PIK3CA mutational status in tissue and plasma as a prognostic biomarker in HR+/HER2- breast cancer.

INTRODUCTION: Hotspots (HS) mutations in the PIK3CA gene may lead to poorer oncological outcomes and endocrine resistance in advanced breast cancer (BC), but their prognostic role in early-stage disease remains controversial. The overall agreement within plasma and tissue methods has not been well explored. Our aim was to correlate tissue and plasma approaches and to analyze the prognostic impact of PIK3CA mutations (PIK3CAm) in HR+/HER2- BC. METHODS: A retrospective and unicentric analysis of PIK3CA mutational status in tissue and plasma samples by Cobas®PIK3CA Mutation Kit in patients with HR+/HER2- BC. RESULTS: We analyzed 225 samples from 161 patients with luminal BC. PIK3CA mutations were identified in 62 patients (38.5%), of which 39.6% were found in tissue and 11.8% in plasma. In advanced disease, plasma and tissue correlation rate was performed in 64 cases, with an overall agreement of 70.3%. Eighty patients were treated with CDK4/6 inhibitors + endocrine therapy. We observed a moderately worse progression-free survival (PFS) in PIK3CAm versus wild-type (WT) (24 m vs. 30 m; HR = 1.39, p = 0.26). A subanalysis was carried out based on exons 9 and 20, which showed a statistically poorer PFS in PIK3CAm exon 9 versus 20 population (9.7 m vs. 30.3 m; HR = 2.84; p = 0.024). Furthermore, detection of PIK3CAm in plasma was linked to a worse PFS vs PIK3CAm detection just in tissue (12.4 vs. 29.3; HR = 2.4; p = 0.08). CONCLUSIONS: Our findings suggest the PIK3CA evaluation in tissue as the diagnostic method of choice, however, additional investigations are required to improve the role of liquid biopsy in the PIK3CA assessment. PIK3CAm show worse outcomes in advanced luminal BC, especially in exon 9 mutation carriers, despite visceral involvement, prior exposure to endocrine therapy or detection of PIK3CAm in plasma, with an unclear prognosis in early-stage disease. Nonetheless, this should be validated in a prospective cohort study.

The impact of advanced paternal age on the live birth rate in patients undergoing Assisted Reproduction treatment: Findings from an analysis at a public reproductive center in Brazil.

OBJECTIVE: Human reproduction presents a challenge for our species, as evidenced by the escalating rates of infertility. This trend has prompted inquiries into diverse strategies aimed at mitigating infertility and enhancing conception rates. Despite the extensive research on advanced maternal age as a risk factor for reproductive outcomes, paternal age has historically garnered comparatively less attention. The aim of this study was to assess the impact of paternal age on embryos and its subsequent repercussions on fertilization rate, biochemical pregnancy, clinical pregnancy, and live birth rate in individuals undergoing assisted reproductive treatment in a public reproductive center located in Brazil. METHODS: This investigation adopted a retrospective cohort, cross-sectional, analytical design, utilizing the analysis of secondary data, covering the period from July 2015 to July 2021. RESULTS: A total of 350 couples grappling with infertility and undergoing intrauterine insemination (IUI), in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI) were included in the analysis. Examination of age groups revealed a notable correlation between the ages of women and men (correlation coefficient R=0.12, p<0.0001). In the analysis of IVF techniques, a discernible trend towards a negative correlation with paternal age was observed, signifying that higher paternal age was linked to lower fertilization rates (p=0.004). CONCLUSIONS: Advanced paternal age significantly impacts full-term birth rates in IVF procedures, emphasizing the need for preconception public health advisories that underscore the risks associated with delaying parenthood for both men and women, particularly among those necessitating assisted reproductive techniques.

Commercial propolis extracts from Apis mellifera, Scaptotrigona bipunctata and Melipona quadrifasciata: quality control, antioxidant and antiglycemic potential.

