Inflammation is a key event in hypertensive organ damage, and TNF-α and IL-1β are elevated in hypertension. In this study, we evaluated the effects of TNF-α and IL-1β elevation on hypertensive cardiac damage by treatment with a bifunctional inflammatory inhibitor, TNF receptor 2-fragment crystalization-IL-1 receptor antagonist (TFI), which can neutralize these 2 cytokines simultaneously. A mouse hypertension model of angiotensin II (Ang II) infusion (1500 ng/kg·min for 7 d) was induced in wild-type mice. TNF-α and IL-1β were inhibited by TFI administration (5 mg/kg, every other day), the effects of inhibition on cardiac damage were examined, and its mechanism on inflammatory infiltration was further studied in vivo and in vitro. Ang II infusion induced cardiac injury, including increased macrophage infiltration, expression of inflammatory cytokines (IL-12, IL-6, etc), and cardiac fibrosis, such as elevated α-smooth muscle actin, collagen I, and TGF-β expression. Importantly, the Ang II-induced cardiac injury was suppressed by TFI treatment. Moreover, TFI reduced the expression of adhesion molecules (intercellular adhesion molecule-1 and vascular cell adhesion molecule-1) and monocyte chemotactic protein-1 expression in Ang II-treated hearts. Additionally, blockade of TNF-α and IL-1β by TFI reduced monocyte adherence to endothelia cell and macrophage migration. This study demonstrates that blocking TNF-α and IL-1β by TFI prevents cardiac damage in response to Ang II, and targeting these 2 cytokines simultaneously might be a novel tool to treat hypertensive heart injury.

The Tinnitus Research Consortium (TRC) issued a Request for Proposals in 2003 to develop a new tinnitus outcome measure that would: (1) be highly sensitive to treatment effects (validated for "responsiveness"); (2) address all major dimensions of tinnitus impact; and (3) be validated for scaling the negative impact of tinnitus. A grant was received by M. Meikle to conduct the study. In that observational study, all of the TRC objectives were met, with the final 25-item Tinnitus Functional Index (TFI) containing eight subscales. The study was published in 2012, and since then the TFI has received increasing international use and is being translated into at least 14 languages. The present study utilized data from a randomized controlled trial (RCT) that involved testing the efficacy of "telephone tinnitus education" as intervention for bothersome tinnitus. These data were used to confirm results from the original TFI study. Overall, the TFI performed well in the RCT with Cohen's d being 1.23. There were large differences between the eight different subscales, ranging from a mean 13.2-point reduction (for the Auditory subscale) to a mean 26.7-point reduction (for the Relaxation subscale). Comparison of TFI performance was made with the Tinnitus Handicap Inventory. All of the results confirmed sensitivity of the TFI along with its subscales. This article is part of a Special Issue entitled.

A complex regulatory cascade is required for normal cardiac development, and many aspects of this network are conserved from Drosophila to mammals. In Drosophila, the seven-up (svp) gene, an ortholog of the vertebrate chick ovalbumin upstream promoter transcription factors (COUP-TFI and II), is initially activated in the cardiac mesoderm and is subsequently restricted to cells forming the cardiac inflow tracts. Here, we investigate svp regulation in the developing cardiac tube. Using bioinformatics, we identify a 1007-bp enhancer of svp which recapitulates its entire expression in the embryonic heart and other mesodermal derivatives, and we show that this enhancer is initially activated by the NK homeodomain factor Tinman (Tin) via two conserved Tin binding sites. Mutation of the Tin binding sites significantly reduces enhancer activity both during normal development and in response to ectopic Tin. This is the first identification of an enhancer for the complex svp gene, demonstrating the effectiveness of bioinformatics tools in assisting in unraveling transcriptional regulatory networks. Our studies define a critical component of the svp regulatory cascade and place gene regulatory events in direct apposition to the formation of critical cardiac structures.

