Melipona scutellaris Geopropolis: Chemical Composition and Bioactivity.

Geopropolis has been used in traditional medicine for centuries. In this study, the botanical origin, physicochemical profile, and biological activities of geopropolis from Melipona scutellaris harvested during rainy and dry seasons were investigated. Palynological analysis identified over 50 pollen types, with Schinus terebinthifolius and Cecropia being the predominant types. The analytical results were in line with those reported in the literature. Rainy-season geopropolis exhibited higher total phenol and flavonoid content (determined using High Performance Liquid Chromatography-25.13% and 3.92%, respectively) compared to the dry season (19.30% and 2.09%); the major peaks (naringin, gallic acid, and catechin) were similar among samples. Antioxidant capacity was assessed via DPPH, reducing power, and ß-carotene/linoleic acid discoloration assays. Rainy-season samples displayed superior antioxidant activity across methods. Antimicrobial effects were determined using microdilution, while the impact on the cholinesterase enzyme was quantified using 5-thio-2-nitrobenzoic acid accumulation. Anti-inflammatory and antimutagenic activities were assessed through hyaluronidase enzyme inhibition and by utilizing Saccharomyces cerevisiae ATCC-20113 cells. Both samples exhibited anti-inflammatory and antimutagenic properties. Moreover, a significant inhibition of acetylcholinesterase was observed, with IC50 values of 0.35 µg/mL during the rainy season and 0.28 µg/mL during the dry season. Additionally, the geopropolis displayed antimicrobial activity, particularly against Staphylococcus aureus. These findings suggest the therapeutic potential of M. scutellaris geopropolis in the context of inflammatory, oxidative, and infectious diseases.

Biosynthesis of Metal and Metal Oxide Nanoparticles Using Microbial Cultures: Mechanisms, Antimicrobial Activity and Applications to Cultural Heritage.

Nanoparticles (1 to 100 nm) have unique physical and chemical properties, which makes them suitable for application in a vast range of scientific and technological fields. In particular, metal nanoparticle (MNPs) research has been showing promising antimicrobial activities, paving the way for new applications. However, despite some research into their antimicrobial potential, the antimicrobial mechanisms are still not well determined. Nanoparticles' biosynthesis, using plant extracts or microorganisms, has shown promising results as green alternatives to chemical synthesis; however, the knowledge regarding the mechanisms behind it is neither abundant nor consensual. In this review, findings from studies on the antimicrobial and biosynthesis mechanisms of MNPs were compiled and evidence-based mechanisms proposed. The first revealed the importance of enzymatic disturbance by internalized metal ions, while the second illustrated the role of reducing and negatively charged molecules. Additionally, the main results from recent studies (2018-2022) on the biosynthesis of MNPs using microorganisms were summarized and analyzed, evidencing a prevalence of research on silver nanoparticles synthesized using bacteria aiming toward testing their antimicrobial potential. Finally, a synopsis of studies on MNPs applied to cultural heritage materials showed potential for their future use in preservation.

The impact of sound in people's behaviour in outdoor settings: A study using virtual reality and eye-tracking.

This paper presents an analysis of space perception and how visual cues, such as landmarks and sound, are perceived and impact people's behaviour while exploring a given outdoor space. The primary goal of the research is to investigate how auditory sensations and visual stimuli influence people's behaviour in outdoor built environments. Our technique compares people's perception of the built environment in different conditions: the real world and a replicated virtual world. As a case study, a university campus was used, and four experimental conditions were designed. The study followed a between-subjects design, and the data collection included gaze data acquired from an eye-tracking device as well as self-reports. The study concludes that sound influences human behaviour in such settings. More specifically conclusions are that: i) human behaviour in virtual replications of the real space, including both visual and sound stimuli, is tendentially more similar to human behaviour in the real world than in simulations omitting sound; and ii) there is a difference in human behaviour when people explore the same virtually replicated outdoor space, by varying the presence of sound. This study is particularly useful for researchers working on the comparison between human behaviour in virtual and real environments, related to visual and sound stimuli.

Response to the letter to the editor: "Score risk scale as a prognostic factor after sudden sensorineural hearing loss".

We appreciate the opportunity to comment the observations on our paper entitled "Score risk scale as a prognostic factor after sudden sensorineural hearing loss", by Capuano et al. This letter highlights several important points, including the role of hyperbaric oxygen therapy and the possible association between patent foramen ovale and sudden sensorineural hearing loss (SSHL). Further research is needed to strengthen the association between cardiovascular risk and SSHL. We thank the authors for their insights into our paper and for adding their experience and observations on the potential role of cardiovascular risk in the etiology of SSHL.