Quality control of propolis plays a pivotal role in ensuring the appropriate concentrations of active compounds, limiting unwanted substances, verifying authenticity, and adhering to regulatory standards. This study aimed to assess the identity and quality standards, the individual phenolic composition (LC-ESI-MS/MS), and the antioxidant and antiglycemic potential of commercial propolis extracts (CPEs) from Apis mellifera, Scaptotrigona bipunctata, and Melipona quadrifasciata bees. CPEs met wax content and oxidation activity criteria, surpassing minimum thresholds for total phenolic content (TPC) and flavonoid content (TFC), although stingless bee CPE did not test positive for 10% lead acetate. CPEs exhibited antioxidant and potential antiglycemic activities. Epicatechin among the thirty-three identified phenolics, showed significant correlation with TPC, DPPH, ABTS, and EC50 values of α-amylase enzyme. These promising attributes underscore the potential health benefits of commercial propolis extracts from Apis mellifera and stingless bees for further medicinal and nutritional applications.

Erythropoietin effects on cryopreserved/transplanted cat ovarian tissue: A comparison of two incubation methods.

Many feline species are currently threatened with extinction. Therefore, germplasm bank establishment has become imperative. However, cryoinjury and ischemia-reperfusion injury pose significant obstacles to both cryopreservation and xenotransplantation. In this regard, erythropoietin (Epo) represents a potential alternative strategy due to its properties. This study aimed to assess the incubation of domestic cat ovarian tissue in Epo, both before and after cryopreservation, and investigate its effectiveness in promoting revascularization following xenotransplantation. Sixteen ovaries from 8 healthy cats were sliced following elective bilateral ovariohysterectomy (OHE). Subsequently, 8 fragments measuring 3 mm³ each were obtained from the cortical region of each ovary. The fragments were allocated into 3 treatment groups: Cryo group, fragments were cryopreserved, thawed and immediately transplanted; Cryo + Epo group, fragments were first cryopreserved in nitrogen, thawed, incubated in Epo (100 IU) for 2h and transplanted; and the Epo + Cryo group, in which fragments were first incubated in Epo (100 IU) for 2h, cryopreserved, thawed and immediately transplanted. The fragments were then xenotransplanted into the dorsal subcutaneous region of ovariectomized female nude mice and retrieved at 7, 14, 21, and 28 days post-transplantation. The results indicated that Epo effectively enhanced follicular survival, preservation of viability, and tissue revascularization. The Epo + Cryo group displayed better revascularization rates on D14 and D21 post-transplantation and an increase in primordial and growing follicles on D28, the Cryo + Epo group exhibited significantly more follicles on D14 and D21, with fewer degenerated follicles.

Nutritional Composition, Simulated Digestion and Biological Activities of Campomanesia xanthocarpa Fruit.

Gabirobeira fruits are known for their rich nutrient content and bioactive phytochemical compounds that contribute to significant biological activities. Despite these attributes, the antioxidant potential and stability of phenolic compounds in gabiroba by-products after digestion have yet to be studied. The objective of this work was to evaluate the physical-chemical composition, antibacterial activity, α-amylase, and α-glucosidase inhibitory effects, as well as the in vitro digestibility of total phenolic compounds, total flavonoids, and antioxidant activity of powder and extract from gabiroba to valorize these byproducts. The gabiroba powder had low moisture, high carbohydrate and fiber content. The extraction using 80% ethanol demonstrated higher antioxidant, antibacterial, α-amylase, and α-glucosidase inhibition activities compared to the 12% ethanol and water extracts. Catechin and ferulic acid were the major phenolic compounds identified by HPLC-DAD. After digestion, both the powder and the gabiroba extract exhibited a bioaccessibility of more than 30% for total phenolic compounds and antioxidant activity during the gastric phase. However, the dry ethanol extract displayed higher total phenolic compounds after both the gastric and intestinal phases compared to the flour. Processing gabiroba into powder and extract is a promising approach to fully utilize this seasonal fruit, minimize waste, concentrate health-beneficial compounds, and valorize a by-product for use as a functional ingredient and raw material within the food and pharmaceutical industries.

Exposure to microcrystallized cellulose affects the health of tadpoles and sheds light on the threat these materials pose to amphibians.

