Further Insights on Honey and Propolis from Gerês (Portugal) and Their Bioactivities: Unraveling the Impact of Beehive Relocation.

Propolis, a bee product, is known for its variability of chemical and bioactive profiles. However, Portuguese propolis from Gerês, normally obtained by mixing propolis from three places-Bugalho, Felgueiras and Toutelo-has shown similar chemical and biological profiles over the years. Recently, a new propolis place-Roca-was added to the apiary to replace Bugalho, lost to the 2017 wildfires, hence questioning the previously claimed constancy of Gerês propolis. To unravel to what extent the beehive relocation affected this constancy, we studied different Gerês propolis samples collected in three consecutive years (2017-2019) composed of different combinations of source places. Two honey samples, collected before (2017) and after (2018) the occurrence of the wildfire, were also investigated. Total phenolics, flavonoids and ortho-diphenols contents were determined and the antioxidant and antimicrobial activities were evaluated, using the DPPH assay and the agar dilution method, respectively. Although both antimicrobial and antioxidant activities were generally in the ranges usually obtained from Gerês propolis, some variations were detected for the samples, with different compositions when compared to previous years. This work reinforces the importance of the consistency of a combination of several factors for the protection and preservation of the flora near the hives, providing bee products with more constant chemical and biological profiles over the years.

Machine Learning-Assisted Optimization of Drug Combinations in Zeolite-Based Delivery Systems for Melanoma Therapy.

Two independent artificial neural network (ANN) models were used to determine the optimal drug combination of zeolite-based delivery systems (ZDS) for cancer therapy. The systems were based on the NaY zeolite using silver (Ag+) and 5-fluorouracil (5-FU) as antimicrobial and antineoplastic agents. Different ZDS samples were prepared, and their characterization indicates the successful incorporation of both pharmacologically active species without any relevant changes to the zeolite structure. Silver acts as a counterion of the negative framework, and 5-FU retains its molecular integrity. The data from the A375 cell viability assays, involving ZDS samples (solid phase), 5-FU, and Ag+ aqueous solutions (liquid phase), were used to train two independent machine learning (ML) models. Both models exhibited a high level of accuracy in predicting the experimental cell viability results, allowing the development of a novel protocol for virtual cell viability assays. The findings suggest that the incorporation of both Ag and 5-FU into the zeolite structure significantly potentiates their anticancer activity when compared to that of the liquid phase. Additionally, two optimal AgY/5-FU@Y ratios were proposed to achieve the best cell viability outcomes. The ZDS also exhibited significant efficacy against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus); the predicted combination ratio is also effective against S. aureus, underscoring the potential of this approach as a therapeutic option for cancer-associated bacterial infections.

Valorization of plant by-products in the biosynthesis of silver nanoparticles with antimicrobial and catalytic properties.

Biosynthesis based on natural compounds has emerged as a sustainable approach for the production of metallic nanoparticles (MNP). The main objective of this study was to biosynthesize stable and multifunctional silver nanoparticles (AgNP) using different plant by-products as reducers and capping agents. Extracts obtained from Eucalyptus globulus, Pinus pinaster, Citrus sinensis, Cedrus atlantica and Camellia sinensis by-products, were evaluated. From all plant by-products tested, aqueous extract of eucalyptus leaves (EL), green tea (GT) and black tea (BT) were selected due to their higher antioxidant phenolic content and were individually employed as reducers and capping agents to biosynthesize AgNP. The green AgNP showed zeta potential values of -31.8 to -36.3 mV, with a wide range of particle sizes (40.6 to 86.4 nm), depending on the plant extract used. Green AgNP exhibited an inhibitory effect against various pathogenic bacteria, including Gram-negative (P. putida, E. coli, Vibrio spp.) and Gram-positive (B. megaterium, S. aureus, S. equisimilis) bacteria with EL-AgNP being the nanostructure with the greatest antimicrobial action. EL-AgNP showed an excellent photodegradation of indigo carmine (IC) dye under direct sunlight, with a removal percentage of up to 100% after 75 min. A complete cost analysis revealed a competitive total cost range of 8.0-9.0 €/g for the biosynthesis of AgNP.

Necrosis of the Tongue as a Late Complication of Radiotherapy.

