Influence of Aldehyde Dehydrogenase Inhibition on Stemness of Endometrial Cancer Stem Cells.

Endometrial cancer is one of the most common gynaecological malignancies. Although often diagnosed at an early stage, there is a subset of patients with recurrent and metastatic disease for whom current treatments are not effective. Cancer stem cells (CSCs) play a pivotal role in triggering tumorigenesis, disease progression, recurrence, and metastasis, as high aldehyde dehydrogenase (ALDH) activity is associated with invasiveness and chemotherapy resistance. Therefore, this study aimed to evaluate the effects of ALDH inhibition in endometrial CSCs. ECC-1 and RL95-2 cells were submitted to a sphere-forming protocol to obtain endometrial CSCs. ALDH inhibition was evaluated through ALDH activity and expression, sphere-forming capacity, self-renewal, projection area, and CD133, CD44, CD24, and P53 expression. A mass spectrometry-based proteomic study was performed to determine the proteomic profile of endometrial cancer cells upon N,N-diethylaminobenzaldehyde (DEAB). DEAB reduced ALDH activity and expression, along with a significant decrease in sphere-forming capacity and projection area, with increased CD133 expression. Additionally, DEAB modulated P53 expression. Endometrial cancer cells display a distinct proteomic profile upon DEAB, sharing 75 up-regulated and 30 down-regulated proteins. In conclusion, DEAB inhibits ALDH activity and expression, influencing endometrial CSC phenotype. Furthermore, ALDH18A1, SdhA, and UBAP2L should be explored as novel molecular targets for endometrial cancer.

AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells.

The non-homologous end joining pathway is vital for repairing DNA double-strand breaks (DSB), with DNA-dependent protein kinase (DNA-PK) playing a critical role. Altered DNA damage response (DDR) in chronic (CML) and acute myeloid leukemia (AML) offers potential therapeutic opportunities. We studied the therapeutic potential of AZD-7648 (DNA-PK inhibitor) in CML and AML cell lines. This study used two CML (K-562 and LAMA-84) and five AML (HEL, HL-60, KG-1, NB-4, and THP-1) cell lines. DDR gene mutations were obtained from the COSMIC database. The copy number and methylation profile were evaluated using MS-MLPA and DDR genes, and telomere length using qPCR. p53 protein expression was assessed using Western Blot, chromosomal damage through cytokinesis-block micronucleus assay, and γH2AX levels and DSB repair kinetics using flow cytometry. Cell density and viability were analyzed using trypan blue assay after treatment with AZD-7648 in concentrations ranging from 10 to 200 µM. Cell death, cell cycle distribution, and cell proliferation rate were assessed using flow cytometry. The cells displayed different DNA baseline damage, DDR gene expressions, mutations, genetic/epigenetic changes, and p53 expression. Only HEL cells displayed inefficient DSB repair. The LAMA-84, HEL, and KG-1 cells were the most sensitive to AZD-7648, whereas HL-60 and K-562 showed a lower effect on density and viability. Besides the reduction in cell proliferation, AZD-7648 induced apoptosis, cell cycle arrest, and DNA damage. In conclusion, these results suggest that AZD-7648 holds promise as a potential therapy for myeloid leukemias, however, with variations in drug sensitivity among tested cell lines, thus supporting further investigation to identify the specific factors influencing sensitivity to this DNA-PK inhibitor.

Short Peripheral Venous Catheters Contamination and the Dangers of Bloodstream Infection in Portugal: An Analytic Study.

Peripheral venous catheters (PVCs) are the most used vascular access devices in the world. However, failure rates remain considerably high, with complications such as PVC-related infections posing significant threats to patients' well-being. In Portugal, studies evaluating the contamination of these vascular medical devices and characterizing the associated microorganisms are scarce and lack insight into potential virulence factors. To address this gap, we analyzed 110 PVC tips collected in a large tertiary hospital in Portugal. Experiments followed Maki et al.'s semi-quantitative method for microbiological diagnosis. Staphylococcus spp. were subsequently studied for the antimicrobial susceptibility profile by disc diffusion method and based on the cefoxitin phenotype, were further classified into strains resistant to methicillin. Screening for the mecA gene was also done by a polymerase chain reaction and minimum inhibitory concentration (MIC)-vancomycin as determined by E-test, proteolytic and hemolytic activity on skimmed milk 1% plate and blood agar, respectively. The biofilm formation was evaluated on microplate reading through iodonitrotetrazolium chloride 95% (INT). Overall, 30% of PVCs were contaminated, and the most prevalent genus was Staphylococcus spp., 48.8%. This genus presented resistance to penicillin (91%), erythromycin (82%), ciprofloxacin (64%), and cefoxitin (59%). Thus, 59% of strains were considered resistant to methicillin; however, we detected the mecA gene in 82% of the isolates tested. Regarding the virulence factors, 36.4% presented α-hemolysis and 22.7% ß-hemolysis, 63.6% presented a positive result for the production of proteases, and 63.6% presented a biofilm formation capacity. Nearly 36.4% were simultaneously resistant to methicillin and showed expression of proteases and/or hemolysins, biofilm formation, and the MIC to vancomycin were greater than 2 µg/mL. Conclusion: PVCs were mainly contaminated with Staphylococcus spp., with high pathogenicity and resistance to antibiotics. The production of virulence factors strengthens the attachment and the permanence to the catheter's lumen. Quality improvement initiatives are needed to mitigate such results and enhance the quality and safety of the care provided in this field.

