Molecular genetics study of deafness in Brazil: 8-year experience

Hereditary hearing loss is a complex disorder that involves a large number of genes. In developed countries, 1 in 1,000 children is born with deafness severe enough to require special education services, and about 60% of the cases of isolated deafness have a genetic origin. Although more than 100 genes for hearing loss are known currently, only a few are routinely tested in the clinical practice. In this study, we present our findings from the molecular diagnostic screening of the GJB2 and GJB3 genes, del(GJB6-D13S1,830) and del(GJB6-D13S1,854) deletions in the GJB6 gene, Q829X mutation in the otoferlin gene (OTOF) and, the A1,555G and A7,445G mutations in the mitochondrial genome over an 8-year period. Mutations analysis in the previously mentioned genes and mutations was performed on 645 unrelated Brazilian patients with hearing loss who fell into two different testing groups. Different mutations in the GJB2 gene were responsible for most of cases studied, but deletions in the GJB6 gene as well as mitochondrial mutations were also found. While most cases of hearing loss in this country are due to environmental factors, the genetic etiology of deafness will increasingly be determined as more genetic tests become available.     

Heparin-interacting sites of bovine lactoferrin are involved in anti-adenovirus activity

Lactoferrin, a member of the transferrin family of approximately 80 kDa, consists of a single polypeptide chain folded in two symmetric, globular lobes (N- and C-lobes), each able to bind one ferric ion. This glycoprotein, found in physiological fluids of mammals, plays an important role in immune regulation and in defense mechanisms against bacteria, fungi, parasites, and viruses. Although the antiviral activity of lactoferrin is one of the major biological functions of such protein, the mechanism of action is still under debate. We have investigated both the role of tryptic fragments of bovine lactoferrin and the mechanism of lactoferrin antiviral effect toward adenovirus infection in HEp-2 cells. The results obtained demonstrated that the anti-adenovirus activity of lactoferrin is mediated by the N-terminal half of the protein as the N-lobe was able to inhibit adenovirus infection, even if at lower extent than undigested lactoferrin, whereas C-lobe was ineffective. The results also showed that the anti-adenovirus action of lactoferrin and of its N-terminal peptide lactoferricin took place on virus attachment to cell membrane, mainly through competition for common glycosaminoglycan receptors. The data provide evidence that the anti-adenovirus activity of lactoferrin is mediated mainly by the cluster of positive charges at the N-terminus of whole molecule and that the N-terminal peptide lactoferricin alone is sufficient to prevent infection.     

Genetic relationship between Streptococcus pneumoniae isolates from nasopharyngeal and cerebrospinal fluid of two infants with Pneumococcal Meningitis

The molecular epidemiology of Streptococcus pneumoniae isolates from carriage and cerebrospinal fluid (CSF) concurrently recovered from the same individual has not yet been reported. By using pulsed-field gel electrophoresis, we demonstrated the genetic linkage among strains from CSF and nasopharynges of two children with pneumococcal meningitis.     

HIV-1 diversity among inmates of Italian prisons

In Italy, the prevalence of non-B HIV-1 subtypes ranges reportedly from 5.4% to 12.6%, yet there are no data on their circulation in prisons, where the prevalence of HIV infection is high. A retrospective study was conducted to evaluate the circulation of non-B subtypes and to characterize their determinants in five Italian prisons. To this end an aliquot of samples of blood was taken in the period 2001-2006 from all 262 HIV-positive inmates in whom antiretroviral treatment had failed. Complete HIV-1 PR and RT regions were sequenced for all samples and subjected to phylogenetic analysis; 250 (95.4%) sequences clustered with subtype B. The non-B subtype was found in 4% of Italian prison inmates and 16.7% of non-Italian prison inmates; the overall percentage increased from 1.8% for inmates infected in 1982-1990 to 4.4% in 1991-1999 and 21.9% in 2000-2006. Factors significantly associated with non-B subtypes were an exposure to other than injecting drug use and a first positive HIV test in 2000-2006. Non-B subtypes were distributed within five monophyletic clades. In all cases but one, it was possible to correlate the history of HIV-exposure to the origin of the clade, with high bootstrap values. In conclusion, although the sample may not be representative of the prison inmate population in Italy, the data suggest strongly that the circulation of non-B subtypes has apparently increased. Non-B subtypes were found to have been associated with heterosexual contact and time of the first HIV-positive test. Knowledge of the different subtypes circulating in prisons may be useful for tracking the epidemiology of HIV infection and for choosing antiretroviral therapy.     

