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BACKGROUND: This study aimed to engineer and optimise a dysbiotic biofilm model to develop in vitro root caries for investigating microbial modulation strategies. The model involved growing complex biofilms from a saliva inoculum collected from four volunteers using two strategies. In the first strategy ("pre-treatment strategy"), bovine root slabs were used, and two natural compounds were incorporated at time 0 of the 10-day biofilm experiment, which included sucrose cycles mimicking the cariogenic environment. In the second strategy ("post-treatment strategy"), mature biofilms were grown in a modified Calgary biofilm device coated with collagen and hydroxyapatite for 7 days and then were exposed to the same natural compounds. The metatranscriptome of each biofilm was then determined and analysed. Collagenase activity was examined, and the biofilms and dentine were imaged using confocal and scanning electron microscopy (SEM). Mineral loss and lesion formation were confirmed through micro-computed tomography (µ-CT). RESULTS: The pH confirmed the cariogenic condition. In the metatranscriptome, we achieved a biofilm compositional complexity, showing a great diversity of the metabolically active microbiome in both pre- and post-treatment strategies, including reads mapped to microorganisms other than bacteria, such as archaea and viruses. Carbohydrate esterases had increased expression in the post-treated biofilms and in samples without sugar cycles, while glucosyltransferases were highly expressed in the presence of sucrose cycles. Enrichment for functions related to nitrogen compound metabolism and organic cyclic component metabolism in groups without sucrose compared to the sucrose-treated group. Pre-treatment of the roots with cranberry reduced microbial viability and gelatinase (but not collagenase) activity (p < 0.05). SEM images showed the complexity of biofilms was maintained, with a thick extracellular polysaccharides layer. CONCLUSIONS: This root caries model was optimized to produce complex cariogenic biofilms and root caries-like lesions, and could be used to test microbial modulation in vitro. Pre-treatments before biofilm development and cariogenic challenges were more effective than post-treatments. The clinical significance lies in the potential to apply the findings to develop varnish products for post-professional tooth prophylaxis, aiming at implementing a strategy for dysbiosis reversal in translational research. Video Abstract.
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Biofilmes , Microbiota , Cárie Radicular , Saliva , Humanos , Cárie Radicular/microbiologia , Saliva/microbiologia , Bovinos , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Dentina/microbiologia , Colagenases/metabolismoRESUMO
Objetivo: Estimar a prevalência de positividade do Teste Tuberculínico (TT) no rastreamento da infecção latente pelo Mycobacterium tuberculosis (ILTB) e identificar fatores associados entre trabalhadores da Atenção Primária à Saúde (APS) e média complexidade no segundo maior município do Estado da Bahia. Métodos: Inquérito epidemiológico, com 370 participantes, que avaliou dados sociodemográficos, ocupacionais, consumo de tabaco, álcool e variáveis relacionadas à tuberculose (TB), no período de abril/2021 a março/2022. Aplicou-se o TT por via intradérmica, com leitura após 72h. Resultados: A prevalência de positividade do TT foi de 41,3% e 17,6% para TT ≥ 5mm e TT ≥ 10mm, respectivamente. Para TT ≥ 5mm, as variáveis associadas foram: possuir cicatriz da vacina Bacilo Calmette-Guérin (BCG) e contato prévio com pessoas com TB. Conclusão: A prevalência de positividade do TT foi elevada entre os participantes do estudo. Fazem-se necessárias medidas de proteção ambiental, coletiva e individual para redução do risco de ILTB.
Objective: To estimate the prevalence of tuberculin test (TT) positivity in screening for Latent Mycobacterium tuberculosis Infection (LTBI) and to identify associated factors in Primary Health Care (PHC) and medium complexity workers in the second largest municipality in the State of Bahia, Brazil. Methods: Epidemiological survey with 370 workers on sociodemographic and occupational data, tobacco and alcohol use and variables related to Tuberculosis (TB), during April 2021 and March 2022. TT was applied intradermally, with reading after 72 hours. Results: The prevalence of positive TT was 41.3% and 17.6% for TT ≥ 5mm and TT ≥ 10mm, respectively. For TT ≥ 5mm, the associated variables were: presence of Bacillus Calmette-Guérin (BCG) vaccine scar and previous contact with TB patients. Conclusion: The prevalence of TST positivity was high among study participants. Environmental, collective and individual protection measures are necessary to reduce the risk of LTBI.