Bioactive Components and Antioxidant and Antibacterial Activities of Different Varieties of Honey: A Screening Prior to Clinical Application.

This study assessed 16 different honey samples in order to select the best one for therapeutic purposes. First, a study of honey's main bioactive compounds was carried out. Then phenolic profiles were determined and specific compounds quantified using a HPLC system coupled to a mass spectrometer. Then, antioxidant activity, by three in vitro methods, and antibacterial activity against reference strains and clinical isolates were evaluated. Great variability among samples was observed regarding ascorbic acid (between 0.34 ± 0.00 and 75.8 ± 0.41 mg/100 g honey; p < 0.001), total phenolic compounds (between 23.1 ± 0.83 and 158 ± 5.37 mg/100 g honey; p < 0.001), and total flavonoid contents (between 1.65 ± 0.11 and 5.93 ± 0.21 mg/100 g honey; p < 0.001). Forty-nine different phenolic compounds were detected, but only 46 of them were quantified by HPLC. The concentration of phenolic compounds and the phenolic profiles varied widely among samples (between 1.06 ± 0.04 and 18.6 ± 0.73 mg/100 g honey; p < 0.001). Antioxidant activity also varied significantly among the samples. All honey varieties exhibited antibacterial activity against both reference and clinical strains (effective concentrations ranged between 0.05 and 0.40 g/mL depending on the honey sample and bacteria tested). Overall, samples with better combinations of bioactive properties were avocado and chestnut honeys.

Antioxidant, photoprotective and inhibitory activity of tyrosinase in extracts of Dalbergia ecastaphyllum.

Dalbergia ecastaphyllum is a native Brazil plant with importance for beekeeping, and widely used in folk medicine. For the first time, the extracts of this plant were assessed for the presence of hydrophilic and lipophilic antioxidants, as well as inhibition of tyrosinase, free radicals scavenging and sunscreen protection. The antioxidant activity was evaluated by free radical scavenging (DPPH) and ß-carotene bleaching assay. The tyrosinase inhibitory activity was evaluated and calculated the EC50. The photoprotective activity was measured using different concentrations of D. ecastaphyllum extracts. The Sun Protection Factor (SPF) of the samples was higher than 6, and the sample from Ilhéus showed the most pronounced photoprotective effect. Sample from Canavieiras presented the highest antioxidant activity by free radical scavenging DPPH and ß-carotene bleaching method, with 92.41% and 48.34%, respectively. All samples inhibited the tyrosinase, especially the sample from Prado that was most effective (124.62 µg.mL-1). Significant negative correlation was found between flavonoid contents and inhibition of tyrosinase. The overall results provide relevant information about the Dalbergia ecastaphyllum species, indicating as potential material to cosmetic and pharmaceutical industry.

Use of Copaifera multijuga for acute corneal repair after chemical injury: A clinical, histopathological and toxicogenetic study.

Copaiba oil is widely used in medicine, but there are no reports regarding its application in ophthalmology. Therefore, the objective of this study was to evaluate the clinical, histopathological and toxicogenetic effects of eye drops containing 0.1 and 0.5% of Copaifera multijuga Hayne oil on superficial corneal ulcers induced with alkali in the left eye of rats. For histological analysis, the percent reduction in ulcers and thickness of the corneal epithelium and stroma were evaluated 48 and 72 h after ulcer induction. Additionally, neovascularization and polymorphonuclear infiltration were classified in the stroma. The bone marrow micronucleus test was used for toxicogenetic assessment. None of the animals exhibited clinical signs of immediate ocular discomfort after instillation and the eye drops were harmless to the ocular surface. There was a significant difference in percent ulcer reduction and corneal stroma thickness between animals treated with the C. multijuga eye drops and untreated animals with corneal injury and the negative control, respectively, suggesting a healing effect of the oleoresin. Analysis of the thickness of the corneal epithelium at the two time points showed that the eye drops formulated did not significantly reduce the damage caused by alkali. The same was observed for the treatments with the reference drugs. No difference in stromal neovascularization or inflammatory infiltration was observed between the treated groups. The toxicogenetic results revealed the absence of cytotoxicity and genotoxicity of the treatments. In conclusion, the C. multijuga eye drops did not cause damage to the ocular surface under the present experimental conditions and corneal epithelization was similar to the conventional treatments. These results indicate that eye drops containing C. multijuga oleoresin are a promising option for the treatment of superficial keratitis.

A novel approach for honey pollen profile assessment using an electronic tongue and chemometric tools.