ESR1 and APOE gene polymorphisms, serum lipids, and hormonal replacement therapy

OBJECTIVES: The risks and benefits of hormone replacement therapy (HRT) are, at least in part, mediated by the metabolic individuality of women. Therefore, we investigated the association between polymorphisms at the estrogen receptor 1 gene (ESR1) and at the apolipoprotein E gene (APOE) with lipid and lipoprotein levels in order to verify whether these concentrations are modulated by these gene variants in women with different hormonal status. METHODS: One hundred and eighteen postmenopausal women using oral HRT with estrogen or estrogen plus progestagen (HRT+, mean age=56+/-6.7 years, 39-75 years) and 167 postmenopausal women that were not on HRT (HRT-, mean age=58+/-9.8 years, 38-85 years) participated in the study. The polymorphisms were genotyped by PCR-RFLP methods. RESULTS: No significant effect of ESR1 genotypes or haplotypes and ESR1*HRT interactions were detected on lipid levels in two-way analysis of variance. Postmenopausal women HRT nonusers carriers of the APOE*4 allele had higher T-chol and LDL-C levels than postmenopausal women HRT nonusers carriers of the APOE*3 and APOE*2 allele. T-chol and LDL-C concentrations in postmenopausal users of HRT that were APOE*4 carriers were similar to those in postmenopausal women nonusers of HRT homozygotes for APOE*3 and APOE*2 carriers. A significant APOE*4/HRT interaction was detected on T-chol and LDL-C levels by multiple regression analysis. CONCLUSION: The results from this study suggest that the HRT influence on T-chol and LDL-C levels is modulated by APOE isoforms but not by ESR1 polymorphisms.     

Sleep disturbance scale for children: translation, cultural adaptation, and validation

IntroductionThe Sleep Disturbance Scale for Children (SDSC) is a 26-item instrument for evaluating sleep among children aged 3–18 years. It differentiates among conditions such as disorders of initiating and maintaining sleep, sleep breathing disorders, disorders of arousal, sleep–wake transition disorders, excessive somnolence, and sleep hyperhydrosis. The aim of this study was to translate, culturally adapt, and validate it for Brazilian Portuguese.MethodThe study was carried out in two phases: (1) forward translation, back translation, pretesting, and calculation of sample size; (2) validation: reliability (Chronbach’s alpha), convergent analysis (Pearson correlation), and discriminatory validity (comparing the scores of the test with the results of polysomnography). One hundred children, aged 3–18 years, accompanied by their parents and/or guardians participated in the phases. PSG studies have been done to calculate the sample size and validation.ResultsThe scale instructions and items were adapted regarding semantic, experiential, conceptual, and cultural equivalence validation. The scale structure related to visual communication was also adapted to Brazilian population preference and habits, and this resulted in a chart with clear instructions and easy recognition of the statements and possible responses. Reliability analysis showed values greater than 0.55. There has been reasonable convergent validity. Discriminatory validity using the PSG study for positive sleep-disordered breathing (SDB) was 8.9, attesting discriminatory validity only for SDB. The three questions of the scale can screen SDB.ConclusionThe SDSC was translated, adapted and validated for Brazilian Portuguese, and it presented internal consistency and convergent and discriminatory validity. It can be used in population-based studies in order to screen for sleep-disordered breathing in children.     

Micro-shear bond strength and surface micromorphology of a feldspathic ceramic treated with different cleaning methods after hydrofluoric acid etching

Objective

The aim of this study was to evaluate the effect of feldspathic ceramic surface cleaning on micro-shear bond strength and ceramic surface morphology.

Material and Methods

Forty discs of feldspathic ceramic were prepared and etched with 10% hydrofluoric acid for 2 minutes. The discs were randomly distributed into five groups (n=8): C: no treatment, S: water spray + air drying for 1 minute, US: immersion in ultrasonic bath for 5 minutes, F: etching with 37% phosphoric acid for 1 minute, followed by 1-minute rinse, F+US: etching with 37% phosphoric acid for 1 minute, 1-minute rinse and ultrasonic bath for 5 minutes. Composite cylinders were bonded to the discs following application of silane and hydrophobic adhesive for micro-shear bond strength testing in a universal testing machine at 0.5 mm/min crosshead speed until failure. Stereomicroscopy was used to classify failure type. Surface micromorphology of each treatment type was evaluated by scanning electron microscopy at 500 and 2,500 times magnification.

Results

One-way ANOVA test showed no significant difference between treatments (p=0.3197) and the most common failure types were cohesive resin cohesion followed by adhesive failure. Micro-shear bond strength of the feldspathic ceramic substrate to the adhesive system was not influenced by the different surface cleaning techniques. Absence of or less residue was observed after etching with hydrofluoric acid for the groups US and F+US.

Conclusions

Combining ceramic cleaning techniques with hydrofluoric acid etching did not affect ceramic bond strength, whereas, when cleaning was associated with ultrasound, less residue was observed.     

