Texture-based classification of lung disease patterns in chronic hypersensitivity pneumonitis and comparison to clinical outcomes.

Computer-aided detection algorithms applied to CT lung imaging have the potential to objectively quantify pulmonary pathology. We aim to develop an automatic classification method based on textural features able to classify healthy and pathological patterns on CT lung images and to quantify the extent of each disease pattern in a group of patients with chronic hypersensitivity pneumonitis (cHP), in comparison to pulmonary function tests (PFTs).27 cHP patients were scanned via high resolution CT (HRCT) at full-inspiration. Regions of interest (ROIs) were extracted and labeled as normal (NOR), ground glass opacity (GGO), reticulation (RET), consolidation (C), honeycombing (HB) and air trapping (AT). For each ROI, statistical, morphological and fractal parameters were computed. For automatic classification, we compared two classification methods (Bayesian and Support Vector Machine) and three ROI sizes. The classifier was therefore applied to the overall CT images and the extent of each class was calculated and compared to PFTs. Better classification accuracy was found for the Bayesian classifier and the 16x16 ROI size: 92.1±2.7%. The extent of GGO, HB and NOR significantly correlated with forced vital capacity (FVC) and the extent of NOR with carbon monoxide diffusing capacity (DLCO).Clinical Relevance- Texture analysis can differentiate and objectively quantify pathological classes in the lung parenchyma and may represent a quantitative diagnostic tool in cHP.

WFS1 and non-syndromic low-frequency sensorineural hearing loss: a novel mutation in a Portuguese case.

Low-frequency sensorineural hearing loss (LFSNHL) is an unusual type of HL in which frequencies at 2,000 Hz and below are predominantly affected. Most of the families with LFSNHL carry missense mutations in WFS1 gene, coding for wolframin. A Portuguese patient aged 49, reporting HL since her third decade of life, and also referring tinnitus, was shown to display bilateral moderate LFSNHL after audiological evaluation. Molecular analysis led to the identification of a novel mutation, c.511G>A (p.Asp171Asn), found in heterozygosity in the exon 5 of the WFS1 gene, and changing the aspartic acid at position 171 to an asparagine, in the extracellular N-terminus domain of the wolframin protein. This novel mutation wasn't present either in 200 control chromosomes analyzed or in the hearing proband's half-brother, and it had not been reported in 1000 Genomes, Exome Variant Server, HGMD or dbSNP databases. No mutations were found in GJB2 and GJB6 genes. Multi-alignment of 27 wolframin sequences from mammalian species, against the human wolframin sequence in ConSurf, indicated a conservation score corresponding to 7 in a 1-9 color scale where 9 is conserved and 1 is variable. In addition, the mutation p.Asp171Asn was predicted to be damaging and possibly damaging by SIFT and Polyphen-2, respectively. The auditory phenotype of this patient could thus be due to the novel mutation p.Asp171Asn. Further functional characterization might enable to elucidate in which way the change in the residue 171, as other changes introduced by LFSNHL-associated mutations previously described, leads to this type of HL.

Assessing Noncoding Sequence Variants of GJB2 for Hearing Loss Association.

Involvement of GJB2 noncoding regions in hearing loss (HL) has not been extensively investigated. However, three noncoding mutations, c.-259C>T, c.-23G>T, and c.-23+1G>A, were reported. Also, c.-684_-675del, of uncertain pathogenicity, was found upstream of the basal promoter. We performed a detailed analysis of GJB2 noncoding regions in Portuguese HL patients (previously screened for GJB2 coding mutations and the common GJB6 deletions) and in control subjects, by sequencing the basal promoter and flanking upstream region, exon 1, and 3'UTR. All individuals were genotyped for c.-684_-675del and 14 SNPs. Novel variants (c.-731C>T, c.-26G>T, c.*45G>A, and c.*985A>T) were found in controls. A hearing individual homozygous for c.-684_-675del was for the first time identified, supporting the nonpathogenicity of this deletion. Our data indicate linkage disequilibrium (LD) between SNPs rs55704559 (c.*168A>G) and rs5030700 (c.*931C>T) and suggest the association of c.[*168G;*931T] allele with HL. The c.*168A>G change, predicted to alter mRNA folding, might be involved in HL.

Dilution and magnification effects on image analysis applications in activated sludge characterization.