Members of the chicken ovalbumin upstream promoter-transcription factor (COUP-TF) subfamily of orphan nuclear receptors, which minimally includes COUP-TFI and ARP1, are highly expressed in brain and are generally considered to be constitutive repressors of transcription. We have used a yeast two-hybrid system to isolate proteins expressed in brain that interact with ARP1. One of the proteins isolated in this screen was Ear2, another orphan receptor that has been suggested to be a member of the COUP-TF subfamily. Here we demonstrate that ARP1 and Ear2 form heterodimers in solution and on directly repeated response elements with high efficiency and a specificity differing from that of homodimeric complexes composed of either receptor. ARP1 and Ear2 were observed to interact in mammalian cells, and the tissue distribution of Ear2 transcripts was found to overlap precisely with the expression pattern of ARP1 in several mouse tissues and embryonal carcinoma cell lines. Heterodimeric interactions between ARP1 and Ear2 may define a distinct pathway of orphan receptor signaling.

Disability is an important health outcome for older persons; it is associated with impaired quality of life, future hospitalization, and mortality. Disability also places a high burden on health care professionals and health care systems. Disability is regarded as an adverse outcome of physical frailty. The main objective of this study was to assess the predictive validity of the eight individual self-reported components of the physical frailty subscale of the TFI for activities of daily living (ADL) and instrumental activities of daily living (IADL) disability. This longitudinal study was carried out with a sample of Dutch citizens. At baseline the sample consisted at 429 people aged 65 years and older and a subset of all respondents participated again two and a half years later (N=355, 83% response rate). The respondents completed a web-based questionnaire comprising the TFI and the Groningen Activity Restriction Scale (GARS) for measuring disability. Five components together (unintentional weakness, weakness, poor endurance, slowness, low physical activity), referring to the phenotype of Fried et al., predicted disability, even after controlling for previous disability and other background characteristics. The other three components of the physical frailty subscale of the TFI (poor balance, poor hearing, poor vision) together did not predict disability. Low physical activity predicted both total and ADL disability, and slowness both total and IADL disability. In conclusion, self-report assessment using the physical subscale of the TFI aids the prediction of future ADL and IADL disability in older persons two and a half years later.

Hyperfunctioning adrenocortical adenomas produce excessive amounts of various corticosteroids due to dysregulated expression of steroidogenic enzymes. Since no genetic mutations in steroidogenic enzyme genes have been identified as yet, the dysregulated expression at the transcription level may be crucial. Chicken ovalbumin upstream promoter-transcription factors (COUP-TFs) and steroidogenic factor-1 (SF-1) play key roles in the transcriptional regulation of steroidogenic P450 genes. Transfection studies showed that SF-1 activated and COUP-TFs repressed the transcription of bovine CYP17 gene promoter from the CRS2 element in a mutually exclusive manner in Y-1 cells. The results indicate that COUP-TFs negatively regulate the transcriptional activity of SF-1, a steroidogenic cell-specific activator of various steroidogenic P450 genes. Expression of both COUP-TFI and COUP-TFII was significantly decreased in the cortisol-producing adenomas, in which CYP17 was drastically overexpressed, indicating that decreased expression of COUP-TFs play a key role in overexpression of CYP17 in this type of tumors. We then screened for COUP-TFI-interacting proteins from a cortisol-producing adenoma cDNA library using a yeast two-hybrid system and identified a novel RING finger-containing protein which can function as a coregulator for COUP-TFI. Notably, COUP-TFI activated rather than repressed several target genes including the human CYP11B2 gene promoter, the results of which were opposite to those of the CYP17 promoter. The bifunctional activities of COUP-TFI may be derived from the promoter context and our newly identified COUP-TFI coregulator.

Our previous studies demonstrated regulation of the human LH receptor (hLHR) promoter by nuclear orphan receptors EAR2, EAR3/COUP-TFI (repression), and TR4 (activation) through a direct-repeat motif (hDR). The current studies investigated the differential binding of orphan receptors to rat (rLHR) and hLHR promoters, and their modulation of rLHR gene transcription in rat granulosa cells. The rLHR DR with one nucleotide difference from hDR at its core sequence mediated inhibition of the rLHR transcription, to which EAR2 and EAR3/COUP-TFI but not TR4 bound. The A/C mismatch was responsible for the lack of TR4 binding and function, but had no effect on EAR2 and EAR3/COUP-TFI. EAR2 and EAR3/COUP-TF bound to the rLHR DR with lower affinity than to the hDR, and exhibited lesser inhibitory capacity. This difference resulted from the lack of a guanine in the rDR, which is present 3' next to the hDR core. These studies have identified sequence-specific requirements for the binding of EAR2, EAR3/COUP-TFI, and TR4 to the DRs that explain their differential regulation of rat and human LHR genes. In addition, hCG treatment significantly reduced the inhibition of rLHR gene in granulosa cells and also decreased EAR2 and EAR3/COUP-TFI protein levels. These results indicate that hormonally regulated expression of EAR2 and EAR3/COUP-TFI contributes to gonadotropin-induced derepression of LHR promoter activity in granulosa cells.

