Reliability of nasal auscultation and their relation to middle ear condition – a clinical interest to pediatrics respiratory physiotherapy

Background: Acute respiratory infections are usual in children under three years old occurring in upper respiratory tract, having an impact on child and caregiver’s quality of life predisposing to otitis media or bronchiolitis. There are few valid and reliable measures to determine the child’s respiratory condition and to guide the physiotherapy intervention. Aim: To assess the intra and inter rater reliability of nasal auscultation, to analyze the relation between sounds’ classification and middle ear’s pressure and compliance as well as with the Clinical Severity Score. Methods: A cross-sectional observational study was composed by 125 nursery children aged up to three years old. Tympanometry, pulmonary and nasal auscultation and application of Clinical Severity Score were performed to each child. Nasal auscultation sounds’ were recorded and sent to 3 blinded experts, that classified, as “obstructed” and “unobstructed”, with a 48 hours interval, in order to analyze inter and intra rater reliability. Results: Nasal auscultation revealed a substantial inter and intra rater reliability (=0,749 and evaluator A - K= 0,691; evaluator B - K= 0,605 and evaluator C - K= 0,724, respectively). Both ears’ pressure was significantly lower in children with an "unobstructed" nasal sound when compared with an “obstructed” nasal sound (t=-3,599, p<0,001 in left ear; t=-2,258, p=0,026 in right ear). Compliance in both ears was significantly lower in children with an "obstructed" nasal sound when compared with “unobstructed” nasal sound (t=-2,728, p=0,007 in left ear; t=-3,830, p<0,001 in right ear). There was a statistically significant association between sounds’ classification and tympanograms types in both ear’s (=11,437, p=0,003 in left ear; =13,535, p=0,001 in right ear). There was a trend to children with an "unobstructed" nasal sound that had a lower clinical severity score when compared with “obstructed” children. Conclusion: It was observed a good intra and substantial inter reliability for nasal auscultation. Nasal auscultation sounds’ classification was related to middle ears’ pressure and compliance.

Intervenção em fisioterapia numa prótese total do ombro

Introdução: A artroplastia total do ombro é largamente aceite como tratamento para uma variedade de patologias do ombro com dor persistente e perda acentuada da funcionalidade. Em termos globais, os resultados obtidos após a intervenção cirúrgica são bastante bons, parecendo estar, no entanto, condicionados com a patologia subjacente e a qualidade da coifa dos rotadores. Objectivo: Pretende-se com este trabalho descrever e avaliar a eficácia de um plano de intervenção numa prótese total do ombro com integridade da coifa dos rotadores. Metodologia: Paciente sexo feminino, 61 anos, reformada, com dor acentuada no ombro esquerdo desde há três anos, submetida a uma cirurgia de colocação de prótese total do ombro. Apresentava limitação da amplitude articular, diminuição da força, alteração do ritmo escapulo-umeral e ligeira incapacidade funcional. Cinco semanas após a cirurgia, iniciou sessões diárias de fisioterapia durante 12 semanas. A intervenção incluiu a aplicação de modalidades anti-álgicas, mobilização articular, fortalecimento muscular e reeducação neuromuscular. Foi avaliada em quatro momentos: antes da intervenção (M1), 4 (M2), 7 (M3) e 12 (M4) semanas após. Resultados: A dor diminuiu de 4 para 0 (END) no final de 7 semanas e as amplitudes articulares passivas aproximaram-se dos valores definidos pelo protocolo com a flexão/elevação anterior a evoluir de 90º para 130º; a abdução/elevação lateral de 80º para 110º e a rotação medial de 35º para 70º. A força muscular inicial era de 3 nos grupos musculares flexores, extensores e rotadores do ombro atingindo no final grau 4. O ritmo escapulo-umeral demonstrou assincronia durante a elevação do ombro na primeira avaliação, ficando quase normalizado no final das sessões. Quanto à funcionalidade, antes da intervenção o score obtido foi de 30.8 atingindo um valor de 11.6 no final das sessões de tratamento. Conclusão: Após 12 semanas de tratamento, os objectivos delineados foram praticamente atingidos, uma vez que a utente já não refere dor no ombro e apresenta uma boa capacidade funcional, apesar de apresentar ligeiras diferenças nas amplitudes articulares e força muscular esperadas, apontando para o sucesso do protocolo de tratamento aplicado.

