Quality of life after total laryngectomy: impact of different vocal rehabilitation methods in a middle income country.

INTRODUCTION: The impact of advanced laryngeal cancer and its extensive surgical treatments cause significant morbidity for these patients. Total laryngectomy impacts essential functions such as breathing, communication and swallowing, and may influence the quality of life as well as affecting the social life of laryngeal cancer patients. OBJECTIVE: Describe the quality of life and analyze the factors associated with the reduced quality of life in patients who have undergone total laryngectomy. METHOD: Observational cross-sectional study was carried out to evaluate the quality of life of patients who had undergone total laryngectomy due to laryngeal cancer. The fourth version of the UW-QOL Quality of Life Assessment Questionnaire from Washington University, validated for Portuguese, was used. RESULTS: The study population was 95 patients, and the mean composite score of the QOL was 80.4. In the subjective domains the majority of the patients (38.9%) reported they felt much better at present compared to the month before being diagnosed with cancer. When questioned about how they evaluated their health-related quality of life, there was a predominance of those who considered it good (43.2%), and most considered they had a good quality of life (46.3%) considering personal well-being. The overall quality of life was considered good to excellent by 83.2% of the patients. Patients with tracheoesophageal prosthesis reported a better quality of life, compared to patients using an electrolarynx or esophageal voice. CONCLUSION: The high mean value of the composite score for quality of life revealed that the patients assessed their quality of life positively. The absence of vocal emission was the only variable associated with a lower quality of life within the composite score according to the UW-QOL questionnaire.

Cytogenetic characterization of species of the family Heptapteridae (Teleostei: Siluriformes) from the Cuiabá and Ivaí River Basins, Brazil.

We cytogenetically characterized three species of Heptapteridae (Pimelodella sp, Pimelodella taenioptera, and Imparfinis schubarti) by investigating the distribution of constitutive heterochromatin and nucleolar organizer regions by silver nitrate impregnation (Ag-NOR) and fluorescence in situ hybridization. Pimelodella sp showed had a diploid number (2n) = 46 chromosomes, 26m + 10sm + 10st, and FN = 92; P. taenioptera, 2n = 52 chromosomes, 26m + 22sm + 4st, and FN = 104; and I. schubarti, 2n = 58 chromosomes, 28m + 28sm + 2st, and FN = 116. The two Pimelodella species had Ag-NORs sites on the submetacentric pair 14, located on the short arm in terminal position. In I. schubarti, the Ag-NORs sites were in an interstitial position on the long arm of the metacentric pair 1. C-banding revealed that Pimelodella sp contained a small amount of constitutive heterochroma-tin, whereas P. taenioptera contained a higher number of heterochromatic regions, in the pericentromeric, interstitial, and telomeric positions. I. schubarti had markers in centromeric and telomeric regions of a few chromosomes, and a large pericentromeric block on pair 1. Fluorochrome chromomycin A3 (CMA3) staining revealed positive signals on pair 14 in both Pimelodella species. Treatment with 4ꞌ,6-diamidino- 2-phenylindole (DAPI) revealed no markings in P. taenioptera, but an interstitial marking on the long arm of pair 14 in Pimelodella sp. In I. schubarti, positive signals of CMA3 were detected in the first pair, but negative signals were detected for DAPI staining. These results con-tribute to the karyotypic description of the less-studied species in the Brazilian Midwest.

Karyotype structure of Hypostomus cf. plecostomus (Linnaeus, 1758) from Tapajós River basin, Southern Amazon: occurrence of sex chromosomes (ZZ/ZW) and their evolutionary implications.

