Widespread introduced species dominate the urban tree assemblage on the endemic-rich tropical island of São Tomé.

The Afrotropics are experiencing some of the fastest urbanisation rates on the planet but the impact of city growth on their rich and unique biodiversity remains understudied, especially compared to natural baselines. Little is also known about how introduced species influence ß-diversity in these contexts, and how patterns coincide with native ranges of species. Here we investigated how tree assemblages of the endemic-rich Afrotropical island of São Tomé differed between urban, rural and natural zones. These were primarily characterised by urban greenspaces, shade plantations, and old-growth forests, respectively. Based on 81 transects, we assessed biodiversity metrics of endemic, native and introduced species. Tree abundance and species richness were highest in the natural zone, where the composition was most different from the urban zone. The tree community of the rural zone was the most uneven and had the least variation among transects, representing the lowest ß-diversity. The urban zone was dominated by introduced species (57.7%), while the natural zone hosted almost exclusively native species (93.3%), including many endemics (26.1%). The biogeographic realms that species originated from were particularly diverse in the urban zone, with few species from the Afrotropics. In contrast to native and endemic trees, introduced trees were clearly associated with urban and rural expansion, as they were much more abundant and species-rich in these zones than in the natural zone, facilitating biotic homogenisation. These findings highlight how urban and rural environments are affecting the native tree flora of São Tomé, and the need for conservation measures geared towards globally threatened and endemic tree species. Importantly, these require the protection of natural forests, despite the rising land demands for settlements and agriculture. Ultimately, such action to conserve endemic trees will contribute to global efforts to prevent further biodiversity declines.

Design, synthesis, docking studies and bioactivity evaluation of 1,2,3-triazole eugenol derivatives.

Aim: The design, synthesis, docking studies and evaluation of the in vitro antifungal and cytotoxic properties of eugenol (EUG) containing 1,2,3-triazole derivatives are reported. Most of the derivatives have not been reported.Materials & methods: The EUG derivatives were synthesized, molecular docked and tested for their antifungal activity.Results: The compounds showed potent antifungal activity against Trichophyton rubrum, associated with dermatophytosis. Compounds 2a and 2i exhibited promising results, with 2a being four-times more potent than EUG. The binding mode prediction was similar to itraconazole in the lanosterol-14-α-demethylase wild-type and G73E mutant binding sites. Additionally, the pharmacokinetic profile prediction suggests good gastrointestinal absorption and potential oral administration.Conclusion: Compound 2a is a promising antifungal agent against dermatophytosis caused by T. rubrum.

[Box: see text].

Impact of educational intervention on understanding health recommendations after liver transplantation.

OBJECTIVES: to evaluate the impact of educational intervention on understanding health recommendations after liver transplantation. METHODS: randomized and prospective clinical trial, with 68 liver transplant recipients in two institutions. The level of understanding was assessed using a statement agreement scale and the understanding score was classified. Chi-square test was used to compare groups. RESULTS: the level of understanding was reasonable in 77.9% of patients, 73.5% in the Control Group and 82.3% in the Intervention Group (p=0.399). For topics covered after educational action, there were more than 80% correct answers regarding nutrition, frequent hydration, usage and function of immunosuppressants. However, there were less than 10% correct answers regarding hand hygiene, contact with animals and crowds of people. The use of the patient's audio, visual and tactile resources led to improved understanding of skin care (p=0.014). CONCLUSIONS: the level of understanding acquired regarding health recommendations was only reasonable.

Design and Synthesis of Eugenol Derivatives Bearing a 1,2,3-Triazole Moiety for Papaya Protection against Colletotrichum gloeosporioides.

A series of 19 novel eugenol derivatives containing a 1,2,3-triazole moiety was synthesized via a two-step process, with the key step being a copper(I)-catalyzed azide-alkyne cycloaddition reaction. The compounds were assessed for their antifungal activities against Colletotrichum gloeosporioides, the causative agent of papaya anthracnose. Triazoles 2k, 2m, 2l, and 2n, at 100 ppm, were the most effective, reducing mycelial growth by 88.3, 85.5, 82.4, and 81.4%, respectively. Molecular docking calculations allowed us to elucidate the binding mode of these derivatives in the catalytic pocket of C. gloeosporioides CYP51. The best-docked compounds bind closely to the heme cofactor and within the channel access of the lanosterol (LAN) substrate, with crucial interactions involving residues Tyr102, Ile355, Met485, and Phe486. From such studies, the antifungal activity is likely attributed to the prevention of substrate LAN entry by the 1,2,3-triazole derivatives. The triazoles derived from natural eugenol represent a novel lead in the search for environmentally safe agents for controlling C. gloeosporioides.