Lipid content of frozen fish: comparison of different extraction methods and variability during freezing storage

In this work, a microwave-assisted extraction (MAE) methodology was compared with several conventional extraction methods (Soxhlet, Bligh & Dyer, modified Bligh & Dyer, Folch, modified Folch, Hara & Radin, Roese-Gottlieb) for quantification of total lipid content of three fish species: horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), and sardine (Sardina pilchardus). The influence of species, extraction method and frozen storage time (varying from fresh to 9 months of freezing) on total lipid content was analysed in detail. The efficiencies of methods MAE, Bligh & Dyer, Folch, modified Folch and Hara & Radin were the highest and although they were not statistically different, differences existed in terms of variability, with MAE showing the highest repeatability (CV = 0.034). Roese-Gottlieb, Soxhlet, and modified Bligh & Dyer methods were very poor in terms of efficiency as well as repeatability (CV between 0.13 and 0.18).

Accessing indoor fungal contamination using conventional and molecular methods in Portuguese poultries

Epidemiological studies showed increased prevalence of respiratory symptoms and adverse changes in pulmonary function parameters in poultry workers, corroborating the increased exposure to risk factors, such as fungal load and their metabolites. This study aimed to determine the occupational exposure threat due to fungal contamination caused by the toxigenic isolates belonging to the complex of the species of Aspergillus flavus and also isolates fromAspergillus fumigatus species complex. The study was carried out in seven Portuguese poultries, using cultural and molecularmethodologies. For conventional/cultural methods, air, surfaces, and litter samples were collected by impaction method using the Millipore Air Sampler. For the molecular analysis, air samples were collected by impinger method using the Coriolis μ air sampler. After DNA extraction, samples were analyzed by real-time PCR using specific primers and probes for toxigenic strains of the Aspergillus flavus complex and for detection of isolates from Aspergillus fumigatus complex. Through conventional methods, and among the Aspergillus genus, different prevalences were detected regarding the presence of Aspergillus flavus and Aspergillus fumigatus species complexes, namely: 74.5 versus 1.0% in the air samples, 24.0 versus 16.0% in the surfaces, 0 versus 32.6% in new litter, and 9.9 versus 15.9%in used litter. Through molecular biology, we were able to detect the presence of aflatoxigenic strains in pavilions in which Aspergillus flavus did not grow in culture. Aspergillus fumigatus was only found in one indoor air sample by conventional methods. Using molecular methodologies, however, Aspergillus fumigatus complex was detected in seven indoor samples from three different poultry units. The characterization of fungal contamination caused by Aspergillus flavus and Aspergillus fumigatus raises the concern of occupational threat not only due to the detected fungal load but also because of the toxigenic potential of these species.

Suspected acute toxoplasmosis in pregnant women

OBJECTIVE: To determine the prevalence of reagent serology for suspected acute toxoplasmosis in pregnant women and to describe clinical, laboratory and therapeutic profiles of mothers and their children. METHODS: A retrospective study was conducted with IgM-anti-Toxoplasma gondii-reagent pregnant women and their children who attended the public health system in the state of Paraná, Southern Brazil, from January 2001 to December 2003. Information were obtained from clinical, laboratory (ELISA IgM/IgG) and ultrasonographic data and from interviews with the mothers. To test the homogeneity of the IgM indices in relation to the treatment used, the Pearson's Chi-square test was applied. Comparisons were considered significant at a 5% level. RESULTS: Two hundred and ninety (1.0%) cases of suspected IgM-reagent infection were documented, with a prevalence of 10.7 IgM-reagent women per 1,000 births. Prenatal care started within the first 12 weeks for 214/290; 146/204 were asymptomatic. Frequent complaints included headaches, visual disturbance and myalgia. Ultrasonography revealed abnormalities in 13 of 204 pregnancies. Chemoprophylaxis was administered to 112/227; a single ELISA test supported most decisions to begin treatment. Pregnant women with IgM indices =2.000 tended to be treated more often. Among exposed children, 44/208 were serologically followed up and all were IgG-reagent, and three IgM-reagent cases showed clinical symptoms. CONCLUSIONS: The existence of pregnant women with laboratorially suspected acute toxoplasmosis who were not properly followed up, and of fetuses that were not adequately monitored, shows that basic aspects of the prenatal care are not being systematically observed. There is need of implementing a surveillance system of pregnant women and their children exposed to T. gondii.

