Nuclear Pores Promote Lethal Prostate Cancer by Increasing POM121-Driven E2F1, MYC, and AR Nuclear Import.

Nuclear pore complexes (NPCs) regulate nuclear-cytoplasmic transport, transcription, and genome integrity in eukaryotic cells. However, their functional roles in cancer remain poorly understood. We interrogated the evolutionary transcriptomic landscape of NPC components, nucleoporins (Nups), from primary to advanced metastatic human prostate cancer (PC). Focused loss-of-function genetic screen of top-upregulated Nups in aggressive PC models identified POM121 as a key contributor to PC aggressiveness. Mechanistically, POM121 promoted PC progression by enhancing importin-dependent nuclear transport of key oncogenic (E2F1, MYC) and PC-specific (AR-GATA2) transcription factors, uncovering a pharmacologically targetable axis that, when inhibited, decreased tumor growth, restored standard therapy efficacy, and improved survival in patient-derived pre-clinical models. Our studies molecularly establish a role of NPCs in PC progression and give a rationale for NPC-regulated nuclear import targeting as a therapeutic strategy for lethal PC. These findings may have implications for understanding how NPC deregulation contributes to the pathogenesis of other tumor types.

Pharmacokinetics of cefiderocol during extracorporeal membrane oxygenation: A case report.

Patients with extracorporeal membrane oxygenation (ECMO) support do frequently receive broad-spectrum antibiotics, due to the high frequency of infection by multidrug resistant microorganisms. The extracorporeal circuit can alter the pharmacokinetics (PK) of administered drugs, and in the case of antibiotics this may lead to treatment failure. Cefiderocol is a new cephalosporin that exhibits excellent in vitro activity against many multidrug-resistant (MDR) microorganisms, but there is no published data about the modifications of its PK in patients with ECMO support. Herein we report the results of a pharmacokinetic investigation of cefiderocol in a critically ill patient receiving extracorporeal respiratory support.

sFlt-1/PlGF Ratio at 24 Weeks Gestation in Twin Pregnancies as a Predictor of Preeclampsia or Fetal Growth Restriction.

INTRODUCTION: The objective was to elucidate if the sFlt-1/PlGF ratio at 24 weeks in twin pregnancies could be useful to select patients who subsequently develop diseases related to placental dysfunction, such as preeclampsia or fetal growth restriction (FGR). METHODS: This was a prospective study among all twin pregnancies followed up at a tertiary hospital. The sFlt-1/PlGF ratio was determined at 24 weeks. RESULTS: A total of 108 patients with a twin gestation were included. Pregnant women who developed preeclampsia and/or FGR displayed a significantly higher sFlt-1/PlGF ratio at 24 weeks, compared to those who did not develop these diseases (20.3 vs. 4.3, p = 0.002). The mean sFlt-1/PlGF ratio was not significantly different between patients who subsequently developed preeclampsia compared with those that developed FGR (29.8 vs. 18.45, p = 0.42). A sFlt-1/PlGF ratio ≥17 at 24 weeks is associated with a significant increase in the frequency of preeclampsia (odds ratio, 37.13 [95% confidence interval, 4.78-288.25]; p = 0.002), and FGR (odds ratio, 39.58 [95% confidence interval, 6.31-248.17]; p < 0.001). The addition of maternal characteristics and mean pulsatility index of the uterine arteries to the sFlt-1/PlGF ratio at 24 weeks enhances the identification of patients who develop preeclampsia or FGR. CONCLUSION: The sFlt-1/PlGF ratio at 24 weeks in twin pregnancies, combined with the mean pulsatility index of the uterine arteries and maternal characteristics, could select patients who develop preeclampsia or FGR. These patients might benefit from a close follow-up in order to avoid maternal-fetal adverse outcomes.

Urine Extracellular Vesicle GATA2 mRNA Discriminates Biopsy Result in Men with Suspicion of Prostate Cancer.