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The limited therapeutic options for fungal infections and the increased incidence of fungal strains resistant to antifungal drugs, especially Candida spp., require the development of new antifungal drugs and strategies. Histone deacetylase inhibitors (HDACi), like vorinostat, have been studied in cancer treatment and have antifungal effects, acting alone or synergistically with classical antifungals. Here we investigated the antifungal activity of two novel sustainable HDACi (LDT compounds) based on vorinostat structure. Molecular docking simulation studies reveal that LDT compounds can bind to Class-I HDACs of Candida albicans, C. tropicalis, and Cryptococcus neoformans, which showed similar binding mode to vorinostat. LDT compounds showed moderate activity when tested alone against fungi but act synergistically with antifungal azoles against Candida spp. They reduced biofilm formation by more than 50% in C. albicans (4 µg/mL), with the main action in fungal filamentation. Cytotoxicity of the LDT compounds against RAW264.7 cells was evaluated and LDT536 demonstrated cytotoxicity only at the concentration of 200 µmol/L, while LDT537 showed IC50 values of 29.12 µmol/L. Our data indicated that these sustainable and inexpensive HDACi have potential antifungal and antibiofilm activities, with better results than vorinostat, although further studies are necessary to better understand the mechanism against fungal cells.
Fungal infections are neglected diseases that affect more than a billion people worldwide. Some histone deacetylase inhibitors can act against fungal cells. Our data reveal that HDACi LDT536 and LDT537 have potential antibiofilm and antifungal activities.
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Phenolic lipids components of the cashew nutshell liquid (CNSL) have molecular structures capable of chemical signalling that regulate gene expression, metabolism and inflammation. This study sets out to assess how CNSL derivatives impact oral bacteria, from an antibacterial and anti-collagenolytic perspective, as well as its biocompatibility with dental pulp stem cells. Two hemi-synthetic saturated CNSL derivative compounds were selected (LDT11-Anacardic Acids-derivative and LDT409-cardanol-derivative). Bacteriostatic activity was tested against Streptococcus mutans and Veillonella parvula. Antimicrobial capacity against preformed S. mutans biofilms was investigated using a collagen-coated Calgary Biofilm Device and confocal microscopy. Clostridium histolyticum, P. gingivalis and S. mutans biofilms were used to assess anti-collagenolytic activity. Biocompatibility with human dental pulp stromal cells (HDPSCs) was investigated (MTT for viability proportion, LDH assays for cell death rate). LDTs inhibited the bacterial growth, as well as partially inhibited bacterial collagenases in concentrations higher than 5 µg/mL. Dose-response rates of biofilm cell death was observed (LDT11 at 20, 50, 100 µg/mL = 1.0 ± 0.4, 0.7 ± 0.3, 0.6 ± 0.03, respectively). Maximum cytotoxicity was 30%. After 1 week, LDT409 had no HDPSCs death. HDPSCs viability was decreased after 24 h of treatment with LDT11 and LDT409, but recovered at 72 h and showed a massive increase in viability and proliferation after 1 week. LDTs treatment was associated with odontoblast-like morphology. In conclusion, LDT11 multifunctionality and biocompatibility, stimulating dental pulp stem cells proliferation and differentiation, indicates a potential as a bio-based dental material for regenerative Dentistry. Its potential as a bacterial collagenases inhibitor to reduce collagen degradation in root/dentinal caries can be further explored.