The FSH receptor (FSHR) is selectively expressed in the granulosa and Sertoli cells in a development-dependent manner. Little is known regarding how the regulatory factors balance expression of this gene in ovarian cycles or spermatogenic stages. We have used the ovine FSHR promoter as a model system and identified a third regulatory element (RE-3) located at -197 to -171 of the strongest promoter. Gel mobility shift and antibody supershift assays demonstrated that nuclear factors c-Fos/c-Jun, steroidogenic factor-1 (SF-1), upstream stimulatory factor-1/2 (USF-1/2), and chicken ovalbumin upstream promoter transcription factor-1/2 (COUP-TFI/II) potentially bound to RE-3. We have also extended our previous observations by showing that a sequence containing an E-box was not only bound by USF proteins but also recognized by COUP-TF orphan receptors. Functional studies demonstrated that USF-1/2, c-Fos/c-Jun, and SF-1 were activators, whereas COUP-TFs were repressors. Our studies indicated that RE-3 mediated SF-1 activation as well as phorbol 12-myristate 13-acetate stimulation, whereas COUP-TFs inhibited AP-1, USFs, and SF-1 activation. We also demonstrated that both COUP-TF-binding sites in the core promoter were required for the bipartite elements to oppose their competitor binding. These data suggest a mechanism by which positive and negative regulators compete for the common regulatory elements, providing antagonistic pathways that might govern the expression of FSHR in gonadal cells.

Chicken ovalbumin upstream promoter-transcription factor (COUP-TF), DAX-1, and steroidogenic factor-1 (SF-1) are orphan members of the nuclear hormone receptor superfamily. COUP-TF and DAX-1 have been shown to negatively regulate the transcriptional activity of SF-1, a steroidogenic cell-specific activator of various steroidogenic cytochrome P450 genes. We therefore examined the expression levels and immunolocalization of COUP-TF, DAX-1, and SF-1 in human adrenal gland (NL) and adrenocortical adenomas, and compared the results with CYP17 expression levels and its enzyme activities to study their potential correlation with adrenocortical steroidogenesis. In NL (n = 10), expressions of COUP-TF, DAX-1, and SF-1 were detected in the nuclei of adrenocortical cells, but not in the medulla. In cortisol-producing adenomas causing Cushing syndrome (CS, n = 20), CYP17 expression was upregulated (298 +/- 2% vs NL 98 +/- 4%), whereas expression levels of both COUP-TFs (COUP-TFI, 52 +/- 5% vs NL 98 +/- 4%; COUP-TFII, 18 +/- 4% vs NL 98 +/- 4%) and DAX-1 (42 +/- 4% vs NL 100 +/- 4%) were reduced. In deoxycorticosterone-producing adenomas (DOC, n = 2), on the other hand, CYP17 expression was extremely reduced (8 and 12% vs NL 98 +/- 4%), whereas DAX-1 expression increased markedly (350 and 360% vs NL 100 +/- 4%). Expression levels of SF-1 did not differ between NL (100 +/- 8%) and CS (106 +/- 10%), but its expression appeared to be decreased in DOC (25 and 20%). These results showed CYP17 expression to be upregulated and downregulated in CS and DOC, respectively, in a manner reciprocal to that of its repressors, COUP-TF and/or DAX-1. In summary, the results indicate that co-localization of COUP-TF, DAX-1, and SF-1 in NL was lost in adrenocortical tumors and that these orphan receptors play an important role in the regulation of steroidogenesis in human adrenals.