PURPOSE: Prostate specific antigen has limited performance in detecting prostate cancer. The transcription factor GATA2 is expressed in aggressive prostate cancer. We analyzed the predictive value of urine extracellular vesicle GATA2 mRNA alone and in combination with a multigene panel to improve detection of prostate cancer and high risk disease. MATERIALS AND METHODS: GATA2 mRNA was analyzed in matched extracellular vesicles isolated from urines before and after prostatectomy (16) and paired urine and tissue prostatectomy samples (19). Extracellular vesicle GATA2 mRNA performance to distinguish prostate cancer and high grade disease was tested in training (52) and validation (165) cohorts. The predictive value of a multigene score including GATA2, PCA3 and TMPRSS2-ERG (GAPT-E) was tested in both cohorts. RESULTS: Confirming its prostate origin, urine extracellular vesicle GATA2 mRNA levels decreased significantly after prostatectomy and correlated with prostate cancer tissue GATA2 mRNA levels. In the training and validation cohort GATA2 discriminated prostate cancer (AUC 0.74 and 0.66) and high grade disease (AUC 0.78 and 0.65), respectively. Notably, the GAPT-E score improved discrimination of prostate cancer (AUC 0.84 and 0.72) and high grade cancer (AUC 0.85 and 0.71) in both cohorts when compared with each biomarker alone and PT-E (PCA3 and TMPRSS2-ERG). A GAPT-E score for high grade prostate cancer would avoid 92.1% of unnecessary prostate biopsies, compared to 61.9% when a PT-E score is used. CONCLUSIONS: Urine extracellular vesicle GATA2 mRNA analysis improves the detection of high risk prostate cancer and may reduce the number of unnecessary biopsies.

Tumour initiating cells and IGF/FGF signalling contribute to sorafenib resistance in hepatocellular carcinoma.

OBJECTIVE: Sorafenib is effective in hepatocellular carcinoma (HCC), but patients ultimately present disease progression. Molecular mechanisms underlying acquired resistance are still unknown. Herein, we characterise the role of tumour-initiating cells (T-ICs) and signalling pathways involved in sorafenib resistance. DESIGN: HCC xenograft mice treated with sorafenib (n=22) were explored for responsiveness (n=5) and acquired resistance (n=17). Mechanism of acquired resistance were assessed by: (1) role of T-ICs by in vitro sphere formation and in vivo tumourigenesis assays using NOD/SCID mice, (2) activation of alternative signalling pathways and (3) efficacy of anti-FGF and anti-IGF drugs in experimental models. Gene expression (microarray, quantitative real-time PCR (qRT-PCR)) and protein analyses (immunohistochemistry, western blot) were conducted. A novel gene signature of sorafenib resistance was generated and tested in two independent cohorts. RESULTS: Sorafenib-acquired resistant tumours showed significant enrichment of T-ICs (164 cells needed to create a tumour) versus sorafenib-sensitive tumours (13 400 cells) and non-treated tumours (1292 cells), p<0.001. Tumours with sorafenib-acquired resistance were enriched with insulin-like growth factor (IGF) and fibroblast growth factor (FGF) signalling cascades (false discovery rate (FDR)<0.05). In vitro, cells derived from sorafenib-acquired resistant tumours and two sorafenib-resistant HCC cell lines were responsive to IGF or FGF inhibition. In vivo, FGF blockade delayed tumour growth and improved survival in sorafenib-resistant tumours. A sorafenib-resistance 175 gene signature was characterised by enrichment of progenitor cell features, aggressive tumorous traits and predicted poor survival in two cohorts (n=442 patients with HCC). CONCLUSIONS: Acquired resistance to sorafenib is driven by T-ICs with enrichment of progenitor markers and activation of IGF and FGF signalling. Inhibition of these pathways would benefit a subset of patients after sorafenib progression.

Attention to emotion through a go/no-go task in children with oppositionality and callous-unemotional traits.

