NR2F1 regulates regional progenitor dynamics in the mouse neocortex and cortical gyrification in BBSOAS patients.

The relationships between impaired cortical development and consequent malformations in neurodevelopmental disorders, as well as the genes implicated in these processes, are not fully elucidated to date. In this study, we report six novel cases of patients affected by BBSOAS (Boonstra-Bosch-Schaff optic atrophy syndrome), a newly emerging rare neurodevelopmental disorder, caused by loss-of-function mutations of the transcriptional regulator NR2F1. Young patients with NR2F1 haploinsufficiency display mild to moderate intellectual disability and show reproducible polymicrogyria-like brain malformations in the parietal and occipital cortex. Using a recently established BBSOAS mouse model, we found that Nr2f1 regionally controls long-term self-renewal of neural progenitor cells via modulation of cell cycle genes and key cortical development master genes, such as Pax6. In the human fetal cortex, distinct NR2F1 expression levels encompass gyri and sulci and correlate with local degrees of neurogenic activity. In addition, reduced NR2F1 levels in cerebral organoids affect neurogenesis and PAX6 expression. We propose NR2F1 as an area-specific regulator of mouse and human brain morphology and a novel causative gene of abnormal gyrification.

Sequential Multilocus Repetitive Transcranial Magnetic Stimulation for Treatment of Tinnitus With and Without Comorbid Major Depressive Disorder.

OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) is a promising treatment for tinnitus, although outcomes are highly variable. We previously described a multilocus sequential rTMS treatment protocol for tinnitus involving stimulation of both prefrontal and auditory targets. In this study, we report results using this approach in an open-label treatment study of tinnitus with and without comorbid major depressive disorder (MDD). MATERIALS AND METHODS: Forty patients with chronic tinnitus (mean age 56 years, ten female) and with (n = 17) or without (n = 23) MDD received multilocus rTMS administered sequentially to 1) left dorsolateral prefrontal cortex, followed by 2) auditory cortex (Heschel's gyrus). Patients completed weekly self-report ratings using the Tinnitus Functional Index (TFI) and Tinnitus Handicap Inventory, and patients with MDD completed the Inventory of Depressive Symptomatology Self-Report 30-item. RESULTS: Patients showed significant mean improvement in tinnitus at sessions 5 (mean TFI improvement 6.8 points ± 12.2, p = 0.002) and 10 (mean improvement 9.2 points ± 14.1, p = 0.002), with 48% of patients responding within ten treatment sessions. Responders were significantly older than nonresponders (61.5 ± 15 years vs 51.3 ± 16 years), and there was a trend toward decreased likelihood of response in subjects with comorbid MDD compared with subjects without comorbidity (odds ratio = 0.28, p = 0.06). Patients with comorbid MDD reported significantly less improvement after ten sessions than did those with tinnitus alone (4.3 ± 10.3 vs 14.7 ± 15.0 points, p = 0.04). Post hoc analyses suggested that the comorbid group achieved improvement comparable to that of the tinnitus-only group after 30 treatments. CONCLUSIONS: Patients showed significant improvement in tinnitus from multilocus sequential rTMS treatment, and those with tinnitus alone improved more quickly. Those with depression who continued rTMS through a full 30-session course further improved, indicating that tinnitus with comorbid MDD may respond with extended treatment.

Prediction of Tinnitus Treatment Outcomes Based on EEG Sensors and TFI Score Using Deep Learning.

Tinnitus is a hearing disorder that is characterized by the perception of sounds in the absence of an external source. Currently, there is no pharmaceutical cure for tinnitus, however, multiple therapies and interventions have been developed that improve or control associated distress and anxiety. We propose a new Artificial Intelligence (AI) algorithm as a digital prognostic health system that models electroencephalographic (EEG) data in order to predict patients' responses to tinnitus therapies. The EEG data was collected from patients prior to treatment and 3-months following a sound-based therapy. Feature selection techniques were utilised to identify predictive EEG variables with the best accuracy. The patients' EEG features from both the frequency and functional connectivity domains were entered as inputs that carry knowledge extracted from EEG into AI algorithms for training and predicting therapy outcomes. The AI models differentiated the patients' outcomes into either therapy responder or non-responder, as defined by their Tinnitus Functional Index (TFI) scores, with accuracies ranging from 98%-100%. Our findings demonstrate the potential use of AI, including deep learning, for predicting therapy outcomes in tinnitus. The research suggests an optimal configuration of the EEG sensors that are involved in measuring brain functional changes in response to tinnitus treatments. It identified which EEG electrodes are the most informative sensors and how the EEG frequency and functional connectivity can better classify patients into the responder and non-responder groups. This has potential for real-time monitoring of patient therapy outcomes at home.