Microbial interactions among Gardnerella, Prevotella and Fannyhessea prior to incident bacterial vaginosis: protocol for a prospective, observational study.

INTRODUCTION: The aetiology of bacterial vaginosis (BV), a biofilm-associated vaginal infection, remains unknown. Epidemiologic data suggest that it is sexually transmitted. BV is characterised by loss of lactic acid-producing lactobacilli and an increase in facultative and strict anaerobic bacteria. Gardnerella spp are present in 95%-100% of cases; Gardnerella vaginalis has been found to be more virulent than other BV-associated bacteria (BVAB) in vitro. However, G. vaginalis is found in women with normal vaginal microbiota and colonisation is not sufficient for BV development. We hypothesise that Gardnerella spp initiate BV biofilm formation, but incident BV (iBV) requires incorporation of other key BVAB (ie, Prevotella bivia, Fannyhessea vaginae) into the biofilm that alter the transcriptome of the polymicrobial consortium. This study will investigate the sequence of microbiologic events preceding iBV. METHODS AND ANALYSIS: This study will enrol 150 women aged 18-45 years with normal vaginal microbiota and no sexually transmitted infections at a sexual health research clinic in Birmingham, Alabama. Women will self-collect twice daily vaginal specimens up to 60 days. A combination of 16S rRNA gene sequencing, qPCR for Gardnerella spp, P. bivia and F. vaginae, and broad range 16S rRNA gene qPCR will be performed on twice daily vaginal specimens from women with iBV (Nugent score 7-10 on at least 2 consecutive days) and controls (with comparable age, race, contraceptive method and menstrual cycle days) maintaining normal vaginal microbiota to investigate changes in the vaginal microbiota over time for women with iBV. Participants will complete daily diaries on multiple factors including sexual activity. ETHICS AND DISSEMINATION: This protocol is approved by the University of Alabama at Birmingham Institutional Review Board (IRB-300004547) and written informed consent will be obtained from all participants. Findings will be presented at scientific conferences and published in peer-reviewed journals as well as disseminated to providers and patients in communities of interest.

Optimized bacterial absolute quantification method by qPCR using an exogenous bacterial culture as a normalization strategy in triple-species BV-like biofilms.

Quantitative Polymerase Chain Reaction (qPCR) is a widely used method in molecular biology to quantify target DNA sequences. Despite its accuracy, there are important experimental controls that should be considered to avoid biased results. One of them is gDNA loss during extraction, which is higher among samples with lower bacterial concentrations. Improvement in qPCR quantification procedures is mandatory to obtain reproducible and accurate results. Herein, we report an improved qPCR method for bacterial quantification of Gardnerella vaginalis, Prevotella bivia, and Fannyhessea vaginae, three key-bacterial vaginosis (BV)-associated bacteria (BVAB) thought to play important roles in the pathogenesis of this common vaginal infection. The formation of a mature biofilm on vaginal epithelial cells is an unique feature of BV and, despite over 60 years of research, the exact etiology of BV remains unknown. Here, we optimized a qPCR method that accurately quantified triple-species biofilms containing these key BVAB, after the addition of an exogenous bacterial control containing a fixed concentration of Escherichia coli, prior to gDNA extraction. This improved method minimized and normalized the inherent losses associated with bacterial centrifugation, which allows better sensitivity at lower bacterial concentrations.

A pragmatic trial of a family-centered approach to childhood obesity treatment: Rationale and study design.

