Redox proteomics in melanoma cells: An optimized protocol.

Cancer development and progression are intimately related with post-translational protein modifications, e.g., highly reactive thiol moiety of cysteines enables structural rearrangements resulting in redox biological switches. In this context, redox proteomics techniques, such as 2D redox DIGE, biotin switch assay and OxIcat are fundamental tools to identify and quantify redox-sensitive proteins and to understand redox mechanisms behind thiol modifications. Given the great variability in redox proteomics protocols, problems including decreased resolution of peptides and low protein amounts even after enrichment steps may occur. Considering the biological importance of thiol's oxidation in melanoma, we adapted the biotin-switch assay technique for melanoma cells in order to overcome the limitations and improve coverage of detected proteins.

Trastuzumab deruxtecan in human epidermal growth factor receptor 2-positive breast cancer brain metastases: a systematic review and meta-analysis.

BACKGROUND: Trastuzumab deruxtecan (T-DXd) has shown promising results in patients with breast cancer brain metastases (BCBMs). We conducted a systematic review and meta-analysis to evaluate the effectiveness and safety of T-DXd in the human epidermal growth factor receptor 2 (HER2)-positive BCBM population. PATIENTS AND METHODS: We searched PubMed, Embase, and Cochrane Library databases as well as American Society of Clinical Oncology (ASCO), European Society for Medical Oncology (ESMO), and San Antonio Breast Cancer Symposium (SABCS) websites for clinical trials (CTs) and observational studies evaluating T-DXd in patients with HER2-positive BCBM. Heterogeneity was assessed with I2 statistics. Random effects models were used for all statistical analyses, which were carried out using R software (version 4.2.2). RESULTS: Ten studies were included, six CTs (n = 189) and four observational studies (n = 130), with a total of 319 patients. The median progression-free survival was 15 months [95% confidence interval (CI) 13.9-16.1 months]. The objective response rate (ORR) was 61% (95% CI 52% to 70%), and the intracranial (IC)-ORR was 61% (95% CI 54% to 69%). No significant differences in ORR and IC-ORR were observed between CTs and observational studies (P = 0.31 and 0.58, respectively). The clinical benefit rate (CBR) was 80% (95% CI 52% to 94%), and the IC-CBR was 70% (95% CI 54% to 82%). The ORR was 68% (95% CI 57% to 77%) in the subgroup of patients with stable BMs and 60% (95% CI 48%-72%) in patients with active BM, with no significant difference between groups (P = 0.35). CONCLUSIONS: Our systematic review and meta-analysis supports the IC activity of T-DXd in patients with stable BM and active BM. TRIAL REGISTRATION: International Prospective Register of Systematic Reviews (PROSPERO) under the protocol number CRD42023422589.

Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model

Abstract This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Resumo Este estudo avaliou o efeito do óleo volátil de Alpinia zerumbet (OVAz) na expressão do gene da caveolina-1 e na fibrose muscular. Os ratos foram imobilizados para induzir a fibrose do músculo gastrocnêmio, e foram tratados com OVAz. A qualidade do colágeno foi avaliada com histologia e à expressão do gene caveolina-1 (CAV-1) foi avaliada usando qPCR. A análise histomorfológica indicou uma redução significativa no perímetro, largura e intensidade do colágeno nos grupos tratados. Os resultados dos níveis de expressão sugeriram diminuição nos grupos de lesão e em dois grupos de tratamento (0,0115 µg/g e 0,009 µg/g). No entanto, com a menor concentração (0,0065 µg/g), não foi observada diferença significativa, apresentando níveis semelhantes aos encontrados em tecido saudável. O uso do OVAz foi eficaz para reverter as alterações do colágeno causadas pela fibrose, e sua menor concentração apresentou uma possível tendência de aumento na expressão do CAV-1. Portanto, os resultados mostraram que o OVAz tem potencial para ser uma alternativa não invasiva e de baixo custo para auxiliar no tratamento da fibrose muscular.

Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model / Caracterização química e efeitos do óleo volátil de Alpinia zerumbet na qualidade da deposição do colágeno e expressão da caveolina-1 em modelo murino de fibrose muscular

Abstract This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Resumo Este estudo avaliou o efeito do óleo volátil de Alpinia zerumbet (OVAz) na expressão do gene da caveolina-1 e na fibrose muscular. Os ratos foram imobilizados para induzir a fibrose do músculo gastrocnêmio, e foram tratados com OVAz. A qualidade do colágeno foi avaliada com histologia e à expressão do gene caveolina-1 (CAV-1) foi avaliada usando qPCR. A análise histomorfológica indicou uma redução significativa no perímetro, largura e intensidade do colágeno nos grupos tratados. Os resultados dos níveis de expressão sugeriram diminuição nos grupos de lesão e em dois grupos de tratamento (0,0115 µg/g e 0,009 µg/g). No entanto, com a menor concentração (0,0065 µg/g), não foi observada diferença significativa, apresentando níveis semelhantes aos encontrados em tecido saudável. O uso do OVAz foi eficaz para reverter as alterações do colágeno causadas pela fibrose, e sua menor concentração apresentou uma possível tendência de aumento na expressão do CAV-1. Portanto, os resultados mostraram que o OVAz tem potencial para ser uma alternativa não invasiva e de baixo custo para auxiliar no tratamento da fibrose muscular.

Encefalopatia toxica secundaria a vigabatrina: Reporte de caso / Toxic encephalopathy secondary to vigabatrin: Case report

El síndrome de West es una encefalopatía epiléptica caracterizada por espasmos en flexión, hipsarritmia en el electroencefalograma y retraso en el neurodesarrollo. Reportamos el caso de una paciente de 11 meses con diagnóstico de Síndrome de West y encefalopatía tóxica secundaria al uso de vigabatrina

West syndrome is an epileptic encephalopathy characterized by flexing spasms, hypsarritmia in the electroencephalogram and delayed neurodevelopment. We report an 11-month-old patient with a diagnosis of West syndrome and toxic encephalopathy secondary to the use of vigabatrin

Automated, High-Throughput Screening of Hybrid Elastin-like Polypeptide/Polysaccharide Multilayer Film Deposition.

The self-assembled layer-by-layer technique has attracted a great deal of attention as a method for engineering bio-functional surfaces under mild chemical conditions. The production of multilayer films, starting from newly designed building blocks, may be laborious, considering the inherent limitations for anticipating how minimal changes in the macromolecular composition may impact both film deposition and performance. This paper presents an automated, high-throughput approach to depositing polyelectrolyte multilayers (PEMs) in multiwell plates, enabling the screening of nearly 100 film formulations in the same process. This high-throughput layer-by-layer (HT-LbL) method runs in an affordable, fully commercial platform using Python-coded routines that can be easily adapted for the materials science lab settings. The HT-LbL system was validated by investigating the deposition of polysaccharide-based films in multiwell plates, probing the absorbance signal of ionically stained polyelectrolyte multilayers (PEMs) prepared in one single batch. The HT-LbL method was also used to investigate the deposition of PEMs with a small library of genetically engineered elastin-like polypeptides (ELPs) with different levels of ionizable and hydrophobic amino acid residues. The deposition of ELP/chitosan films was assessed based on the signal of fluorescently labeled species (chitosan or ELP-mCherry), demonstrating that both electrostatic and hydrophobic residues are essential for film buildup. The growth and surface properties of ELP-mCherry/chitosan films also seemed susceptible to the assembly pH, forming a higher film growth and a rougher and more hydrophobic surface for both polyelectrolytes deposited under a low ionization degree. Overall, this study illustrates the challenge of predicting the growth and properties of multilayer films and how the HT-LbL can accelerate the development of multilayer films that demand high levels of testing and optimization.

Influence of HER2 expression on prognosis in metastatic triple-negative breast cancer-results from an international, multicenter analysis coordinated by the AGMT Study Group.