Nowadays the main honey producing countries require accurate labeling of honey before commercialization, including floral classification. Traditionally, this classification is made by melissopalynology analysis, an accurate but time-consuming task requiring laborious sample pre-treatment and high-skilled technicians. In this work the potential use of a potentiometric electronic tongue for pollinic assessment is evaluated, using monofloral and polyfloral honeys. The results showed that after splitting honeys according to color (white, amber and dark), the novel methodology enabled quantifying the relative percentage of the main pollens (Castanea sp., Echium sp., Erica sp., Eucaliptus sp., Lavandula sp., Prunus sp., Rubus sp. and Trifolium sp.). Multiple linear regression models were established for each type of pollen, based on the best sensors' sub-sets selected using the simulated annealing algorithm. To minimize the overfitting risk, a repeated K-fold cross-validation procedure was implemented, ensuring that at least 10-20% of the honeys were used for internal validation. With this approach, a minimum average determination coefficient of 0.91 ± 0.15 was obtained. Also, the proposed technique enabled the correct classification of 92% and 100% of monofloral and polyfloral honeys, respectively. The quite satisfactory performance of the novel procedure for quantifying the relative pollen frequency may envisage its applicability for honey labeling and geographical origin identification. Nevertheless, this approach is not a full alternative to the traditional melissopalynologic analysis; it may be seen as a practical complementary tool for preliminary honey floral classification, leaving only problematic cases for pollinic evaluation.

Microbiological Assessment, Nutritional Characterization and Phenolic Compounds of Bee Pollen from Mellipona mandacaia Smith, 1983.

This study aims to assess the microbiological parameters and the chemical composition of 21 samples of stingless bee pollen (Melipona mandacaia) from two regions of Bahia, Brazil (João Dourado and Uibaí), with particular emphasis on the nutritional value, total phenols and flavonoids and fatty acids composition. Regarding the microbiological quality, the studied microorganisms (moulds and yeasts, coliforms, Escherichia coli, Staphylococcus aureus, Salmonella sp., psychrotrophic and sulfite-reducing Clostridia) were absent in all samples. On the other hand, the values obtained for the aerobic mesophilic microorganism ranged from 11.0 ± 1.0 to 1.32 ± 1.2 cfu∙g-1 (JD samples) and from 282 ± 3.82 to 688 ± 10.1 cfu∙g-1 (U samples). The nutritional parameters (moisture, ash, water activity, pH, total acidity, protein, fiber, total phenolic, flavonoids and reducing sugars) were within the stipulated by law, except for pH and moisture content, which presented superior and inferior values, respectively. Polyunsaturated fatty acids (54.1%) were significantly higher than saturated (42.18%) and monounsaturated (3.71%). It was found that the bee pollen is safe from the microbiological point of view and has a good nutritional quality. The influence of the geographical origin on the assessed parameters was evident, especially concerning the fatty acid profile.

Endodontics pastes formulated with copaiba oil: action on oral microbiota and dentin bridge formation in dogs / Pastas endodônticas formuladas com óleo de copaíba: ação na microbiota bucal e formação de ponte dentinária em cães

This study analysed the effect of pastes formulated with calcium hydroxide P.A. and different vehicles (saline solution - paste A and Copaifera langsdorffii Desfon oil - paste B) on oral microorganisms and dentin bridge formation in dogs. The antimicrobial action of the pastes and their components was analysed by the minimum inhibitory concentration in agar gel technique. The components were diluted and tested on fifteen standard strains of microorganisms associated with endodontic diseases. The microorganisms were cultivated and after incubation data was analysed using One-Way ANOVA and Turkey's test (P≤0.05). Four superior incisors of ten animals were used to evaluate dentin bridge formation. Two incisors were capped with paste A (GA) and two with paste B (GB). After 90 days, the teeth were extracted for histological analysis and the degree of dentin bridge formation evaluated. Data was analysed by the Kruskal-Wallis test (P<0.05). The pastes and their components were classified in the following decreasing order of antimicrobial action: calcium hydroxide P.A., paste A, paste B and Copaifera langsdorffii Desfon oil. Calcium hydroxide P.A. showed significantly higher antimicrobial action than the pastes or their vehicles. No significant difference was observed between the two pastes in dentin bridge formation. Based on the microorganisms studied, it can be concluded that the pastes analysed showed similar antimicrobial potential but differed significantly from their individual components. No significant difference was observed in dentin bridge formation between the different pastes tested.