Differential expression of the eukaryotic release factor 3 (eRF3/GSPT1) according to gastric cancer histological types

Background: There are now several lines of evidence to suggest that protein synthesis and translation factors are involved in the regulation of cell proliferation and cancer development. Aims: To investigate gene expression patterns of eukaryotic releasing factor 3 (eRF3) in gastric cancer. Methods: RNA was prepared from 25 gastric tumour biopsies and adjacent non-neoplastic mucosa. Real time TaqMan reverse transcription polymerase chain reaction (RT-PCR) was performed to measure the relative gene expression levels. DNA was isolated from tumour and normal tissues and gene dosage was determined by a quantitative real time PCR using SYBR Green dye. Results: Different histological types of gastric tumours were analysed and nine of the 25 tumours revealed eRF3/GSPT1 overexpression; moreover, eight of the 12 intestinal type carcinomas analysed overexpressed the gene, whereas eRF3/GSPT1 was overexpressed in only one of the 10 diffuse type carcinomas (Kruskal-Wallis Test; p , 0.05). No correlation was found between ploidy and transcript expression levels of eRF3/GSPT1. Overexpression of eRF3/GSPT1 was not associated with increased translation rates because the upregulation of eRF3/GSPT1 did not correlate with increased eRF1 levels. Conclusions: Overexpression of eRF3/GSPT1 in intestinal type gastric tumours may lead to an increase in the translation efficiency of specific oncogenic transcripts. Alternatively, eRF3/GSPT1 may be involved in tumorigenesis as a result of its non-translational roles, namely (dis)regulating the cell cycle, apoptosis, or transcription.

Role of the wider Azores region as a nursery ground for North Atlantic blue shark (Prionace glauca)

Tese de Doutoramento, Ciências do Mar (Ecologia Marinha)

Mould and yeast identification in archival settings: preliminary results on the use of traditional methods and molecular biology options in Portuguese archives

This project was developed to fully assess the indoor air quality in archives and libraries from a fungal flora point of view. It uses classical methodologies such as traditional culture media – for the viable fungi – and modern molecular biology protocols, especially relevant to assess the non-viable fraction of the biological contaminants. Denaturing high-performance liquid chromatography (DHPLC) has emerged as an alternative to denaturing gradient gel electrophoresis (DGGE) and has already been applied to the study of a few bacterial communities. We propose the application of DHPLC to the study of fungal colonization on paper-based archive materials. This technology allows for the identification of each component of a mixture of fungi based on their genetic variation. In a highly complex mixture of microbial DNA this method can be used simply to study the population dynamics, and it also allows for sample fraction collection, which can, in many cases, be immediately sequenced, circumventing the need for cloning. Some examples of the methodological application are shown. Also applied is fragment length analysis for the study of mixed Candida samples. Both of these methods can later be applied in various fields, such as clinical and sand sample analysis. So far, the environmental analyses have been extremely useful to determine potentially pathogenic/toxinogenic fungi such as Stachybotrys sp., Aspergillus niger, Aspergillus fumigatus, and Fusarium sp. This work will hopefully lead to more accurate evaluation of environmental conditions for both human health and the preservation of documents.

Determination of the radiochemical purity of 99mTc-Tetrofosmin: comparision between five methods

GOAL: The manufacturing and distribution of strips of instant thin - layer chromatography with silica gel (ITLC - SG) (reference method) is currently discontinued so there is a need for an alternative method f or the determination of radiochemical purity (RCP) of 99m Tc - tetrofosmin. This study aims to compare five alternative methods proposed by the producer to determine the RCP of 99m Tc - tetrofosmin. METHODS: Nineteen vials of tetrofosmin were radiolabelled with 99m Tc and the percentages of the RCP were determined. Five different methods were compared with the standard RCP testing method (ITLC - SG, 2x20 cm): Whatman 3MM (1x10 cm) with acetone and dichloro - methane (method 1); Whatman 3MM (1x1 0 cm) with ethyl acetate (method 2); aluminum oxide - coated plastic thin - layer chromatography (TLC) plate (1x10 cm) and ethanol (method 3); Whatman 3MM (2x20 cm) with acetone and dichloro - methane (method 4); solid - phase extraction method C18 cartridge (meth od 5). RESULTS: The average values of RCP were 95,30% ± 1,28% (method 1), 93,95 ± 0,61% (method 2), 96,85% ± 0,93% (method 3), 92,94% ± 0,99% (method 4) and 96,25% ± 2,57% (method 5) (n=12 each), and 93,15% ± 1,13% for the standard method (n=19). There we re statistical significant differences in the values obtained for methods 1 (P=0,001), 3 (P=0,000) and 5 (P=0,004), and there were no statistical significant differences in the values obtained for methods 2 (P=0,113) and 4 (P=0,327). CONCLUSION: From the results obtained, methods 2 and 4 showed a higher correlation with the standard method. Unlike method 4, method 2 is less time - consuming than the reference method and can overcome the problems associated with the solvent toxicity. The remaining methods (1, 3 and 5) tended to overestimate RCP value compared to the standard method.