The increasing use of cellulose-based materials (CBMs) has provided beneficial applications in different sectors. However, its release into environments may represent an ecological risk, therefore demanding that ecotoxicological studies be conducted to understand the risks (current and future) of CBM pollution. Thus, we evaluated the possible effects of microcrystalline cellulose (CMs) in Physalaemus cuvieri tadpoles. After seven days of exposure to CMs (at 58.29 and 100 mg/L), the animals were subjected to behavioral evaluation, and different biomarkers (biometric and biochemical) were evaluated. Although our data do not point to a neurotoxic effect of CMs (inferred by the absence of behavioral changes and changes in AChE and BChE activity), animals exposed to CMs showed differences in body condition. Furthermore, we noticed an increase in the frequency of erythrocyte nuclear abnormalities and DNA damage, which were correlated with the ingestion of CMs. We noticed that the antioxidant activity of tadpoles exposed to CMs (inferred by SOD, CAT, and DPPH radical scavenging activity) was insufficient to control the increase in ROS and MDA production. Furthermore, exposure to CMs induced a predominant Th2-specific immune response, marked by suppressed IFN-γ and increased IL-10 levels, with a consequent reduction in NO levels. Principal component analysis and IBRv-2 indicate, in general, a primarily more toxic response to animals exposed to the highest CM concentration. Therefore, our study evidence that CMs affect the health of P. cuvieri tadpoles and sheds light on the threat these materials pose to amphibians.

Insulin-like growth factor (IGF) performance in ovarian function and applications in reproductive biotechnologies.

The role of the insulin-like growth factor (IGF) system has attracted close attention. The activity of IGF binding proteins (IGFBPs) within the ovary has not been fully elucidated to date. These proteins bind to IGF with an equal, or greater, affinity than to the IGF1 receptor, thus being in the main position to regulate IGF signalling, in addition to extending the half-life of IGFs within the bloodstream and promoting IGF storage in specific tissue niches. IGF1 has an important part in cell proliferation, differentiation and apoptosis. Considering the importance of IGFs in oocyte maturation, this review sought to elucidate aspects including: IGF production mechanisms; constituent members of their family and their respective functions; the role that these factors play during folliculogenesis, together with their functions during oocyte maturation and apoptosis, and their performance during luteal development. This review also explores the role of IGFs in biotechnological applications, focusing specifically on animal genetic gain.

Expression of tumoral GSK3-ß, PD-L1, and CD8 cell density in urothelial carcinomas, association with tumor grade and overall survival.

Bladder cancer is the most common malignancy in the urinary tract, and is biologically and clinically quite heterogeneous. Around 90% of diagnoses are made in the 6th decade, being more prevalent in males. The programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) axis play a putative role in immune checkpoint and as a means through which cancer evades the immune system. Inhibition of the glicogênio synthase kinase (GSK) 3 leads to the downregulation of PD-1 via upregulation of the transcription factor Tbet. The use of biomarkers PD-L1 and GSK-3ß and evaluation of the immune infiltrate have very promising correlations with urothelial carcinoma prognosis and treatment prediction. OBJECTIVE: To investigate the protein expression of PD-L1 and GSK-3ß and the CD8-positive immune infiltrates in bladder carcinomas. MATERIALS AND METHODS: This was a cross-sectional study of 140 samples of urothelial carcinomas from 2015 to 2018. Automated digitally assisted scoring and conventional analyses of the markers of GSK-3ß (27C10), CD8 (7103ß) and PDL-1 (22c3), were reviewed by two pathologists independently and a histologic score was calculated. The density of CD8 was also measured. RESULTS: The immunoexpression of GSK-3ß (91%) was presented in most samples, PD-L1 in 62.9% and CD8 cells present in 46.3% of cases. When analyzed in conjunction, the levels of GSK-3ß and PD-L1 (P = 0.033), and CD8 and PD-L1 (P<0.002) showed significant correlations. No significant associations were observed between GSK-3ß and CD8. The positivity of GSK-3ß and PD-L1 was predominant in high-grade tumors. CONCLUSION: Despite the tumor microenvironment heterogeneity, the expression of CD8, GSK-3ß and PDL1 could be valuable and GSK-3ß could be a potential target in advanced bladder cancer, especially in the context of immunotherapy.