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Anacardium , Cárie Dentária , Anacardium/química , Antibacterianos/farmacologia , Biofilmes , Odontologia , Humanos , Lipídeos , Streptococcus mutansRESUMO
Alzheimer's disease (AD) represents a global problem, with an estimation of the majority of dementia patients in low- and middle-income countries by 2050. Thus, the development of sustainable drugs has attracted much attention in recent years. In light of this, taking inspiration from the HDAC inhibitor vorinostat (1), we develop the first HDAC inhibitors derived from cashew nut shell liquid (CNSL), an inexpensive agro-food waste material. CNSL derivatives 8 and 9 display a HDAC inhibitory profile similar to 1, together with a more promising safety for 9 compared to 1. Moreover, both compounds and particularly 9 were able to effectively modulate glial cell-induced inflammation and to revert the pro-inflammatory phenotype. All these results demonstrate that the use of inexpensive food waste materials could be successfully applied for the development of accessible and sustainable drug candidates for the treatment of AD.
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Arylpiperazines 2-11 were synthesized, and their biological profiles at α1-adrenergic receptors (α1-ARs) assessed by binding assays in CHO cells expressing human cloned subtypes and by functional experiments in isolated rat vas deferens (α1A), spleen (α1B), and aorta (α1D). Modifications at the 1,3-benzodioxole and phenyl phamacophoric units resulted in the identification of a number of potent compounds (moderately selective with respect to the α1b-AR), in binding experiments. Notably, compound 7 (LDT451) showed a subnanomolar pKi of 9.41 towards α1a-AR. An encouragingly lower α1B-potency was a general trend for all the series of compounds, which showed α1A/D over α1B selectivity in functional assays. If adequately optimized, such peculiar selectivity could have relevance for a potential LUTS/BPH therapeutic application.
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Antagonistas de Receptores Adrenérgicos alfa 1/síntese química , Antagonistas de Receptores Adrenérgicos alfa 1/farmacologia , Desenho de Fármacos , Piperazinas/síntese química , Piperazinas/farmacologia , Receptores Adrenérgicos alfa 1/metabolismo , Antagonistas de Receptores Adrenérgicos alfa 1/química , Antagonistas de Receptores Adrenérgicos alfa 1/metabolismo , Animais , Células CHO , Linhagem Celular Tumoral , Técnicas de Química Sintética , Clonagem Molecular , Cricetinae , Cricetulus , Humanos , Masculino , Simulação de Acoplamento Molecular , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Piperazinas/química , Piperazinas/metabolismo , Conformação Proteica , Ratos , Receptores Adrenérgicos alfa 1/química , Receptores Adrenérgicos alfa 1/genética , Transdução de Sinais/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
Cardanol is a phenolic lipid component of cashew nut shell liquid (CNSL), obtained as the byproduct of cashew nut food processing. Being a waste product, it has attracted much attention as a precursor for the production of high-value chemicals, including drugs. On the basis of these findings and in connection with our previous studies on cardanol derivatives as acetylcholinesterase (AChE) inhibitors, we designed a novel series of analogues by including a protonable amino moiety belonging to different systems. Properly addressed docking studies suggested that the proposed structural modifications would allow the new molecules to interact with both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE, thus being able to act as dual binding inhibitors. To disclose whether the new molecules showed the desired profile, they were first tested for their cholinesterase inhibitory activity towards EeAChE and eqBuChE. Compound 26, bearing an N-ethyl-N-(2-methoxybenzyl)amine moiety, showed the highest inhibitory activity against EeAChE, with a promising IC50 of 6.6 µM, and a similar inhibition profile of the human isoform (IC50 = 5.7 µM). As another positive feature, most of the derivatives did not show appreciable toxicity against HT-29 cells, up to a concentration of 100 µM, which indicates drug-conform behavior. Also, compound 26 is capable of crossing the blood-brain barrier (BBB), as predicted by a PAMPA-BBB assay. Collectively, the data suggest that the approach to obtain potential anti-Alzheimer drugs from CNSL is worth of further pursuit and development.