Harvesting the Power of Green Synthesis: Gold Nanoparticles Tailored for Prostate Cancer Therapy.

Biosynthetic gold nanoparticles (bAuNPs) present a promising avenue for enhancing bio-compatibility and offering an economically and environmentally responsible alternative to traditional production methods, achieved through a reduction in the use of hazardous chemicals. While the potential of bAuNPs as anticancer agents has been explored, there is a limited body of research focusing on the crucial physicochemical conditions influencing bAuNP production. In this study, we aim to identify the optimal growth phase of Pseudomonas aeruginosa cultures that maximizes the redox potential and coordinates the formation of bAuNPs with increased efficiency. The investigation employs 2,6-dichlorophenolindophenol (DCIP) as a redox indicator. Simultaneously, we explore the impact of temperature, pH, and incubation duration on the biosynthesis of bAuNPs, with a specific emphasis on their potential application as antitumor agents. Characterization of the resulting bAuNPs is conducted using ATR-FT-IR, TEM, and UV-Vis spectroscopy. To gain insights into the anticancer potential of bAuNPs, an experimental model is employed, utilizing both non-neoplastic (HPEpiC) and neoplastic (PC3) epithelial cell lines. Notably, P. aeruginosa cultures at 9 h/OD600 = 1, combined with biosynthesis at pH 9.0 for 24 h at 58 °C, produce bAuNPs that exhibit smaller, more spherical, and less aggregated characteristics. Crucially, these nanoparticles demonstrate negligible effects on HPEpiC cells while significantly impacting PC3 cells, resulting in reduced viability, migration, and lower IL-6 levels. This research lays the groundwork for the development of more specialized, economical, and ecologically friendly treatment modalities.

Platinum-Triggered Bond-Cleavage of Pentynoyl Amide and N-Propargyl Handles for Drug-Activation.

The ability to create ways to control drug activation at specific tissues while sparing healthy tissues remains a major challenge. The administration of exogenous target-specific triggers offers the potential for traceless release of active drugs on tumor sites from antibody-drug conjugates (ADCs) and caged prodrugs. We have developed a metal-mediated bond-cleavage reaction that uses platinum complexes [K2PtCl4 or Cisplatin (CisPt)] for drug activation. Key to the success of the reaction is a water-promoted activation process that triggers the reactivity of the platinum complexes. Under these conditions, the decaging of pentynoyl tertiary amides and N-propargyls occurs rapidly in aqueous systems. In cells, the protected analogues of cytotoxic drugs 5-fluorouracil (5-FU) and monomethyl auristatin E (MMAE) are partially activated by nontoxic amounts of platinum salts. Additionally, a noninternalizing ADC built with a pentynoyl traceless linker that features a tertiary amide protected MMAE was also decaged in the presence of platinum salts for extracellular drug release in cancer cells. Finally, CisPt-mediated prodrug activation of a propargyl derivative of 5-FU was shown in a colorectal zebrafish xenograft model that led to significant reductions in tumor size. Overall, our results reveal a new metal-based cleavable reaction that expands the application of platinum complexes beyond those in catalysis and cancer therapy.

Piper Essential Oils Inhibit Rhizopus oryzae Growth, Biofilm Formation, and Rhizopuspepsin Activity.

Piper is the largest genus of the Piperaceae family. The species of this genus have diverse biological activities and are used in pharmacopeia throughout the world. They are also used in folk medicine for treatment of many diseases in several countries including Brazil, China, India, Jamaica, and Mexico. In Brazil, Piper species are distributed throughout the national territory, making this genus a good candidate for biological activity screening. During our studies with Piper essential oils, we evaluated its activity against Rhizopus oryzae, the main agent of mucormycosis. The main compounds of seven Piper essential oils analyzed were Piper callosum-safrole (53.8%), P. aduncum-dillapiole (76.0%), P. hispidinervum-safrole (91.4%), P. marginatum-propiopiperone (13.2%), P. hispidum-γ-terpinene (30.9%), P. tuberculatum-(E)-caryophyllene (30.1%), and Piper sp.-linalool (14.6%). The minimum inhibitory concentration of Piper essential oils against R. oryzae ranged from 78.12 to >1250 µg/mL. The best result of total inhibition of biofilm formation was obtained with Piper sp. starting from 4.88 µg/mL. Considering the bioactive potential of EOs against planktonic cells and biofilm formation of R. oryzae could be of great interest for development of antimicrobials for therapeutic use in treatment of fungal infection.