Adesão ao preservativo masculino: realidade comportamental e conhecimento de universitários / Adherence to the male condom: behavioral reality and knowledge of university students

Homens e mulheres saberem a função do preservativo na prevenção de Infecções Sexualmente Transmissíveis e gravidez, várias situações dificultam o seu uso, entre elas, a ideia de que o preservativo interfere no prazer. OBJETIVO: Descrever o conhecimento e comportamentos de universitários sobre a adesão ao preservativo masculino. METODO-LOGIA: Estudo exploratório descritivo, quantitativo, com 136 estudantes de uma Universidade Pública. RESULTADOS: A média de idade foi de 20 anos, maior parte são heterossexuais (76,5%); a maioria teve a primeira relação sexual aos 17 anos, sendo que 33,5% não usaram camisinha, 85% utilizam camisinha para evitar gravidez, 92,5% para prevenção de Infecções Sexualmente Transmissíveis, 48,5% deixaram de ter relação sexual com penetração por não ter preservativo; 84% não utilizaram por não conseguir colocar e 54% tinham camisinha e não utilizou. CONCLUSÃO: Evidenciou a necessidade de repensar a educação sexual para instrumentalizar quanto a percepção, comportamento e adesão ao preservativo.(AU)

Men and women recognize condoms' role in preventing both Sexually Transmitted Infec-tions and pregnancy. However, various obstacles, such as concerns about reduced pleasure, impede their use. OBJECTIVE: To describe the knowledge and behaviors of university students regarding adherence to male condoms. METHODOLOGY: This was a descriptive, quantitative exploratory study involving 136 students from a public university. RESULTS: The average age was 20 years, with most identifying as heterosexual (76.5%). A significant portion (33.5%) did not use condoms. Furthermore, 85% used con-doms for pregnancy prevention, 92.5% for Sexually Transmitted Infections prevention, while 48.5% ceased sexual activity due to a lack of condoms. Additionally, 84% faced challenges with putting on condoms, and 54% possessed unused condoms. CONCLU-SION: These findings highlight the necessity to reevaluate sex education, providing indi-viduals with the tools to comprehend, modify their behavior, and enhance adherence to condom usage.(AU)

Hombres y mujeres conocen el papel del condón en la prevención de las Infecciones de Transmisión Sexual y el embarazo, varias situaciones dificultan su uso, entre ellas la idea de que el condón interfiere con el placer. OBJETIVO: Describir los conocimientos y comportamientos de estudiantes universitarios respecto de la adherencia al condón masculino. METODOLOGÍA: Estudio descriptivo, cuantitativo, exploratorio con 136 estudiantes de una Universidad Pública. RESULTADOS: La edad promedio fue de 20 años, la mayoría son heterosexuales (76,5%); la mayoría tuvo su primera relación sexual a los 17 años, el 33.5% no utilizó condón, el 85% utilizó condón para evitar el embarazo, el 92.5% para prevenir Infecciones de Transmisión Sexual, el 48.5% dejó de tener relaciones sexuales con penetración debido a no tener condón; El 84% no lo usó porque no pudo colocarlo y el 54% tenía condón pero no lo usó. CONCLUSIÓN: Se destacó la necesidad de repensar la educación sexual para brindar herramientas sobre percepción, comportamiento y adherencia al uso del condón.(AU)

Facile Formation of Sulfurized Nanorod-Like ZnO/Zn(OH)2 and Hierarchical Flower-Like γ-Zn(OH)2 /ϵ-Zn(OH)2 from a Green Synthesis and Application as Luminescent Solar Concentrator.