BACKGROUND: Family-based behavioral treatment (FBT) is an effective intensive health behavior and lifestyle treatment for obesity reduction in children and adolescents, but families have limited access. The purpose of this randomized, pragmatic, comparative effectiveness trial was to examine changes in child relative weight in a 12-month, enhanced standard of care (eSOC) intervention combined with FBT (eSOC+FBT) vs. eSOC alone. METHODS: Children aged 6 to 15 years with obesity, and their primary caregiver, were recruited from primary care clinics. Families were randomized 1:1 to eSOC, a staged approach led by the primary care provider that gradually intensified dependent on a child's response to care and aligns with the American Medical Association guidelines, or the eSOC+FBT arm, which included regular meetings with a health coach for healthy eating, physical activity, positive parenting strategies, and managing social and environmental cues. Both treatments align with the 2023 American Academy of Pediatrics clinical practice guidelines. Assessments occurred at baseline, midpoint (month 6), end-of-intervention (month 12), and follow-up (month 18). Primary outcome was change from baseline to 12 months in child percent overweight (percentage above the median body mass index in the general US population normalized for age and sex). Secondary outcomes were parent weight, child psychosocial factors, heterogeneity of treatment effects, and cardiometabolic risk factors. Exploratory outcomes assessed reach, effectiveness, adoption, implementation, and maintenance. CONCLUSION: This pragmatic trial will generate evidence for the comparative effectiveness of implementing two guidelines-based approaches in primary care for obesity reduction in children and adolescents. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03843424.

An Indirect Fluorescence Microscopy Method to Assess Vaginal Lactobacillus Concentrations.

Lactobacillus species are the main colonizers of the vaginal microbiota in healthy women. Their absolute quantification by culture-based methods is limited due to their fastidious growth. Flow cytometry can quantify the bacterial concentration of these bacteria but requires the acquisition of expensive equipment. More affordable non-culturable methods, such as fluorescence microscopy, are hampered by the small size of the bacteria. Herein, we developed an indirect fluorescence microscopy method to determine vaginal lactobacilli concentration by determining the correlation between surface area bacterial measurement and initial concentration of an easily cultivable bacterium (Escherichia coli) and applying it to lactobacilli fluorescence microscopy counts. In addition, vaginal lactobacilli were quantified by colony-forming units and flow cytometry in order to compare these results with the indirect method results. The colony-forming-unit values were lower than the results obtained from the other two techniques, while flow cytometry and fluorescence microscopy results agreed. Thus, our developed method was able to accurately quantify vaginal lactobacilli.

Validation of remote child weight and height measurements within a weight management trial.

OBJECTIVE: The aim of this substudy within the Treatment Efforts Addressing Child Weight Management by Unifying Patients, Parents, and Providers (TEAM UP) pragmatic clinical trial was to compare the validity of anthropometric measurements collected remotely versus in person (≤7 days apart) among youth with obesity who were 6 to 15 years of age. METHODS: Child (n = 37) weight and height were measured in person by a trained data assessor. These were compared with measurements taken remotely by the child's parent with live videoconferencing observation by a study data assessor. In-person and remote measurements were compared using Bland-Altman plots, Pearson correlations, and two one-sided paired t tests. A priori bounds of acceptability were set at ±0.68 kg to allow for typical weight fluctuations within the 7-day comparison period. RESULTS: Measurements were highly correlated (height: r = 0.991, p < 0.0001; weight: r = 0.999; p = 0.03). For height, two one-sided t tests for upper, t(36) = 3.95, and lower, t(36) = -2.63, bounds (-1, 1) revealed an overall p = 0.006; absolute error was 3.5 cm. For weight, two one-sided t tests for upper, t(36) = 1.93, and lower, t(36) = -7.91, bounds (-0.68, 0.68) revealed an overall p = 0.03; absolute error was 1.7 kg. CONCLUSIONS: The present findings support the utility and interpretation of remotely assessed weight management outcomes for both research and clinical purposes. These procedures may offer greater accessibility to evidence-based measurement.