Effects of 7-hydroxycalamenene isolated from Croton cajucara essential oil on growth, lipid content and ultrastructural aspects of Rhizopus oryzae.

The leaves and bark of Croton cajucara, a shrub from the Amazon region, have been used in folk medicine to treat diabetes, malaria, and gastrointestinal and liver disorders. The essential oil from the leaves, rich in linalool, presented antileishmanial and antimicrobial activities. A chemotype of this species was found with an essential oil rich in 7-hydroxycalamenene. During our studies of the C. cajucara essential oil, we isolated 7-hydroxycalamenene at > 98 % purity. The minimum inhibitory concentration of 7-hydroxycalamenene against Absidia cylindrospora, Cunninghamella elegans, Mucor circinelloides, Mucor circinelloides f. circinelloides, Mucor mucedo, Mucor plumbeus, Mucor ramosissimus, Rhizopus microsporus, Rhizopus oryzae, and Syncephalastrum racemosum ranged from 19.53 to 2500 µg/mL. The reference drug used, amphotericin B, presented a minimum inhibitory concentration ranging from 0.085 µg/mL to 43.87 µg/mL. 7-Hydroxycalamenene also altered spore differentiation and total lipid content. Ultrastructural analysis by transmission electron microscopy showed significant alterations in the cellular structure of R. oryzae.

Inhibitory effect of linalool-rich essential oil from Lippia alba on the peptidase and keratinase activities of dermatophytes.

Abstract Lippia alba (Miller) N.E. Brown is an aromatic plant known locally as "Erva-cidreira-do-campo" that has great importance in Brazilian folk medicine. The aim of our study was to evaluate the antidermatophytic potential of linalool-rich essential oil (EO) from L. alba and analyze the ability of this EO to inhibit peptidase and keratinase activities, which are important virulence factors in dermatophytes. The minimum inhibitory concentrations (MICs) of L. alba EO were 39, 156 and 312 µg/mL against Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum, respectively. To evaluate the influence of L. alba EO on the proteolytic and keratinolytic activities of these dermatophytes, specific inhibitory assays were performed. The results indicated that linalool-rich EO from L. alba inhibited the activity of proteases and keratinases secreted from dermatophytes, and this inhibition could be a possible mechanism of action against dermatophytes. Due to the effective antidermatophytic activity of L. alba EO, further experiments should be performed to explore the potential of this linalool-rich EO as an alternative antifungal therapy.

[A Cognitive Behavioural-Based Intervention in Reducing Test Anxiety in High School Students: A Pilot Cluster Randomized Controlled Trial]. / Intervenção de Base Cognitivo-Comportamental na Redução da Ansiedade Perante os Testes em Alunos do Ensino Secundário: Estudo Piloto Aleatorizado e Controlado Por Clusters.

INTRODUCTION: School-age test anxiety is an important risk factor for school performance. Notwithstanding, few studies seek to identify which strategies are effective in improving test anxiety. The aim of this study was to assess whether a cognitive-behavioural intervention for high school students could significantly reduce test anxiety. MATERIAL AND METHODS: Two-arm, cluster-randomized controlled, unblinded, parallel, trial. Participants were 10th grade students from Alves Martins High School in Viseu, Portugal. Students were randomized at class level to receive a cognitive-behavioural-based intervention combined with mindfulness, psychoeducation, and relaxation techniques, or to a control group with no intervention. Participants' anxiety levels were measured using the Test Anxiety Questionnaire. The analysis of the effect of the intervention was carried out on an intention-to-treat basis at the class level, using multilevel mixed effects models and Bayesian modelling. RESULTS: The intervention had a significant effect in reducing test anxiety (d = 0.81, 95% CI 0.45;1.17, Bayes factor = 31.3). Male gender was an independent risk factor for smaller reductions in anxiety levels. The intervention was more effective in reducing the worry component of test anxiety (d = 0.76, 95% CI 0.41;1.11, Bayes factor = 19.9) than the emotionality component (d = 0.63, 95% CI 0.31;0.95, Bayes factor = 6.6). CONCLUSION: A cognitive-behavioural intervention specifically designed to reduce test anxiety, using a combination of mindfulness, psychoeducation and relaxation techniques, was effective in reducing test anxiety levels. TRIAL REGISTRATION: Retrospectively registered on clinicaltrials.gov (NCT05481099) in 08/01/2022.