Time Course of Priming Effect of TF Inducers on Synergistic TF Expression and Intra-Cellular Gap Formation of Human Vascular Endothelial Cells via the Extrinsic Coagulation Cascade.

Chronic spontaneous urticaria (CSU) is characterized by daily recurring wheal and flare with itch for more than 6 weeks. The extrinsic coagulation system has been shown to be activated in correlation with CSU severity. We have reported that tissue factor (TF), a trigger of the extrinsic coagulation cascade, is synergistically expressed on vascular endothelial cells by simultaneous stimulation with TF inducers (TFI), followed by activation of the extrinsic coagulation cascade and hyper permeability in vitro. However, vascular endothelial cells are not likely to be simultaneously stimulated by multiple TFIs under physiological conditions. Therefore, in order to know whether sequential, rather than simultaneous, stimuli with interval may induce synergistic activation of TF, we investigated the time course of the priming effects of each TFI for synergistic TF expression in vascular endothelial cells (HUVECs). We stimulated HUVECs with a TFI (first stimulation) and then stimulated cells with another TFI at indicated time points (second stimulation) and detected TF expression and activity. The TF expression induced by simultaneous stimulation diminished in a few hours. However, both synergistic enhancement of TF expression and activation level of the coagulation cascade were detected even when the second stimulation was added 18 or 22 h after the first stimulation. Thus, the priming effect of TFI for synergistic TF expression may persist for a half day or longer.

Preconditioned water-fat total field inversion: Application to spine quantitative susceptibility mapping.

PURPOSE: To (a) develop a preconditioned water-fat total field inversion (wfTFI) algorithm that directly estimates the susceptibility map from complex multi-echo gradient echo data for water-fat regions and to (b) evaluate the performance of the proposed wfTFI quantitative susceptibility mapping (QSM) method in comparison with a local field inversion (LFI) method and a linear total field inversion (TFI) method in the spine. METHODS: Numerical simulations and in vivo spine multi-echo gradient echo measurements were performed to compare wfTFI to an algorithm based on disjoint background field removal (BFR) and LFI and to a formerly proposed TFI algorithm. The data from 1 healthy volunteer and 10 patients with metastatic bone disease were included in the analysis. Clinical routine computed tomography (CT) images were used as a reference standard to distinguish osteoblastic from osteolytic changes. The ability of the QSM methods to distinguish osteoblastic from osteolytic changes was evaluated. RESULTS: The proposed wfTFI method was able to decrease the normalized root mean square error compared to the LFI and TFI methods in the simulation. The in vivo wfTFI susceptibility maps showed reduced BFR artifacts, noise amplification, and streaking artifacts compared to the LFI and TFI maps. wfTFI provided a significantly higher diagnostic confidence in differentiating osteolytic and osteoblastic lesions in the spine compared to the LFI method (p = .012). CONCLUSION: The proposed wfTFI method can minimize BFR artifacts, noise amplification, and streaking artifacts in water-fat regions and can thus better differentiate between osteoblastic and osteolytic changes in patients with metastatic disease compared to LFI and the original TFI method.

[German-language versions of the Tinnitus Functional Index : Comparison of the two validated German-language versions of the Tinnitus Functional Index for Switzerland and Germany]. / Deutschsprachige Versionen des Tinnitus Functional Index : Vergleich der beiden validierten deutschsprachigen Versionen des Tinnitus Functional Index für die Schweiz und für Deutschland.

