images were acquired with an intraoral camera (SoproLife®) in WL and FL modes. DR was obtained using an intraoral X-ray and a semi-direct digital system. A total of 780 images were needed, organized in a template, to be later examined by twenty-six dentists invited to compose the study. The Generalized Estimation Equations model was used to compare the proportions of the correct answers between the three methods and the gold standard. When significant, Bonferroni post-hoc test was used to identify differences (α=5%). Results: Most of the examiners were specialists (76.9%) with 14.5 years of experience. All diagnostic methods were similar and showed low agreement (DR 12.7%, WL 16.5%, and FL 16.5%) compared with gold standard caries diagnostic scores. Regarding treatment decisions, mean agreement for all diagnostic methods was higher (43.2%; p<0.001), and among all methods, WL (48.1%) and FL (51.2%) modes performed better than DR (30.4%, p<0.001). Conclusion: SoproLife® images could help clinicians to propose rational, minimally invasive treatments for occlusal caries lesions.

Interchangeability of medications and biopharmaceutical implication of taking drugs with fluids other than water: ibuprofen case study

Abstract The aim of this study was to compare the dissolution properties of ibuprofen solid oral dosage forms commercially available in Bosnia and Herzegovina and to estimate the influence of dissolution medium composition on the drug release. Eight products (A-H) were subjected to in vitro dissolution test using experimental conditions described in USP42-NF37. Dissolution properties of one selected product were examined in the presence of alcohol (22.2% v/v) and fruit juice (22.2% v/v). Products marked B-H complied with the pharmacopeial criteria. Dissolution profile of product B was similar with dissolution profiles of products D, E, F and G and similarity was also found between products A-D, C-G, D-G and E-F. Drug release from most of the examined preparations fitted best to the Weibull kinetic model. In the presence of alcohol in the medium, higher amount of ibuprofen was dissolved. Contrary, ibuprofen dissolved in the presence of fruit juice was significantly lower. Differences in the dissolution profiles of investigated preparations suggest that their interchangeability should be additionally considered and demonstrated with in vivo bioequivalence studies. Presence of different substances in the medium can affect dissolution properties of ibuprofen, emphasizing the importance of the patient's compliance.

Bioactivity evaluation of least explored traditionally acclaimed medicinally potent herb Nanorrhinum ramosissimum(Wall. ) Betsche

Abstract Present study analysed the therapeutic potential of traditionally acclaimed medicinal herb Nanorrhinum ramosissimum, using plant parts extracted with different solvents (10 mg/mL). Shoot extracts exhibited comparatively better antimicrobial properties, in comparison to root extracts. Total phenolic content was estimated, to ascertain its dependency on antioxidant properties of plant extracts. Antioxidant assay revealed promising results in comparison to IC50 value of standard ascorbic acid (52.2±0.07 µg/mL), for methanolic extracts of shoot (61.07±0.53 µg/mL and 64.33±0.33 µg/mL) and root (76.705±0.12 µg/mL and 89.73±0.28 µg/ mL) for in vivo and in vitro regenerants respectively. Correlation coefficient R2 values ranged between 0.90-0.95, indicating a positive correlation between phenolic contents and antioxidant activity. Plant extracts were also able to inhibit DNA oxidative damage again indicating their antioxidative potential. Antidiabetic potential was confirmed by alpha amylase inhibition assay where shoot methanolic extracts (invivo, in vitro) exhibited the best IC50 values (54.42±0.16 µg/mL, 66.09±0.12 µg/mL) in comparison to standard metformin (41.92±0.08 µg/mL). Ethanolic extracts of roots (in vitro, invivo) exhibited the relative IC50 values (88.97±0.32µg/mL,96.63±0.44 µg/mL) indicating that shoot parts had a better alpha amylase inhibition property; thus proving the herb's bioactive potential and its prospective therapeutic source for curing various ailments.

Prediction of the Impact of CYP2C19 Polymorphism on Drug-Drug Interaction between Voriconazole and Tacrolimus Using Physiologically-Based Pharmacokinetic Modelling

Abstract Voriconazole increases tacrolimus blood concentration significantly when coadministrated. The recommendation of reducing tacrolimus to 1/3 in voriconazole package insert seems not to be satisfactory in clinical practice. In vitro studies demonstrated that the magnitude of inhibition depends on the concentration of voriconazole, while voriconazole exposure is determined by the genotype status of CYP2C19. CYP2C19 gene polymorphism challenges the management of drug-drug interactions(DDIs) between voriconazole and tacrolimus. This work aimed to predict the impact of CYP2C19 polymorphism on the DDIs by using physiologically based pharmacokinetics (PBPK) models. The precision of the developed voriconazole and tacrolimus models was reasonable by evaluating the pharmacokinetic parameters fold error, such as AUC0-24, Cmax and tmax. Voriconazole increased tacrolimus concentration immediately in all population. The simulated duration of DDIs disappearance after voriconazole withdrawal were 146h, 90h and 66h in poor metabolizers (PMs), intermediate metabolizers (IMs) and extensive metabolizers(EMs), respectively. The developed and optimized PBPK models in this study can be applied to assit the dose adjustment for tacrolimus with and without voriconazole.