BACKGROUND: There is debate about whether the difficulties that children with different degrees of oppositionality (ODD) and callous-unemotional traits (CU) have in processing emotions are global or specific. The aim of this study is to identify difficulties in recognizing emotion (happiness, anger, sadness and fear) through a go/no-go task in children with different levels of ODD and CU traits. METHOD: A total of 320 8-year-old children were assessed through questionnaires filled out by teachers about oppositional defiant symptoms and CU traits and were then distributed into four groups: LowCU-HighODD, HighCU-LowODD, HighCU-HighODD and a comparison group (LowCU-LowODD). RESULTS: The analyses of variance comparing the 4 groups showed that the two groups with high ODD were less accurate than the control group in recognizing the emotion when the stimuli expressed happiness, fear or neutral emotion. The HighCU-HighODD group differed in the quality of the response (correct/wrong responses) but not in the reaction time in relation to the comparison group. The LowCU-HighODD group was faster to respond to emotions than the comparison group. IMPLICATIONS: The results show that the deficit in emotion processing is not restricted to specific distressing emotions such as fear or sadness, but they point to a global impairment in emotion processing in children scoring high in the constructs studied. The results also suggest that the difficulties that children with combined CU traits and oppositional conduct problems have in processing emotions are more of an emotional rather than an attentional nature.

Enrichment of Cells with Cancer Stem Cell-Like Markers in Relapses of Chemoresistant Patients with Locally Advanced Head and Neck Squamous Cell Carcinoma.

BACKGROUND: Patients with head and neck squamous cell carcinoma (HNSCC) present different responses to chemotherapy and radiotherapy. One explanation may be the differences in the individual rates of stem cell-like cells. METHODS: We included patients with HNSCC and tumor progression or relapse. Tumor samples were obtained before and after primary chemotherapy, and immunohistochemical analyses were performed for CD44, HLA class I (HLA-I), pancytokeratin, and phosphorylated epidermal growth factor receptor (p-EGFR). Differences in expression between the first and second specimens were assessed. RESULTS: Expression between the first and second specimens varied as follows: CD44 increased by 14.67% (95% confidence interval, CI: 6.94 to 22.40; p < 0.01); HLA-I decreased by 16.72% (95% CI: -23.87 to -9.47; p < 0.01); pancytokeratin decreased by 24.91% (95% CI: -32.8 to -17.7; p < 0.01), and p-EFGR expression decreased by 12.30% (95% CI: -20.61 to -3.98; p < 0.005). CONCLUSIONS: Among patients with HNSCC, there is an enrichment of cells with stem-like markers in relapsed tumors when compared with the primary tumor. This finding should be considered when developing treatment strategies.

The discriminative capacity of CBCL/1½-5-DSM5 scales to identify disruptive and internalizing disorders in preschool children.

This paper studies the discriminative capacity of CBCL/1½-5 (Manual for the ASEBA Preschool-Age Forms & Profiles, University of Vermont, Research Center for Children, Youth, & Families, Burlington, 2000) DSM5 scales attention deficit and hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), anxiety and depressive problems for detecting the presence of DSM5 (DSM5 diagnostic and statistical manual of mental disorders, APA, Arlington, 2013) disorders, ADHD, ODD, Anxiety and Mood disorders, assessed through diagnostic interview, in children aged 3-5. Additionally, we compare the clinical utility of the CBCL/1½-5-DSM5 scales with respect to analogous CBCL/1½-5 syndrome scales. A large community sample of 616 preschool children was longitudinally assessed for the stated age group. Statistical analysis was based on ROC procedures and binary logistic regressions. ADHD and ODD CBCL/1½-5-DSM5 scales achieved good discriminative ability to identify ADHD and ODD interview's diagnoses, at any age. CBCL/1½-5-DSM5 Anxiety scale discriminative capacity was fair for unspecific anxiety disorders in all age groups. CBCL/1½-5-DSM5 depressive problems' scale showed the poorest discriminative capacity for mood disorders (including depressive episode with insufficient symptoms), oscillating into the poor-to-fair range. As a whole, DSM5-oriented scales generally did not provide evidence better for discriminative capacity than syndrome scales in identifying DSM5 diagnoses. CBCL/1½-5-DSM5 scales discriminate externalizing disorders better than internalizing disorders for ages 3-5. Scores on the ADHD and ODD CBCL/1½-5-DSM5 scales can be used to screen for DSM5 ADHD and ODD disorders in general populations of preschool children.