Hypostomus is a group of fish with numerical and struc-tural karyotypic variability. Among them, only six species, three of which belong to the Amazon basin, show a sex chromosome. In this study, we present the karyotype structure of Hypostomus cf. plecos-tomus from the Teles Pires river basin in the municipality of Alta Flo-resta, MT. The species has 2n = 68 and the karyotype formula 14m+ 24sm+ 14st+ 16a [fundamental number (FN) = 120] in males and 15m+ 24sm+14st+15a (FN = 121) in females and sex chromosomes ZZ/ZW. Argyrophilic nucleolar organizer regions (AgNORs) were identified in two pairs of chromosomes at different positions: short arm of the pair 21and long arm of the pair 27, matching the signals displayed by 18S FISH and indicating multiple NORs. Analysis of band C detected few blocks of constitutive heterochromatin in the pericentromeric regions of most chromosomes and the telomeric regions of some pairs, includ-ing the nucleolar pair 21. However, large blocks on the long arm of the nucleolar pair 27 still stood out. GC-rich heterochromatin (CMA3) was visualized only coincidently with nucleolar sites. Mapping of 5S rDNA sites with FISH revealed markings in eight chromosomes, demonstrat-ing synteny between the 18S and 5S sites. The data obtained for H. cf. plecostomus are important for taxonomic studies of this Amazon com-plex "H. plecostomus group". The occurrence of sex chromosomes in Amazon species of Hypostomus suggests an evolutionary event that is independent of other species in the group.

Karyotypic diversity of the armoured catfish genus Hypostomus (Siluriformes: Loricariidae) in the context of its occurrence and distribution.

This study taxonomically reviewed the specimens studied by Artoni & Bertollo (1996) and assimilated species of Hypostomus into three groups according to their cytogenetic characteristics, vagility and occurrence environments.

Short- and long-term theory-based predictors of physical activity in women who participated in a weight-management program.

This study analyzed psychosocial predictors of the Theory of Planned Behavior (TPB) and Self-Determination Theory (SDT) and evaluated their associations with short- and long-term moderate plus vigorous physical activity (MVPA) and lifestyle physical activity (PA) outcomes in women who underwent a weight-management program. 221 participants (age 37.6 ± 7.02 years) completed a 12-month SDT-based lifestyle intervention and were followed-up for 24 months. Multiple linear regression analyses tested associations between psychosocial variables and self-reported short- and long-term PA outcomes. Regression analyses showed that control constructs of both theories were significant determinants of short- and long-term MVPA, whereas affective and self-determination variables were strong predictors of short- and long-term lifestyle PA. Regarding short-term prediction models, TPB constructs were stronger in predicting MVPA, whereas SDT was more effective in predicting lifestyle PA. For long-term models, both forms of PA were better predicted by SDT in comparison to TPB. These results highlight the importance of comparing health behavior theories to identify the mechanisms involved in the behavior change process. Control and competence constructs are crucial during early adoption of structured PA behaviors, whereas affective and intrinsic sources of motivation are more involved in incidental types of PA, particularly in relation to behavioral maintenance.

Polymorphisms of the nucleolus organizing regions in Loricaria cataphracta (Siluriformes, Loricariidae) of the upper Paraguay River basin indicate an association with transposable elements.

A cytogenetic analysis of Loricaria cataphracta revealed a diploid number of 2n = 64 chromosomes, distributed as 12 metacentric + 8 submetacentric + 2 subtelocentric + 42 acrocentric, with a fundamental number of 86. Analysis of the nucleolus organizing region (NOR) using silver nitrate impregnation and fluorescence in situ hybridization (18S rDNA probe) techniques showed intra-population chromosomal polymorphism that could be classified into five different patterns (I to V), involving four pairs of chromosomes (8, 9, 12, and 13). In pattern I, the NOR was located in pair 12, whereas in pattern II, the NOR was detected in pair 8; these two patterns were characterized as a simple-NOR system. A multiple NOR system was evident in the other patterns (III, IV, and V). In pattern III, the NOR was located in only one of the homologs of pairs 12 and 8, and in patterns IV and V, the NOR was observed in pair 12 and in only one of the homologs of pairs 9 and 13, respectively. In addition, C-band analysis also showed this pattern of variation, and characterized a polymorphism in relation to the constitutive heterochromatin; the composition of this region was GC-rich (positive CMA3) and 4',6-diamidino-2-phenylindole negative. Transposition of NOR sites for mobile elements is suggested to explain this polymorphism.

Polymorphisms of the promoter and exon 3 of the receptor for advanced glycation end products (RAGE) in Euro- and Afro-Brazilians.