Estrogen-responsive genes are regulated by altering the balance of estrogen receptor (ER) interaction with transcription activators and inhibitors. Here we examined the role of ER ligand on ER interaction with the Chicken Ovalbumin Upstream Promoter Transcription Factor (COUP-TF) orphan nuclear receptor. COUP-TF binding to half-site estrogen response elements (EREs) was increased by the addition of estradiol (E2) -liganded ER (E2-ER), but not by ER liganded with the antiestrogens 4-hydroxytamoxifen (4-OHT-ER) or tamoxifen aziridine (TAz-ER). ER did not bind to single half-sites. Conversely, COUP-TF enhanced the ERE binding of purified E2-ER, but did not affect TAz-ER-ERE binding. In contrast, only antiestrogens enhanced direct interaction between ER and COUP-TF as assessed by GST pull-down assays. Identical results were obtained using either purified bovine or recombinant human ERalpha. Co-immunoprecipitation assays showed that ER and COUP-TF interact in extracts from MCF-7 and ERalpha-transfected MDA-MB-231 cells. Here we document that ER ligand impacts COUP-TF-ER interaction. COUP-TF interaction is mediated by the DNA binding and ligand-binding domains of ER. We suggest that changes in ER conformation induced by DNA binding reduce ER-COUP-TF interaction. Transient transfection of human MCF-7 breast cancer cells with a COUP-TFI expression vector repressed E2-induced luciferase reporter gene expression from single or multiple tandem copies of a consensus ERE. COUP-TFI stimulated 4-OHT-induced luciferase activity from a minimal ERE. Alone, COUP-TFI increased transcription from ERE half-sites or a single ERE in a sequence-dependent manner. These data provide evidence that the ERE sequence and its immediate flanking regions influence whether COUP-TF enhances, inhibits, or has no effect on ER ligand-induced ERE reporter gene expression and that COUP-TFI activates gene transcription from ERE half-sites. We suggest that COUP-TFI plays a role in mitigating estrogen-responsive gene expression.

OBJECTIVE

The aim of this study was to evaluate micro-shear bond strength and morphological analysis of a self-etching primer adhesive system to fluorosed enamel.

METHODS

Extracted human molars were classified according to the severity of fluorosis using Thylstrup and Fejerskov index into four groups (TFI: 0, normal; 1-3, mild fluorosis; 3-6, moderate fluorosis; 6-7, severe fluorosis) and divided into following two sub-groups. For the first sub-group, a self-etching primer adhesive system was applied to the ground enamel surfaces and the other sub-group was conditioned with 37% phosphoric acid for 30s prior to application of the same adhesive system. Teeth were then restored with a resin composite, stored for 24h in water and micro-shear bond strengths were measured. After shear testing, the fracture modes were observed under a laser scanning microscope. Morphological study of etching patterns and adhesive interface was done under a scanning electron microscope (SEM). The data were analyzed using two-way ANOVA and Sheffe test (p=0.05).

RESULTS

No statistically significant difference was there between the different degrees of fluorosis in each sub-group. However, significant difference in the bond strengths between phosphoric acid etching and self-etching was found in moderate and severe groups. The SEM observations showed at the resin-enamel interface, thick resin tag like extensions penetrated around 3.5 microm into the enamel etched with phosphoric acid, while self-etching primer created, 1 microm lamina like penetration.

CONCLUSIONS

Severity of fluorosis affects the micro-shear bond strength of a self-etching bonding system to fluorosed enamel.

