Prognostic Value of Liquid-Biopsy-Based Biomarkers in Upper Tract Urothelial Carcinoma.

Currently, there are no reliable prognostic factors to determine which upper tract urothelial carcinoma (UTUC) patients will progress after radical nephroureterectomy (RNU). We aim to evaluate whether liquid-biopsy-based biomarkers (circulating tumor cells (CTCs), cell-free DNA (cfDNA), and circulating tumor DNA (ctDNA)) were able to predict clinical outcomes in localized UTUC patients undergoing RNU. Twenty patients were prospectively enrolled between 2021 and 2023. Two blood samples were collected before RNU and three months later. CTCs and cfDNA were isolated and evaluated using the IsoFlux system and Quant-iT PicoGreen dsDNA kit, respectively. Droplet digital PCR was performed to determine ctDNA status. Cox regression analysis was performed on CTCs, cfDNA, and ctDNA at two different follow-up time points to examine their influence on tumor progression and cancer-specific survival (CSS). During a median follow-up of 18 months, seven (35%) patients progressed and three (15%) died. Multivariate analysis demonstrated that cfDNA levels three months after RNU are a significant predictor of tumor progression (HR = 1.085; p = 0.006) and CSS (HR = 1.168; p = 0.029). No associations were found between CTC enumeration and ctDNA status with any of the clinical outcomes evaluated. The evaluation of cfDNA levels in clinical practice could improve the disease management of UTUC patients.

The transcription of the main gene associated with Treacher-Collins syndrome (TCOF1) is regulated by G-quadruplexes and cellular nucleic acid binding protein (CNBP).

Treacle ribosome biogenesis factor 1 (TCOF1) is responsible for about 80% of mandibular dysostosis (MD) cases. We have formerly identified a correlation between TCOF1 and CNBP (CCHC-type zinc finger nucleic acid binding protein) expression in human mesenchymal cells. Given the established role of CNBP in gene regulation during rostral development, we explored the potential for CNBP to modulate TCOF1 transcription. Computational analysis for CNBP binding sites (CNBP-BSs) in the TCOF1 promoter revealed several putative binding sites, two of which (Hs791 and Hs2160) overlap with putative G-quadruplex (G4) sequences (PQSs). We validated the folding of these PQSs measuring circular dichroism and fluorescence of appropriate synthetic oligonucleotides. In vitro studies confirmed binding of purified CNBP to the target PQSs (both folded as G4 and unfolded) with Kd values in the nM range. ChIP assays conducted in HeLa cells chromatin detected the CNBP binding to TCOF1 promoter. Transient transfections of HEK293 cells revealed that Hs2160 cloned upstream SV40 promoter increased transcription of downstream firefly luciferase reporter gene. We also detected a CNBP-BS and PQS (Dr2393) in the zebrafish TCOF1 orthologue promoter (nolc1). Disrupting this G4 in zebrafish embryos by microinjecting DNA antisense oligonucleotides complementary to Dr2393 reduced the transcription of nolc1 and recapitulated the craniofacial anomalies characteristic of Treacher Collins Syndrome. Both cnbp overexpression and Morpholino-mediated knockdown in zebrafish induced nolc1 transcription. These results suggest that CNBP modulates the transcriptional expression of TCOF1 through a mechanism involving G-quadruplex folding/unfolding, and that this regulation is active in vertebrates as distantly related as bony fish and humans. These findings may have implications for understanding and treating MD.

Tumor-Agnostic Circulating Tumor DNA Testing for Monitoring Muscle-Invasive Bladder Cancer.

Circulating tumor DNA (ctDNA) has recently emerged as a real-time prognostic and predictive biomarker for monitoring cancer patients. Here, we aimed to ascertain whether tumor-agnostic ctDNA testing would be a feasible strategy to monitor disease progression and therapeutic response in muscle-invasive bladder cancer (MIBC) patients after radical cystectomy (RC). Forty-two MIBC patients who underwent RC were prospectively included. Blood samples from these patients were collected at different follow-up time points. Two specific mutations (TERT c.1-124C>T and ATM c.1236-2A>T) were analyzed in the patients' plasma samples by droplet digital PCR to determine their ctDNA status. During a median follow-up of 21 months, 24% of patients progressed in a median of six months. ctDNA status was identified as a prognostic biomarker of tumor progression before RC and 4 and 12 months later (HR 6.774, HR 3.673, and HR 30.865, respectively; p < 0.05). Lastly, dynamic changes in ctDNA status between baseline and four months later were significantly associated with patient outcomes (p = 0.045). In conclusion, longitudinal ctDNA analysis using a tumor-agnostic approach is a potential tool for monitoring MIBC patients after RC. The implementation of this testing in a clinical setting could improve disease management and patients' outcomes.