The receptor for advanced glycation end products (RAGE or AGER), a member of the immunoglobulin superfamily, is involved in pathologies such as atherosclerosis and diabetes. Over 50 SNPs were reported for RAGE, among which were the promoter region polymorphisms -429T>C (rs1800625), -374T>A (rs1800624) and a 63-bp deletion (-407 to -345 bp), all related to increased RAGE expression. Additionally, in the exon 3, a putative site of binding ligands, the missense variation G82S (rs2070600) was associated with skin disorders in patients with diabetes. We have determined allele, genotype and haplotype frequencies of RAGE polymorphisms -429T>C, -374T>A, 63-bp deletion and G82S in Euro-Brazilians (n = 108) and Afro-Brazilians (n = 91), characterized according to the predominant ancestry of the individuals. The allele frequencies for Euro- and Afro-Brazilians were as follows: -429C, 12.5% vs. 12.1% (P = 0.90); -374A, 31.5% vs. 26.2% (P = 0.25); 63del, 0.0% vs. 3.8% (P = 0.004); and 82S, 1.9% vs. 0.6% (P = 0.24). Absolute linkage disequilibrium was found between the promoter polymorphisms -429T>C and -374T>A plus the 63-bp deletion (D'=1.000; P < 0.0001). The haplotype frequencies differed (P = 0.003) between Euro- and Afro-Brazilians. Our results showed that the frequencies of the 63-bp deletion were higher in Afro-Brazilians, while the other analysed polymorphisms were similarly distributed in the studied populations. The -374T>A plus 63-bp deletion polymorphism captures more than 80% of the haplotypic variation in the studied population.

Low prevalence of glucokinase gene mutations in gestational diabetic patients with good glycemic control.

Glucokinase (GCK) plays a key role in glucose homeostasis. Gestational diabetes mellitus increases the risk of gestational complications in pregnant women and fetuses. We screened for mutations in coding and flanking regions of the GCK gene in pregnant women with or without gestational diabetes in a Brazilian population. A sample of 200 pregnant women classified as healthy (control, N = 100) or with gestational diabetes (N = 100) was analyzed for mutations in the GCK gene. All gestational diabetes mellitus patients had good glycemic control maintained by diet alone and no complications during pregnancy. Mutations were detected by single-strand conformation polymorphism and DNA sequencing. Thirteen of the 200 subjects had GCK gene mutations. The mutations detected were in intron 3 (c.43331A>G, new), intron 6 (c.47702T>C, rs2268574), intron 9 (c.48935C>T, rs2908274), and exon 10 (c.49620G>A, rs13306388). None of these GCK mutations were found to be significantly associated with gestational diabetes mellitus. In summary, we report a low frequency of GCK mutations in a pregnant Brazilian population and describe a new intronic variation (c.43331A>G, intron 3). We conclude that mutations in GCK introns and in non-translatable regions of the GCK gene do not affect glycemic control and are not correlated with gestational diabetes mellitus.

Prion-like Doppel gene polymorphisms and scrapie susceptibility in Portuguese sheep breeds.

The establishment of an association between prion protein gene (PRNP) polymorphisms and scrapie susceptibility in sheep has enabled the development of breeding programmes to increase scrapie resistance in the European Union. Intense selection for PRNP genotype may lead to correlated selection for genes linked to PRNP. We intended to investigate if any association exists between genetic variation in prion-like protein Doppel gene (PRND) and scrapie susceptibility, determined through PRNP genotyping. Sampling included 460 sheep from eight Portuguese breeds and the PRND gene coding region was analysed by multiple restriction fragment-single strand conformation polymorphism (MRF-SSCP), whereas PRNP genotyping was carried out by primer extension. A synonymous substitution (c.78G>A) was detected in codon 26 of the PRND gene, in all breeds except Churra Mondegueira. Linkage disequilibrium was found between the PRND and PRNP loci (P = 0.000). Specifically, PRND was monomorphic in the 45 animals with the more resistant ARR/ARR PRNP genotype (P = 0.003), whereas a higher frequency of PRND heterozygotes (GA) was associated with ARQ/AHQ (P = 0.029). These results constitute preliminary evidence of an association between a polymorphism in the PRND gene and scrapie susceptibility, and indicate that the possibility of undesirable consequences from widespread selection for PRNP genotype on genetic diversity and reproduction traits needs to be further investigated.