A common MicroRNA signature consisting of miR-133a, miR-139-3p, and miR-142-3p clusters bladder carcinoma in situ with normal umbrella cells.

miRNAs are small noncoding RNAs with critical roles in a large variety of biological processes such as development and tumorigenesis. miRNA expression profiling has been reported to be a powerful tool to classify tissue samples, including cancers, based on their developmental lineage. In this study, we have profiled the expression of miRNAs in bladder carcinoma in situ (CIS) and distinct cell compartments of the normal bladder, namely umbrella and basal-intermediate urothelial cells, as well as the muscularis propria. We identified several miRNAs differentially expressed between umbrella and basal-intermediate cells (miR-133a, miR-139-3p, miR-142-3p, miR-199b-5p, and miR-221). In situ hybridization confirmed the expression of miR-133a and miR-139-3p in umbrella cells, and miR-142-3p in basal-intermediate cells. Strikingly, miRNA expression levels of CIS most closely resembled the miRNA profile of umbrella cells. Finally, we examined well-established umbrella and basal-intermediate cell immunohistochemical biomarkers in an independent series of CIS samples. Again, this analysis revealed the significant expression of umbrella-specific markers in CIS when compared to non-CIS lesions. Overall, our studies represent a comprehensive and accurate description of the different miRNAs expressed in CIS tumors and three distinct histological areas of the urinary bladder. Notably, this study provides evidence of the possible origin relationship between CIS and normal umbrella cells.

Prevalence of DSM-IV disorders, comorbidity and impairment in 3-year-old Spanish preschoolers.

PURPOSE: The goal is to examine the prevalence, comorbidity and impairment of DSM-IV disorders in 3-year-old children from the Spanish general population. METHOD: A sample of 1,341 3-year-old preschoolers were randomly selected and screened for a double-phase design. In total, 622 families were assessed with a diagnostic semi-structured interview and functional impairment measures. RESULTS: Prevalence of any diagnosis was 29.9%, the most prevalent disorders being primary insomnia (11.7%) and oppositional defiant disorder (ODD) (6.9%). There were no sex differences in the prevalence. One-third of the families had sought professional help for the child's symptoms, and 9.4% received treatment (4.4% psychological and 2.1% pharmacological). After controlling for other comorbidities, ADHD was significantly associated with ODD, CD, insomnia and social phobia; ODD was associated with CD, separation anxiety, specific phobia and major depression. Diagnostic categories were associated with impairment, family burden, seeking professional help and receiving treatment. A diagnosis was more frequent in children of low socioeconomic status, born outside Spain, from one-parent families, with younger parents and with parents of lower educational level. CONCLUSIONS: Psychopathology, comorbidity and associated factors are very frequent from age three, suggesting a need for efforts of detection, prevention and treatment in the different societies.

The yin and yang of chromosomal instability in prostate cancer.

Metastatic prostate cancer remains an incurable lethal disease. Studies indicate that prostate cancer accumulates genomic changes during disease progression and displays the highest levels of chromosomal instability (CIN) across all types of metastatic tumours. CIN, which refers to ongoing chromosomal DNA gain or loss during mitosis, and derived aneuploidy, are known to be associated with increased tumour heterogeneity, metastasis and therapy resistance in many tumour types. Paradoxically, high CIN levels are also proposed to be detrimental to tumour cell survival, suggesting that cancer cells must develop adaptive mechanisms to ensure their survival. In the context of prostate cancer, studies indicate that CIN has a key role in disease progression and might also offer a therapeutic vulnerability that can be pharmacologically targeted. Thus, a comprehensive evaluation of the causes and consequences of CIN in prostate cancer, its contribution to aggressive advanced disease and a better understanding of the acquired CIN tolerance mechanisms can translate into new tumour classifications, biomarker development and therapeutic strategies.

Psychometric properties of the Strengths and Difficulties Questionnaire(3-4) in 3-year-old preschoolers.