Introdução: A ansiedade face aos testes é um importante fator condicionante da performance escolar. Contudo, são escassos os estudos que procuram identificar quais as estratégias eficazes na sua melhoria. Este estudo teve como objetivo testar a eficácia de uma intervenção cognitivo-comportamental na redução da ansiedade face aos testes em alunos do ensino secundário. Material e Métodos: Estudo experimental, aleatorizado por clusters (turmas), controlado, sem ocultação, com dois grupos paralelos, com alunos do 10.º ano da Escola Secundária Alves Martins em Viseu, Portugal. Os alunos foram aleatorizados ao nível da turma para receber uma combinação de técnicas cognitivas e comportamentais, de mindfulness, psicoeducação e técnicas de relaxamento, ou para um grupo controlo sem intervenção. Os níveis de ansiedade dos participantes foram medidos através do Questionário de Ansiedade face aos Testes. A análise do efeito da intervenção foi realizada na base de intenção de tratar ao nível da turma recorrendo a modelos de efeitos mistos multinível e modelação bayesiana. Resultados: A intervenção teve um efeito significativo na redução da ansiedade face aos testes (d = 0,81, IC 95% 0,45;1,17, fator de Bayes = 31,3). Pertencer ao sexo masculino revelou-se um fator de risco independente para uma menor redução nos níveis de ansiedade. O efeito da intervenção foi mais pronunciado na redução da preocupação face aos testes (d = 0,76, IC 95% 0,41;1,11, fator de Bayes = 19,9) quando comparado com a emocionalidade (d = 0,63, IC 95% 0,31;0,95, fator de Bayes = 6,6). Conclusão: Uma intervenção especificamente desenhada para reduzir a ansiedade face aos testes, usando uma combinação de técnicas cognitivas e comportamentais de mindfulness, psicoeducação e técnicas de relaxamento, foi eficaz na redução dos níveis de ansiedade. Registo do Estudo: Estudo registado a posteriori (registo retrospetivo) em ClinicalTrials.gov com o número NCT05481099 em 01/08/2022.

Macrophages directly kill bladder cancer cells through TNF signaling as an early response to BCG therapy.

The Bacillus Calmette-Guérin (BCG) vaccine is the oldest cancer immunotherapeutic agent in use. Despite its effectiveness, its initial mechanisms of action remain largely unknown. Here, we elucidate the earliest cellular mechanisms involved in BCG-induced tumor clearance. We developed a fast preclinical in vivo assay to visualize in real time and at single-cell resolution the initial interactions among bladder cancer cells, BCG and innate immunity using the zebrafish xenograft model. We show that BCG induced the recruitment and polarization of macrophages towards a pro-inflammatory phenotype, accompanied by induction of the inflammatory cytokines tnfa, il1b and il6 in the tumor microenvironment. Macrophages directly induced apoptosis of human cancer cells through zebrafish TNF signaling. Macrophages were crucial for this response as their depletion completely abrogated the BCG-induced phenotype. Contrary to the general concept that macrophage anti-tumoral activities mostly rely on stimulating an effective adaptive response, we demonstrate that macrophages alone can induce tumor apoptosis and clearance. Thus, our results revealed an additional step to the BCG-induced tumor immunity model, while providing proof-of-concept experiments demonstrating the potential of this unique model to test innate immunomodulators.

Sialoglycoproteins in morphological distinct stages of Mucor polymorphosporus and their influence on phagocytosis by human blood phagocytes.