BACKGROUND: Two validated German-language versions of the Tinnitus Functional Index (TFI) exist, one for Switzerland and one for Germany. The TFI is considered to be a possible new standard questionnaire for evaluation of tinnitus severity and tinnitus treatment. OBJECTIVE: Considering the standardization taking place in tinnitus evaluation, our aim was to compare the two German-language TFI versions and to recommend only one TFI version in the German-speaking area. MATERIALS AND METHODS: The two German-language TFI versions were compared in a multicenter and randomized online questionnaire study with a crossover design. RESULTS: The total score of the two TFI versions did not differ in the total population. However, when further divided in terms of population and order of presentation of the TFI versions, there were significant differences in some cases, albeit with only moderate effect sizes. This suggests that the two versions are slightly different but still comparable. In factor analysis for the TFI version for Germany, in the entire population as well as in the Swiss population, six factors could be extracted. In contrast, for the German and Swiss TFI versions, only five factors could be extracted in the German population, and for the Swiss TFI version, only five factors in the Swiss population. CONCLUSION: The two German-language versions of the TFI are well comparable with each other. However, the factor analysis rather argues for use of the TFI version for Germany in the entire German-speaking region.

COUP-TFI/Nr2f1 Orchestrates Intrinsic Neuronal Activity during Development of the Somatosensory Cortex.

The formation of functional cortical maps in the cerebral cortex results from a timely regulated interaction between intrinsic genetic mechanisms and electrical activity. To understand how transcriptional regulation influences network activity and neuronal excitability within the neocortex, we used mice deficient for Nr2f1 (also known as COUP-TFI), a key determinant of primary somatosensory (S1) area specification during development. We found that the cortical loss of Nr2f1 impacts on spontaneous network activity and synchronization of S1 cortex at perinatal stages. In addition, we observed alterations in the intrinsic excitability and morphological features of layer V pyramidal neurons. Accordingly, we identified distinct voltage-gated ion channels regulated by Nr2f1 that might directly influence intrinsic bioelectrical properties during critical time windows of S1 cortex specification. Altogether, our data suggest a tight link between Nr2f1 and neuronal excitability in the developmental sequence that ultimately sculpts the emergence of cortical network activity within the immature neocortex.

A comparison of the severity of tinnitus in patients with and without hearing loss using the tinnitus functional index (TFI).

OBJECTIVE: The objective of this study was to compare the severity of tinnitus in tinnitus patients with and without hearing loss. DESIGN AND STUDY SAMPLE: 73 tinnitus patients were included in this study at an audiology clinic in Amman, Jordan. Participants were assigned to two groups according to their hearing status. The severity of tinnitus was evaluated using the Tinnitus Functional Index questionnaire. All participants were interviewed, followed by an otoscopic examination, pure tone audiometry, and tests for admittance and tinnitus matching. RESULTS: The normal hearing group included 34 participants (46.6%) whose TFI scores were divided as follows: mild annoyance (17), significant annoyance (14), and severe annoyance (3). The sensorineural loss group included 39 participants (53.4%) with mild annoyance (11), significant annoyance (12), and severe annoyance (16). A statistically significant association was found between hearing status and the severity of tinnitus using a Chi-Squared test (x2 = 0.487, p = 0.007). There was no association between tinnitus severity and age or gender. CONCLUSION: Tinnitus severity was significantly worse in tinnitus patients with a hearing loss than tinnitus patients with normal hearing thresholds. This should be taken into consideration when clinicians are planning counselling and management protocols for individual patients.

COUP-TFI mitotically regulates production and migration of dentate granule cells and modulates hippocampal Cxcr4 expression.

Development of the dentate gyrus (DG), the primary gateway for hippocampal inputs, spans embryonic and postnatal stages, and involves complex morphogenetic events. We have previously identified the nuclear receptor COUP-TFI as a novel transcriptional regulator in the postnatal organization and function of the hippocampus. Here, we dissect its role in DG morphogenesis by inactivating it in either granule cell progenitors or granule neurons. Loss of COUP-TFI function in progenitors leads to decreased granule cell proliferative activity, precocious differentiation and increased apoptosis, resulting in a severe DG growth defect in adult mice. COUP-TFI-deficient cells express high levels of the chemokine receptor Cxcr4 and migrate abnormally, forming heterotopic clusters of differentiated granule cells along their paths. Conversely, high COUP-TFI expression levels downregulate Cxcr4 expression, whereas increased Cxcr4 expression in wild-type hippocampal cells affects cell migration. Finally, loss of COUP-TFI in postmitotic cells leads to only minor and transient abnormalities, and to normal Cxcr4 expression. Together, our results indicate that COUP-TFI is required predominantly in DG progenitors for modulating expression of the Cxcr4 receptor during granule cell neurogenesis and migration.