Introduction: Irradiation to treat head and neck cancer, causing chronic tissue damage, is associated with the development of vascular disease. Interest has risen over the effects of radiotherapy on major vessels, due to its high morbidity and mortality rate. However, small-vessel disease has been poorly studied and described. Case Report: We present a case of a patient with bilateral necrosis of the anterior third of the tongue, occurring 3 years after chemoradiotherapy treatment for squamous cell carcinoma of the floor of the mouth. Contrast-enhanced CT scan showed multiple areas of stenosis concerning both external carotid arteries and their branches, and total opacification of lingual arteries. Conservative management was performed, with auto-amputation on the fifth day, which allowed healing by secondary intention. Conclusions: Necrosis of the tongue appears as a rare late complication of radiotherapy, possibly due to its acceleration effect on the atherosclerosis process. Following small-vessel disease, one can assume a higher potential risk of major-vessel disease, highlighting the importance of a routine assessment and prophylaxis of thrombotic events.

Modelling the In Vitro Growth of Phytopathogenic Filamentous Fungi and Oomycetes: The Gompertz Parameters as Robust Indicators of Propolis Antifungal Action.

Propolis is a resinous mixture produced by honeybees, mainly from plant exudates. With a rich chemical composition including many phenolic compounds, mostly responsible for its biological properties, namely antimicrobial ones, propolis may be a promising alternative to synthetic pesticides. The study of propolis from the south of Portugal and of its potential against phytopathogenic agents are still very recent and different methodological approaches hinder a comparison of efficacies. In this context, we aimed to test the value of a mathematical model for the multiparametric characterization of propolis' antifungal action on solid medium assays. An ethanol extract (EE) of a propolis sample harvested in 2016 from Alves (A16) was characterized in terms of phenolic composition and antimicrobial potential against five phytopathogenic species. A16.EE (500-2000 µg/mL) inhibited the mycelial growth of all the species, with Phytophthora cinnamomi and Biscogniauxia mediterranea being the most susceptible and Colletotrichum acutatum being the least affected. The Gompertz mathematical model proved to be a suitable tool for quantitatively describing the growth profiles of fungi and oomycetes, and its parameters exhibit a high level of discrimination. Our results reveal that propolis extracts may have potential applications beyond traditional uses, particularly within the agri-food sector, allowing beekeepers to make their businesses more profitable and diversified.

Examination of Raw Samples and Ethanol Extracts of Gerês Propolis Collected in Different Years.

Propolis, a natural resin created by bees, has garnered significant attention from both the scientific community and industry due to an impressive range of bioactivities. Nonetheless, the intrinsic variability in its chemical composition and bioactive profiles has been hindering propolis' full potential use. We previously showed that ethanol extracts (EEs) of a Portuguese propolis sample (Gerês) collected over four consecutive years displayed similar chemical and biological profiles, a constancy never documented before. However, the characteristics of the unprocessed samples of Gerês propolis were never described. Hence, the central objective of this study is to assess the quality parameters of unprocessed propolis samples collected from Gerês (G), over a four-year period (2019-2022), alongside the analysis of the chemical composition and bioactivities of the EEs prepared with the same raw samples. The ash, wax, balsam and water contents of the unprocessed samples-G19 to G22-showed minor fluctuations, likely attributed to uncontrollable natural events impacting the propolis source and collection process. On the other hand, the antimicrobial and antioxidant activities of all the four ethanol extracts (G19.EE-G22.EE) consistently align with prior studies. Furthermore, the Gerês propolis extracts showed remarkable uniformity in chemical composition parameters too, particularly concerning total polyphenol, flavonoid and ortho-diphenol contents. In summary, our research reinforces the beneficial properties of propolis and show that extracts' bioactivities remain within the reference ranges for Gerês propolis, despite minor differences in unprocessed samples, suggesting a consistent action over time. Thus, this work could be instrumental towards the establishment of standard parameters for propolis applications, offering valuable insights to this field of propolis research.

Phenolic Compounds Contribution to Portuguese Propolis Anti-Melanoma Activity.