Assessment of aggressive bladder cancer mutations in plasma cell-free DNA.

Background and aims: The spatial and temporal genetic heterogeneity of bladder cancer (BC) makes challenging to find specific drivers of metastatic disease, thus preventing to determine those BC patients at high risk of tumor progression. Our aim was to identify DNA mutations providing aggressive behavior to bladder tumors and analyze them in patients' cell-free DNA (cfDNA) during their follow-up after radical cystectomy (RC) in order to monitor tumor evolution. Methods: Six BC patients who underwent RC and presented disease progression during their follow-up were included. Next-generation sequencing was used to determine somatic mutations in several primary tumor and metastatic specimens from each patient. Shared DNA mutations between primary bladder tumor and metastatic sites were identified in cfDNA samples through droplet digital PCR. Results: Besides BC genetic heterogeneity, specific mutations in at least one of these genes -TERT, ATM, RB1, and FGFR3- were found in primary tumors and their metastases in all patients. These mutations were also identified in the patients' cfDNA at different follow-up time points. Additionally, the dynamic changes of these mutations in cfDNA allowed us to determine tumor evolution in response to treatment. Conclusion: The analysis of BC mutations associated with poor prognosis in plasma cfDNA could be a valuable tool to monitor tumor evolution, thus improving the clinical management of BC patients.

Cell-Free DNA as a Prognostic Biomarker for Monitoring Muscle-Invasive Bladder Cancer.

Cell-free DNA (cfDNA) has recently emerged as a real-time biomarker for diagnosis, monitoring and prediction of therapy response in tumoral disease. Here, we evaluated cfDNA as a prognostic biomarker for monitoring muscle-invasive bladder cancer (MIBC) patients at different follow-up time points. Blood samples from 37 MIBC patients who underwent radical cystectomy (RC) were collected at cystectomy and 1, 4, 12 and 24 months later. Plasma cfDNA amount and fragmentation patterns were determined. Four mutations were analyzed in cfDNA to detect circulating tumor DNA (ctDNA) during patient follow-up. During a median follow-up of 36 months, 46% of patients progressed; median time to progression was 10 months. cfDNA levels and ctDNA status four months after RC were identified as independent prognostic biomarkers of tumor progression (HR 5.290; p = 0.033) and cancer-specific survival (HR 4.199; p = 0.038), respectively. Furthermore, ctDNA clearance four months after RC was significantly associated with patients' clinical outcomes. In conclusion, cfDNA levels and ctDNA status four months after RC have prognostic implications in MIBC patients. In addition, cfDNA monitoring is useful to predict patient outcomes after RC. cfDNA analysis in the clinical setting could greatly improve MIBC patient management.

Multicentric validation of diagnostic tests based on BC-116 and BC-106 urine peptide biomarkers for bladder cancer in two prospective cohorts of patients.

BACKGROUND: Non-invasive urine-based biomarkers can potentially improve current diagnostic and monitoring protocols for bladder cancer (BC). Here we assess the performance of earlier published biomarker panels for BC detection (BC-116) and monitoring of recurrence (BC-106) in combination with cytology, in two prospectively collected patient cohorts. METHODS: Of the 602 patients screened for BC, 551 were found eligible. For the primary setting, 73 patients diagnosed with primary BC (n = 27) and benign urological disorders, including patients with macroscopic haematuria, cystitis and/or nephrolithiasis (n = 46) were included. In total, 478 patients under surveillance were additionally considered (83 BC recurrences; 395 negative for recurrence). Urine samples were analysed with capillary electrophoresis-mass spectrometry. The biomarker score was estimated via support vector machine-based software. RESULTS: Validation of BC-116 biomarker panel resulted in 89% sensitivity and 67% specificity (AUCBC-116 = 0.82). A diagnostic score based on cytology and BC-116 resulted in good (AUCNom116 = 0.85) but not significantly better performance (P = 0.5672). A diagnostic score including BC-106 and cytology was evaluated (AUCNom106 = 0.82), significantly outperforming both cytology (AUCcyt = 0.72; P = 0.0022) and BC-106 (AUCBC-106 = 0.67; P = 0.0012). CONCLUSIONS: BC-116 biomarker panel is a useful test for detecting primary BC. BC-106 classifier integrated with cytology showing >95% negative predictive value, might be useful for decreasing the number of cystoscopies during surveillance.