A unique insight into the MiRNA profile during genital chlamydial infection.

BACKGROUND: Genital C. trachomatis infection may cause pelvic inflammatory disease (PID) that can lead to tubal factor infertility (TFI). Understanding the pathogenesis of chlamydial complications including the pathophysiological processes within the female host genital tract is important in preventing adverse pathology. MicroRNAs regulate several pathophysiological processes of infectious and non-infectious etiologies. In this study, we tested the hypothesis that the miRNA profile of single and repeat genital chlamydial infections will be different and that these differences will be time dependent. Thus, we analyzed and compared differentially expressed mice genital tract miRNAs after single and repeat chlamydia infections using a C. muridarum mouse model. Mice were sacrificed and their genital tract tissues were collected at 1, 2, 4, and 8 weeks after a single and repeat chlamydia infections. Histopathology, and miRNA sequencing were performed. RESULTS: Histopathology presentation showed that the oviduct and uterus of reinfected mice were more inflamed, distended and dilated compared to mice infected once. The miRNAs expression profile was different in the reproductive tissues after a reinfection, with a greater number of miRNAs expressed after reinfection. Also, the number of miRNAs expressed each week after chlamydia infection and reinfection varied, with weeks eight and one having the highest number of differentially expressed miRNAs for chlamydia infection and reinfection respectively. Ten miRNAs; mmu-miR-378b, mmu-miR-204-5p, mmu-miR-151-5p, mmu-miR-142-3p, mmu-miR-128-3p, mmu-miR-335-3p, mmu-miR-195a-3p, mmu-miR-142-5p, mmu-miR-106a-5p and mmu-miR-92a-3p were common in both primary chlamydia infection and reinfection. Pathway analysis showed that, amongst other functions, the differentially regulated miRNAs control pathways involved in cellular and tissue development, disease conditions and toxicity. CONCLUSIONS: This study provides insights into the changes in miRNA expression over time after chlamydia infection and reinfection, as well as the pathways they regulate to determine pathological outcomes. The miRNAs networks generated in our study shows that there are differences in the focus molecules involved in significant biological functions in chlamydia infection and reinfection, implying that chlamydial pathogenesis occurs differently for each type of infection and that this could be important when determining treatments regime and disease outcome. The study underscores the crucial role of host factors in chlamydia pathogenesis.

Molecular control of two novel migratory paths for CGE-derived interneurons in the developing mouse brain.

GABAergic interneurons are highly heterogeneous and originate in the subpallium mainly from the medial (MGE) and caudal (CGE) ganglionic eminences according to a precise temporal sequence. MGE-derived cells disperse dorsally and migrate towards all regions of the cortex, but little is known about how CGE-derived cells reach their targets during development. Here, we unravel the existence of two novel CGE caudo-rostral migratory streams, one located laterally (LMS) and the other one more medially (MMS), that, together with the well-known caudal migratory stream (CMS), contribute to populate the neocortex, hippocampus and amygdala. These paths appear in a precise temporal sequence and express a distinct combination of transcription factors, such as SP8, PROX1, COUP-TFI and COUP-TFII. By inactivating COUP-TFI in developing interneurons, the lateral and medial streams are perturbed and expression of SP8 and COUP-TFII affected. As a consequence, adult mutant neocortices have laminar-specific alterations of distinct cortical interneuron subtypes. Overall, we propose that the existence of spatially and temporally regulated migratory paths in the subpallium contributes to the laminar distribution and specification of distinct interneuron subpopulations in the adult brain.

Transcription factors COUP-TFI and COUP-TFII are required for the production of granule cells in the mouse olfactory bulb.