Melanoma is the deadliest type of skin cancer, with about 61,000 deaths annually worldwide. Late diagnosis increases mortality rates due to melanoma's capacity to metastasise rapidly and patients' resistance to the available conventional therapies. Consequently, the interest in natural products as a strategy for drug discovery has been emerging. Propolis, a natural product produced by bees, has several biological properties, including anticancer effects. Propolis from Gerês is one of the most studied Portuguese propolis. Our group has previously demonstrated that an ethanol extract of Gerês propolis collected in 2018 (G18.EE) and its fractions (n-hexane, ethyl acetate, and n-butanol) decrease melanoma cell viability. Out of all the fractions, G18.EE-n-BuOH showed the highest potential as a melanoma pharmacological therapy. Thus, in this work, G18.EE-n-BuOH was fractioned into 17 subfractions whose effect was evaluated in A375 BRAF-mutated melanoma cells. The subfractions with the highest cytotoxic activity were analysed by UPLC-DAD-ESI/MSn in an attempt to understand which phenolic compounds could account for the anti-melanoma activity. The compounds identified are typical of the Gerês propolis, and some of them have already been linked with antitumor effectiveness. These results reaffirm that propolis compounds can be a source of new drugs and the isolation of compounds could allow its use in traditional medicine.

Predictive factors of revision endoscopic sinus surgery.

PURPOSE: Despite endoscopic sinus surgery (ESS) being the standard best treatment for chronic rhinosinusitis with nasal polyps (CRSwNP), there still remains a high recurrence rate of nasal polyps after ESS. The aim of this study was to analyze the impact of ESS in the treatment of CRSwNP and identify possible predictors for revision surgery. METHODS: We conducted a cross-sectional retrospective study of patients with CRSwNP submitted to ESS between the years 2013 and 2021. RESULTS: We analyzed 517 cases, of which 15.1% were submitted to revision ESS. The presence of hyposmia/anosmia, asthma, intolerance/allergy to non-steroid inflammatory drugs, non-steroidal anti-inflammatory drug exacerbated respiratory disease, and bilateral polyposis were more frequent in patients who needed revision surgery. Initial values of peripheral eosinophilia and Lund-Mackay scores were significantly higher in this sub-group of patients. Lund-Mackay score was the only independent risk factor for revision surgery. CONCLUSIONS: Factors like peripheral eosinophilia and Lund-Mackay score were associated with a higher probability of revision ESS. Applying these concepts to the clinical practice could help predict the evolution of patient's pathology in the post-operative period and identify in advance those cases that may need a multidisciplinary approach.

Design of a postpartum hemorrhage and transfusion risk calculator.

Background: Postpartum hemorrhage is the major cause of maternal deaths due to childbirth and also responsible for maternal morbidity. Objectives: In this study we set out to look the incidence of postpartum hemorrhage in our population, to identify the most important risk factors for postpartum hemorrhage and thus develop a predictive risk calculator for postpartum hemorrhage and transfusion. Study design: data was taken from patients who presented vaginal delivery or cesarean section from January 1 to December 31, 2016, the variables taken into account as risk factors were as follows: Gestational age, history of chronic or gestational hypertension, preeclampsia, previous abortions, parity, previous cesarean section, placenta previa, labor time, and postpartum hemorrhage as the event of interest. An objective quantification was performed on a weight scale in grams for the estimation of bleeding, considering postpartum hemorrhage those with >500 ml in vaginal delivery and >1000 ml of blood loss in cesarean section. Subsequently, a predictive risk calculator was developed using the Naïve Bayes algorithm. Results: A success rate of 58% was obtained in the identification of patients at high risk of hemorrhage, and 36% for transfusion, with a sensitivity of 50.7% and specificity of 64.06%, identifying as risk factors for postpartum hemorrhage gestational age between 35 and 40 weeks, hypertension and preeclampsia, previous cesarean section, duration of labor <1 h or more than 10 h, placenta previa and previous history of postpartum hemorrhage. Conclusion: A postpartum hemorrhage risk calculator has been designed, which due to its improved accuracy after incorporation of data becomes a useful tool that will require a larger study population to improve its performance in clinical practice and more similar studies to validate it.

Essential Oils Encapsulated in Zeolite Structures as Delivery Systems (EODS): An Overview.