The properties of activated sludge systems can be characterized using image analysis procedures. When these systems operate with high biomass content, accurate sludge characterization requires samples to be diluted. Selection of the best image acquisition magnification is directly related to the amount of biomass screened. The aim of the present study was to survey the effects of dilution and magnification on the assessment of aggregated and filamentous bacterial content and structure using image analysis procedures. Assessments of biomass content and structure were affected by dilutions. Therefore, the correct operating dilution requires careful consideration. Moreover, the acquisition methodology comprising a 100× magnification allowed data on aggregated and filamentous biomass to be determined and smaller aggregates to be identified and characterized, without affecting the accuracy of lower magnifications regarding biomass representativeness.

Validity of the WHO VAW study instrument for estimating gender-based violence against women.

OBJECTIVE: To validate the instrument of the World Health Organization Violence Against Women (WHO VAW) study on psychological, physical and sexual violence against women perpetrated by intimate partners. METHODS: This was a cross-sectional study conducted in several countries between 2000 and 2003, including Brazil. Representative random samples of women aged 15-49 years with intimate partners were selected, living in the city of São Paulo (n = 940) and in the Zona da Mata, Pernambuco (n = 1,188), southeastern and northeastern regions, respectively. Exploratory factor analysis on questions relating to violence was performed (four psychological, six physical and three sexual questions), with varimax rotation and creation of three factors. Cronbach's alpha was calculated to analyze the internal consistency. To validate through extreme groups, mean scores (0 to 13 points) for violence were tested in relation to the following outcomes: self-rated health, daily activities, presence of discomfort or pain, suicidal ideation or attempts, heavy alcohol consumption and presence of common mental disorders. RESULTS: Three factors were defined, with similar accumulated variance (0.6092 in São Paulo and 0.6350 in the Zona da Mata). For São Paulo, the first factor was determined by physical violence, the second by sexual violence and the third by psychological violence. For the Zona da Mata, the first factor was formed by psychological violence, the second by physical violence and the third by sexual violence. Cronbach's alpha coefficients were 0.88 in São Paulo and 0.89 in the Zona da Mata. The mean scores for violence were significantly higher for less favorable outcomes, with the exception of suicide attempts in São Paulo. CONCLUSIONS: The instrument was shown to be adequate for estimating gender-based violence against women perpetrated by intimate partners and can be used in studies on this subject. It has high internal consistency and a capacity to discriminate between different forms of violence (psychological, physical and sexual) perpetrated in different social contexts. The instrument also characterizes the female victim and her relationship with the aggressor, thereby facilitating gender analysis.

Validade do instrumento WHO VAW STUDY para estimar violência de gênero contra a mulher / Validez de instrumento para estimar violencia de género contra la mujer / Validity of the WHO VAW study instrument for estimating gender-based violence against women

OBJETIVO: Validar o instrumento do estudo World Health Organization Violence Against Women (WHO VAW) sobre violência psicológica, física e sexual por parceiros íntimos contra mulheres. MÉTODOS: Estudo transversal realizado em vários países entre 2000 e 2003, inclusive Brasil. Selecionaram-se amostras aleatórias e representativas de mulheres de 15-49 anos com parceiros íntimos, residentes na cidade de São Paulo, SP, (n = 940) e na Zona da Mata de Pernambuco (n = 1.188). Realizou-se análise fatorial exploratória das perguntas sobre violências (quatro psicológicas, seis físicas e três sexuais), com rotação varimax e criação de três fatores. Calculou-se alfa de Cronbach para análise da consistência interna. Para a validação por grupos extremos, médias de escores (zero a 13 pontos) de violência foram testadas em relação aos desfechos: auto-avaliação de saúde, atividades diárias, presença de dor ou desconforto, ideação e tentativa de suicídio, grande consumo de álcool e presença de transtorno mental comum. RESULTADOS: Foram definidos três fatores com variância acumulada semelhante (0,6092 em São Paulo e 0,6350 na Zona da Mata)...