Neural stem cells (NSCs) persist in the adult mammalian subventricular zone (SVZ) of the lateral ventricle. Primary NSCs generate rapidly dividing intermediate progenitor cells, which in turn generate neuroblasts that migrate along the rostral migratory stream (RMS) to the olfactory bulb (OB). Here, we have examined the role of the COUP-TFI and COUP-TFII orphan nuclear receptor transcription factors in mouse OB interneuron development. We observed that COUP-TFI is expressed in a gradient of low rostral to high caudal within the postnatal SVZ neural stem/progenitor cells. COUP-TFI is also expressed in a large number of migrating neuroblasts in the SVZ and RMS, and in mature interneurons in the OB. By contrast, very few COUP-TFII-expressing (+) cells exist in the SVZ-RMS-OB pathway. Conditional inactivation of COUP-TFI resulted in downregulation of tyrosine hydroxylase expression in the OB periglomerular cells and upregulation of COUP-TFII expression in the SVZ, RMS and OB deep granule cell layer. In COUP-TFI/COUP-TFII double conditional mutant SVZ, cell proliferation was increased through the upregulation of the proneural gene Ascl1. Furthermore, COUP-TFI/II-deficient neuroblasts had impaired migration, resulting in ectopic accumulation of calretinin (CR)+ and NeuN+ cells, and an increase in apoptotic cell death in the SVZ. Finally, we found that most Pax6+ and a subset of CR+ granular cells were lost in the OB. Taken together, these results suggest that COUP-TFI/II coordinately regulate the proliferation, migration and survival of a subpopulation of Pax6+ and CR+ granule cells in the OB.

Gradient COUP-TFI Expression Is Required for Functional Organization of the Hippocampal Septo-Temporal Longitudinal Axis.

The hippocampus (HP), a medial cortical structure, is subdivided into a distinct dorsal (septal) and ventral (temporal) portion, which is separated by an intermediate region lying on a longitudinal curvature. While the dorsal portion is more dedicated to spatial navigation and memory, the most ventral part processes emotional information. Genetic factors expressed in gradient during development seem to control the size and correct positioning of the HP along its longitudinal axis; however, their roles in regulating differential growth and in supporting its anatomical and functional dissociation remain unexplored. Here, we challenge the in vivo function of the nuclear receptor COUP-TFI (chicken ovalbumin upstream promoter transcription factor 1) in controlling the hippocampal, anatomical, and functional properties along its longitudinal axis. Loss of cortical COUP-TFI function results in a dysmorphic HP with altered shape, volume, and connectivity, particularly in its dorsal and intermediate regions. Notably, topographic inputs from the entorhinal cortex are strongly impaired in the dorsal portion of COUP-TFI mutants. These severe morphological changes are associated with selective spatial learning and memory impairment. These findings identify a novel transcriptional regulator required in the functional organization along the hippocampal septo-temporal axis supporting a genetic basis of the hippocampal volumetric growth with its final shape, circuit, and type of memory function.

Tinnitus functional index: validation of the German version for Switzerland.

BACKGROUND: Different standardized questionnaires are used to assess tinnitus severity, making comparisons across studies difficult. These questionnaires are also used to measure treatment-related changes in tinnitus although they were not designed for this purpose. To solve these problems, a new questionnaire - the Tinnitus Functional Index (TFI) - has been established. The TFI is highly responsive to treatment-related change and promises to be the new gold standard in tinnitus evaluation. The aim of the current study was to validate a German version of the TFI for a German-speaking population in Switzerland. METHODS: At the ENT department of the University Hospital Zurich, 264 subjects completed an online survey including the German version for Switzerland of TFI, Tinnitus Handicap Inventory (THI), Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI) and sociodemographic variables. Internal consistency of the TFI was calculated with Cronbach's alpha coefficient. Pearson correlation coefficients were used for the test-retest reliability of the TFI and to investigate convergent and discriminant validity between the THI and the BDI and BAI, respectively. Factor analysis was assessed using a principal component analysis with oblique rotation. The different factors extracted were then compared with the original questionnaire. RESULTS: The German version of the TFI for Switzerland showed an excellent internal consistency (Cronbach's alpha of 0.97) and an excellent test-retest reliability of 0.91. The convergent validity with THI was high (r = 0.86). The discriminant validity with BAI and BDI showed moderate results (BAI: r = 0.60 and BDI: r = 0.65). In the factor analysis only five factors with one main factor could be extracted instead of eight factors as described in the original version. Nevertheless, relations to the original eight subscales could be demonstrated. CONCLUSION: The German version of the TFI for Switzerland is a suitable instrument for measuring the impact of tinnitus. The reliability and validity of this version are comparable with the original version of the TFI. Although this study showed only five factors in the factor analysis, relations to the original eight subscales were identified. Therefore, the German version of the TFI for Switzerland can deliver relevant information regarding the different tinnitus domains. TRIAL REGISTRATION: Clinical trial registration number on clinicaltrial.gov: NCT01837368 .

