MAP kinases and NF-kappaB collaborate to induce ICAM-1 gene expression in the early phase of adenovirus infection

Replication-defective adenoviruses (Ad) utilized as vectors for gene transfer are known to induce an inflammatory and immune response upon exposure to respiratory cells in vitro and in vivo. Among the different mediators of inflammation, we recently demonstrated that a replication-defective Ad serotype 5, deleted in the early genes E1 and E3 (Ad.CFTR), induces the proinflammatory intercellular adhesion molecule 1 (ICAM-1) in A549 respiratory cells in vitro and in lung portions of nonhuman primates in vivo, Gene Ther. 5, 131-136). More recently, we described the involvement of the nuclear factor kappaB (NF-kappaB) in the induction of ICAM-1 upon 24 h of exposure of the same Ad5-derived vector, Gene Ther. 8, 1436-1442). Here we investigated whether the early phase of virus-cell interaction is sufficient to stimulate ICAM-1 upregulation. A549 cells were exposed to wild-type Ad5 (Ad5), to Ad.CFTR, and to Ad5 inactivated by incubation at 56 degrees C (Ad5/56 degrees C). Ad5, Ad.CFTR, and Ad5/56 degrees C activated NF-kappaB and increased ICAM-1 mRNA levels within 4 h after exposure. The role of the mitogen-activated protein kinases (MAPKs) on the ICAM-1 mRNA induction was studied. ICAM-1 mRNA upregulation was inhibited upon incubation with several chemicals, namely, the ERK1/2 inhibitors PD98059 and AG1288 (by 98 and 67%, respectively), of the p38/MAPK pathway SB203580 (by 50%), of the JNK pathway dimethylaminopurine (by 83%), and of the NF-kappaB parthenolide (by 96%). Ad5 and Ad5/56 degrees C stimulated ERK1/2, p38/MAPK, and JNK1 starting 10 min and peaking 20-30 min after exposure. The present results indicate a link between the activation of the three major MAPK pathways, NF-kappaB, and the upregulation of ICAM-1 gene expression evoked by Ad5 in the very initial phase of infection.     

Validation of the Portuguese version of the King's Health Questionnaire for urinary incontinent women

OBJECTIVE: To translate into Portuguese and evaluate the condition-specific quality of life King's Health Questionnaire (KHQ) for female urinary incontinence. METHODS: Two Brazilian translators, aware of the aim of the project, prepared two versions of the KHQ into Portuguese, which were back-translated into English by two other English translators. The differences were harmonized and pre-tested in a pilot study. The final version of the KHQ and the "Short-Form Health Survey" (SF-36), which has already been translated and validated into Portuguese were simultaneously administered to 156 and 119 women respectively. KHQ's psychometric properties such as reliability (internal consistency and retest) and construct validity were tested. A retest was performed within 2 weeks from the start date. RESULTS: The cultural adjustment process resulted in no changes in the KHQ Portuguese version, although for low schooling patients the questionnaire had to be read by the researcher during face-to-face interview. For all other patients, the KHQ was self-administered. KHQ's standardized Cronbach's alpha was 0.87 and when assessed by domains ranged from 0.49 to 0.92. Reliability measured by intraclass correlation (ICC) was considered moderate to strong for all domains and the severity measure scale ranged from 0.53 to 0.81. Pearson's correlation coefficient between KHQ and SF-36 was considered weak to moderate in the majority of the related domains, ranging from -0.27 to -0.53. CONCLUSIONS: The KHQ Portuguese version was translated and adjusted for Brazilian women with urinary incontinence complaints. It represents an important tool for the assessment of incontinent women in clinical trials.     

Validation of the "International Consultation on Incontinence Questionnaire -- Short Form" (ICIQ-SF) for Portuguese

OBJECTIVE: To translate into and validate for Portuguese the "International Consultation on Incontinence Questionnaire -- Short Form" (ICIQ-SF), a condition-specific quality-of-life questionnaire for patients with urinary incontinence. METHODS: Two Brazilians independently translated the original ICIQ-SF into Portuguese. These two translations were harmonized, and then checked by independent back-translation by two native English speakers. The harmonized translation was pre-tested in a pilot study on 20 patients. The final version of the ICIQ-SF in Portuguese was applied to 123 consecutive patients aged 16 or over (29 males and 94 females) with a complaint of urinary incontinence, who had sought the Department of Urogynecology and Uroneurology of the School of Medical Sciences of Unicamp. The Portuguese version of the King's Health Questionnaire (KHQ) was also applied to the same group. The psychometric properties of the questionnaire, such as reliability and construct validity were assessed. RESULTS: The median age was 53 years (range: 16 to 86). The mean retest interval for the ICIQ-SF was 14.37 days (range: 6 to 41). No changes from the original format of the ICIQ-SF were observed at the end of the process of translation and cultural adaptation. The internal consistency was high (0.88), as measured by the Cronbach alpha coefficient. The test-retest value was considered moderate to strong, as measured by the weighted Kappa index (range: 0.72 to 0.75) and Pearson correlation coefficient (0.89). The correlation between the ICIQ-SF and KHQ was considered to be moderate to good for most items (range: 0.44 to 0.77). The evaluation of the construct and concurrent validity was also satisfactory and statistically significant. CONCLUSIONS: The "International Consultation on Incontinence Questionnaire" (ICIQ-SF) was successfully translated into Portuguese and validated for application to Brazilian female and male patients complaining of urinary incontinence, with satisfactory reliability and construct validity.     