Impact of educational intervention on understanding health recommendations after liver transplantation / El impacto de una intervención educativa sobre la comprensión de las recomendaciones sanitarias stras un trasplante de hígado / Impacto de intervenção educativa na compreensão sobre recomendações para saúde após transplante de fígado

ABSTRACT Objectives: to evaluate the impact of educational intervention on understanding health recommendations after liver transplantation. Methods: randomized and prospective clinical trial, with 68 liver transplant recipients in two institutions. The level of understanding was assessed using a statement agreement scale and the understanding score was classified. Chi-square test was used to compare groups. Results: the level of understanding was reasonable in 77.9% of patients, 73.5% in the Control Group and 82.3% in the Intervention Group (p=0.399). For topics covered after educational action, there were more than 80% correct answers regarding nutrition, frequent hydration, usage and function of immunosuppressants. However, there were less than 10% correct answers regarding hand hygiene, contact with animals and crowds of people. The use of the patient's audio, visual and tactile resources led to improved understanding of skin care (p=0.014). Conclusions: the level of understanding acquired regarding health recommendations was only reasonable.

RESUMEN Objetivos: evaluar el impacto de una intervención educativa sobre la comprensión de las recomendaciones sanitarias tras un trasplante de hígado. Métodos: se trata de un ensayo clínico prospectivo aleatorizado con 68 pacientes trasplantados de hígado de dos instituciones. Se evaluó el nivel de conocimiento mediante una escala de acuerdo con las afirmaciones y se clasificó la puntuación de la comprensión. Se utilizó la prueba de chi-cuadrado para comparar los grupos. Resultados: el nivel de comprensión fue razonable en el 77,9% de los pacientes, con el 73,5% del Grupo de Control y el 82,3% del Grupo de Intervención (p=0,399). En cuanto a los temas tratados tras la actividad educativa, más del 80% tenía conocimiento sobre alimentación, hidratación frecuente, uso y función de los inmunosupresores. Sin embargo, hubo menos de un 10% de respuestas correctas sobre la higiene de las manos, el contacto con animales y las aglomeraciones de personas. La utilización de los recursos auditivos, visuales y táctiles del paciente mejoró la comprensión del cuidado de la piel (p=0,014). Conclusiones: el nivel de comprensión adquirido sobre las recomendaciones sanitarias fue solo razonable.

RESUMO Objetivos: avaliar o impacto de intervenção educativa na compreensão sobre recomendações para saúde após transplante de fígado. Métodos: ensaio clínico randomizado prospectivo, com 68 transplantados de fígado em duas instituições. Avaliou-se o nível de compreensão por meio de escala de concordância de afirmativas e classificou-se o escore de compreensão. Utilizou-se teste qui-quadrado para comparação dos grupos. Resultados: o nível de compreensão foi razoável em 77,9% dos pacientes, sendo 73,5% no Grupo Controle e 82,3% no Grupo Intervenção (p=0,399). Para temas abordados após ação educativa, houve mais de 80% de acertos sobre alimentação, hidratação frequente, uso e função do imunossupressor. Entretanto, houve menos de 10% de acertos sobre higienização das mãos, contato com animais e aglomerações de pessoas. A utilização dos recursos áudio, visual e tátil do paciente ocasionou melhora na compreensão sobre cuidado com a pele (p=0,014). Conclusões: o nível de compreensão adquirido às recomendações para saúde foi apenas razoável.

A new cyclodipeptide from Tetragonisca angustula honey active against Neospora caninum and in silico study.

A new cyclic natural compound formed by succinic acid and two alanine amino acid units was isolated from the Tetragonisca angustula honey extract. The chemical structure of 1 was established based on spectroscopic data analysis, including one- (1H and 13C NMR) and two-dimensional NMR techniques (1H-1H-COSY, HSQC and HMBC). A primary culture model previously infected with Neospora caninum was used to evaluate 1 for two time intervals (24 and 72 h), showing a reduction (40-56%) of the number of tachyzoites in the first 24 h and until 72 h, a dose-dependent reduction in parasite proliferation (25-50%). Glial cells treated with 1 did not demonstrate toxicity at concentrations up to 25 ug/mL. Treated and infected cultures showed an increase in NO when compared to control cells in 24 h and 72 h. In silico studies suggest that the new compound may affect DNA synthesis and impair -protein production.

Accurate Absolute Quantification of Bacterial Populations in Mixed Cultures by qPCR.