COUP-TFs and eye development.

Recent studies reveal that COUP-TF genes are essential for neural development, cardiovascular development, energy metabolism and adipogenesis, as well as for organogenesis of multiple systems. In this review, we mainly describe the COUP-TF genes, molecular mechanisms of COUP-TF action, and their crucial functions in the morphogenesis of the murine eye. Mutations of COUP-TF genes lead to the congenital coloboma and/or optic atrophy in both mouse and human, indicating that the study on COUP-TFs and the eye will benefit our understanding of the etiology of human ocular diseases. This article is part of a Special Issue entitled: Nuclear receptors in animal development.

Efficacy of Internet-Based Therapies for Tinnitus: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: Tinnitus presents a major public health challenge, impacting quality of life. With conventional therapies being often time-consuming and costly, interest in Internet-based treatments, such as auditory treatments and Internet-based cognitive behavioral therapy, has grown due to their improved patient adherence. This meta-analysis aims to review existing scientific literature to assess the effectiveness of Internet-based therapies (IBTs) in treating tinnitus. METHODS: Studies up to February 2024 using the Tinnitus Functional Index (TFI), Tinnitus Handicap Inventory (THI), or Tinnitus Reactions Questionnaire (TRQ) to monitor tinnitus before and after IBTs were searched in PubMed, Google Scholar, Web of Science, and the Cochrane Central Register of Controlled Trials. Variation of the score with time was analyzed and a comparison was made with non-IBT studies. Treatment effects were analyzed using Cohen's d model. RESULTS: A total of 14 articles were considered, with a total of 1574 patients. Significant improvements in questionnaire scores were noted post-treatment. In the IBT group, THI and TFI decreased by 17.97 and 24.56 points, respectively (Cohen's d THI: 0.85; TFI: 0.80). In the control group, THI and TFI decreased by 13.7 and 4.25 points, respectively (Cohen's d THI: 0.55; TFI: 0.10). CONCLUSIONS: Internet-based therapies showed reliable effectiveness, possibly due to improved patient compliance, accessibility, cost-effectiveness, and customization.

Research on domain ontology construction based on the content features of online rumors.

Online rumors are widespread and difficult to identify, which bring serious harm to society and individuals. To effectively detect and govern online rumors, it is necessary to conduct in-depth semantic analysis and understand the content features of rumors. This paper proposes a TFI domain ontology construction method, which aims to achieve semantic parsing and reasoning of the rumor text content. This paper starts from the term layer, the frame layer, and the instance layer, and based on the reuse of the top-level ontology, the extraction of core literature content features, and the discovery of new concepts in the real corpus, obtains the core classes (five parent classes and 88 subclasses) of the rumor domain ontology and defines their concept hierarchy. Object properties and data properties are designed to describe relationships between entities or their features, and the instance layer is created according to the real rumor datasets. OWL language is used to encode the ontology, Protégé is used to visualize it, and SWRL rules and pellet reasoner are used to mine and verify implicit knowledge of the ontology, and judge the category of rumor text. This paper constructs a rumor domain ontology with high consistency and reliability.

Evaluating fit indices in a multilevel latent growth model with unbalanced design: a Monte Carlo study.

This study informed researchers about the performance of different level-specific and target-specific model fit indices in the Multilevel Latent Growth Model (MLGM) with unbalanced design. As the use of MLGMs is relatively new in applied research domain, this study helped researchers using specific model fit indices to evaluate MLGMs. Our simulation design factors included three levels of number of groups (50, 100, and 200) and three levels of unbalanced group sizes (5/15, 10/20, and 25/75), based on simulated datasets derived from a correctly specified MLGM. We evaluated the descriptive information of the model fit indices under various simulation conditions. We also conducted ANOVA to calculated the extent to which these fit indices could be influenced by different design factors. Based on the results, we made recommendations for practical and theoretical research about the fit indices. CFI- and TFI-related fit indices performed well in the MLGM and could be trustworthy to use to evaluate model fit under similar conditions found in applied settings. However, RMSEA-related fit indices, SRMR-related fit indices, and chi square-related fit indices varied by the factors included in this study and should be used with caution for evaluating model fit in the MLGM.