Application of Perinatal Derivatives on Oncological Preclinical Models: A Review of Animal Studies.

The increasing cancer incidence has certified oncological management as one of the most critical challenges for the coming decades. New anticancer strategies are still needed, despite the significant advances brought to the forefront in the last decades. The most recent, promising therapeutic approaches have benefitted from the application of human perinatal derivatives (PnD), biological mediators with proven benefits in several fields beyond oncology. To elucidate preclinical results and clinic outcomes achieved in the oncological field, we present a narrative review of the studies resorting to animal models to assess specific outcomes of PnD products. Recent preclinical evidence points to promising anticancer effects offered by PnD mediators isolated from the placenta, amniotic membrane, amniotic fluid, and umbilical cord. Described effects include tumorigenesis prevention, uncontrolled growth or regrowth inhibition, tumor homing ability, and adequate cell-based delivery capacity. Furthermore, PnD treatments have been described as supportive of chemotherapy and radiological therapies, particularly when resistance has been reported. However, opposite effects of PnD products have also been observed, offering support and trophic effect to malignant cells. Such paradoxical and dichotomous roles need to be intensively investigated. Current hypotheses identify as explanatory some critical factors, such as the type of the PnD biological products used or the manufacturing procedure to prepare the tissue/cellular treatment, the experimental design (including human-relevant animal models), and intrinsic pathophysiological characteristics. The effective and safe translation of PnD treatments to clinical practice relies on the collaborative efforts of all researchers working with human-relevant oncological preclinical models. However, it requires proper guidelines and consensus compiled by experts and health workers who accurately describe the methodology of tissue collection, PnD isolation, manufacturing, preservation, and delivery to the final user.

Ring-Fused meso-Tetraarylchlorins as Auspicious PDT Sensitizers: Synthesis, Structural Characterization, Photophysics, and Biological Evaluation.

Novel 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine-fused meso-tetraarylchlorins, with different degrees of hydrophilicity (with methyl ester, hydroxymethyl, and carboxylic acid moieties), have been synthesized and their photophysical characterization as well as in vitro photocytotoxicity assessment against human melanoma and esophageal and bladder carcinomas was carried out. An integrated analysis of the photosensitizers' performance, considering the singlet oxygen generation data, cell internalization, and intracellular localization, allowed to establish relevant structure-photoactivity relationships and the rationalization of the observed photocytotoxicity. In the diacid and monoalcohol series, chlorins derived from meso-tetraphenylporphyrin proved to be the most efficient photodynamic therapy agents, showing IC50 values of 68 and 344 nM against A375 cells, respectively. These compounds were less active against OE19 and HT1376 cells, the diacid chlorin with IC50 values still in the nano-molar range, whereas the monohydroxymethyl-chlorin showed significantly higher IC50 values. The lead di(hydroxymethyl)-substituted meso-tetraphenylchlorin confirmed its remarkable photoactivity with IC50 values below 75 nM against the studied cancer cell lines. Subcellular accumulation of this chlorin in the mitochondria, endoplasmic reticulum, and plasma membrane was demonstrated.

Applications of Photodynamic Therapy in Endometrial Diseases.

Photodynamic therapy (PDT) is a medical procedure useful for several benign conditions (such as wound healing and infections) and cancer. PDT is minimally invasive, presents few side effects, good scaring, and is able to minimal tissue destruction maintaining organ anatomy and function. Endoscopic access to the uterus puts PDT in the spotlight for endometrial disease treatment. This work systematically reviews the current evidence of PDT's potential and usefulness in endometrial diseases. Thus, this narrative review focused on PDT applications for endometrial disease, including reports regarding in vitro, ex vivo, animal, and clinical studies. Cell lines and primary samples were used as in vitro models of cancer, adenomyosis and endometrioses, while most animal studies focused the PDT outcomes on endometrial ablation. A few clinical attempts are known using PDT for endometrial ablation and cancer lesions. This review emphasises PDT as a promising field of research. This therapeutic approach has the potential to become an effective conservative treatment method for endometrial benign and malignant lesions. Further investigations with improved photosensitisers are highly expected.