This research endeavors to overcome the significant challenge of developing materials that simultaneously possess photostability and photosensitivity to UV-visible irradiation. Sulfurized nanorod (NR)-like ZnO/Zn(OH)2 and hierarchical flower-like γ-Zn(OH)2 /ϵ-Zn(OH)2 were identified from XRD diffraction patterns and Raman vibrational modes. The sulfurized material, observed by FEG-SEM and TEM, showed diameters ranging from 10 and 40 nm and lengths exceeding 200 nm. The S2- ions intercalated Zn2+ , modulating NRs to dumbbell-like microrods. SAED and HRTEM illustrated the atomic structure in (101) crystal plane. Its direct band gap of 3.0 eV was attributed to the oxygen vacancies, which also contribute to the deep-level emissions at 422 and 485 nm. BET indicated specific surface area of 4.4 m2 g-1 and pore size as mesoporosity, which are higher compared to the non-sulfurized analogue. These findings were consistent with the observed photocurrent, photostability and photoluminescence (PL), further supporting the suitability of sulfurized NR-like ZnO/Zn(OH)2 as a promising candidate for Luminescent solar concentrators (LSC)-photovoltaic (PV) system.

Cellulose microcrystalline: A promising ecofriendly approach to control Culex quinquefasciatus larvae.

The growing use of synthetic chemical compounds/substances in vector control of mosquitoes, associated with their adverse effects on the environment and non-target organisms, has demanded the development of eco-friendly alternatives. In this context, this study aimed to evaluate the larvicidal action of different cellulose microcrystalline (CMs) concentrations and investigate their toxicity mechanisms in Culex quinquefasciatus fourth instar larvae as a model species. Probit analysis revealed that the median lethal concentrations (LC50) for 24 h and 36 h exposure were 100 and 58.29 mg/L, respectively. We also showed that such concentrations induced a redox imbalance in the larvae, marked by an increase in the production of reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS), as well as a reduction in the activity of superoxide dismutase (SOD) and catalase (CAT). Furthermore, different alterations in the external morphology of the larvae were associated with the ingestion of CMs. On the other hand, exposure of adult zebrafish (Danio rerio) to LC5024h and LC5036h for seven days did not induce any behavioral changes or alterations mutagenic, genotoxic, biochemical, or in the production of cytokines IFN-γ and IL-10. Thus, taken together, our study demonstrates for the first time that the use of CMs can constitute a promising strategy in the control of C. quinquefasciatus larvae, combining insecticidal efficiency with an "eco-friendly" approach in the fight against an important mosquito vector of several human diseases.

Effects of erythropoietin on ischaemia-reperfusion when administered before and after ovarian tissue transplantation in mice.

RESEARCH QUESTION: Is the optimal timing for administering erythropoietin to minimize ischaemic injury in ovarian tissue transplantation before ovary removal for cryopreservation and subsequent transplantation or after transplantation? DESIGN: Thirty Swiss mice (nu/nu) were divided into three groups: treatment control group (n = 10); erythropoietin before harvesting group (EPO-BH) (n = 10) and erythropoietin after transplantation group (EPO-AT) (n = 10). Animals underwent bilateral ovariohysterectomy and their hemiovaries were cryopreserved by slow freezing. At the same time, previously cryopreserved hemiovaries were transplanted subcutaneously in the dorsal region. Erythropoietin (250 IU/kg) and sterile 0.9% saline solution were administered every 12/12 h over 5 consecutive days in the EPO-AT and EPO-BH groups, respectively. RESULTS: Administration of erythropoietin in the EPO-AT group improved the viability of ovarian follicles, reducing degeneration and increasing the number of morphologically normal growing follicles at 14 days after transplantation compared with the EPO-BH group (P = 0.002). This group also showed higher percentages of proliferative follicles at 7 days after transplantation (P ≤ 0.03), increased blood vessel count (P ≤ 0.03) and greater tissue area occupied by blood vessels at days 7 and 14 after transplantation (P ≤ 0.03), compared with hormone administration before cryopreservation (EPO-BH group) and the treatment control group. Additionally, treatment with erythropoietin before or after transplantation reduced fibrotic areas at 7 days after transplantation (P = 0.004). CONCLUSION: Erythropoietin treatment after transplantation reduced ischaemic damage in transplanted ovarian tissue, increased angiogenesis, maintenance of ovarian follicle proliferation and reduced fibrosis areas in the grafted tissue.

Toxins from Animal Venoms as a Potential Source of Antimalarials: A Comprehensive Review.