Endoscopic ultrasound assessment and tissue acquisition of mediastinal and abdominal lymph nodes

The differential diagnosis between benign and malignant lymph nodes (LNs) has a fundamental role in the characterization and staging of malignant conditions, as well as in subsequent patients’ management. All imaging modalities (i.e. computed tomography and magnetic resonance imaging) rely mainly on size; endoscopic ultrasound (EUS) criteria based on B-mode evaluation and Doppler features fail to adequately characterize with high specificity LNs nature. The introduction of EUS-elastography and contrast-enhanced harmonic EUS are useful techniques to increase the diagnostic yield in identifying metastatic LNs, to identify which suspicious LN should require pathological characterization and, finally, to target tissue acquisition. EUS-guided tissue acquisition (EUS-TA) is increasingly being used for diagnosing lymphadenopathy whenever the characterization modifies patients’ subsequent management and when no superficial LN is accessible. Since target therapy are currently available (i.e. lung cancer, breast cancer), EUS-TA of malignant LNs could be required to identify tumor biology. In this field, both fine needle aspiration and biopsy needles are able to guarantee accurate results with almost perfect specificity and sub-optimal sensitivity. We finally propose a diagnostic algorithm based on most recent, high-level evidence for the diagnostic approach to suspected LNs assessment.     

Dimerization and nuclear entry of mPER proteins in mammalian cells

Nuclear entry of circadian oscillatory gene products is a key step for the generation of a 24-hr cycle of the biological clock. We have examined nuclear import of clock proteins of the mammalian period gene family and the effect of serum shock, which induces a synchronous clock in cultured cells. Previously, mCRY1 and mCRY2 have been found to complex with PER proteins leading to nuclear import. Here we report that nuclear translocation of mPER1 and mPER2 (1) involves physical interactions with mPER3, (2) is accelerated by serum treatment, and (3) still occurs in mCry1/mCry2 double-deficient cells lacking a functional biological clock. Moreover, nuclear localization of endogenous mPER1 was observed in cultured mCry1/mCry2 double-deficient cells as well as in the liver and the suprachiasmatic nuclei (SCN) of mCry1/mCry2 double-mutant mice. This indicates that nuclear translocation of at least mPER1 also can occur under physiological conditions (i.e., in the intact mouse) in the absence of any CRY protein. The mPER3 amino acid sequence predicts the presence of a cytoplasmic localization domain (CLD) and a nuclear localization signal (NLS). Deletion analysis suggests that the interplay of the CLD and NLS proposed to regulate nuclear entry of PER in Drosophila is conserved in mammals, but with the novel twist that mPER3 can act as the dimerizing partner.     

Female double incontinence: prevalence,incidence, and risk factors from the SABE (Health,Wellbeing and Aging) study

Introduction and hypothesis

Double Incontinence (DI) is incontinence of urine and stool and is an extreme manifestation of pelvic floor dysfunction. The objective of this study was to estimate the prevalence and incidence of DI and the risk factors in elderly women in São Paulo, Brazil.

Methods

This was a prospective study in women aged 65 years or older evaluated in 2006 and re-evaluated in 2010. The sample was selected by two-phase stratified sampling with replacement and probability proportional to size. The likelihood ratio test was performed and Cox regression curves were generated to evaluate the equality of survival. Poisson’s regression was used to evaluate risk factors.

Results

This is the first study on the incidence of DI in elderly women. A total of 864 elderly women were interviewed in 2006. The prevalence rate of DI was 4.9%. The incidence rate of DI in the period between 2006 and 2010 was 13.8/1,000 person-years. Associated factors were the presence of chronic obstructive pulmonary disease, hypertension, difficulty with basic activities of daily living (BADL) and instrumental activities of daily living (IADL), polypharmacy and falls in the last year. Poisson’s regression analysis showed that falls in the last year and difficulty with at least three IADL were risk factors for DI.

Conclusions

The incidence of DI seems to be high in this population. Falls in the last year and difficulty with at least three IADL were identified as risk factors. Preventive measures must be implemented with public health policies to prevent increases in DI.