Quantitative PCR (qPCR) is a well-established technique that allows to accurately quantify nucleic acids or proteins, being widely used in several types of biological samples for bacterial load quantification. However, there are many recent studies that do not consider the potential pitfalls involved in key experimental qPCR stages, namely, those related to the extraction and purification of genomic DNA and to the thermal amplification process, that can lead to biased results in mixed cultures. Herein, we outline a proper protocol for bacterial quantification by qPCR, addressing how to overcome the main issues in that methodology.

Synthesis and Fungicide Activity on Asperisporium caricae of Glycerol Derivatives Bearing 1,2,3-Triazole Fragments.

In agriculture, the control of fungal infections is essential to improve crop quality and productivity. This study describes the preparation and fungicidal activity evaluation of 12 glycerol derivatives bearing 1,2,3-triazole fragments. The derivatives were prepared from glycerol in four steps. The key step corresponded to the Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC) click reaction between the azide 4-(azidomethyl)-2,2-dimethyl-1,3-dioxolane (3) and different terminal alkynes (57-91% yield). The compounds were characterized by infrared spectroscopy, nuclear magnetic resonance (1H and 13C), and high-resolution mass spectrometry. The in vitro assessment of the compounds on Asperisporium caricae, that is, the etiological agent of papaya black spot, at 750 mg L-1 showed that the glycerol derivatives significantly inhibited conidial germination with different degrees of efficacy. The most active compound 4-(3-chlorophenyl)-1-((2,2-dimethyl-1,3-dioxolan-4-yl) methyl)-1H-1,2,3-triazole (4c) presented a 91.92% inhibition. In vivo assays revealed that 4c reduced the final severity (70.7%) and area under the disease severity progress curve of black spots on papaya fruits 10 days after inoculation. The glycerol-bearing 1,2,3-triazole derivatives also present agrochemical-likeness properties. Our in silico study using molecular docking calculations show that all triazole derivatives bind favorably to the sterol 14α-demethylase (CYP51) active site at the same region of the substrate lanosterol (LAN) and fungicide propiconazole (PRO). Thus, the mechanism of action of the compounds 4a-4l may be the same as the fungicide PRO, blocking the entrance/approximation of the LAN into the CYP51 active site by steric effects. The reported results point to the fact that the glycerol derivatives may represent a scaffold to be explored for the development of new chemical agents to control papaya black spot.

Tecnologias da informação e comunicação utilizadas por enfermeiros da atenção primária na pandemia de covid-19 / Tecnologías de la información y comunicación utilizadas por enfermeros de atención primaria en la pandemia de covid-19 / Information and communication technologies used by primary care nurses in the covid-19 pandemic

RESUMO Objetivo: identificar as Tecnologias da Informação e Comunicação utilizadas por enfermeiros da atenção primária à saúde durante o processo de trabalho frente à Covid-19. Método: estudo exploratório e qualitativo, por meio de entrevistas semiestruturadas, com 10 enfermeiros da Atenção Primária à Saúde, coletado em fevereiro de 2022. Utilizou-se a análise de conteúdo de Bardin. Resultados: evidenciou-se a inclusão das tecnologias como uma ferramenta de complementação e extensão para desenvolver do processo de trabalho da enfermagem, nesse período. A Tecnologia mais utilizada foi o WhatsApp, com a finalidade de realizar monitoramento, orientações e agendamentos. Discussão: a tecnologia permite ao enfermeiro desenvolver o processo de trabalho, favorecendo ao cuidado em saúde e à comunicação, entre os profissionais e desses com os usuários. Entretanto, há entraves como dificuldade de acesso à internet de qualidade. Considerações finais: as tecnologias da informação se constituem como possibilidades ao enfermeiro para desenvolver a sua prática. Apesar das dificuldades de acesso à internet, a utilização das tecnologias durante a pandemia esteve presente na rotina de todos os enfermeiros, sendo utilizadas para qualificar a comunicação e o acesso às informações necessárias à população.