Malaria is an infectious disease caused by Plasmodium spp. and it is mainly transmitted to humans by female mosquitoes of the genus Anopheles. Malaria is an important global public health problem due to its high rates of morbidity and mortality. At present, drug therapies and vector control with insecticides are respectively the most commonly used methods for the treatment and control of malaria. However, several studies have shown the resistance of Plasmodium to drugs that are recommended for the treatment of malaria. In view of this, it is necessary to carry out studies to discover new antimalarial molecules as lead compounds for the development of new medicines. In this sense, in the last few decades, animal venoms have attracted attention as a potential source for new antimalarial molecules. Therefore, the aim of this review was to summarize animal venom toxins with antimalarial activity found in the literature. From this research, 50 isolated substances, 4 venom fractions and 7 venom extracts from animals such as anurans, spiders, scorpions, snakes, and bees were identified. These toxins act as inhibitors at different key points in the biological cycle of Plasmodium and may be important in the context of the resistance of Plasmodium to currently available antimalarial drugs.

Evaluation of mobile applications focused on the care of patients with anxiety disorders: A systematic review in app stores in Brazil.

OBJECTIVE: To identify and evaluate the quality of mobile apps available in Brazil focused on the care of patients with anxiety disorders. METHODS: A comprehensive search was conducted until October 2021 on Play Store (Android) and Apple Store (iOS) in Brazil, using the terms "anxiety," "phobia," "panic attack," and "social phobia." Two independent authors identified the apps and performed data extraction and quality assessment using the Mobile App Rating Scale (MARS). Pearson's correlation was used to analyze the relationship between user star rating and the quality defined by the MARS instrument. RESULTS: A total of 3,278 potential apps were identified, of which 71 fully met the eligibility criteria. Most apps were made available on the Play Store (91.74%), in English (69.01%), and updated in the last two years (90.14%). Approximately half of the apps (50.70%) did not inform the developer's country and most of them did not report the user star rating (70.42%). The target population was indicated as free by most apps (85.92%), with generalized anxiety disorder being the most addressed disorder (74.65%), followed by panic disorder (33.80%). The three main purposes of the apps were education (83.10%), self-assessment (38.03%), and meditation/breathing (32.39%). Only 31 apps (43.66%) had acceptable quality (above 3.0) and the average total MARS quality score of 2.93 (2.20 to 3.90), with the functionality section receiving the highest score (3.90) and the lowest scoring sections being engagement (2.16). The apps that were rated stars by users (29.58%) showed a negative Pearson correlation (ρ = -0.100), evidencing a difference in the user's evaluation and that performed using the MARS instrument. CONCLUSION: Gaps in the quality of apps focused on the care of patients with anxiety disorders were evidenced since most were classified as having low quality through the MARS instrument. Thus, users are recommended to use these apps with caution.

First report on the toxicity of SARS-CoV-2, alone and in combination with polyethylene microplastics in neotropical fish.

The COVID-19 pandemic has caused unprecedented negative impacts in the modern era, including economic, social, and public health losses. On the other hand, the potential effects that the input of SARS-CoV-2 in the aquatic environment from sewage may represent on non-target organisms are not well known. In addition, it is not yet known whether the association of SARS-CoV-2 with other pollutants, such as microplastics (MPs), may further impact the aquatic biota. Thus, we aimed to evaluate the possible ecotoxicological effects of exposure of male adults Poecilia reticulata, for 15 days, to inactivated SARS-CoV-2 (0.742 pg/L; isolated SARS.CoV2/SP02.2020.HIAE.Br) and polyethylene MP (PE MPs) (7.1 × 104 particles/L), alone and in combination, from multiple biomarkers. Our data suggest that exposure to SARS-CoV-2 induced behavioral changes (in the open field test), nephrotoxic effect (inferred by the increase in creatinine), hepatotoxic effect (inferred by the increase in bilirubin production), imbalance in the homeostasis of Fe, Ca, and Mg, as well as an anticholinesterase effect in the animals [marked by the reduction of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity]. On the other hand, exposure to PE MPs induced a genotoxic effect (assessed by the comet assay), as well as an increase in enzyme activity alpha-amylase, alkaline phosphatase, and carboxylesterases. However, we did not show synergistic, antagonistic, or additive effects caused by the combined exposure of P. reticulata to SARS-CoV-2 and PE MPs. Principal component analysis (PCA) and values from the "Integrated Biomarker Response" index indicate that exposure to SARS-CoV-2 was determinant for a more prominent effect in the evaluated animals. Therefore, our study sheds light on the ecotoxicity of the new coronavirus in non-target organisms and ratifies the need for more attention to the impacts of COVID-19 on aquatic biota.