OBJECTIVE: To validate the instrument of the World Health Organization Violence Against Women (WHO VAW) study on psychological, physical and sexual violence against women perpetrated by intimate partners. METHODS: This was a cross-sectional study conducted in several countries between 2000 and 2003, including Brazil. Representative random samples of women aged 15-49 years with intimate partners were selected, living in the city of São Paulo (n = 940) and in the Zona da Mata, Pernambuco (n = 1,188), southeastern and northeastern regions, respectively. Exploratory factor analysis on questions relating to violence was performed (four psychological, six physical and three sexual questions), with varimax rotation and creation of three factors. Cronbach's alpha was calculated to analyze the internal consistency. To validate through extreme groups, mean scores (0 to 13 points) for violence were tested in relation to the following outcomes: self-rated health, daily activities, presence of discomfort or pain, suicidal ideation or attempts, heavy alcohol consumption and presence of common mental disorders. RESULTS: Three factors were defined, with similar accumulated variance (0.6092 in São Paulo and 0.6350 in the Zona da Mata)...

OBJETIVO: Validar preguntas sobre violencia psicológica, física y sexual por parejas íntimas contra mujeres. MÉTODOS: Estudio transversal, coordinado por la Organización Mundial de la Salud, realizado en varios países (2000-2003), inclusive Brasil. Se seleccionaron muestras aleatorias y representativas de mujeres de 15-49 años con parejas íntimas, residentes, en la ciudad de Sao Paulo (en Sureste de Brasil, n=940) y en la Zona de Mata de Pernambuco (en Noreste de Brasil, n=1188). Se realizó análisis factorial exploratorio de las preguntas sobre violencias (cuatro psicológicas, seis físicas y tres sexuales), con rotación varimax y elaboración de tres factores. Se calculó alfa de Cronbach para análisis de la consistencia interna. Para la validación por grupos extremos, promedios de escores (cero a 13 puntos) de violencia fueron evaluadas con relación a los resultados: auto-evaluación de salud, actividades diarias, presencia de dolor o incomodidad, concepción de idea e intento de suicidio, grande consumo de alcohol y presencia de trastorno mental común. RESULTADOS: Fueron definidos tres factores con varianza acumulada semejante (0,6092 en Sao Paulo y 0,6350 en la Zona de Mata)...

A Comparison between bright field and phase-contrast image analysis techniques in activated sludge morphological characterization.

Different approaches using microscopy image analysis procedures were employed for characterization of activated sludge systems. The approaches varied mainly on the type of visualization and acquisition method used for collection of data. In this context, this study focused on the comparison of the two most common acquisition methods: bright field and phase-contrast microscopy. Images were acquired from seven different wastewater treatment plants for a combined period of two years. Advantages and disadvantages of each acquisition technique and the results are discussed. Bright field microscopy proved to be more simple and inexpensive and provided the best overall results.

Correlation between sludge settling ability and image analysis information using partial least squares.

In the last years there has been an increase on the research of the activated sludge processes, and mainly on the solid-liquid separation stage, considered of critical importance, due to the different problems that may arise affecting the compaction and the settling of the sludge. Furthermore, image analysis procedures are, nowadays considered to be an adequate method to characterize both aggregated and filamentous bacteria, and increasingly used to monitor bulking events in pilot plants. As a result of that, in this work, image analysis routines were developed in Matlab environment, allowing the identification and characterization of microbial aggregates and protruding filaments. Moreover, the large amount of activated sludge data collected with the image analysis implementation can be subsequently treated by multivariate statistical procedures such as PLS. In the current work the implementation of image analysis and PLS techniques has shown to provide important information for better understanding the behavior of activated sludge processes, and to predict, at some extent, the sludge volume index. As a matter of fact, the obtained results allowed explaining the strong relationships between the sludge settling properties and the free filamentous bacteria contents, aggregates size and aggregates morphology, establishing relevant relationships between macroscopic and microscopic properties of the biological system.

Monitoring of activated sludge settling ability through image analysis: validation on full-scale wastewater treatment plants.

In recent years, a great deal of attention has been focused on the research of activated sludge processes, where the solid-liquid separation phase is frequently considered of critical importance, due to the different problems that severely affect the compaction and the settling of the sludge. Bearing that in mind, in this work, image analysis routines were developed in Matlab environment, allowing the identification and characterization of microbial aggregates and protruding filaments in eight different wastewater treatment plants, for a combined period of 2 years. The monitoring of the activated sludge contents allowed for the detection of bulking events proving that the developed image analysis methodology is adequate for a continuous examination of the morphological changes in microbial aggregates and subsequent estimation of the sludge volume index. In fact, the obtained results proved that the developed image analysis methodology is a feasible method for the continuous monitoring of activated sludge systems and identification of disturbances.