Prognostic Gene Expression-Based Signature in Clear-Cell Renal Cell Carcinoma.

The inaccuracy of the current prognostic algorithms and the potential changes in the therapeutic management of localized ccRCC demands the development of an improved prognostic model for these patients. To this end, we analyzed whole-transcriptome profiling of 26 tissue samples from progressive and non-progressive ccRCCs using Illumina Hi-seq 4000. Differentially expressed genes (DEG) were intersected with the RNA-sequencing data from the TCGA. The overlapping genes were used for further analysis. A total of 132 genes were found to be prognosis-related genes. LASSO regression enabled the development of the best prognostic six-gene panel. Cox regression analyses were performed to identify independent clinical prognostic parameters to construct a combined nomogram which includes the expression of CERCAM, MIA2, HS6ST2, ONECUT2, SOX12, TMEM132A, pT stage, tumor size and ISUP grade. A risk score generated using this model effectively stratified patients at higher risk of disease progression (HR 10.79; p < 0.001) and cancer-specific death (HR 19.27; p < 0.001). It correlated with the clinicopathological variables, enabling us to discriminate a subset of patients at higher risk of progression within the Stage, Size, Grade and Necrosis score (SSIGN) risk groups, pT and ISUP grade. In summary, a gene expression-based prognostic signature was successfully developed providing a more precise assessment of the individual risk of progression.

Prevalence of workplace violence against health care workers in hospital and pre-hospital settings: An umbrella review of meta-analyses.

Introduction: Workplace violence (WPV) is associated with adverse consequences for patients and health care workers (HCWs). The aim of this study was to assess the prevalence of WPV against HCWs in the hospital and pre-hospital settings. Methods: Using PRISMA guidelines, data resources including Scopus, PubMed, Web of Science, and Google Scholar were used for the search. The searches were conducted without any time limit until the end of December 2021. The random-effects model was used for this meta-analysis. I 2 index was used to examine heterogeneity and the Egger test was used to examine publication bias. Results: Of the 255 studies identified, 14 studies entered the umbrella review. The overall prevalence was as follows: WPV (58.7%); physical violence (20.8%); verbal violence (66.8%); and sexual harassment (10.5%). Conclusion: The prevalence of WPV and its types against HCWs is relatively high. WPV is associated with physical, psychological, and occupational consequences. Measures should be taken to reduce the consequences of WPV.

Identifying Chirality in Line Drawings of Molecules Using Imbalanced Dataset Sampler for a Multilabel Classification Task.

Chirality, the ability of some molecules to exist as two non-superimposable mirror images, profoundly influences both chemistry and biology. Advances in deep learning enable the automatic recognition of chemical structure diagrams, however, studies on discovering the molecule chirality are scarce and the machine-readable molecular representations are not always sufficient to fully support the encoding of this important property. Here, we pretrained networks on a ChEMBL+ dataset (79641 molecules) and fine-tuned them for the binary classification of chirality (achiral/chiral) or multilabel chirality type classifications (none/centre/axial/planar). To address the label combination imbalanced problem in the multilabel task, the study proposed a Formulated Imbalanced Dataset Sampler (FIDS) to sample a formulated amount of minority label combinations on top of the training set. On a 10-fold cross validation experiment using our CHIRAL dataset (1142 manually curated molecules), our models achieved up to an accuracy of 90 % in the binary task. In the multilabel task incorporated with FIDS, the overall performance increases from 87 % to 89 % and the accuracy per label combination can attained up to a 50 % increase. Through the study of heatmaps, our work also exemplified the potential of deep neural network to make predictions based on the actual location of chirality elements.