Utility of CYP2D6 copy number variants as prognostic biomarker in localized anal squamous cell carcinoma.

BACKGROUND: Anal squamous cell carcinoma (ASCC) is an infrequent tumor whose treatment has not changed since the 1970s. The aim of this study is the identification of biomarkers allowing personalized treatments and improvement of therapeutic outcomes. METHODS: Forty-six paraffin tumor samples from ASCC patients were analyzed by whole-exome sequencing. Copy number variants (CNVs) were identified and their relation to disease-free survival (DFS) was studied and validated in an independent retrospective cohort of 101 ASCC patients from the Multidisciplinary Spanish Digestive Cancer Group (GEMCAD). GEMCAD cohort proteomics allowed assessing the biological features of these tumors. RESULTS: On the discovery cohort, the median age was 61 years old, 50% were males, stages I/II/III: 3 (7%)/16 (35%)/27 (58%), respectively, median DFS was 33 months, and overall survival was 45 months. Twenty-nine genes whose duplication was related to DFS were identified. The most representative was duplications of the CYP2D locus, including CYP2D6, CYP2D7P, and CYP2D8P genes. Patients with CYP2D6 CNV had worse DFS at 5 years than those with two CYP2D6 copies (21% vs. 84%; p < .0002, hazard ratio [HR], 5.8; 95% confidence interval [CI], 2.7-24.9). In the GEMCAD validation cohort, patients with CYP2D6 CNV also had worse DFS at 5 years (56% vs. 87%; p = .02, HR = 3.6; 95% CI, 1.1-5.7). Mitochondria and mitochondrial cell-cycle proteins were overexpressed in patients with CYP2D6 CNV. CONCLUSIONS: Tumor CYP2D6 CNV identified patients with a significantly worse DFS at 5 years among localized ASCC patients treated with 5-fluorouracil, mitomycin C, and radiotherapy. Proteomics pointed out mitochondria and mitochondrial cell-cycle genes as possible therapeutic targets for these high-risk patients. PLAIN LANGUAGE SUMMARY: Anal squamous cell carcinoma is an infrequent tumor whose treatment has not been changed since the 1970s. However, disease-free survival in late staged tumors is between 40% and 70%. The presence of an alteration in the number of copies of CYP2D6 gene is a biomarker of worse disease-free survival. The analysis of the proteins in these high-risk patients pointed out mitochondria and mitochondrial cell-cycle genes as possible therapeutic targets. Therefore, the determination of the number of copies of CYP2D6 allows the identification of anal squamous carcinoma patients with a high-risk of relapse that could be redirected to a clinical trial. Additionally, this study may be useful to suggest new treatment strategies to increase current therapy efficacy.

Exposure of adult zebrafish (Danio rerio) to SARS-CoV-2 at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish.