Possible origins of B chromosomes in Rineloricaria pentamaculata (Loricariidae, Siluriformes) from the Paraná River basin.

We made a cytogenetic study of Rineloricaria pentamaculata from the Tauá Stream, in the Pirapó River sub-basin in Paraná State, Brazil, focused on the occurrence and origins of the B chromosomes. The diploid number varied from 2n = 56 to 2n = 59, due to the presence of 0 to 3 B microchromosomes of the acrocentric type, which were observed in 92.3% of the specimens (N = 12). These chromosomes were totally heterochromatic, with the C banding technique, and there were inter- and intraindividual numerical differences. Meiotic cells in metaphase I had 28 bivalent chromosomes and 0 to 3 univalent chromosomes. We suggest that the B microchromosomes are centric fragments originated from chromosome rearrangements.

The functional polymorphisms -429T>C and -374T>A of the RAGE gene promoter are not associated with gestational diabetes in Euro-Brazilians.

The receptor for advanced glycation end products (RAGE or AGER) is a multiligand member of the immunoglobulin superfamily. RAGE is expressed in several tissues, including human myometrium, chorionic villi and placenta. Advanced glycation end products are the best studied ligands of RAGE; they have pro-inflammatory actions in human gestational tissues, increasing oxidative stress and the release of cytokines and prostaglandins. We investigated the association of RAGE gene promoter polymorphisms -429T>C (rs1800625) and -374T>A (rs1800624) with gestational diabetes. A sample of 750 unrelated European origin pregnant Brazilian women were classified as nondiabetic (control group, N = 600) or having gestational diabetes (N = 150) according to American Diabetes Association 2009 criteria. Genotyping was performed by PCR-RFLP. The frequencies of the rare alleles -429C (6.3 versus 9.1%) and -374A (26 versus 30%) were not significantly different between the gestational diabetes patients and healthy pregnant women. Also, the -429T>C and -374T>A polymorphisms were not associated with body mass index, lipid profile, fasting glycemia, HbA1C, or insulin requirement. We found that functional promoter polymorphisms of the RAGE gene were not associated with gestational diabetes or its complications in these Euro-Brazilian patients.

Difficulty in detecting low levels of polymyxin resistance in clinical Klebsiella pneumoniae isolates: evaluation of Rapid Polymyxin NP test, Colispot Test and SuperPolymyxin medium.

Polymyxins are important therapeutic options for treating infections, mainly those caused by carbapenem-resistant Klebsiella pneumoniae. Specific chemical characteristics of polymyxins make it difficult to perform antimicrobial susceptibility testing, especially within the clinical laboratory. Here we aimed to evaluate the performance of three phenotypic methods: Rapid NP Polymyxin Test, ColiSpot test and the SuperPolymyxin medium. To accomplish this, 170 non-duplicate clinical K. pneumoniae isolates were analysed (123 colistin-resistant and 47 susceptible). The sensitivity and specificity obtained for Rapid Polymyxin NP Test, Colispot and SuperPolymyxin medium were, respectively, 90% and 94%, 74% and 100%, and 82% and 85%. Very major errors occurred more frequently in low-level colistin-resistant isolates (MICs 4 and 8 µg/mL). Rapid Polymyxin NP proved to be a method capable of identifying colistin-resistant strains in acceptable categorical agreement. However, major errors and very major errors of this method were considered unacceptable for colistin-resistance screening. Although the Colispot test is promising and easy to perform and interpret, the results did not reproduce well in the isolates tested. The colistin-containing selective medium (SuperPolymyxin) showed limitations, including quantification of mucoid colonies and poor stability. Nevertheless, Colispot and SuperPolymyxin medium methods did not present acceptable sensitivity, specificity and categorical agreement. It is essential to use analytical tools that faithfully reproduce bacterial resistance in vitro, especially in last-line drugs, such as polymyxins, when misinterpretation of a test can result in therapeutic ineffectiveness.