Prognostic implication of TERT promoter mutation and circulating tumor cells in muscle-invasive bladder cancer.

PURPOSE: Current clinical prognostic factors are not accurate enough to identify and monitor those muscle-invasive bladder cancer (MIBC) patients at high risk of progression after radical cystectomy (RC). Here, we determined genetic alterations in the tumor and circulating tumor cell (CTC) enumeration to find biomarkers useful for the management of MIBC after RC. METHODS: Thirty-nine MIBC patients undergoing RC were included. Tumoral tissue DNA was analyzed by next generation sequencing. CTCs were isolated from blood collected before RC and one, four and 12 months later. RESULTS: Sixteen (41%) patients progressed in a median time of 8.5 months and 11 (69%) of these patients harbored the TERT c.-124C > T mutation. All progressive patients harboring the TERT c.-124C > T mutation presented a significant increase in CTC number 12 months after RC compared to those without the mutation. Additionally, CTC number at 12 months was identified as an independent prognostic biomarker for tumor progression and cancer specific survival (CSS). Ten (63%) progressive patients showed an increment of CTC number with a median anticipation period of four months compared with imaging techniques. CONCLUSIONS: The TERT c.-124C > T mutation could be considered a biomarker of aggressivity. CTC enumeration is a useful tool for identifying MIBC patients at high risk of progression and CSS after RC and for detecting tumor progression earlier than imaging techniques.

Clinicopathological and Molecular Prognostic Classifier for Intermediate/High-Risk Clear Cell Renal Cell Carcinoma.

The probability of tumor progression in intermediate/high-risk clear cell renal cell carcinoma (ccRCC) is highly variable, underlining the lack of predictive accuracy of the current clinicopathological factors. To develop an accurate prognostic classifier for these patients, we analyzed global gene expression patterns in 13 tissue samples from progressive and non-progressive ccRCC using Illumina Hi-seq 4000. Expression levels of 22 selected differentially expressed genes (DEG) were assessed by nCounter analysis in an independent series of 71 ccRCCs. A clinicopathological-molecular model for predicting tumor progression was developed and in silico validated in a total of 202 ccRCC patients using the TCGA cohort. A total of 1202 DEGs were found between progressive and non-progressive intermediate/high-risk ccRCC in RNAseq analysis, and seven of the 22 DEGs selected were validated by nCounter. Expression of HS6ST2, pT stage, tumor size, and ISUP grade were found to be independent prognostic factors for tumor progression. A risk score generated using these variables was able to distinguish patients at higher risk of tumor progression (HR 7.27; p < 0.001), consistent with the results obtained from the TCGA cohort (HR 2.74; p < 0.002). In summary, a combined prognostic algorithm was successfully developed and validated. This model may aid physicians to select high-risk patients for adjuvant therapy.

The Prevalence of Post-traumatic Stress Disorder Among Health Care Workers During the COVID-19 Pandemic: An Umbrella Review and Meta-Analysis.

Introduction: Frontline health care workers (HCWs) have had an increased risk of developing health problems during the COVID-19 pandemic. In addition to physical illness, they have experienced mental health challenges, including post-traumatic stress disorder (PTSD). The aim of this study is to investigate the prevalence of PTSD among HCWs during the COVID-19 pandemic via an umbrella review and meta-analysis. Methods: This study was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline to perform a systematic literature search using various medical databases (Web of science, PubMed, Scopus, Cochrane, ProQuest, Science Direct, Embase, and Google scholar). The search included all articles published through the first of January 2020 the end of March 2021. The systematic review and meta-analysis studies that reported the prevalence of PTSD among health care workers were included in the study, and studies that reported the prevalence of PTSD in normal people or other epidemics were excluded. The random effects model was used to perform a meta-analysis, and the I 2 index was used to evaluate heterogeneity among studies. Publication bias was assessed using the Egger test. Data was analyzed using STATA (version 14) software. Results: The initial literature search yielded 145 studies. After excluding duplicates and assessing the quality of the studies, 7 studies were selected for meta-analysis. The results showed that the overall prevalence of PTSD among HCWs during the COVID-19 pandemic was 13.52% (95% CI: 9.06-17.98, I 2 = 65.5%, p = 0.008). Conclusion: There is a high prevalence of PTSD among frontline HCWs during the COVID-19 pandemic. It is important to invest in efforts to screen HCWs for mental health disorders such as PTSD and provide them with mental health support.