BACKGROUND: Triple-negative breast cancer (TNBC) is associated with poor prognosis, and new treatment options are urgently needed. About 34%-39% of primary TNBCs show a low expression of human epidermal growth factor receptor 2 (HER2-low), which is a target for new anti-HER2 drugs. However, little is known about the frequency and the prognostic value of HER2-low in metastatic TNBC. PATIENTS AND METHODS: We retrospectively included patients with TNBC from five European countries for this international, multicenter analysis. Triple-negativity had to be shown in a metastatic site or in the primary breast tumor diagnosed simultaneously or within 3 years before metastatic disease. HER2-low was defined as immunohistochemically (IHC) 1+ or 2+ without ERBB2 gene amplification. Survival probabilities were calculated by the Kaplan-Meier method, and multivariable hazard ratios (HRs) were estimated by Cox regression models. RESULTS: In total, 691 patients, diagnosed between January 2006 and February 2021, were assessable. The incidence of HER2-low was 32.0% [95% confidence interval (CI) 28.5% to 35.5%], with similar proportions in metastases (n = 265; 29.8%) and primary tumors (n = 425; 33.4%; P = 0.324). The median overall survival (OS) in HER2-low and HER2-0 TNBC was 18.6 and 16.1 months, respectively (HR 1.00; 95% CI 0.83-1.19; P = 0.969). Similarly, in multivariable analysis, HER2-low had no significant impact on OS (HR 0.95; 95% CI 0.79-1.13; P = 0.545). No difference in prognosis was observed between HER2 IHC 0/1+ and IHC 2+ tumors (HR 0.89; 95% CI 0.69-1.17; P = 0.414). CONCLUSIONS: In this large international dataset of metastatic TNBC, the frequency of HER2-low was 32.0%. Neither in univariable nor in multivariable analysis HER2-low showed any influence on OS.

Estilo de Decisão e Planificação dos treinadores do Special Olympics Portugal: Influência da formação formal e não formal / El estilo de decisiones y de planificación de los entrenadores de Special Olympics Portugal: la influencia de la formación formal y no formal / Decision and Planning Style of Special Olympics Portugal coaches: the influence of formal and non-formal training

O Special Olympics Portugal é uma das principais organizações nacionais para o desenvolvimento do desporto para atletas com deficiência intelectual. Os treinadores desempenhem um papel relevante no desenvolvimento dos atletas, sendo os estilos de decisão e planificação aspetos essenciais para adequar o treino às necessidades especificas dos atletas. Estas prerrogativas são pouco exploradas cientificamente quanto aos treinadores deste tipo de atletas. O estudo tem como objetivo identificar os estilos de decisão e planificação do treinador do Special Olympics Portugal. Pretende ainda analisar a relação da formação formal e não formal com os estilos de decisão e planificação. A investigação é descritiva e de corte transversal dos treinadores do Special Olympics Portugal, com a participação de 50 sujeitos. Foram utilizados dois questionários: o Questionário dos Estilos de Decisão no Desporto e o Questionário dos Estilos de Planificação no Desporto. Os resultados revelam que os treinadores do Special Olympics Portugal possuem um amplo espetro de estilos de atuação no treino, predominando os estilos de decisão democrático e planificação flexível. Esta situação implica que os treinadores desta área são mais reflexivos, necessitando de adaptar as suas metodologias para trabalhar com os atletas e procurar novas soluções, de forma a adaptarem o treino às necessidades efetivas dos desportistas. (AU)

Special Olympics Portugal is one of the main national organizations for the development of sport for athletes with intellectual disabilities. Coaches play a relevant role in the development of athletes, and the decision and planning styles are essential aspects to adapt the training to the athletes' specific needs. These prerogatives are scientifically little explored related with the coaches of this type of athletes. The study aims to identify the decision and planning styles of the Special Olympics Portugal coaches. It also intends to analyze the relationship of formal and non-formal training with the decision and planning styles. The research is descriptive and cross-sectional of the Special Olympics Portugal coaches, with the participation of 50 subjects. Two questionnaires were used: the Questionnaire of Decision Styles in Sport and the Questionnaire ofPlanning Styles in Sport. The results show that the Special Olympics Portugal coaches have a wide spectrum of styles of action in training, predominantly the democratic decision style and flexible planning. This situation implies that the coaches in this area are more reflective, and need to adapt their methodologies to work with the athletes and seek new solutions, in order to adapt the training to the effective needs of the athletes. (AU)