Essential oils (EO) obtained from plants have proven industrial applications in the manufacturing of perfumes and cosmetics, in the production and flavoring of foods and beverages, as therapeutic agents in aromatherapy, and as the active principles or excipients of medicines and pharmaceutics due to their olfactory, physical-chemical, and biological characteristics. On behalf of the new paradigm of a more natural and sustainable lifestyle, EO are rather appealing due to their physical, chemical, and physiological actions in human beings. However, EO are unstable and susceptible to degradation or loss. To tackle this aspect, the encapsulation of EO in microporous structures as zeolites is an attractive solution, since these host materials are cheap and non-toxic to biological environments. This overview provides basic information regarding essential oils, including their recognized benefits and functional properties. Current progress regarding EO encapsulation in zeolite structures is also discussed, highlighting some representative examples of essential oil delivery systems (EODS) based on zeolites for healthcare applications or aromatherapy.

An Insight on the Biomedical Potential of Portuguese Propolis from Gerês.

Osteoarthritis (OA), a progressive degenerative disease of weight-bearing joints, is the second leading cause of disability in the world. Despite all the advances and research over the last years, none of the proposed strategies has been effective in generating functional and long-lasting tissue. Due to the high prevalence of OA and the urgent need for an effective and successful treatment, interest in natural products as anti-inflammatory agents, such as propolis and its components, has emerged. In this work, we estimate the biomedical potential of Portuguese propolis, evaluating the in vitro antioxidant and anti-inflammatory effects of single hydroalcoholic extracts prepared with propolis from Gerês sampled over a five-year period (2011-2015) (G.EE70 and G.EE35). The in vivo and in vitro anti-inflammatory potential of the hydroalcoholic extract of mixtures of the same samples (mG.EE70 and mG.EE35) was evaluated for the first time too. DPPH• radical scavenging and superoxide anion scavenging assays showed the strong antioxidant potential of both hydroalcoholic extracts, either prepared from single propolis samples or from the mixtures of the same samples. Results also revealed an anti-inflammatory effect of mG.EE35, both in vitro by inhibiting BSA denaturation and in vivo in the OA-induced model by improving mechanical hyperalgesia as well as the gait pattern parameters. Results further support the use of propolis blends as a better and more efficient approach to take full advantage of the bioactive potential of propolis.

Propolis Efficacy: The Quest for Eco-Friendly Solvents.

Propolis, a natural product made by bees with resins and balsams, is known for its complex chemical composition and remarkable bioactivities. In this study, propolis extraction was studied seeking extracts with strong bioactivities using less orthodox solvents, with some derived from apiary products. For that, a propolis sample collected from Gerês apiary in 2018 (G18) was extracted by maceration with six different solvents: absolute ethanol, ethanol/water (7:3), honey brandy, mead, propylene glycol and water. The solvent influence on the chemical composition and antioxidant and antimicrobial activities of the extracts was investigated. Antioxidant potential was assessed by the DPPH free-radical-scavenging assay and the antimicrobial activity by the agar dilution method. Chemical composition of the extracts was determined in vitro by three colorimetric assays: total ortho-diphenols, phenolics and flavonoids contents and the LC-MS technique. To our knowledge, this is the first time that solvents such as honey brandy and mead have been studied. Honey brandy showed considerable potential to extract propolis active compounds able to inhibit the growth of bacteria such as the methicillin-sensitive Staphylococcus aureus and Propionibacterium acnes (MIC values of 100 and 200 µg/mL, respectively) and the fungi Candida albicans and Saccharomyces cerevisiae (MIC = 500 µg/mL, for both). Mead extracts displayed high antioxidant capacity (EC50 = 1.63 ± 0.27 µg/mL) and great activity against resistant bacteria such as the methicillin-resistant Staphylococcus aureus and Escherichia coli (MIC = 750 µg/mL, for both). The production of such solvents made from beehive products further promotes a diversification of apiary products and the exploration of new applications using eco-friendly solutions.

Mixing Propolis from Different Apiaries and Harvesting Years: Towards Propolis Standardization?