RESUMEN Objetivo: identificar las Tecnologías de la Información y Comunicación utilizadas por enfermeros de la atención primaria de salud durante el proceso de trabajo frente a Covid-19. Método: estudio exploratorio y cualitativo, por medio de entrevistas semiestructuradas, con 10 enfermeros de la Atención Primaria de Salud, recolectado en febrero de 2022. Se utilizó el análisis de contenido de Bardin. Resultados: se evidenció la inclusión de las tecnologías como una herramienta de complementación y extensión para desarrollar el proceso de trabajo de enfermería, en ese período. La Tecnología más utilizada fue WhatsApp, con la finalidad de realizar monitoreo, orientaciones y planificaciones. Discusión: la tecnología permite al enfermero desarrollar el proceso de trabajo, favoreciendo el cuidado en salud y la comunicación, entre los profesionales y de estos con los usuarios. No obstante, existen obstáculos como la dificultad de acceso a Internet de calidad. Consideraciones finales: las tecnologías de la información se constituyen como posibilidades al enfermero para desarrollar su práctica. A pesar de las dificultades de acceso a Internet, la utilización de las tecnologías durante la pandemia estuvo presente en la rutina de todos los enfermeros y se utilizó para cualificar la comunicación y el acceso a las informaciones necesarias a la población.

ABSTRACT Objective: to identify the Information and Communication Technologies used by primary health care nurses during the work process against Covid-19. Method: exploratory and qualitative study, through semi-structured interviews, with 10 nurses from Primary Health Care, collected in February 2022. Bardin content analysis was used. Results: the inclusion of technologies as a tool of complementation and extension to develop the nursing work process in this period was evidenced. The most used technology was WhatsApp, for the purpose of monitoring, guidance and scheduling. Discussion: technology allows nurses to develop the work process, favoring health care and communication between professionals and users. However, there are obstacles such as difficulty in accessing quality internet. Final thoughts: information technologies constitute as possibilities for nurses to develop their practice. Despite the difficulties in accessing the internet, the use of technologies during the pandemic was present in the routine of all nurses, being used to qualify communication and access to the information needed by the population.

Assessing recovery rates of distinct exogenous controls for gDNA extraction efficiency using phenol-chloroform or silica-column based extractions.

Assessment of genomic DNA (gDNA) extraction efficiency is required for accurate bacterial quantification by qPCR. Exogenous DNA molecules are often added after bacterial cultures are lysed, but before DNA purification steps, to determine extraction efficiency. Herein we found that different exogenous DNA controls have different recovery rates, suggesting distinct DNA extraction efficiencies. Recovery rates are also affected by the gDNA extraction method being more affected in silica-based columns than in phenol-chloroform extraction. Overall, we determined that the use of long DNA fragments, such as gDNA, as exogenous controls have a higher recovery rate than use of smaller size DNA molecules.

DNA extraction leads to bias in bacterial quantification by qPCR.

Quantitative PCR (qPCR) has become a widely used technique for bacterial quantification. The affordability, ease of experimental design, reproducibility, and robustness of qPCR experiments contribute to its success. The establishment of guidelines for minimum information for publication of qPCR experiments, now more than 10 years ago, aimed to mitigate the publication of contradictory data. Unfortunately, there are still a significant number of recent research articles that do not consider the main pitfalls of qPCR for quantification of biological samples, which undoubtedly leads to biased experimental conclusions. qPCR experiments have two main issues that need to be properly tackled: those related to the extraction and purification of genomic DNA and those related to the thermal amplification process. This mini-review provides an updated literature survey that critically analyzes the following key aspects of bacterial quantification by qPCR: (i) the normalization of qPCR results by using exogenous controls, (ii) the construction of adequate calibration curves, and (iii) the determination of qPCR reaction efficiency. It is primarily focused on original papers published last year, where qPCR was applied to quantify bacterial species in different types of biological samples, including multi-species biofilms, human fluids, and water and soil samples. KEY POINTS: • qPCR is a widely used technique used for absolute bacterial quantification. • Recently published papers lack proper qPCR methodologies. • Not including proper qPCR controls significantly affect experimental conclusions.

Synthesis of cinnamic acid ester derivatives with antiproliferative and antimetastatic activities on murine melanoma cells.