We provide the first validation data on the Strengths and Difficulties Questionnaire (SDQ(3-4)), a brief screening tool for behavioral and emotional problems, in preschool children. Parents of a community sample of 1341 Spanish 3-year-olds and teachers of a sample of 622 children responded to the SDQ(3-4) and different measures of psychopathology. Confirmatory factor analysis yielded adequate fit of the model to the original structure. Internal consistency (omega coefficient) for total scores was .87 for parents and .91 for teachers. Convergent validity of SDQ(3-4)-parents' reports with Achenbach's taxonomy and diagnostic interview was good, but low for SDQ(3-4)-teachers' reports. The SDQ(3-4) showed predictive accuracy for discriminating use of mental health services and functional impairment. This is the first work presenting empirical evidence of the reliability and validity of the parents' and teachers' SDQ(3-4) for preschoolers. The SDQ(3-4) presents acceptable psychometric properties for use in the identification of preschool children who might have behavioral or emotional problems.

Advances in the management of muscle-invasive bladder cancer through risk prediction, risk communication, and novel treatment approaches.

Although level I evidence supports the use of neoadjuvant cisplatin-based chemotherapy followed by radical cystectomy for the management of patients with muscle-invasive bladder cancer (MIBC), these treatment modalities are utilized in only a subset of patients. The reasons for lack of implementation of these treatment standards are multiple; patients may be considered ineligible for cisplatin or too old for safe cystectomy. Better means of determining a patient's probability of recurrence with surgery alone, or likelihood of benefit with neoadjuvant chemotherapy, are clearly needed. Models have been developed to individualize estimates of non-organ-confined disease based on pretreatment variables. It is critical that clinicians are able to effectively communicate complex risk-related data to patients to facilitate a shared medical decision.

Inventory of callous-unemotional traits in a community sample of preschoolers.

The purpose of this study was to test the factor structure of the Inventory of Callous-Unemotional Traits (ICU; Frick, 2004 ) and to study the relation between the derived dimensions and external variables in a community sample of preschool children. A total of 622 children 3 and 4 years of age were assessed with a semistructured diagnostic interview, the ICU, and other questionnaires on psychopathology, temperament, and executive functioning, completed by parents and teachers. Confirmatory factor analysis derived from teachers' ICU responses yielded three dimensions: Callousness, Uncaring, and Unemotional. Callousness and Uncaring subscale scores correlated with the specific scales related to aggressive behavior, temperament, executive functioning, and conduct problems. The ICU scale scores discriminated cross-sectionally oppositional defiant disorder (ODD) and conduct disorder (CD) diagnoses, aggressive and nonaggressive symptoms of CD, use of services, and ODD/CD-related family burden. Longitudinally, Callousness subscale score at age 3 predicted ODD or CD diagnosis at age 4. Unemotional was not associated with aggressive measures, but it was linked to anxiety disorders cross-sectionally and longitudinally. Callous-Unemotional traits contributed significantly to predicting disruptive behavior disorders controlling for sex, temperament, and executive functioning (predictive accuracy between 3 and 5%). The ICU is a promising questionnaire for identifying early Callous and Uncaring traits in preschool years that may help in the identification of a subset of preschool children who might have severe behavioral problems.

Measuring the popularity of football players with Google Trends.

Google Trends is a valuable tool for measuring popularity since it collects a large amount of information related to Google searches. However, Google Trends has been underused by sports analysts. This research proposes a novel method to calculate several popularity indicators for predicting players' market value. Google Trends was used to calculate six popularity indicators by requesting information about two football players simultaneously and creating popularity layers to compare players of unequal popularity. In addition, as the main idea is to obtain the popularity indicators of all players on the same scale, a cumulative conversion factor was used to rescale these indicators. The results show that the proposed popularity indicators are essential to predicting a player's market value. In addition, using the proposed popularity indicators decreases the transfer fee prediction error for three different models that are fitted to the data using the multiple linear regression, random forest, and gradient boosting machine methods. The popularity indicator Min, which is a robust reflection of the popularity that represents a player's popularity during the periods when they are less popular, is the most important popularity indicator, with a significant effect on the market value. This research provides practical guidance for developing and incorporating the proposed indicators, which could be applied in sports analytics and in any study in which popularity is relevant.