Nursing Students' Perceptions on Healthcare-Associated Infection Control and Prevention Teaching and Learning Experience in Portugal.

Healthcare-associated infections (HAI) are one of the major concerns worldwide, posing significant challenges to healthcare professionals' education and training. This study intended to measure nursing students' perceptions regarding their learning experiences on HAI prevention and control. In the first phase of the study, a cross-sectional and descriptive study with a convenience sample composed of undergraduate nursing students from Portugal, Spain, Poland, and Finland was conducted to develop the InovSafeCare questionnaire. In the second phase, we applied the InovSafeCare scale in a sample of nursing students from two Portuguese higher education institutions to explore which factors impact nursing students' adherence to HAI prevention and control measures in clinical settings. In phase one, the InovSafeCare questionnaire was applied to 1326 students internationally, with the instrument presenting adequate psychometric qualities with reliability results in 14 dimensions. During phase two, the findings supported that Portuguese nursing students' adherence to HAI prevention and control measures is influenced not only by the curricular offerings and resources available in academic settings, but also by the standards conveyed by nursing tutors during clinical placements. Our findings support the need for a dedicated curricular focus on HAI prevention and control learning, not only through specific classroom modules, innovative resources, and pedagogical approaches, but also through a complementary and coordinated liaison between teachers and tutors in academic and clinical settings.

Nursing Students' Perceptions on Healthcare-Associated Infection Control and Prevention Teaching and Learning Experience: Development and Validation of a Scale in Four European Countries.

Healthcare-associated infections are one of the major concerns worldwide. This study presents the development and the validation process of the InovSafeCare scale and aimed at identifying and measuring the ecosystem variables related to healthcare-associated infection (HCAI) prevention and control practices in European nurse students. Qualitative and quantitative approaches were used to (1) elaborate an item pool related to the educational environment, the healthcare setting environment, and the attitudes, beliefs, and performance of the nursing students regarding HCAI prevention and control and (2) analyze psychometric properties of the scale using factor analysis. The validated InovSafeCare scale was applied to undergraduate nursing students of five European Higher Education Institutions. The partial least square structural equation modeling (PLS-SEM) method with SMART-PLS3 software was used. The study sample consists of 657 nursing students, who responded a self-report inventory. From the analyzed data were identified 14 factors. The InovSafeCare scale reveals good validity and reliability of the dimensions in different European countries.

Peripherally Inserted Central Catheter Placement in a Cardiology Ward: A Focus Group Study of Nurses' Perspectives.

Intravenous therapy administration through peripheral venous catheters is one of the most common nursing procedures performed in clinical contexts. However, peripherally inserted central catheters (PICC) remain insufficiently used by nurses and can be considered a potential alternative for patients who need aggressive intravenous therapy and/or therapy for extended periods. The purpose of this study was to understand nurses' perspectives about PICC implementation in their clinical practice. As part of an action-research project, three focus groups were developed in June 2019 with nineteen nurses of a cardiology ward from a Portuguese tertiary hospital. From the content analysis, two main categories emerged: 'nursing practices' and 'patients'. Nurses considered PICC beneficial for their clinical practice because it facilitates maintenance care and catheter replacement rates. Moreover, nurses suggested that, since there is a need for specific skills, the constitution of vascular access teams, as recommended by international guidelines, could be an advantage. Regarding patient benefits, nurses highlighted a decrease in the number of venipunctures and also of patient discomfort, which was associated with the number of peripheral venous catheters. Infection prevention was also indicated. As an emerging medical device used among clinicians, peripherally inserted central catheters seem to be essential to clinical practice.

Personal and technological skills to coach people with noncommunicable diseases: development and validation of a scale for nursing students.