Global demand for safe, effective and natural products has been increasing in parallel with consumers' concerns about personal and environmental health. Propolis, a traditional and potentially medicinal product with several health benefits, is a beehive product with a worldwide reputation. However, despite the bioactivities reported, the low productivity and high chemical heterogeneity have been extensively hampering broader industrial uses. To assist in overcoming some of these problems, we prepared and characterized mixtures of ethanol extracts of a heterogeneous propolis sample (Pereiro) collected over a five-year period (2011-2015) and, additionally, we mixed two different propolis samples from distinct regions of Portugal (Pereiro and Gerês), also harvested at different times. An investigation of the antimicrobial and antioxidant properties, as well as characterization of the chemical composition of the eleven propolis blends were performed in this work. The antioxidant and antimicrobial activities of such blends of propolis samples, either from different localities and/or different years, were maintained, or even enhanced, when a comparison of the individual extracts was conducted. The differences in the chemical composition of the original propolis samples were also diluted in the mixtures. The results reemphasize the great potential of propolis and suggest that mixing different samples, regardless of provenance or harvesting date, can contribute to propolis standardization while simultaneously increasing its availability and adding value to this beehive byproduct.

Selective Cytotoxicity of Portuguese Propolis Ethyl Acetate Fraction towards Renal Cancer Cells.

Renal cell carcinoma is the most lethal cancer of the urological system due to late diagnosis and treatment resistance. Propolis, a beehive product, is a valuable natural source of compounds with bioactivities and may be a beneficial addition to current anticancer treatments. A Portuguese propolis sample, its fractions (n-hexane, ethyl acetate, n-butanol and water) and three subfractions (P1-P3), were tested for their toxicity on A498, 786-O and Caki-2 renal cell carcinoma cell lines and the non-neoplastic HK2 kidney cells. The ethyl acetate fraction showed the strongest toxicity against A498 (IC50 = 0.162 µg mL-1) and 786-O (IC50 = 0.271 µg mL-1) cells. With similar toxicity against 786-O, P1 (IC50 = 3.8 µg mL-1) and P3 (IC50 = 3.1 µg mL-1) exhibited greater effect when combined (IC50 = 2.5 µg mL-1). Results support the potential of propolis and its constituents as promising coadjuvants in renal cell carcinoma treatment.

Disease-Specific Vs Non-Specific Questionnaires on Septoplasty Outcomes.

Introduction: Validated questionnaires are a valuable tool in medical practice. The role of septoplasty in improving patients' non-nasal symptoms and their quality of life is still controversial. The aim of this study was to determine the differences in outcome after septoplasty measured by a disease-specific questionnaire vs a general QoL questionnaire. Materials and Methods: A total of 50 patients underwent septoplasty and completed the SNOT-22 and the SF-36v2 questionnaires preoperatively and at 6 months post-op. Pre-op, post-op and variation for each domain in both questionnaires were calculated and compared with a measure of self-reported health transition (Question 2 of SF-36v2) and with the Minimal important difference (MID) for our sample. Results: SNOT-22 scores significantly improved for each specific question and for the total score. SF-36v2 showed a significant improvement in scores for mental domains (Mental Health, Role Emotional, and Vitality) but less so for the physical domains. MID for our sample was 4.2 points. Patients with variations greater than 4.2 in SNOT-22 total score (74%) revealed significantly better variations in Physical Function, General Health, Social Function and Vitality. Conclusions: Validated questionnaires are a fundamental tool for assessing outcomes of commonly performed surgeries in ENT. Disease-specific questionnaire showed improvement in scores for the majority of patients. The general QoL showed improvement only in Mental Domains. This may suggest that the impact of septoplasty in patient's general health might not be significant.

Portuguese Propolis Antitumoral Activity in Melanoma Involves ROS Production and Induction of Apoptosis.