Foi avaliada a ação de pastas formuladas com hidróxido de cálcio P.A. e diferentes veículos (solução fisiológica - pasta A e óleo de Copaifera langsdorffii Desfon - pasta B) sobre microrganismos bucais e formação de ponte dentinária em cães. A ação antimicrobiana das pastas e de seus componentes individuais foi avaliada pela técnica da concentração inibitória mínima pela diluição em ágar. Os materiais foram diluídos e avaliados contra quinze cepas padrão de microrganismos relacionados a doenças endodônticas. Os microrganismos foram cultivados e, após a incubação, os dados foram analisados (Anova One-Way, Tukey, P≤0,05). Para a avaliação da formação de ponte dentinária, quatro incisivos superiores de dez animais foram tratados, sendo dois capeados com pasta A (GA) e dois com a pasta B (GB). Após 90 dias, os dentes foram extraídos para obtenção de cortes histológicos, com o objetivo de se avaliar o grau de formação de ponte dentinária. Os dados obtidos foram submetidos à análise estatística (Kruskal-Wallis, P<0,05). Segundo a ação antimicrobiana, os materiais foram classificados em ordem decrescente: hidróxido de cálcio P.A., pasta A, pasta B e óleo de Copaifera langsdorffii Desfon, com diferenças estatisticamente significantes entre o hidróxido de cálcio P.A., as pastas e os veículos. Ao final do estudo, observou-se que as duas pastas avaliadas foram semelhantes quanto à formação de ponte dentinária. Considerando-se os microrganismos selecionados, conclui-se que as pastas avaliadas foram semelhantes entre si em termos de potencial antimicrobiano, diferindo dos componentes individuais. Em relação à formação de ponte dentinária, não foram observadas diferenças significantes entre os dentes tratados com as diferentes pastas.

Practical procedure for discriminating monofloral honey with a broad pollen profile variability using an electronic tongue.

Colour and floral origin are key parameters that may influence the honey market. Monofloral light honey are more demanded by consumers, mainly due to their flavour, being more valuable for producers due to their higher price when compared to darker honey. The latter usually have a high anti-oxidant content that increases their healthy potential. This work showed that it is possible to correctly classify monofloral honey with a high variability in floral origin with a potentiometric electronic tongue after making a preliminary selection of honey according their colours: white, amber and dark honey. The results showed that the device had a very satisfactory sensitivity towards floral origin (Castanea sp., Echium sp., Erica sp., Lavandula sp., Prunus sp. and Rubus sp.), allowing a leave-one-out cross validation correct classification of 100%. Therefore, the E-tongue shows potential to be used at analytical laboratory level for honey samples classification according to market and quality parameters, as a practical tool for ensuring monofloral honey authenticity.

Single-cultivar extra virgin olive oil classification using a potentiometric electronic tongue.

Label authentication of monovarietal extra virgin olive oils is of great importance. A novel approach based on a potentiometric electronic tongue is proposed to classify oils obtained from single olive cultivars (Portuguese cvs. Cobrançosa, Madural, Verdeal Transmontana; Spanish cvs. Arbequina, Hojiblanca, Picual). A meta-heuristic simulated annealing algorithm was applied to select the most informative sets of sensors to establish predictive linear discriminant models. Olive oils were correctly classified according to olive cultivar (sensitivities greater than 97%) and each Spanish olive oil was satisfactorily discriminated from the Portuguese ones with the exception of cv. Arbequina (sensitivities from 61% to 98%). Also, the discriminant ability was related to the polar compounds contents of olive oils and so, indirectly, with organoleptic properties like bitterness, astringency or pungency. Therefore the proposed E-tongue can be foreseen as a useful auxiliary tool for trained sensory panels for the classification of monovarietal extra virgin olive oils.

Bioassays against pinewood nematode: assessment of a suitable dilution agent and screening for bioactive essential oils.

Acetone was investigated and found to be an appropriate alternative to Triton X-100 as a solvent of essential oils in bioassays aimed to investigate their effects on pinewood nematode (Bursaphelenchus xylophilus) mortality. Therefore it was used as dilution agent to screen the effectiveness of fifty two essential oils against this pest. Thirteen essential oils were highly effective, resulting in more than 90% pinewood nematode mortality at 2 mg/mL, with six of them resulting in 100% mortality. LC100 values ranged between 0.50 mg/mL and 0.83 mg/mL for the essential oils of Origanum vulgare and Satureja montana, respectively. Essential oils were submitted to gas chromatography and gas chromatography-mass spectrometry analysis and their chemical composition established. Data from essential oils with 100% mortality at 2 mg/mL and other essential oils previously found to have LC100 ≤ 2 mg/mL was combined, their chemical profiles investigated by correspondences analysis plus automatic classification.