The possible role of sialic acids in host cells-fungi interaction and their association with glycoproteins were evaluated using a clinical isolate of the dimorphic fungus Mucor polymorphosporus. Lectin-binding assays with spores and yeast cells denoted the presence of surface sialoglycoconjugates containing 2,3- and 2,6-linked sialylglycosyl groups. Western blotting with peroxidase-labeled Limulus polyphemus agglutinin revealed the occurrence of different sialoglycoprotein types in both cell lysates and cell wall protein extracts of mycelia, spores, and yeasts of M. polymorphosporus. Sialic acids contributed to the surface negative charge of spores and yeast forms as evaluated by adherence to a cationic substrate. Sialidase-treated spores were less resistant to phagocytosis by human neutrophils and monocytes from healthy individuals than control (untreated) fungal suspensions. The results suggest that sialic acids are terminal units of various glycoproteins of M. polymorphosporus, contributing to negative charge of yeasts and spore cells and protecting infectious propagules from destruction by host cells.

Antioxidant and antimicrobial activities of 7-hydroxy-calamenene-rich essential oils from Croton cajucara Benth.

Croton cajucara is a shrub native to the Amazon region locally known as "sacaca". Two morphotypes are known: white and red "sacaca". The essential oils (EO) obtained by hydrodistillation from leaves of the red morphotype were, in general, rich in 7-hydroxycalamenene (28.4%-37.5%). The effectiveness of these EO regarding the antimicrobial activity against pathogenic microorganisms was initially investigated by the drop test method, showing significant inhibition zones. Among the microorganisms tested, the essential oils rich in 7-hydroxycalamenene were more effective against methicillin resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, Mycobacterium tuberculosis, M. smegmatis, Mucor circinelloides and Rhizopus oryzae. The minimum inhibitory concentrations (MIC) of the oils were determined using the broth dilution assay. It was possible to observe that 7-hydroxycalamenene-rich oils presented high antimicrobial activity, with MIC of 4.76 × 10⁻³ µg/mL for MRSA, 4.88 µg/mL for M. tuberculosis, 39.06 µg/mL for M. smegmatis, and 0.152 µg/mL for R. oryzae and 3.63 × 10⁻8 µg/mL for M. circinelloides. The antioxidant activity of this EO suggests that 7-hydroxycalamenene provides more antioxidant activity according with EC(50) less than 63.59 µg/mL. Considering the bioactive potential of EOs and 7-hydroxycalamenene could be of great interest for development of antimicrobials for therapeutic use in treatment of bacterial and fungal infections in humans and/or veterinary practice.

Time-series analyses of land surface temperature changes with Google Earth Engine in a mountainous region.

Studying changes in temperature is fundamental for understanding its interactions with the environment and biodiversity. However, studies in mountainous areas are few, due to their complex formation and the difficulty of obtaining local data. We analysed changes in temperature over time in Montesinho Natural Park (MNP) (Bragança, Portugal), an important conservation area due to its high level of biodiversity. Specifically, we aimed to analyse: i) whether temperature increased in MNP over time, ii) what environmental factors influence the Land Surface Temperature (LST), and iii) whether vegetation is related to changes in temperature. We used annual summer and winter mean data acquired from the Moderate-Resolution Imaging Spectroradiometer (MODIS) datasets/products (e.g. LST, gathered at four different times: 11am, 1pm, 10pm and 2am, Enhance vegetation index - EVI, and Evapotranspiration - ET), available on the cloud-based platform Google Earth Engine between 2003 and 2021). We analysed the dynamics of the temporal trend patterns between the LST and local thermal data (from a weather station) by correlations; the trends in LST over time with the Mann-Kendall trend test; and the stability of hot spots and cold spots of LST with Local Statistics of Spatial Association (LISA) tests. The temporal trend patterns between LST and Air Temperature (Tair) data were very similar (ρ > 0.7). The temperature in the MNP remained stable over time during summer but increased during winter nights. The biophysical indices were strongly correlated with the summer LST at 11am and 1pm. The LISA results identified hot and cold zones that remained stable over time. The remote-sensed data proved to be efficient in measuring changes in temperature over time.