Atherosclerotic involvement of the radial artery in patients with coronary artery disease and its relation with midterm radial artery graft patency and endothelial function

BACKGROUND: The radial artery has recently been proposed as an alternative arterial conduit for surgical myocardial revascularization. This study was conceived to evaluate the degree of atherosclerotic involvement of the radial artery in patients with coronary artery disease and the eventual influence of a subtle degree of preoperative atherosclerosis on the midterm results of radial artery grafts. METHODS AND RESULTS: The intima-media thickness of the radial artery, common carotid artery, and internal thoracic artery was evaluated in 42 coronary artery disease patients and in 26 control patients. All radial arteries were then used for myocardial revascularization; 30 patients submitted to control angiography after 5 years. The mean intima-media thickness was 0.92 +/- 0.22 mm for the common carotid artery, 0.54 +/- 0.16 mm for the internal thoracic artery, 0.55 +/- 0.11 mm for the radial artery in coronary artery disease patients versus 0.79 +/- 0.14 mm, 0.52 +/- 0.11 mm, and 0.56 +/- 0.09 mm, respectively, in control patients (P =.001 only for the common carotid artery). No correlation was found between the intima-media thickness of the carotid, internal thoracic, and radial artery. No correlation was found between the preoperative intima-media thickness of the radial artery and the midterm patency and endothelial-mediated vasodilating capacity of radial artery grafts. CONCLUSION: In coronary artery disease patients, radial artery atherosclerotic involvement is more frequent than that of the gold standard internal thoracic artery but still by far less severe than that of the common carotid artery. The early atherosclerotic signs often observed in the radial artery do not seem to have the potential to influence radial artery graft patency and endothelial function.     

Discovery and cross‐validation of peripheral blood and renal biopsy gene expression signatures from ethnically diverse kidney transplant populations

We determined peripheral blood (PB) and biopsy (Bx) RNA expression signatures in a Brazilian and US cohort of kidney transplant patients. Phenotypes assigned by precise histology were: acute rejection (AR), interstitial fibrosis/tubular atrophy/chronic rejection (CR), excellent functioning transplants (TX), and glomerulonephritis recurrence (GN). Samples were analyzed on microarrays and profiles from each cohort were cross‐validated on the other cohort with similar phenotypes. We discovered signatures for each tissue: (1) AR vs TX, (2) CR vs TX, and (3) GN vs TX using the Random Forests algorithm. We validated biopsies signatures of AR vs TX (area under the curve [AUC] 0.97) and CR vs TX (AUC 0.87). We also validated both PB and Bx signatures of AR vs TX and CR vs TX with varying degrees of accuracy. Several biological pathways were shared between AR and CR, suggesting similar rejection mechanisms in these 2 clinical phenotypes. Thus, we identified gene expression signatures for AR and CR in transplant patients and validated them in independent cohorts of significantly different racial/ethnic backgrounds. These results reveal that there are strong unifying immune mechanisms driving transplant diseases and identified in the signatures discovered in each cohort, suggesting that molecular diagnostics across populations are feasible despite ethnic and environmental differences.     

Effect of surgical revascularization of a right coronary artery tributary of an infarcted nonischemic territory on the outcome of patients with three-vessel disease: a prospective randomized trial

BACKGROUND: We evaluated the in-hospital and long-term effects of surgical grafting of a dominant graftable right coronary artery tributary of an infarcted nonischemic territory in patients with triple-vessel disease who were undergoing coronary artery bypass grafting. METHODS: Of 303 consecutive patients undergoing coronary artery bypass grafting with 3-vessel coronary disease and a dominant right coronary artery tributary of an infarcted nonischemic territory, 154 were randomized to right coronary artery revascularization and 149 to no right coronary artery grafting. In all cases, standard on-pump surgical myocardial revascularization was performed. RESULTS: Overall hospital mortality was 2 of 154 versus 1 of 149 (P =.97); no difference in in-hospital outcome was observed between the 2 groups. At follow-up, cardiac event-free survival was 84 of 152 in the right coronary artery grafting series and 62 of 148 in the non-right coronary artery grafting group (P =.20). However, when the analysis was limited to surviving patients without new scintigraphic evidence of ischemia (to avoid confounding factors derived from ischemia in the left coronary system or right coronary artery graft malfunction), we found that patients who received a right coronary artery graft had fewer cardiac events, a lower incidence of arrhythmia, and less left ventricular dilatation than did the non-right coronary artery revascularized series. CONCLUSIONS: Surgical grafting of a right coronary artery tributary of an infarcted nonischemic territory in patients with 3-vessel coronary artery disease submitted to coronary artery bypass grafting improved late electric stability, ventricular geometry, and event-free survival but did not affect in-hospital or 10-year survival.