Comparative study of different Portuguese samples of propolis: pollinic, sensorial, physicochemical, microbiological characterization and antibacterial activity.

The aim of this work was to study four propolis samples from Trás-os-Montes region of Portugal. The propolis samples' color was different, which pollen analysis showed to be due to different botanical sources: Populus sp., Pinus sp., Quercus sp. and Castanea sativa. The data from physicochemical analysis (moisture, soluble and insoluble solids content, pH, conductivity, ash content, wax, total phenolics and flavonoids content) was treated using multivariate statistical tools as cluster heat map, principal components analysis and linear discriminant analysis with the purpose of classifying the sample accordingly to the botanical/geographical origin. The discriminant analysis was applied with stepwise to select the variables that most contribute to sample identification accordingly to pollinic profile. The cross-validation technique was applied, using the leave-one-out procedure, which showed good prediction capabilities of the samples. Microbiologically, the commercial quality was satisfactory, since the samples didn't contain deterioration or pathogenic microorganisms. All the samples studied presented antimicrobial activity against Methicillin-resistant Staphylococcus aureus, in a dose dependent way. The antimicrobial activity was strictly related to the physicochemical composition. This work will allow connecting a particular chemical propolis type to a specific type of biological activity, what is essential for the use in therapeutic applications.

Comprehensive study of honey with protected denomination of origin and contribution to the enhancement of legal specifications.

In this study the characterization of a total of 60 honey samples with Protected Denomination of Origin (PDO) collected over three harvests (2009-2011, inclusive), from the Northeast of Portugal was carried out based on the presence of pollen, physicochemical and microbiological characteristics. All samples were found to meet the European Legislation, but some didn't meet the requirements of the PDO specifications. Concerning the floral origin of honey, our results showed the prevalence of rosemary (Lavandula pedunculata) pollen. The microbiological quality of all the analyzed samples was satisfactory, since fecal coliforms, sulfite-reducing clostridia and Salmonella were absent, and molds and yeasts were detected in low counts. Significant differences between the results were studied using one-way analysis of variance (ANOVA), followed by Tukey's HSD test. The samples were submitted to discriminant function analysis, in order to determine which variables differentiate between two or more naturally occurring groups (Forward Stepwise Analysis). The variables selected were in this order: diastase activity, pH, reducing sugars, free acidity and HMF. The pollen spectrum has perfect discriminatory power. This is the first study in which a honey with PDO was tested, in order to assess its compliance with the PDO book of specifications.

Physicochemical, microbiological and antimicrobial properties of commercial honeys from Portugal.

The present study aimed to characterize five commercial honeys available in the Portuguese market in respect to their floral origins, physicochemical parameters and microbial safety and commercial quality assessment. Pollen profile, colour, moisture content, ash, acidity, electrical conductivity, pH, reducing sugars, apparent sucrose and HMF were the parameters analysed in each honey sample. Aerobic mesophiles, moulds and yeasts, fecal coliforms and sulphite-reducing clostridia were the microbial contaminants of interest studied. The antimicrobial effect against four fermentative yeasts was determined. Concerning the physicochemical parameters, all honey samples were found to meet European Legislation (EC Directive 2001/110) for all parameters, except for HMF and apparent sucrose. Microbiologically, the commercial quality was considered good and all samples showed to be negative in respect to safety parameters. We also verified that the presence of honey differentially affected the growth of fermentative yeasts under study, depending on the type of yeast, but this growth was not significantly influenced by the type of honey used.

Antioxidant properties, total phenols and pollen analysis of propolis samples from Portugal.

Pollen analysis, total phenols content and antioxidant activity were studied for the first time in Portuguese propolis samples from Bornes and Fundão regions. Total phenols content was determined by colorimetric assay and their amount was of 329 mg/g of GAE in Bornes sample and 151 mg/g of GAE in Fundão propolis. The antioxidant capacity of propolis extracts was assessed through the scavenging effects on DPPH (2,2-diphenyl-1-picrylhydrazyl) and reducing power of iron (III)/ ferricyanide complex assays. A concentration-dependent antioxidative capacity was verified in DPPH and reducing power assays. Low values of EC50 on DPPH scavenging assay were obtained for Bornes and Fundão propolis (of 6.22 microg/mL and 52.00 microg/mL, respectively). For reducing power the values were 9.00 microg/mL, for Bornes propolis, and 55.00 microg/mL, for Fundão propolis. The high activity of propolis from Bornes could be related with their different pollen composition. The results obtained indicate that Portuguese propolis is an important source of total phenols showing antioxidant properties that could be beneficial for human health.