Differentiation of Y chromosome in the X(1)X(1)X(2)X(2)/X(1)X(2)Y sex chromosome system of Hoplias malabaricus (Characiformes, Erythrinidae).

Six populations of Hoplias malabaricus from the Lower Paranapanema River were studied and showed 2n = 40 with metacentric/submetacentric chromosomes for females and 2n = 39 with m/sm chromosomes for males, thereby revealing a multiple sex chromosome system of the X(1)X(1)X(2)X(2)/X(1)X(2)Y type. The Y chromosome was the largest metacentric chromosome in all karyotypes. The heterochromatic blocks were located in a pericentromeric region of all pairs and in telomeric regions in some other pairs. A conspicuous pericentromeric C-band was detected in a single pair, equivalent to the X(1) chromosome. No markers on X(2) chromosomes were detected, but they could be identified by their size. The females displayed the same distribution of C and chromomycin A(3) bands and 18S rDNA sites among the populations. However, the males exhibited differences in both number and position of Giemsa C-bands on the Y chromosome, which were also evident after Chromomycin A(3) banding and FISH with the 18S rDNA probe. These cytogenetic tools allowed for the identification of possible mechanisms involved in the differentiation and evolution of the Y chromosome.

Potency evaluation of antivenoms in Brazil: the national control laboratory experience between 2000 and 2006.

Envenoming from snakebites is an important public health issue in Brazil. In 2005, 28,597 cases were notified (15 cases/100,000 inhabitants), 87.5% due to Bothrops and 9.2% to Crotalus genus. Antivenoms available in Brazil are liquid preparations containing purified equine Fab'2. Since 1987, the National Institute for Quality Control in Health (INCQS/FIOCRUZ) has been testing all lots prior to batch release. Between 2000 and 2006, 619 lots of antivenoms were tested, comprising 2,513,690 ampoules. The potency assay was performed only for bothropic and crotalic antivenoms (485 lots corresponding to 1,866,726 ampoules) due to the unavailability of the other reference venoms. This paper aims to report the last 7-year activities of INCQS on the quality control, batch release and potency evaluation of antivenoms.

Studies of Terminalia catappa L. oil: characterization and biodiesel production.

Since the biodiesel program has been started in Brazil, the investigation of alternative sources of triacylglycerides from species adapted at semi-arid lands became a very important task for Brazilian researchers. Thus we initiated studies with the fruits of the Terminalia catappa L (TC), popularly known in Brazil as "castanhola", evaluating selected properties and chemical composition of the oil, as well any potential application in biodiesel production. The oil was obtained from the kernels of the fruit, with yields around 49% (% mass). Also, its fatty acid composition was quite similar to that of conventional oils. The crude oil of the TC was transesterified, using a conventional catalyst and methanol to form biodiesel. The studied physicochemical properties of the TC biodiesel are in acceptable range for use as biodiesel in diesel engines.

Biopsied and vitrified bovine embryos viability is improved by trans10, cis12 conjugated linoleic acid supplementation during in vitro embryo culture.

Bovine embryos cultured in serum-containing media abnormally accumulate lipids in the cytoplasm. This is well known to contribute to their higher susceptibility to cryopreservation and biopsied embryos are even further susceptible. We aimed to improve in vitro produced (IVP) embryos resistance to micromanipulation and cryopreservation by supplementing serum-containing media with trans-10, cis-12 conjugated linoleic acid (t10, c12 CLA). The effect of t10, c12 CLA on lipid deposition and embryonic development was also tested. After in vitro maturation and fertilization (IVF day=D0), zygotes were cultured on granulosa cells+M199+10% serum+100microM GSH supplemented with 100microM of t10, c12 CLA (CLA group, n=1394) or without supplementation (control group, n=1431). Samples of D7/D8 embryos were observed under Nomarsky microscopy for lipid droplets evaluation while others were biopsied and vitrified (group B-Control, n=24; group B-CLA, n=23). Non-biopsied embryos were also frozen (group NB-Control, n=49; group NB-CLA, n=45). Biopsied cells were used for embryo sex determination. Postwarming embryo survival and viability were determined at 0 and 24h of culture, respectively. Supplementation of t10, c12 CLA did not influence cleavage, embryo sex ratio, D7/D8 embryo rate or morphological quality. CLA embryos had higher number of small lipid droplets (P

Newborn screening: a national public health programme in Brazil.