This study aims to assess the link between fluoride content in groundwater and its impact on dental health in rural communities of the Ethiopian Rift. A total of 148 water samples were collected from two drainage basins within the Main Ethiopian Rift (MER). In the Ziway-Shala basin in particular, wells had high fluoride levels (mean: 9.4±10.5mg/L; range: 1.1 to 68 mg/L), with 48 of 50 exceeding the WHO drinking water guideline limit of 1.5mg/L. Total average daily intake of fluoride from drinking groundwater (calculated per weight unit) was also found to be six times higher than the No-Observed-Adverse-Effects-Level (NOAEL) value of 0.06 mg/kg/day. The highest fluoride levels were found in highly-alkaline (pH of 7 to 8.9) groundwater characterized by high salinity; high concentrations of sodium (Na⁺), bicarbonate (HCO₃⁻), and silica (SiO₂); and low concentrations of calcium (Ca²⁺). A progressive Ca²⁺ decrease along the groundwater flow path is associated with an increase of fluoride in the groundwater. The groundwater quality problem is also coupled with the presence of other toxic elements, such as arsenic (As) and uranium (U). The health impact of fluoride was evaluated based on clinical examination of dental fluorosis (DF) among local residents using the Thylstrup and Fejerskov index (TFI). In total, 200 rural inhabitants between the ages of 7 and 40 years old using water from 12 wells of fluoride range of 7.8-18 mg/L were examined. Signs of DF (TF score of ≥ 1) were observed in all individuals. Most of the teeth (52%) recorded TF scores of 5 and 6, followed by TF scores of 3 and 4 (30%), and 8.4% had TF scores of 7 or higher. Sixty percent of the teeth exhibited loss of the outermost enamel. Within the range of fluoride contents, we did not find any correlation between fluoride content and DF. Finally, preliminary data suggest that milk intake has contributed to reducing the severity of DF. The study highlights the apparent positive role of milk on DF, and emphasizes the importance of nutrition in management efforts to mitigate DF in the MER and other parts of the world.

The Tilburg Frailty Indicator (TFI) is a self-administered questionnaire with a bio-psycho-social integrated approach that measures the degree of frailty in elderly persons. The TFI was developed in the Netherlands and tested in a population of elderly Dutch men and women. The aim of this study was to translate and culturally adapt the TFI to a Danish context, and to test face validity of the Danish version by cognitive interviewing. An internationally recognized procedure was applied as a basis for the translation process. The primary tasks were forward translation, reconciliation, back translation, harmonization and pretest. Pretest and review of the preliminary version by cognitive interviewing, were performed at a local community center and in an acute medical ward at the University Hospital in Aalborg, Denmark respectively. A large agreement regarding meaning of the items in the forward translation and reconciliation process was seen. Minor discrepancies were solved by consensus. Back translation revealed unclear wording in one matter. The harmonization committee agreed on a version for cognitive interviewing after revision of minor issues and thirty-four participants were interviewed. Two issues became evident and these were revised. The cognitive interviews and final lay-out resulted in minor adjustments as text type size, specific font, and lining for optimizing readability. In conclusion, we consider the TFI to be translated in such rigorous manner that the instrument can be further tested in clinical practice. The overall objective of the questionnaire being to identify frailty and improve the interventions relating to frail elderly persons in Denmark.

The mammalian neocortex is subdivided into cytoarchitectural areas with distinct connectivity, gene expression and neural functions. Areal identity is initially specified by rostrocaudal and mediolateral gene expression gradients in neuroepithelial and radial glial progenitors (the 'protomap'). On further differentiation, distinct sets of gene expression gradients arise in intermediate progenitors and postmitotic neurons, and are necessary to implement areal specification. However, it is still unknown whether postmitotic gene expression gradients can determine areal identity independently of protomap gradients. Here we show, by cell type-restricted genetic loss- and gain-of-function, that high levels of postmitotic COUP-TFI (Nr2f1) expression are necessary and sufficient for the development of sensory (caudal) areal identity. Our data indicate a crucial role for postmitotic patterning genes in areal specification and reveal an unexpected plasticity in this process, which may account for complex and evolutionarily novel structures characteristic of the mammalian neocortex.

The most prevalent, curable sexually important diseases are those caused by Chlamydia trachomatis (C. trachomatis) and genital mycoplasmas. An important characteristic of these infections is their ability to cause long-term sequels in upper genital tract, thus potentially affecting the reproductive health in both sexes. Pelvic inflammatory disease (PID), tubal factor infertility (TFI), and ectopic pregnancy (EP) are well documented complications of C. trachomatis infection in women. The role of genital mycoplasmas in development of PID, TFI, and EP requires further evaluation, but growing evidence supports a significant role for these in the pathogenesis of chorioamnionitis, premature membrane rupture, and preterm labor in pregnant woman. Both C. trachomatis and genital mycoplasmas can affect the quality of sperm and possibly influence the fertility of men. For the purpose of this paper, basic, epidemiologic, clinical, therapeutic, and public health issue of these infections were reviewed and discussed, focusing on their impact on human reproductive health.