Telomere length, TERT and shelterin complex proteins in hepatocellular carcinomas expressing "stemness"-related markers.

BACKGROUND & AIMS: Hepatocellular carcinomas (HCCs) expressing "stemness"-related markers have been associated with aggressive biological behavior and poor prognosis. We examined the relationship between "stemness"-related protein expression and telomere length, hTERT and shelterin complex protein expression and chromosomal instability. METHODS: Quantitative fluorescent in situ hybridization for telomere length, immunohistochemistry for K19, EpCAM, CD133, c-kit, HepPar1, hTERT, TRF1, TRF2, POT1, RAP1 and TPP1, and TUNEL assay were performed in 137 HCCs, and array comparative genomic hybridization was performed with 24 HCCs. RESULTS: Telomeres were significantly longer in HCCs expressing "stemness"-related proteins (K19: p < 0.001, EpCAM: p = 0.002, CD133: p = 0.002). On analyzing different tumor cells within EpCAM-expressing HCCs, EpCAM-positive tumor cells showed longer telomeres (1.329 ± 0.246) compared to EpCAM-negative tumor cells (0.996 ± 0.381) within the same HCCs (p = 0.031). Telomeres were significantly longer in HCCs expressing hTERT (p = 0.048) and RAP1 proteins (p = 0.031). K19-expressing HCCs expressed hTERT (p = 0.002), TRF2 (p = 0.001) and TPP1 (p = 0.013) more frequently compared to K19-negative HCCs. EpCAM-positivity was associated with more frequent hTERT (p = 0.028), TPP1 (p = 0.017), TRF2 (p = 0.027) and POT1 (p = 0.004) expression. Copy number alterations were more frequent in K19 and EpCAM-expressing HCCs compared to HCCs without these markers (K19: p = 0.038, EpCAM: p = 0.009). HCCs with longer telomeres were associated with a shorter overall (p = 0.019) and disease-free survivals (p = 0.049), and decreased disease-free survivals were seen in TRF2-positive HCCs (p = 0.018). CONCLUSIONS: HCCs expressing "stemness"-related proteins are characterized by increased telomere length, increased expression of hTERT and shelterin complex proteins, and increased chromosomal instability compared to conventional HCCs. Longer telomeres and TRF2 expression in HCCs are associated with poor patient outcomes.

Translation into Spanish and validation of the Tinnitus Functional Index (TFI).

OBJECTIVE: The objectives of this study were to translate into Spanish, cross-culturally adapt and validate the TFI. MATERIALS AND METHODS: The TFI questionnaire translated into Spanish (Sp-TFI) and cross-culturally adapted following the published guidelines on cross-cultural adaptation of health questionnaires was evaluated using two indicators. Its internal consistency was assessed with Cronbach's α considering the Tinnitus Handicap Inventory (THI) as the gold standard. Further, its test-retest reliability was assessed with intraclass correlation coefficients (ICCs). ICCs were also calculated for the THI and visual analogue scales (VAS) for tinnitus tested and retested in all participants. RESULTS: The mean age of the 18 participants was 45.77 (SD: 11.87) years; 12 were female (66.67 %) and 6 were male (33.33%). Half of the participants experienced tinnitus in their left ear and half in their right. The mean pure-tone average (PTA) in the affected ear was 29.34 (SD: 8.08) dB-HL. Regarding internal consistency and reliability of the Sp-TFI respectively, Cronbach's α was 0.83 and the ICC type (2,1) was 1 (CI: 0.99-1). Among the variables studied, we found the following independent predictors had statistically significant effects on THI score: sex (p < 0.01), PTA (p = 0.03), overall Sp-TFI score (p = 0.02) and Sp-TFI SL, R and A subscale scores (p = 0.03, p = 0.03, and p < 0.01, respectively). CONCLUSION: Based on the internal consistency and reliability results obtained in this study, the cross-culturally adapted Spanish version of the TFI (Sp-TFI) has been validated for use in Spain. LEVEL OF EVIDENCE: 2B: Individual cohort study/low-quality randomized control studies.