Practical Quality Control: Comparision of Methods on the Quantification of Stationary Phases in Paper and Thin-Layer Chromatographic Systems

Introduction: Paper and thin layer chromatography methods are frequently used in Classic Nuclear Medicine for the determination of radiochemical purity (RCP) on radiopharmaceutical preparations. An aliquot of the radiopharmaceutical to be tested is spotted at the origin of a chromatographic strip (stationary phase), which in turn is placed in a chromatographic chamber in order to separate and quantify radiochemical species present in the radiopharmaceutical preparation. There are several methods for the RCP measurement, based on the use of equipment as dose calibrators, well scintillation counters, radiochromatografic scanners and gamma cameras. The purpose of this study was to compare these quantification methods for the determination of RCP. Material and Methods: 99mTc-Tetrofosmin and 99mTc-HDP are the radiopharmaceuticals chosen to serve as the basis for this study. For the determination of RCP of 99mTc-Tetrofosmin we used ITLC-SG (2.5 x 10 cm) and 2-butanone (99mTc-tetrofosmin Rf = 0.55, 99mTcO4- Rf = 1.0, other labeled impurities 99mTc-RH RF = 0.0). For the determination of RCP of 99mTc-HDP, Whatman 31ET and acetone was used (99mTc-HDP Rf = 0.0, 99mTcO4- Rf = 1.0, other labeled impurities RF = 0.0). After the development of the solvent front, the strips were allowed to dry and then imaged on the gamma camera (256x256 matrix; zoom 2; LEHR parallel-hole collimator; 5-minute image) and on the radiochromatogram scanner. Then, strips were cut in Rf 0.8 in the case of 99mTc-tetrofosmin and Rf 0.5 in the case of 99mTc-HDP. The resultant pieces were smashed in an assay tube (to minimize the effect of counting geometry) and counted in the dose calibrator and in the well scintillation counter (during 1 minute). The RCP was calculated using the formula: % 99mTc-Complex = [(99mTc-Complex) / (Total amount of 99mTc-labeled species)] x 100. Statistical analysis was done using the test of hypotheses for the difference between means in independent samples. Results:The gamma camera based method demonstrated higher operator-dependency (especially concerning the drawing of the ROIs) and the measures obtained using the dose calibrator are very sensitive to the amount of activity spotted in the chromatographic strip, so the use of a minimum of 3.7 MBq activity is essential to minimize quantification errors. Radiochromatographic scanner and well scintillation counter showed concordant results and demonstrated the higher level of precision. Conclusions: Radiochromatographic scanners and well scintillation counters based methods demonstrate to be the most accurate and less operator-dependant methods.

Comparative study between two distinct methods for attenuation correction and the use of no-attenuation correction for the assessment of relative uptake of 99mTc-DMSA on pediatric patients

Introduction: Although relative uptake values aren’t the most important objective of a 99mTc-DMSA scan, they are important quantitative information. In most of the dynamic renal scintigraphies attenuation correction is essential if one wants to obtain a reliable result of the quantification process. Although in DMSA scans the absent of significant background and the lesser attenuation in pediatric patients, makes that this attenuation correction techniques are actually not applied. The geometric mean is the most common method, but that includes the acquisition of an anterior (extra) projection, which it is not acquired by a large number of NM departments. This method and the attenuation factors proposed by Tonnesen will be correlated with the absence of attenuation correction procedures. Material and Methods: Images from 20 individuals (aged 3 years +/- 2) were used and the two attenuation correction methods applied. The mean time of acquisition (time post DMSA administration) was 3.5 hours +/- 0.8h. Results: The absence of attenuation correction showed a good correlation with both attenuation methods (r=0.73 +/- 0.11) and the mean difference verified on the uptake values between the different methods were 4 +/- 3. The correlation was higher when the age was lower. The attenuation correction methods correlation was higher between them two than with the “no attenuation correction” method (r=0.82 +/- 0.8), and the mean differences of the uptake values were 2 +/- 2. Conclusion: The decision of not doing any kind of attenuation correction method can be justified by the minor differences verified on the relative kidney uptake values. Nevertheless, if it is recognized that there is a need for an accurate value of the relative kidney uptake, then an attenuation correction method should be used. Attenuation correction factors proposed by Tonnesen can be easily implemented and so become a practical and easy to implement alternative, namely when the anterior projection - needed for the geometric mean methodology – is not acquired.