BACKGROUND: Current international policies converge to the need of empowering patients and families in becoming more autonomous in the self-caring and management of their noncommunicable diseases (NCDs). Given their professional scope, nurses are the most well-positioned health professionals to answer this societal challenge. In the literature, health coaching and the use of information and communication technologies (ICTs) emerge as two still under-used contributions to nursing practice in this regard. Given the lack of instruments and research developed so far in the use of health coaching and ICTs during nurses training, we aim to develop a scale that explores nursing students' perceptions regarding their coaching skills of people with NCDs and the potential role of ICTs in this domain. METHODS: After a comprehensive literature review, an initial items list (n = 39) was delineated and discussed by a panel of international experts. After conceptual and structural consensus, the pre-validated version of the Personal and Technological Skills to coach people with noncommunicable diseases scale (PTSC-NCD scale) was created. Then, the pre-validated PTSC-NCD scale was translated to Portuguese, Finnish, Flemish and Slovenian following Beaton and colleagues' recommendations, and applied to undergraduate nursing students in five European universities. Principal component analysis and reliability analysis were performed in each country through the statistical program Statistical Package for the Social Sciences (version 22.0). All ethical assumptions were complied with throughout this study. RESULTS: 874 nursing students enrolled in the study, predominantly female (71.1%) and with a mean age of 22.4 years (SD = 5.49). After data analysis across international settings, three dimensions emerged: Coaching Centred Personal Skills (F1); Digital Technology Improving Patient-Centred Care (F2); and Digital Technology Improving Relational Skills (F3). All the dimensions showed good reliability (Cronbach's alpha >.80). CONCLUSION: The PTSC-NCD scale evidence good validity and reliability indicators across different international settings.

GuttaFlow® Bioseal Cytotoxicity Assessment: In Vitro Study.

The sealers used for root canal treatment should be biocompatible for the peri-radicular tissues, to evaluate the cytotoxic effects of GuttaFlow® bioseal sealer and to compare them with AH26® epoxy resin. Culture media were conditioned with the GuttaFlow® bioseal and AH26® pellets. MDPC-23 odontoblast cell cultures were treated with conditioned medium and serial dilutions. To evaluate the metabolic activity and cellular viability, the MTT and SRB assays were performed. To determine the production of reactive oxygen species, the DHE and DCF-DA probes were used. Cell cycle and cell-death types were assessed by cytometry, and to evaluate the mineralization capacity, the Alizarin Red S coloration was used. Statistical analysis was performed using analysis of variance (ANOVA) when normality was found and Kruskal-Wallis on the opposite case. For the comparison with normality values, the Student t-test was used. Cells exposed to the GuttaFlow® bioseal conditioned medium maintained high metabolic activities, except at higher concentrations. Likewise, viability was maintained, but a significant decrease was observed after exposure to the highest concentration (p < 0.001), associated with cell death by late apoptosis and necrosis. When cell cultures were exposed to AH26®, metabolic activity was highly compromised, resulting in cell death. An imbalance in the production of peroxides and superoxide anion was observed. GuttaFlow® bioseal showed higher biocompatibility than AH26®.

The flushing procedure in nursing practices: A cross-sectional study with Portuguese and Brazilian nurses.

BACKGROUND: In patients with peripheral intravenous catheters (PIVCs), performing flushing is an essential procedure to maintain catheter patency and prevent complications. These PIVC related complications can lead to premature removal and therapeutics interruption, which implies the need of a new catheterization thus increasing patient discomfort and pain. AIMS: To identify nursing practices related to the flushing procedure, namely: moment(s) of the flushing; the syringe size used; the flush solution, volume and technique; the knowledge and accomplishment of the recommended standards on flushing by nurses. METHODS: A cross-sectional study was conducted between July and December 2017, with Brazilian and Portuguese nurses. An online questionnaire was developed based on the international recommendations on flushing procedure. Descriptive analysis was performed. RESULTS: A total of 76 nurses answered the questionnaire. The majority of nurses (84.2%) performed flushing: the most common technique used was continuous syringe pressure (31.2%), with the push-pause technique being performed by 23.4% of the nurses. Despite the majority performs flushing at four distinct moments (after the PIVC insertion, before, between and after drug delivery), there are inconsistencies in flush solution, volume, and syringe size. The most used volume to perform flushing was 5 mL, filled using normal saline. Despite this, they also recognized the omission of this procedure due to time constrains, no familiarity with the procedure and unavailable material. CONCLUSIONS: This study identified that flushing procedure isn't always performed by nurses in their clinical practice. Also, several inconsistencies were observed between nurses that performed flushing, reflecting the lack of empirical evidence in this area of research.

Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors.