While the multifaceted social, economic, and public health impacts associated with the COVID-19 pandemic are known, little is known about its effects on non-target aquatic ecosystems and organisms. Thus, we aimed to evaluate the potential ecotoxicity of SARS-CoV-2 lysate protein (SARS.CoV2/SP02.2020.HIAE.Br) in adult zebrafish (Danio rerio) at predicted environmentally relevant concentrations (0.742 and 2.226 pg/L), by 30 days. Although our data did not show locomotor alterations or anxiety-like or/and anxiolytic-like behavior, we noticed that exposure to SARS-CoV-2 negatively affected habituation memory and social aggregation of animals in response to a potential aquatic predator (Geophagus brasiliensis). An increased frequency of erythrocyte nuclear abnormalities was also observed in animals exposed to SARS-CoV-2. Furthermore, our data suggest that such changes were associated with a redox imbalance [↑ROS (reactive oxygen species), ↑H2O2 (hydrogen peroxide), ↓SOD (superoxide dismutase), and ↓CAT (catalase)], cholinesterasic effect [↑AChE (acetylcholinesterase) activity], as well as the induction of an inflammatory immune response [↑NO (nitric oxide), ↑IFN-γ (interferon-gamma), and ↓IL-10 (interleukin-10)]. For some biomarkers, we noticed that the response of the animals to the treatments was not concentration-dependent. However, principal component analysis (PCA) and the "Integrated Biomarker Response" index (IBRv2) indicated a more prominent ecotoxicity of SARS-CoV-2 at 2.226 pg/L. Therefore, our study advances knowledge about the ecotoxicological potential of SARS-CoV-2 and reinforces the presumption that the COVID-19 pandemic has negative implications beyond its economic, social, and public health impacts.

Long-term exposure of zebrafish juveniles to carbon nanofibers at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish.

Although carbon-based nanomaterials (CNMs) toxicity has already been demonstrated in some animal models, little is known about the impact of carbon nanofibers (CNFs) on aquatic vertebrates. Thus, we aimed to evaluate the possible effects of long-term exposure of zebrafish (Danio rerio) juveniles (90 days) to CNFs in predicted environmentally relevant concentrations (10 ng/L and 10 µg/L). Our data revealed that exposure to CNFs did not affect the growth and development of the animals, in addition to not having induced locomotor alterations or anxiety-like behavior. On the other hand, we observed that zebrafish exposed to CNFs showed a response deficit to the vibratory stimulus test, alteration in the density of neuromasts recorded in the final ventral region, as well as an increase in thiobarbituric acid reactive substances levels and a reduction in total antioxidant activity, nitric oxide, and acetylcholinesterase activity in the brain. Such data were directly associated with a higher concentration of total organic carbon in the brain, which suggests the bioaccumulation of CNFs. Furthermore, exposure to CNFs induced a picture suggestive of genomic instability, inferred by the increased frequency of nuclear abnormalities and DNA damage in circulating erythrocytes. Although the individual analyses of the biomarkers did not point to a concentration-dependent effect, the principal component analysis (PCA) and the Integrated Biomarker Response Index (IBRv2) indicate a more prominent effect induced by the higher CNFs concentration (10 µg/L). Therefore, our study confirms the impact of CNFs in the studied model (D. rerio) and sheds light on the ecotoxicological risks of these nanomaterials to freshwater fish. Based on the ecotoxicological screening provided by our study, new horizons are opened for investigations into the mechanisms of action of CNFs, which will help understand the magnitude of the impact of these materials on aquatic biota.

Plant-ZnO nanoparticles interaction: An approach to improve guinea grass (Panicum maximum) productivity and evaluation of the impacts of its ingestion by freshwater teleost fish.

We aimed to evaluate the possible effects of the application of zinc oxide nanoparticles [ZnO NPs; 68.96 ± 33.71 nm; at 100 and 500 mg/kg in a soil mixture of the Typic Dystrophic Red Latosol type and sand (2:1 ratio)] in the cultivation of Panicum maximum (until 125 days), using different biomarkers in addition to evaluating the uptake of Zn by the plants. Furthermore, we assessed the possible transfer of ZnO NPs from P. maximum leaves to zebrafish and their potential. Plants cultivated in substrates with ZnO NPs at 500 mg/kg showed reduced germination rate and growth. However, at 100 mg/kg, plants showed higher biomass and productivity, associated with higher Zn uptake, without inducing oxidative and nitrosative stress. Zinc content in zebrafish was not associated with ingesting leaves of P. maximum cultivated in substrate containing ZnCl2 or ZnO NPs or with genotoxic, mutagenic, and biochemical effects. In conclusion, ZnO NPs (at 100 mg/kg) are promising in the cultivation of P. maximum, and their ingestion by zebrafish did not cause changes in the evaluated biomarkers. However, we recommend that studies with other animal models be conducted to comprehensively assess the ecotoxicological hazard associated with applying ZnO NPs in soil.