Perinatal Outcomes of Monochorionic Twin Pregnancies Conceived Naturally Versus through Assisted Reproductive Techniques.

Objective: It has been reported that monochorionic twin pregnancies conceived through assisted reproductive techniques (ART) display a higher risk of second-trimester miscarriage, cesarean delivery, and neonatal death than those conceived naturally. The aim of this study was to compare the perinatal outcomes of monochorionic diamniotic (MCDA) twin pregnancies conceived naturally and through ART in a tertiary hospital. Methods: This was a retrospective cohort study of all MCDA twin pregnancies that received obstetric care and delivered at La Fe University and Polytechnic Hospital between 2015 and 2021. MCDA pregnancies that were referred to the tertiary hospital for specialized management, follow-up, and delivery were also included. The study was approved by The Health Research Institute Hospital La Fe (IIS La Fe). Results: Among the 184 MCDA pregnancies, 149 (81%) had a natural conception, and 35 (19%) were conceived through ART. Patients with an MCDA pregnancy who conceived through ART had a significantly older maternal age (38.0 [35.5-42.5] vs. 32.0 [29.0-36.0], p < 0.001) and an elevated rate of nulliparity (80.0% vs. 50.3%, p = 0.001). Regarding pregnancy complications, MCDA pregnancies through ART were associated with a significantly higher incidence of gestational diabetes (22.9% vs. 2.7%, p < 0.001), hypertensive disorders during pregnancy (22.9% vs. 9.4%, p = 0.04), and other pregnancy complications such as threatened labor or preterm prelabor rupture of membranes (14.3% vs. 36.2%, p = 0.015), than naturally conceived MCDA pregnancies. No differences were found in the incidence of twin-to-twin transfusion syndrome (20% vs. 33.6%, p = 0.155). MCDA pregnancies through natural conception had a greater rate of vaginal delivery than MCDA through ART (16.8% vs. 2.9%, p = 0.032). When adjusted for confounding factors, MCDA pregnancies through ART were only more likely to develop gestational diabetes than those naturally conceived (aOR 7.86, 95% CI 1.55-39.87). No differences were found regarding neonatal outcomes between groups. Conclusions: Compared with naturally conceived MCDA twin pregnancies, those conceived through ART displayed a significantly higher risk of developing gestational diabetes. No differences regarding other pregnancy complications, mode of delivery, or neonatal outcomes were found between groups.

Harnessing transcriptionally driven chromosomal instability adaptation to target therapy-refractory lethal prostate cancer.

Metastatic prostate cancer (PCa) inevitably acquires resistance to standard therapy preceding lethality. Here, we unveil a chromosomal instability (CIN) tolerance mechanism as a therapeutic vulnerability of therapy-refractory lethal PCa. Through genomic and transcriptomic analysis of patient datasets, we find that castration and chemotherapy-resistant tumors display the highest CIN and mitotic kinase levels. Functional genomics screening coupled with quantitative phosphoproteomics identify MASTL kinase as a survival vulnerability specific of chemotherapy-resistant PCa cells. Mechanistically, MASTL upregulation is driven by transcriptional rewiring mechanisms involving the non-canonical transcription factors androgen receptor splice variant 7 and E2F7 in a circuitry that restrains deleterious CIN and prevents cell death selectively in metastatic therapy-resistant PCa cells. Notably, MASTL pharmacological inhibition re-sensitizes tumors to standard therapy and improves survival of pre-clinical models. These results uncover a targetable mechanism promoting high CIN adaptation and survival of lethal PCa.

Master Transcription Factor Reprogramming Unleashes Selective Translation Promoting Castration Resistance and Immune Evasion in Lethal Prostate Cancer.