Special OlympicsPortugal es una de las principales organizaciones nacionales para el desarrollo del deporte para atletas con discapacidad intelectual. Los entrenadores desempeñan un papel relevante en el desarrollo de los deportistas, siendo los estilos de decisión y planificación aspectos esenciales para adaptar el entrenamiento a las necesidades específicas de los deportistas. Estas prerrogativas están poco exploradas científicamente sobre los entrenadores de este tipo de atletas. Este estudio pretende identificar los estilos de decisión y planificación de los entrenadores de Special Olympics Portugal. También pretende analizar la relación entre la formación formal y no formal con los estilos de decisión y planificación. La investigación fue descriptiva y transversal, con la participación de 50 s entrenadores de Special Olympics Portugal. Se utilizaron dos cuestionarios: el Cuestionario de Estilos de Decisión en el Deporte y el Cuestionario de Estilos de Planificación en el Deporte. Los resultados muestran que los entrenadores de Special Olympics Portugal tienen un amplio espectro de estilos de actuación en el entrenamiento, predominando los estilos de decisión democrático y de planificación flexible. Esta situación implica que los entrenadores de este ámbito profesional son más reflexivos, necesitando adaptar sus metodologías para trabajar con estos deportistas especiales y buscar nuevas soluciones, con el fin de adaptar el entrenamiento a las necesidades efectivas de los deportistas. (AU)

Recovery, separation and production of fuel, plastic and aluminum from the Tetra PAK waste to hydrothermal and pyrolysis processes.

The establishment of a method of separation of materials from Tetra Pak waste to obtain products for use as raw material, fuel or other purposes was investigated in this study. First, the feasibility of hydrothermal treatment for the production of a solid fuel (hydrochar) and solid fraction formed by polyethylene and aluminum, called composite was analyzed. The results indicated that hydrothermal treatment performed at 240 °C yield the formation of hydrochar with good properties for its use as fuel and a composite of polyethylene and aluminum. The best conversion and separation of the cardboard and polyethylene/aluminum were obtained using 120 min as operating time. Then, the recovery of the aluminum fraction from the composite by using spent olive oil waste was studied. A partial separation of the composite layers (polyethylene and aluminum) was accomplished with improved aluminum purity for higher operating temperatures. Finally, the operating conditions of the pyrolysis process for the production of a solid (char) and high purity composite (aluminum) were optimized. The characterization results indicated that both char and aluminum resulting from the pyrolysis of the Tetra Pak at 400 °C still have a significant amount of polyethylene while higher purity levels of aluminum can be obtained at temperatures equal of higher than 500 °C.

Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model.

This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Concurrent training remodels the subcutaneous adipose tissue extracellular matrix of people living with HIV: a non-randomized clinical trial.

Evaluate the effect of 12 wks of concurrent training (CT) in the extracellular matrix (ECM) of subcutaneous adipose tissue (SAT) in people living with HIV/AIDS (PLWHA). In the non-randomized clinical trial, 19 participants, 11 healthy (HIV-) and 18 PLWHA under the use of highly active antiretroviral therapy (HAART) for at least 1 year (HIV+). All participants engaged in a moderate-intensity CT program for 12 weeks, 3 times a week. Before and after CT, aerobic and strength performance were assessed, as well as anthropometric and biochemical blood profiles. In addition, SAT biopsies were performed for histologic and morphometric analyses. Statistical analysis was carried out with R Studio, using descriptive and inferential analysis, ANOVA test, and mixed-effect model (P < 0.05). HIV+ showed higher levels of very-low-density lipoproteins and triglycerides and lower levels of high-density lipoproteins at baseline than HIV- (P < 0.05). All groups showed improved aerobic and strength performances (P < 0.05). Both groups showed reduced adipocyte sizes after CT (P < 0.05). Lastly, HIV+ presented smaller adipocytes and higher elastic fiber deposition at baseline and decreased after training only in HIV+, similar to the HIV group. Thus, CT in PLWHA promoted a decrease in the size heterogeneity of adipocytes and elastic fiber deposition, remodeling the ECM, and improving the SAT fibrosis profile. Brazilian Clinical Trials Registry (ensaiosclinicos.gov.br - UTN: U1111-1214-3022). Novelty: Adipose tissue fibrosis is improved by training in people living with HIV. Concurrent training remodels adipose tissue extracellular matrix.