Analysis of Psychosocial Adjustment in the Family During Confinement: Problems and Habits of Children and Youth and Parental Stress and Resilience.

The COVID-19 health crisis has led to a dramatic change in dynamics and habits of families, which may be a factor involved in the development and maintenance of problems and difficulties in children. The present study is a cross-sectional study that aims to describe and analyze the relationship between the difficulties in psychological adjustment and the change of habits of the infant-juvenile population as perceived by their parents and their stress and resilience during the total confinement of the first wave of the COVID-19 pandemic in Spain, as well as analyzing the course of the changes and the relationships between weeks 3 to 6, that is, the score of different participants in each week of the confinement. The sample is comprised of 883 parents of children and adolescents between 3 and 18 years of age. Children's psychological adjustment, children's habits, parental stress, and parental resilience were assessed by parents. The results show that parents perceive a change in the habits and psychological difficulties in their children. At the same time, our results describe parents with a high level of stress and resilience, with differences depending on the children's ages. The time of confinement accentuates the perception of parents about the psychological difficulties of their children and parental stress, as well as a decrease in resilience. These difficulties are reduced when the parent has resilience competencies. These results show that the resilience of parents mediate the relationship between parental stress and psychological problems of their children. These results shows that COVID-19 lockdown had a considerable effect on families, both on children and parents. Some practical implications based on results are provided.

Investigating the Effective Factors of Using Personal Protective Equipment from the Perspective of Nurses Caring for COVID-19 Patients: A Cross-Sectional Study.

Considering the importance of appropriate personal protective equipment (PPE) for preventing COVID-19 transmission, the aim of this study was to investigate the factors affecting the use of PPE from the perspective of the nurses caring for COVID-19 patients. This descriptive cross-sectional study surveyed 240 nurses working in the central COVID-19 hospitals of Arak, Iran. Nurses were enrolled in the study by a convenience sampling method. The data collection tool was a validated questionnaire. Data were analyzed by SPSS 16 software using descriptive statistics, analysis of variance (ANOVA), and independent sample t-test. Environmental (4.24 ± 0.45), personal (4.16 ± 0.42), and organizational (4.04 ± 0.50) factors all contribute significantly to nursing attitudes about PPE use (p < 0.05). The average score, combining all identified factors, was 4.15 ± 0.31. The most influential factor contributing to appropriate use of PPE was environmental, while the least impactful parameters were related to rules and regulations. Environmental factors have the greatest impact on the use of PPE from the perspective of the nurses caring for patients with COVID-19. Managers and healthcare organizations should provide appropriate and adequate PPE to nurses, educate them on proper use, and monitor the process to resolve barriers.

The prevalence of insomnia among health care workers amid the COVID-19 pandemic: An umbrella review of meta-analyses.

BACKGROUND: Health care workers (HCWs) during the COVID-19 pandemic experience numerous psychological problems, including stress and anxiety. These entities can affect their sleep quality and predispose them to insomnia. The aim of the present study was to investigate the prevalence of insomnia among HCWs during the COVID-19 crisis via an umbrella review. METHODS: The PRISMA guideline was used to conduct this review. By searching relevant keywords in databases of Scopus, PubMed, Web of Science, and Google Scholar, studies that reported the prevalence of insomnia among HCWs during the COVID-19 pandemic (January 2020 to the end of January 2021) and had been published in English were identified and evaluated. The random effects model was used for meta-analysis, and the I2 index was used to assess heterogeneity. The Egger test was used to determine publication bias. Based on the results of the primary search, 96 studies were identified, and ultimately 10 eligible studies entered the meta-analysis phase. RESULTS: The results of the umbrella review of meta-analyses showed that the prevalence of insomnia among HCWs during the COVID-19 pandemic was 36.36% (95% CI: 33.36-39.36, I2 = 59.6%, p = 0.006). CONCLUSIONS: The results of this umbrella review of meta-analyses showed a relatively high prevalence of insomnia among HCWs during the COVID-19 pandemic. As insomnia can be associated with other psychological problems, policymakers and health managers should regularly screen HCWs for psychological disorders as well as a possible tendency for suicide. Furthermore, by treating insomnia, one can reduce the incidence of these psychological disorders.