ZAP-70, CD38 and IGHV mutations have all been reported to have prognostic impact in chronic lymphocytic leukemia (CLL), both individually and in paired combinations. We aimed to determine whether the combination of all three factors provided more refined prognostic information concerning the treatment-free interval (TFI) from diagnosis. ZAP-70, CD38 and IGHV mutations were evaluated in 142 patients. Combining all three factors, the ZAP-70-/CD38-/Mutated group showed the longest median TFI (62 months, n = 37), ZAP-70+/CD38+/Unmutated cases the shortest (11 months, n = 37) and cases discordant for>> or = 1 factor, an intermediate TFI (27 months, n = 68) (p = 0.006). Analysis of discordant cases revealed values that were otherwise masked when measuring single prognostic factors. The presence or absence of cytogenetic abnormalities did not explain the variability among discordant cases. Simultaneous analysis of ZAP-70, CD38 and IGHV mutations in CLL provides more discriminatory prediction of TFI than any factor alone.

OBJECTIVE

Several questionnaires are used to survey how tinnitus affects quality of life, making comparisons across studies difficult. The questionnaires also are used to measure treatment outcome but were not designed for this purpose. To address these issues, a new questionnaire has been suggested, the tinnitus functional index (TFI), which is highly responsive to treatment-related change. The current study aim was to translate and validate the TFI for a Dutch-speaking population. Factor analysis was performed to characterize the TFI profile in a large tinnitus population.

METHODS

The questionnaire was translated using a translation-back translation procedure, and 263 patients in the ENT department of Antwerp University Hospital with tinnitus-related complaints completed it. Factor structure was assessed using exploratory analysis with oblique rotation and compared with the original questionnaire. Internal consistency was measured using Cronbach's alpha coefficient. Spearman correlations with the percentage of time aware of the tinnitus and the visual analogue scales (VAS) for maximum tinnitus loudness and mean tinnitus loudness were calculated to investigate convergent validity.

RESULTS

The original eight-factor structure could be confirmed in the Dutch version of the TFI. Internal consistency (a=0.96) and convergent validity showed good results. Statistically significant correlations were found with the VAS for maximum loudness (r=0.59; p<0.001), VAS for mean loudness (r=0.66; p<0.001), and percentage of time aware of tinnitus (r=0.58; p<0.001).

CONCLUSIONS

The Dutch version of the TFI is suitable for measuring in clinical and research settings how tinnitus affects daily life, with psychometric properties in line with the original version.

Induction of the major drug metabolizing enzyme CYP3A4 by xenobiotics contributes to the pronounced interindividual variability of its expression and often results in clinically relevant drug-drug interactions. It is mainly mediated by PXR, which regulates CYP3A4 expression by binding to several specific elements in the 5' upstream regulatory region of the gene. Induction itself shows a marked interindividual variability, whose underlying determinants are only partly understood. In this study, we investigated the role of nuclear receptor binding to PXR response elements in CYP3A4, as a potential non-genetic mechanism contributing to interindividual variability of induction. By in vitro DNA binding experiments, we showed that several nuclear receptors bind efficiently to the proximal promoter ER6 and distal xenobiotic-responsive enhancer module DR3 motifs. TRalpha1, TRbeta1, COUP-TFI, and COUP-TFII further demonstrated dose-dependent repression of PXR-mediated CYP3A4 enhancer/promoter reporter activity in transient transfection in the presence and absence of the PXR inducer rifampin, while VDR showed this effect only in the absence of treatment. By combining functional in vitro characterization with hepatic expression analysis, we predict that TRalpha1, TRbeta1, COUP-TFI, and COUP-TFII show a strong potential for the repression of PXR-mediated activation of CYP3A4 in vivo. In summary, our results demonstrate that nuclear receptor binding to PXR response elements interferes with PXR-mediated expression and induction of CYP3A4 and thereby contributes to the interindividual variability of induction.

OBJECTIVE

To present the translation and validation process of the Portuguese version of the Tilburg Frailty Indicator (TFI).