Inhibition of α-glucosidase by flavonoids of Cymbopogon citratus (DC) Stapf.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves extracts from Cymbopogon citratus (DC) Stapf. are widely used in traditional medicine exhibiting several in vivo biological activities, including antidiabetic. Several flavonoids, including aglycones and glycosides, were reported in this plant and previous studies suggested that flavonoids may interact with targets related to diabetes. AIM OF THE STUDY: Evaluated the hypoglycemic activity of C. citratus flavonoids through α-glucosidase inhibition and assess the structure-activity relationship using molecular docking studies. MATERIAL AND METHODS: An infusion of C. citratus leaves and its flavonoid-rich fraction were prepared. Five flavonoids from this fraction were isolated and structurally characterized by UV spectral analysis with shift reagents, HPLC-PDA-ESI/MSn and 1H NMR. The antidiabetic potential of C. citratus infusion, its flavonoid-rich fraction, glycosylated flavonoids and aglycones was evaluated trough the in vitro inhibition of yeast α-glucosidase. Posteriorly, molecular docking of the tested flavonoids was performed to investigate its possible interactions with the α-glucosidase pocket. RESULTS: The infusion of C. citratus, its flavonoid-rich fraction, luteolin and five flavone glycosides namely, luteolin 6-C-ß-glucopyranoside (isoorientin), luteolin 7-O-neohesperidoside (ionicerin), luteolin 7-O-ß-glucopyranoside (cynaroside), Luteolin 2″-O-rhamnosyl-C-(6-deoxy-ribo-hexos-3-ulosyl) (cassiaoccidentalin B), luteolin 6-C-α-arabinofuranosil-(1→2)-α-L-rhamnopyranoside (kurilesin A) showed higher inhibitory activity than the reference drug. This activity increased by the addition of a sugar moiety. However, the di-glycosides were less active than mono-glycosides. The docking studies showed interactions of sugar moieties and A or B rings with the catalytic pocket mainly through hydrogen bonds. CONCLUSIONS: Our results corroborate the potential of C. citratus as a medicinal plant for the treatment of diabetes and revealed that its flavonoid glycosides has hypoglycemic effect and can be explored as drug candidates to act as α-glucosidase inhibitors in the treatment of diabetes.

Global warming assessment suggests the endemic Brazilian kelp beds to be an endangered ecosystem.

Kelps are canopy-forming brown seaweed sustaining critical ecosystem services in coastal habitats, including shelter, nursery grounds, and providing food resources to a myriad of associated species. This study modeled the fundamental niche of Laminaria abyssalis along the Brazilian continental margin, an endemic species of the South Atlantic, to anticipate potential distributional range shifts under two contrasting scenarios of future environmental changes (RCP2.6 and RCP8.5). The model for fundamental niche predictions considering the "present scenario" has shown a wider potential area than the realized niche (i.e., the area where the species actually occurs) along the Brazilian coast. In both future scenarios, the models have shown niche erosion on the northern portion of the Brazilian coast and niche gains towards the south. In both scenarios, L. abyssalis populations tend to shift to deeper regions of the reef. The restricted range of occurrence (33,000 km2), intense anthropic activities along these beds (e.g., trawling fisheries, oil/gas mining, or removal for agricultural purposes) acting synergically with global warming, may drive this ecosystem to collapse faster than kelp species' ability to adapt. We propose to classify L. abyssalis as Endangered - (EN) under IUCN criteria, and highlight that long-term monitoring of kelp beds is an urgent need to develop effective conservation initiatives to protect such rare and invaluable ecosystem.

Breast cancer in very young women-a multicenter 10-year experience.

BACKGROUND: Breast cancer (BC) is the most prevalent cancer in adult young women in Europe. Although rare, it is one of the leading causes of death in this age group. The aim of this study is to characterize a cohort of young women regarding tumor stage, biology, treatment and survival. PATIENTS AND METHODS: We present a multicenter retrospective analysis of women <35 years of age, diagnosed with BC between 2008 and 2017. A total of 207 patients from five Portuguese centers were included, from whom 172 were eligible for analysis. Data were analyzed using IBM SPPSS statistics. RESULTS: Median age at diagnosis was 31 years. Fifty-one percent of tumors were hormone receptor (HR)-positive/human epidermal growth factor receptor 2 (HER2)-negative, 20% HR-positive/HER2-positive, 8% HR-negative/HER2-positive and 20% triple-negative BC. Twenty-two percent of patients were diagnosed in stage I, 26% stage II, 45% stage III and 6% had de novo metastatic cancer. Thirty-nine percent of patients were treated with neoadjuvant chemotherapy. Mean follow-up time was 64.9 months and overall survival at 5 years, of the entire cohort and metastatic patients, was 86.5% and 26%, respectively. CONCLUSIONS: In our study we found similar population characteristics to other cohorts <35 years of age. To our knowledge, this is one of the largest cohorts in very young women. BC in young women is an important issue and further studies are needed to provide better care and survivorship to patients.