The effect of mandala colouring on anxiety in hospitalized COVID-19 patients: A randomized controlled clinical trial.

COVID-19 significantly affects patients' mental health, triggering a wide range of psychological disorders, including anxiety. The aim of this study was to investigate the effect of mandala colouring on the anxiety of hospitalized COVID-19 patients. In this randomized controlled clinical trial, 70 hospitalized patients with COVID-19 were randomly divided between the intervention and control groups. Standard care was provided for both groups. The intervention group spent 30 min/day for six consecutive days performing mandala colouring. Patient anxiety was measured prior and subsequent to the intervention in both groups using the Spielberger State-Trait Anxiety Inventory. Data were analysed using SPSS software version 25. The mean anxiety score was not significantly different between the two groups before the intervention (P = 0.08). Subsequent to the intervention, the mean anxiety score in the intervention and control groups was 44.05 ± 4.67 and 67.85 ± 6.25, respectively, indicating a statistically significant (P = 0.0001) decrease in the anxiety measured among the intervention group as compared with that of the control group. The results of this study show that 30 min of mandala colouring daily is an effective strategy for reducing anxiety in hospitalized COVID-19 patients. Mandala colouring can complement routine treatment and provides a non-pharmaceutical option for decreasing patient anxiety.

The Effect of Family Presence During Resuscitation and Invasive Procedures on Patients and Families: An Umbrella Review.

INTRODUCTION: The concept of family presence during resuscitation and invasive procedures is a controversial issue and has not been universally adopted by health care providers. Owing to the sheer number of studies in this field, we conducted this umbrella study to provide an overview of this concept with the aim of investigating the impact of family presence on patients, families, and resuscitation and invasive procedures. METHODS: In this review, using the Joanna Briggs Institute levels of evidence umbrella methodology guidelines, the authors searched PubMed, Google Scholar, Embase, MEDLINE, Web of Science, Scopus, and the Cochrane database for systematic review and meta-analysis studies that evaluated the presence of family during resuscitation and invasive procedures without time limit until July 2020. The following key words were used for the search: family presence; family witness; parent presence; parent witness; and resuscitation. RESULTS: A total of 254 articles published between January 1967 and July 2020 were screened. Five articles (1 meta-analysis and 4 systematic reviews) met the inclusion criteria. The review showed that family presence during resuscitation or invasive procedures does not have negative effects on family members, patients, or the resuscitation or invasive intervention process. Family members focus on the patients, not the ongoing treatment. The presence of family members is beneficial for both family members and health care staff. None of the reviewed studies reported a negative effect on family members. DISCUSSION: The presence of parents and other immediate family members during resuscitation and invasive procedures has positive impacts on patients, families, and health care staff.

Differential gene expression profile between progressive and de novo muscle invasive bladder cancer and its prognostic implication.

This study aimed to ascertain gene expression profile differences between progressive muscle-invasive bladder cancer (MIBC) and de novo MIBC, and to identify prognostic biomarkers to improve patients' treatment. Retrospective multicenter study in which 212 MIBC patients who underwent radical cystectomy between 2000 and 2019 were included. Gene expression profiles were determined in 26 samples using Illumina microarrays. The expression levels of 94 genes were studied by quantitative PCR in an independent set of 186 MIBC patients. In a median follow-up of 16 months, 46.7% patients developed tumor progression after cystectomy. In our series, progressive MIBC patients show a worse tumor progression (p = 0.024) and cancer-specific survival (CSS) (p = 0.049) than the de novo group. A total of 480 genes were found to be differently expressed between both groups. Differential expression of 24 out of the 94 selected genes was found in an independent cohort. RBPMC2 and DSC3 were found as independent prognostic biomarkers of tumor progression and CALD1 and LCOR were identified as prognostic biomarkers of CSS between both groups. In conclusion, progressive and de novo MIBC patients show different clinical outcome and gene expression profiles. Gene expression patterns may contribute to predict high-risk of progression to distant metastasis or CSS.