METHODS

A cross-sectional study was designed using a non-probability sample of 252 community-dwelling older adults. Preliminary studies were carried out for face and content validity assessment. Internal consistency, test-retest reliability, construct (convergent/divergent) and criterion validity were subsequently analyzed.

RESULTS

The sample was mainly women (75.8%), with a mean age of 79.2 ± 7.3 years. TFI internal consistency was good (KR-20 = 0.78). Test-retest reliability for the total was also good (r = 0.91), with kappa coefficients showing substantial agreement for most items. TFI physical and social domains correlated as expected with concurrent measures, whereas the TFI psychological domain showed similar correlations with other psychological and physical measures. The TFI showed a good to excellent discrimination ability in regard to frailty criteria, and fair to good ability to predict adverse outcomes.

CONCLUSIONS

The psychometric properties of the TFI seem to be consistently good. These findings provide initial evidence that the Portuguese version is a valid and reliable measure for assessing frailty in the elderly.

BACKGROUND

Zeta-chain associated protein (ZAP)-70 has been proposed as a surrogate marker for immunoglobulin heavy-chain variable region (IgVH) mutation in chronic lymphocytic leukemia (CLL), but it is still not clear whether it is an independent prognostic factor.

METHODS

The authors evaluated ZAP-70 expression by flow cytometry in 201 untreated patients and correlated ZAP-70 levels with CD38 expression, genetic abnormalities detected by fluorescence in situ hybridization (FISH), and the time from diagnosis to first treatment.

RESULTS

Fifty-seven patients (28%) were positive for ZAP-70 >> or = 20%). Positive ZAP-70 status was associated with advanced disease stage, atypical morphology, CD38-positive status, trisomy 12, del(6q), or no detectable abnormalities; negative ZAP-70 status was correlated with del(13q) as a sole abnormality. The treatment-free interval (TFI) was 17.7 months for ZAP-70-positive patients and 44.6 months for ZAP-70-negative patients (P < 0.001). Multivariate analysis in 117 patients identified advanced stage, CD38>> or = 7%, and the absence of del(13q) as a sole abnormality as independent factors for short TFI. Excluding FISH, ZAP-70 status acquired independent prognostic value along with CD38 status. The authors proposed a risk model that combines ZAP-70 and CD38 to identify patients who are likely to progress. When both markers were positive, the TFI was 12 months; when both were negative, the median TFI was 54 months; a median TFI of 26 months was observed in patients who had discordant results (P < 0.00001).

CONCLUSIONS

The current findings suggested that both ZAP-70 and CD38 should be tested prospectively in all patients with early-stage CLL.

CD38 expression and chromosomal abnormalities are novel prognostic factors in chronic lymphocytic leukaemia (CLL). However, their value remains undetermined. CD38 was evaluated in 123 patients and chromosomal aberrations in 111 cases with fluorescence in situ hybridization (FISH). CD38 expression was found in 27% of the cases. In addition, seven out of 32 CD38- patients became CD38+ during evolution of the disease. Chromosomal abnormalities included isolated 13q deletion (40%), 12q trisomy (14%), 11q deletion (without 17p deletion) (14%) and 17p deletion (7%). CD38 expression was significantly associated with Binet stages B and C, atypical morphology and 11q deletion. On univariate analysis of survival estimates, advanced Binet stages, CD38+ phenotype, atypical morphology and 11q or 17p deletions were associated with shorter event-free survival (EFS), treatment-free interval (TFI) and overall survival (OS). Multivariate analysis identified both Binet stages and CD38 as independent prognostic factors with regard to EFS and TFI. However, CD38 appeared as an independent factor for OS when restricted to Binet stage A. Chromosomal aberrations were re-evaluated during evolution in 31 cases. The 17p deletion was the most frequent new chromosomal abnormality (35%) and significantly associated with death (64%). In conclusion, CD38 expression and secondary 17p deletion are important poor prognostic indicators, especially in Binet stage A CLL.