The newborn screening programme started in Brazil (1976) through isolated initiatives, without governmental directions and/or policies. According to Health Ministry (2000) data the coverage was 55% and unevenly distributed. Only 17 out of 27 Brazilian states had more than 30% coverage. Public budgets covered only diagnostic examinations. There were no official data about assistance, patient follow-up or detected disorders. The creation of the National Programme (2001) has provided new perspective for newborn screening (NBS) in the public health system. It has provided important official data and established management and care units for each state: Reference Services in Newborn Screening. The programme screened about 13 million newborns from October 2001 to December 2005. The coverage increased to 80.2% (2005) and 74% of the states presented coverage of over 70%. Within 34 accredited Reference Services in 27 Brazilian states, all provide screening for PKU and CH. Ten of them provide screening for haemoglobinopathies as well, and three of them provide also for CF. The Reference Services altogether count on at least 170 health professionals, such as paediatricians, endocrinologists, nutritionists, psychologists and social workers. They are qualified to assist positive cases, within the policies established by the National Programme. There has been significant increase in NBS coverage and follow-up assuredness, including detected cases before the National Programme (10,935 positive cases) mostly in those regions where the programme did not exist. There has been significant evolution in the Newborn Screening as a Public Health Program in Brazil due to the government's commitment (federal and each component state).

Cytogenetic characterization of Crenicichla (Pisces, Perciformes, Cichlidae) of the Iguaçu River.

Three populations of the genus Crenicichla, namely Crenicichla iguassuensis, Crenicichla sp 1 and Crenicichla sp 2, from the Iguaçu River, were analyzed cytogenetically, and their nucleolus organizer regions, constitutive heterochromatin distribution and chromomycin A3 markings were studied. Karyotype analyses showed a diploid number of 48 chromosomes, made up of 2 metacentric pairs, 2 submetacentric pairs, 7 subtelocentric pairs, and 13 acrocentric pairs for the three Crenicichla species and no sexual chromosome differentiation. Nucleolus organizer regions showed strong interstitial marking on the first chromosome pair, coincident with a constriction presented by Giemsa and positive marking by chromomycin. Although constitutive heterochromatin patterns were also similar, with pericentromeric markings, small differences in the three species could be observed. Crenicichla sp 2 presented some chromosomes with bitelomeric markings absent in Crenicichla iguassuensis and Crenicichla sp 1.

Cytogenetic analysis and description of the sexual chromosome determination system ZZ/ZW of species of the fish genus Serrapinnus (Characidae, Cheirodontinae).

Four populations of Serrapinnus notomelas and one population of Serrapinnus sp.1, both belonging to the subfamily Cheirodontinae, were analyzed by Giemsa and silver nitrate impregnation techniques. We found 2n = 52 chromosomes for all populations, with interspecific differences in the karyotype formula; S. notomelas showed 16 m + 22 sm + 10 st + 4a, with fundamental number (FN) = 100 for males, and 16 m + 23 sm + 10 st + 3a, with FN = 101 for females. Serrapinnus sp.1 had 8m + 16 sm + 4 st + 24 a, with FN = 80 for males, and 8m + 15 sm + 4 st + 25 a, with FN = 79 for females. The difference in FN for the two sexes is due to a pair of heteromorphic chromosomes in the females of both species, which characterizes a ZZ/ZW-type mechanism of chromosome sexual determination. Interspecies differences were also found in nucleolus organizer regions (NORs). A simple NOR system was detected in three of four S. notomelas populations, while Serrapinnus sp.1 had two chromosome pairs with NOR. Although S. notomelas and Serrapinnus sp.1 have the same diploid number, differences in the karyotype structure indicate that these are different species. Apparently there was pericentric inversion during the karyotype evolution of these species.