Cancer stem cells (CSC) are a small population with self-renewal and plasticity which are responsible for tumorigenesis, resistance to treatment and recurrent disease. This population can be identified by surface markers, enzymatic activity and a functional profile. These approaches per se are limited, due to phenotypic heterogeneity and CSC plasticity. Here, we update the sphere-forming protocol to obtain CSC spheres from breast and gynecological cancers, assessing functional properties, CSC markers and protein expression. The spheres are obtained with single-cell seeding at low density in suspension culture, using a semi-solid methylcellulose medium to avoid migration and aggregates. This profitable protocol can be used in cancer cell lines but also in primary tumors. The tridimensional non-adherent suspension culture thought to mimic the tumor microenvironment, particularly the CSC-niche, is supplemented with epidermal growth factor and basic fibroblast growth factor to ensure CSC signaling. Aiming for robust identification of CSC, we propose a complementary approach, combining functional and phenotypic evaluation. Sphere-forming capacity, self-renewal and sphere projection area establish CSC functional properties. Additionally, characterization comprises flow cytometry evaluation of the markers, represented by CD44+/CD24- and CD133, and Western blot, considering ALDH. The presented protocol was also optimized for primary tumor samples, following a sample digestion procedure, useful for translational research.

Impact of an Innovative Securement Dressing and Tourniquet in Peripheral Intravenous Catheter-Related Complications and Contamination: An Interventional Study.

Reusable tourniquets and conventional securement dressings are considered risk factors for the occurrence of reported complications and catheter-related bloodstream infections. This study's purpose is to assess the impact of single-use disposable tourniquets and advanced occlusive polyurethane dressings with reinforced cloth borders on peripheral intravenous catheter (PIVC)-related complications and contamination. A pre- and post-interventional prospective observational study was conducted in a cardiology ward of a tertiary hospital between April 2018 and February 2019. Overall, demographic and clinical data from 156 patients and PIVC-related outcomes were collected (n = 296) as well as PIVC tips for microbiological analysis (n = 90). In the pre-intervention phase (n = 118), complication rates of 62.1% were reported, while 44.1% of the PIVCs were contaminated (n = 34). In the post-intervention phase (n = 178), complication rates decreased to 57.3%, while contamination rates significantly decreased to 17.9% (p = 0.014; n = 56). Through a logistic regression, it was found that the use of innovative technologies reduces the chance of PIVC contamination by 79% (odds ratio (OR): 0.21; 95% confidence interval (CI): 0.05-0.98; p = 0.046). Meanwhile, PIVC-related complications and fluid therapy emerged as predictors for PIVC contamination. Findings suggest that the adoption of these innovative devices in nurses' practice contributes to the significant reduction of PIVC contamination.

Endometrial Cancer Spheres Show Cancer Stem Cells Phenotype and Preference for Oxidative Metabolism.

This study aimed to characterize endometrial cancer regarding cancer stem cells (CSC) markers, regulatory and differentiation pathways, tumorigenicity and glucose metabolism. Endometrial cancer cell line ECC1 was submitted to sphere forming protocols. The first spheres generation (ES1) was cultured in adherent conditions (G1). This procedure was repeated and was obtained generations of spheres (ES1, ES2 and ES3) and spheres-derived cells in adherent conditions (G1, G2 and G3). Populations were characterized regarding CD133, CD24, CD44, aldehyde dehydrogenase (ALDH), hormonal receptors, HER2, P53 and ß-catenin, fluorine-18 fluorodeoxyglucose ([18F]FDG) uptake and metabolism by NMR spectroscopy. An heterotopic model evaluated differential tumor growth. The spheres self-renewal was higher in ES3. The putative CSC markers CD133, CD44 and ALDH expression were higher in spheres. The expression of estrogen receptor (ER)α and P53 decreased in spheres, ERß and progesterone receptor had no significant changes and ß-catenin showed a tendency to increase. There was a higher 18F-FDG uptake in spheres, which also showed a lower lactate production and an oxidative cytosol status. The tumorigenesis in vivo showed an earlier growth of tumours derived from ES3. Endometrial spheres presented self-renewal and differentiation capacity, expressed CSC markers and an undifferentiated phenotype, showing preference for oxidative metabolism.

Epithelial-mesenchymal transition and microRNAs: Challenges and future perspectives in oral cancer.

BACKGROUND: Head and neck cancer is the sixth most common cancer worldwide, with oral squamous cell carcinoma (OSCC) being the most representative type. OSCC is a public health problem with high morbidity and poor survival rate. Epithelial-mesenchymal transition is emerging as a hallmark in OSCC. METHODS: In this study, we described the role of microRNAs in epithelial-mesenchymal transition regulation in OSCC based on a PubMed search using articles published in English between January 1, 2010, and January 31, 2018. RESULTS: MicroRNA's regulatory networks seem to be a hallmark of epithelial-mesenchymal transition in OSCC pathophysiology becoming a growing challenge to design new studies and strategies from biology to clinical applications. CONCLUSION: Therefore, we propose that targeting therapies to epithelial-mesenchymal transition-type cells, namely, coordinating microRNAs and/or hydrophobic drugs, such as conventional therapy, could be a promising strategy to improve the outcomes of patients with OSCC.