Melanoma is the most aggressive and life-threatening skin cancer type. The melanoma genome is the most frequently mutated, with the BRAF mutation present in 40-60% of melanoma cases. BRAF-mutated melanomas are characterized by a higher aggressiveness and progression. Adjuvant targeted treatments, such as BRAF and MEK inhibitors, are added to surgical excision in BRAF-mutated metastatic melanomas to maximize treatment effectiveness. However, resistance remains the major therapeutic problem. Interest in natural products, like propolis, for therapeutic applications, has increased in the last years. Propolis healing proprieties offer great potential for the development of novel cancer drugs. As the activity of Portuguese propolis has never been studied in melanoma, we evaluated the antitumoral activity of propolis from Gerês (G18.EE) and its fractions (n-hexane, ethyl acetate (EtOAc), and n-butanol) in A375 and WM9 melanoma cell lines. Results from DPPH•/ABTS• radical scavenging assays indicated that the samples had relevant antioxidant activity, however, this was not confirmed in the cell models. G18.EE and its fractions decreased cell viability (SRB assay) and promoted ROS production (DHE/Mitotracker probes by flow cytometry), leading to activation of apoptotic signaling (expression of apoptosis markers). Our results suggest that the n-BuOH fraction has the potential to be explored in the pharmacological therapy of melanoma.

Potential of propolis antifungal activity for clinical applications.

The high incidence of skin diseases of microbial origin along with the widespread increase of microbial resistance demand for therapeutic alternatives. Research on natural compounds has been opening new perspectives for the development of new therapies with health-positive impacts. Propolis, a resinous mixture produced by honeybees from plant exudates, is widely used as a natural medicine since ancient times, mainly due to its antimicrobial properties. More recently, antioxidant, anti-tumour, anti-inflammatory, hepatoprotective and immunomodulatory activities were also reported for this natural product, highlighting its high potential pharmacological interest. In the present work, an extensive review of the main fungi causing skin diseases as well as the effects of natural compounds, particularly propolis, against such disease-causing micro-organisms was organized and compiled in concise handy tables. This information allows to conclude that propolis is a highly effective antimicrobial agent suggesting that it could be used as an alternative skin treatment against pathogenic micro-organisms and also as a cosmeceutical component or as a source of bioactive ingredients.

Propolis antibacterial and antioxidant synergisms with gentamicin and honey.

AIMS: Assess the antibacterial and antioxidant effects of ethanol extracts of Portuguese propolis samples when combined with gentamicin, a widely used aminoglycoside antibiotic, or with honey, collected from the same apiary as propolis. METHODS AND RESULTS: Using the agar dilution method and DPPH free radical scavenging assay, antimicrobial and antioxidant synergistic effects between propolis and gentamicin or honey were assessed. Synergism between propolis and gentamicin was observed for all the tested extracts and against all the indicator bacteria, with particular interest against the Methicillin-Resistant Staphylococcus aureus (MRSA) with a threefold decrease of the gentamicin MIC if mixed with 25 µg ml-1 propolis. Likely to propolis and gentamicin, mixtures of sub-MIC concentrations of propolis and honey enhanced the antibacterial action of each individual natural product against the majority of the strains. However, propolis antioxidant capacity decreased along with higher honey content in the mixture. CONCLUSIONS: Propolis has a strong synergistic effect when combined with gentamicin, allowing the reduction of the therapeutic dose of this drug. Propolis and honey mixtures also display a stronger antibacterial effect than the activity exhibited by each sample when tested individually, whereas the high antioxidant capacity of propolis seems to be not affected when in combination with honey. SIGNIFICANCE AND IMPACT OF STUDY: When mixed with honey, propolis antioxidant potential is maintained, or just slightly reduced up to 1:1 dilution, and show synergistic antibacterial effects, allowing to optimize the use of this usually scarce natural resource. Also, considering the antibiotic resistance problem, natural beehive products, alone or in the mixture, are promising alternatives to retard the outbreak of microbial resistance.

Acute vestibular syndrome and hearing loss mimicking labyrinthitis as initial presentation of multiple sclerosis.

Acute vestibular syndrome is most often caused by vestibular neuritis or stroke, although demyelinating diseases may be responsible for 4% of all AVS episodes. The authors present the case of a previously healthy 17-year-old female patient complaining of spontaneous vertigo and right-sided hearing loss. Otoneurological examination suggested a peripheral vestibular cause and video head impulse test revealed a reduced vestibulo-ocular reflex gain. The presence of sensorineural hearing loss raised the suspicion of a central cause and prompted imaging evaluation. A brain MRI evidenced demyelinating lesions in the right middle cerebellar peduncle and the patient was ultimately diagnosed with Multiple Sclerosis.