Signaling rewiring allows tumors to survive therapy. Here we show that the decrease of the master regulator microphthalmia transcription factor (MITF) in lethal prostate cancer unleashes eukaryotic initiation factor 3B (eIF3B)-dependent translation reprogramming of key mRNAs conferring resistance to androgen deprivation therapy (ADT) and promoting immune evasion. Mechanistically, MITF represses through direct promoter binding eIF3B, which in turn regulates the translation of specific mRNAs. Genome-wide eIF3B enhanced cross-linking immunoprecipitation sequencing (eCLIP-seq) showed specialized binding to a UC-rich motif present in subsets of 5' untranslated regions. Indeed, translation of the androgen receptor and major histocompatibility complex I (MHC-I) through this motif is sensitive to eIF3B amount. Notably, pharmacologic targeting of eIF3B-dependent translation in preclinical models sensitizes prostate cancer to ADT and anti-PD-1 therapy. These findings uncover a hidden connection between transcriptional and translational rewiring promoting therapy-refractory lethal prostate cancer and provide a druggable mechanism that may transcend into effective combined therapeutic strategies. SIGNIFICANCE: Our study shows that specialized eIF3B-dependent translation of specific mRNAs released upon downregulation of the master transcription factor MITF confers castration resistance and immune evasion in lethal prostate cancer. Pharmacologic targeting of this mechanism delays castration resistance and increases immune-checkpoint efficacy. This article is featured in Selected Articles from This Issue, p. 2489.

Distinct expression profiles of p63 variants during urothelial development and bladder cancer progression.

The TP63 gene, a member of the TP53 tumor suppressor gene family, can be expressed as at least six isoforms due to alternative promoter use and alternative splicing. The lack of p63 isoform-specific antibodies has limited the analysis of the biological significance of p63. We report a novel set of well-defined antibodies to examine p63 isoforms in mouse and human urothelium during embryogenesis and tumor progression, respectively. We provide evidence that basal and intermediate urothelial cells express p63 isoforms, with the TAp63 variant the first to be detected during development, whereas umbrella cells are characterized by a p63-negative phenotype. Notably, we report that p63-null mice develop a bladder with an abnormal urothelium, constituted by a single layer of cells that express uroplakin II and low molecular weight cytokeratins, consistent with an umbrella cell phenotype. Finally, analysis of 202 human bladder carcinomas revealed a new categorization of invasive tumors into basal-like (positive for ΔNp63 and high molecular weight cytokeratins and negative for low molecular weight cytokeratins) versus luminal-like (negative for ΔNp63 and high molecular weight cytokeratins and positive for low molecular weight cytokeratins) phenotypes, with ΔNp63 expression associated with an aggressive clinical course and poor prognosis. This study highlights the relevance of p63 isoforms in both urothelial development and bladder carcinoma progression, with ΔNp63 acting as an oncogene in certain invasive bladder tumors.

Dimensions of oppositional defiant disorder in 3-year-old preschoolers.

BACKGROUND: To test the factor structure of oppositional defiant disorder (ODD) symptoms and to study the relationships between the proposed dimensions and external variables in a community sample of preschool children. METHOD: A sample of 1,341 3-year-old preschoolers was randomly selected and screened for a double-phase design. In total, 622 families were assessed with a diagnostic semi-structured interview and questionnaires on psychopathology, temperament and executive functioning completed by parents and teachers. RESULTS: Using categorical and dimensional symptoms of ODD it was possible to confirm, cross-informant and cross-method, distinct dimensions for defining the structure of ODD: one made up of irritable and headstrong and the other of negative affect, oppositional behaviour and antagonistic behaviour. Specific associations with DSM-IV disorders were found, and irritable was associated with anxiety disorders, whereas headstrong was associated with disruptive disorders, including aggressive and non-aggressive CD symptoms. Also, negative affect was associated with anxiety disorders and non-aggressive CD symptoms, oppositional behaviour with disruptive disorders and aggressive CD symptoms, and antagonistic behaviours with disruptive disorders and, in boys, with mood disorders. The dimensions correlated with specific scales of psychopathology, temperament and executive functioning. CONCLUSIONS: Oppositional defiant disorder is a heterogeneous disorder from preschool age. Different dimensions, with moderate to acceptable reliability and convergent and discriminant validity with other psychological constructs, can be identified early in life.