The developmental expression patterns of the nuclear orphan receptors COUP-TFs (chicken ovalbumin upstream promoter-transcription factors) have been correlated to neurogenesis in several animal species. Nevertheless, the role of COUP-TFs in neurogenesis remains unknown. We have studied the functional involvement of COUP-TFI in retinoic acid (RA)-induced neuronal differentiation of P19 embryonal carcinoma cells through two complementary approaches: 1) deregulated expression of COUP-TFI, and 2) inactivation of endogenous COUP-TFs by means of a dominant-negative COUP-TFI mutant. Low levels of wild-type (wt)COUP-TFI transgene expression did not inhibit neural cell fate and primarily enhanced neuron outgrowth from RA-treated P19 aggregates. In contrast, high COUP-TFI expression impeded the neuronal differentiation of P19 cells induced with RA, resulting in cell cultures lacking neurons. This morphological effect was correlated to an elevated level of E-cadherin mRNA. The dominant-negative COUP-TFI mutant induced cell packing after RA treatment and inhibited neurite extension and neuron outgrowth from aggregates. A RGD peptide interference assay indicated that endogenous COUP-TFs could favor migration of neurons through an integrin-dependent mechanism. Accordingly, vitronectin mRNA levels were shown to be up-regulated by COUP-TFI by RT-PCR analysis, and COUP-TFI stimulated the mouse vitronectin promoter activity in transient transfection assays. Taken together, these data indicate that COUP-TFI is not simply a global repressor of retinoid functions, but shows a high selectivity for regulating genes involved in cellular adhesion and migration processes that are particularly important for neuronal differentiation.

OBJECTIVE

Surgical cytoreduction is an integral therapeutic modality for patients with epithelial ovarian (EOC), fallopian tube (FTC), or primary peritoneal (PPC) cancer in the primary setting. The role of surgical cytoreduction in the recurrent setting is not clearly defined and remains controversial. The objective of this study was to assess this potential survival benefit in a large cohort of patients with a long follow-up period.

METHODS

We performed a retrospective chart review of all patients with recurrent EOC, FTC, or PPC who underwent tertiary cytoreduction at our institution from 2/98 to 2/08. Disease-specific survival (DSS) was calculated from the time of tertiary cytoreduction to death or last follow-up. Univariate and multivariate analyses were used to analyze outcomes and to identify potential prognostic factors.

RESULTS

A total of 77 patients were identified, of which 38 (49%) have died of disease. The median time from secondary to tertiary cytoreduction was 25.7 months (range, 4.1-99.4 months). The median follow-up after tertiary cytoreduction was 28.9 months (range, 0.7-123.7 months), with a median DSS for the entire cohort of 47.7 months (95% CI, 25.5-69.9 months). On univariate analysis, residual disease after tertiary cytoreduction and TFI were found to be significant prognostic factors. On multivariate analysis, only residual disease after tertiary cytoreduction retained prognostic significance (P<0.001).

CONCLUSIONS

Tertiary surgical cytoreduction may offer a survival benefit in a highly select group of patients with recurrent EOC, FTC, or PPC. This benefit appears to be greatest in patients in whom a complete gross resection can be achieved.

This study aims to assess the psychometric properties of the Brazilian version of the TFI, an instrument that identifies frailty in elderly individuals. We interviewed 219 individuals aged 60 or older, living in the community. Individuals were predominantly female (52.5%) and mean age was 70.5 (±7.9) years. In order to assess test-retest reliability, 101 individuals were re-interviewed by the same observer within seven to ten days after the first interview. The internal consistency of the instrument was assessed using Cronbach's alpha. To assess construct validity, we used established alternative measures for the items that constitute the TFI, such as: body mass index (BMI), timed up and go (TUG) test, whisper test, Snellen test, upper extremity strength clinical test and mini-mental state examination (MMSE). The test-retest reliability showed high percent agreement for all the items of the instrument, with values ranging from 63% to 100%. Test-retest reliabilities were good (total TFI score r=0.88; physical domain r=0.88; psychological domain r=0.88; and social domain r=0.67). Internal consistency reliability of the Brazilian version was satisfactory (Cronbach's alpha=0.78). The correlations between TFI items and their corresponding measures were consistent except for one item (related to "ability to deal with problems"), demonstrating both convergent and divergent construct validity of the TFI and its items. After the completion of all stages of transcultural adaptation, the Brazilian version of the TFI proved to be well suited for assessing frailty in the elderly population of Brazil.