Assessment of nursing students and nurses' orientation towards patient-centeredness

Background - Being patient centered is a core value for nursing. Patient centered-care has been related to patient and health provider satisfaction, better health outcomes, higher quality of care and more efficient health care delivery. Objectives - The purpose was to assess the orientation adopted by nurses and students in patient care, using The Patient-Practitioner Orientation Scale, as well as to compare the results between resident nurses and students from different academic years. Settings - Public School of Nursing and a Central Hospital, in Lisbon (Portugal). Participants - Students in the first, second and fourth year of nursing school and nurses participated in the study. Methods - For data collection, we used The Patient-Practitioner Orientation Scale (European Portuguese version), an instrument designed to measure individual preferences toward the dimension of caring a sharing in health professional-patient relationship. Students and nurses also filled out two additional questions about their perception of competence in technical and communication skills. Additional demographic information was also collected, including gender, age, academic year and length of professional experience. Results - A total of 525 students (84.7% female) and 108 nurses (77.8% female) participated in this study. In general, caring sub-scores, measuring the preference of about attending to patient emotional aspects, were higher than sharing sub-scores, measuring beliefs about giving information and perceiving patient as a member of the health team. Students were significantly more patient-centered throughout their nursing education (p<0.001). Comparing to students in the second and fourth academic years (p<0.001) nurses' scores were significantly lower both in total PPOS and in caring and sharing subscales. Conclusions - These results reinforce the idea that patient centeredness may be developed in academic context. The scores obtained highlight the importance of studies that aim to identify factors that may explain the decrease of patient centeredness in professional practice.

Efeitos de um programa prolongado de exercício na tolerância ao esforço de indivíduos com antecedentes de síndrome coronário agudo

Objectivos: Este estudo teve por objectivo verificar a influência de um programa prolongado de exercício físico em parâmetros de aptidão física e cardiovasculares avaliados em prova de esforço máximo, em sujeitos com diagnóstico recente de síndrome coronário agudo. Métodos: A amostra, constituída por 50 sujeitos, foi distribuída por um grupo experimental (n=25) e por um grupo controlo (n=25), tendo o grupo experimental ficado sujeito a um programa de exercício físico regular durante 52 semanas. Todos os indivíduos realizaram duas provas de esforço máximas (PEM) em tapete rolante, uma no início e a outra no final do protocolo experimental, tendo a primeira prova sido realizada 2-3 meses após o evento cardíaco. Nas PEM foram registados parâmetros de aptidão física (velocidade máxima, inclinação máxima do tapete e tempo de prova), dos indicadores metabólicos (METs máximo) e parâmetros cardiovasculares (frequência cardíaca repouso e máxima durante a prova, tensão arterial de repouso e máxima durante a prova e o duplo produto máximo e em repouso). Resultados: Comparativamente à primeira prova, na segunda PEM verificou-se no grupo experimental um aumento significativo (p<0,05) dos valores absolutos dos indicadores de aptidão física, com uma percentagem de variação destes parâmetros significativamente superior à do grupo controlo. No entanto, apesar do melhor desempenho físico evidenciado pelo grupo experimental, não se verificaram diferenças significativas entre os dois grupos no que respeita aos parâmetros cardiovasculares. Conclusão: Os resultados permitem concluir que o programa de exercícios se revelou vantajoso para os indivíduos do grupo experimental, tendo-lhes aumentado a tolerância ao esforço físico máximo sem uma aparente sobrecarga cardiovascular adicional.

Efficiency of sampling methods and effort to assess arthropod diversity in Azorean native forests

The BALA project (Biodiversity of Arthropods of Laurisilva of the Azores) is a research initiative to quantify the spatial distribution of arthropod biodiversity in native forests of the Azores archipelago. Arthropods were collected using a combination of two techniques, targeting epigean (ground dwelling) and canopy (arboreal) arthropods: pitfall traps (with Turquin and Ethylene solutions) and beating samples (using the three most dominant plant species). A total of 109 transects distributed amongst 18 forest fragments in seven of the nine Azorean islands were used in this study. The performance of alternative sampling methods and effort were tested. No significant differences were found in the accumulated number of species captured whether an alternative method was used or whether another transect with similar effort was established in another location within the same fragment. A combination of Ethylene and Turquin traps captured more species per individual, Turquin and beating captured more species per sample, and Turquin captured more species per unit time. An optimization exercise was performed and we found that the protocol applied during recent years is very close to optimal, allowing its future replication with confidence. The minimum combinations of sampling effort and methods, in order to monitor or to inventory diversity, taking into account different proportions of sample completeness are discussed.