Preformulation screening of lipids using solubility parameter concept in conjunction with experimental research to develop ceftriaxone loaded nanostructured lipid carriers

Abstract Development of ceftriaxone loaded nanostructured lipid carriers to increase permeability of ceftriaxone across uninflamed meninges after parenteral administration. Lipids were selected by theoretical and experimental techniques and optimization of NLCs done by response surface methodology using Box-Behnken design. The Δδt for glyceryl monostearate and Capryol90 were 4.39 and 2.92 respectively. The drug had maximum solubility of 0.175% (w/w) in glycerol monostearate and 2.56g of Capryol90 dissolved 10mg of drug. The binary mixture consisted of glyceryl monostearate and Capryol90 in a ratio of 70:30. The optimized NLCs particle size was 130.54nm, polydispersity index 0.28, % entrapment efficiency 44.32%, zeta potential -29.05mV, and % drug loading 8.10%. In vitro permeability of ceftriaxone loaded NLCs was 5.06x10-6 cm/s; evidently, the NLCs pervaded through uninflamed meninges, which, was further confirmed from in vivo biodistribution studies. The ratio of drug concentration between brain and plasma for ceftriaxone loaded NLCs was 0.29 and that for ceftriaxone solution was 0.02. With 44.32% entrapment of the drug in NLCs the biodistribution of ceftriaxone was enhanced 7.9 times compared with that of ceftriaxone solution. DSC and XRD studies revealed formation of imperfect crystalline NLCs. NLCs improved permeability of ceftriaxone through uninflamed meninges resulting in better management of CNS infections.

Neuroprotective potential of the Amazonian fruits Euterpe oleracea Mart. and Paullinia cupana Kunth

Abstract Acai (Euterpe oleracea Mart.) and guarana (Paullinia cupana Kunth) are native species from the Amazon Forest that in folk medicine are used to treat several diseases due to their anti-inflammatory and antioxidant properties. This review brings together findings from different studies on the potential neuroprotective effects of acai and guarana, highlighting the importance of the conservation and sustainable exploitation of the Amazon Forest. A bibliographic survey in the PubMed database retrieved indexed articles written in English that focused on the effects of acai and guarana in in vitro and in vivo models of neurodegenerative diseases. In general, treatment with either acai or guarana decreased neuroinflammation, increased antioxidant responses, ameliorated depression, and protected cells from neurotoxicity mediated by aggregated proteins. The results from these studies suggest that flavonoids, anthocyanins, and carotenoids found in both acai and guarana have therapeutic potential not only for neurodegenerative diseases, but also for depressive disorders. In addition, acai and guarana show beneficial effects in slowing down the physiological aging process. However, toxicity and efficacy studies are still needed to guide the formulation of herbal medicines from acai and guarana.

Antioxidant, Antidiabetic and Lipid Profiling of Spermadicyton Suaveolens in Streptozotocin (STZ) Induced Diabetic Rats

ABSTRACT Diabetes is a life-threatening disease, and currently available synthetic medicines for treating diabetes are associated with various side effects. Therefore, there is an unmet need to develop herbal remedies against diabetes as an alternative to synthetic medicines. Although local healers use the roots of Spermadicyton suaveolens (SS) to manage diabetes, there is negligible research to validate its antidiabetic properties. The present investigation aims to the assess the antioxidant, antidiabetic, and antihyperlipidemic potential of the ethanolic extract of S. Suaveolen's roots (EESS) on streptozotocin (STZ) induced diabetic rats. The extract was screened for in vitro antioxidant and antidiabetic activity. The in vivo antidiabetic potential of EESS (at 200 and 400 mg/kg) was studied on STZ-induced diabetic rats for 20 days. The EESS displayed significant (p<0.05) antidiabetic and antioxidant properties. The administration of 200 mg/kg and 400 mg/kg EESS in STZ-induced diabetic rats significantly reduced hyperglycemia, and restored antioxidant enzymes and lipid profile-a high density lipoprotein (HDL) increased by the administration of a single dose of streptozotocin. Thus, EESS could be a promising herbal medicine in the treatment of diabetes and hyperlipidemia