Melanoma is the most aggressive skin cancer, and its incidence has continued to rise during the past decades. Conventional treatments present severe side effects in cancer patients, and melanoma can be refractory to commonly used anticancer drugs, which justify the efforts to find new potential anti-melanoma drugs. An alternative to promote the discovery of new pharmacological substances would be modifying chemical groups from a bioactive compound. Here we describe the synthesis of seventeen compounds derived from cinnamic acid and their bioactivity evaluation against melanoma cells. The compound phenyl 2,3-dibromo-3-phenylpropanoate (3q) was the most effective against murine B16-F10 cells, as observed in cytotoxicity and cell migration assays. Simultaneously, this compound showed low cytotoxic activity on non-tumor cells. At the highest concentration, the compound 3q was able to trigger apoptosis, whereas, at lower concentrations, it affected the cell cycle and melanoma cell proliferation. Furthermore, cinnamate 3q impaired cell invasion, adhesion, colonization, and actin polymerization. In conclusion, these results highlight the antiproliferative and antimetastatic potential of cinnamic acid derivatives on melanoma.

Accurate qPCR quantification in polymicrobial communities requires assessment of gDNA extraction efficiency.

qPCR absolute quantification requires the assessment of PCR reaction efficiency. This is achieved by performing serial dilutions of gDNA. Herein, we demonstrate that when quantifying bacterial load in mixed samples, reaction efficiency should not be used as a calibration curve since gDNA isolation efficiency is neglected, significantly impacting quantification.

Headspace volatolome of peel flours from citrus fruits grown in Brazil.

Citrus fruit peel comprises a pleasant mix of volatile compounds together with fibers, nutrients, and bioactive compounds. Therefore, it has great potential for use as a food ingredient. Studies evaluating the volatile composition of citrus peel flours are limited for most citruses. The goal of this study was to characterize, by HS-SPME/GC-MS, the volatile profile of citrus peel flours made from fruits commonly grown in Brazil. Two composite samples of ten types of citrus peel flours from consecutive harvests were evaluated. 69 volatile compounds were assigned, 49 in Tahiti acid lime, 49 in Sicilian lemon, 37 in Persian lime, 34 in Italian tangerine and oval kumquat, 33 in Valencia orange, 32 in Baia orange and round kumquat, 28 in Blood-of-Mombuca orange and 26 in Lima orange. 26 major compounds represented 93-99% of the total chromatogram peak area. Terpenic compounds were predominant in all samples, especially monoterpenes (about 48-97% of the total chromatogram peak area), while lower proportions of aldehydes (0.2-16.1%), monoterpene alcohols (0.4-11.8%) and esters (0.0-7.7%) were observed. Even though a few compounds like limonene, ß-myrcene, linalool, α-pinene and valencene were detected in all citrus, volatile compounds followed specific patterns in the different citruses, with a clear distinction among them, especially between lemon flours and the remaining flours. The variety of volatile profiles and singular specific volatolomic signatures in citrus peels can be explored for different applications related to food flavoring and preservation, and promotion of good health. These aspects should be thoroughly investigated in future studies.

Hydroxycinnamic acid-spermidine amides from Tetragonisca angustula honey as anti-Neospora caninum: In vitro and in silico studies.

Tetragonisca angustula honey was fractioned in a SiO2 column to furnish three fractions (A-C) in which four hydroxycinnamic acid-Spermidine amides (HCAAs), known as N', N″, N‴-tris-p-coumaroyl spermidine, N', N″-dicaffeoyl, N‴-coumaroyl spermidine, N', N″, N‴-tris-caffeoyl spermidine and N', N″-dicaffeoyl and N‴-feruloyl spermidine were identified in the fractions B and C by electrospray ionization tandem mass spectrometry. A primary culture model previously infected with Neospora caninum (72 h) was used to evaluate the honey fractions (A-C) for two-time intervals: 24 and 72 h. Parasitic reduction ranged from 38% on fraction C (12.5 µg/ml), after 24 h, to 54% and 41% with fractions B and C (25 µg/ml) after 72 h of treatment, respectively. Additionally, HCAAs did not show any cell toxicity for 24 and 72 h. For infected cultures (72 h), the active fractions B (12.5 µg/ml) and C (25 µg/ml) decreased their NO content. In silico studies suggest that HCAAs may affect the parasite's redox pathway and improve the oxidative effect of NO released from infected cells. Here, we presented for the first time, that HCAAs from T. angustula honey have the potential to inhibit the growth of N. caninum protozoa.