A novel M163L mutation in connexin 26 causing cell death and associated with autosomal dominant hearing loss.

Mutations in GJB2 gene (encoding connexin 26) are the most common cause of hereditary non-syndromic sensorineural hearing loss (NSSHL) in different populations. The majority of GJB2 mutations are recessive, but a few dominant mutations have been associated with hearing loss either isolated or associated with skin disease. We describe a novel dominant pathogenic GJB2 mutation, identified in a Portuguese family affected with bilateral mild/moderate high-frequency NSSHL. In vitro functional studies demonstrate that the mutant protein (p.M163L) has defective trafficking to the plasma membrane and is associated with increased cell death.

A7445G mtDNA mutation present in a Portuguese family exhibiting hereditary deafness and palmoplantar keratoderma.

Mitochondrial DNA (mtDNA) A7445G point mutation has been shown to be responsible for familial nonepidermolytic palmoplantar keratoderma (NEPPK) associated with deafness without any additional features. To date, only a few cases have been described. We report a Portuguese pedigree presenting an inherited combination of NEPPK and sensorineural deafness compatible with maternal transmission. Clinical expression and age of onset of NEPPK and deafness were variable. Normal expression patterns of epidermal keratins and filaggrin, intercellular junction proteins including connexin 26, loricrin and cornified envelope proteins, were observed. Molecular analysis revealed that all the affected members, previously screened for Cx26 mutations with negative results, presented the mtDNA A7445G point mutation in the homoplasmic form. To our knowledge, this is the fifth family in whom inherited NEPPK and hearing loss are related to this mitochondrial mutation.

[Endoscopic imaging techniques in the diagnosis of laryngeal carcinoma and its precursor lesions]. / Endoskopische bildgebende Verfahren in der Diagnostik des Kehlkopfkarzinoms und seiner Vorstufen.

BACKGROUND: In order to improve preoperative diagnostic work-up in treatment of patients with laryngeal cancer and its precursor lesions additional endoscopical imaging techniques have been developed: 1. Autofluorescence endoscopy; 2. Contact endoscopy; 3. Endoscopic high-frequency ultrasound. These imaging techniques are used during microlaryngoscopy to get further information about tumor extension and differentiation. This paper describes the diagnostic potential of these imaging techniques in the evaluation of cancerous lesions of the larynx. MATERIAL AND METHODS: Patients in different stages of laryngeal dysplasia, carcinoma in situ and laryngeal cancer were examined by means of the previous mentioned imaging techniques during microlaryngoscopy (Autofluorescence endoscopy [n = 38], contact endoscopy [n = 323], endoscopic high-frequency ultrasound [n = 60]) and the results were compared to pathohistological findings. In autofluorescence endoscopy cancerous mucosa was illuminated using blue filtered light (380-460 nm) to obtain autofluorescence for optical demarcation of the lesion. Contact endoscopy was performed after staining of the laryngeal mucosa with methylene blue (1%). Two different endoscopes with 60 x and 150 x magnification were used. In both techniques a video image was achieved by using a xenon light source and a special video camera to register autofluorescence. The endoscopical high-frequency ultrasound examination was performed after flooding the larynx with 0.9% saline. Newly developed ultrasound catheters with frequencies between 10 to 20 MHz were inserted in the laryngeal lumen and moved in a standardized pattern during the examination. RESULTS: During the autofluorescence examination of the endolaryngeal mucosa precancerous and cancerous lesions showed a red to violet fluorescence outlined against the light green autofluorescence of the normal mucosa. Hyperplastic hyperkeratotic epithelium revealed a higher intensity of light green or even whitish autofluorescence compared to normal mucosa autofluorescence. After staining the vocal cords with methylene blue, it was possible to observe the cells, nuclei and cytoplasm of the laryngeal mucosa and their different grades of abnormality using the specially developed contact endoscopes. Endoscopic high-frequency ultrasound (10 to 20 MHz) was able to measure the vertical extension of laryngeal carcinomas bigger than 3 mm in size. The involvement of the thyroid cartilage or the anterior commissure could be visualized. Preoperatively, the critical T2 stage could be evaluated more precisely. In precancerous lesions and microinvasive cancer ultrasound added no additional Information to the microlaryngoscopical picture. CONCLUSION: Autofluorescence, contact endoscopy as well as endoscopic high-frequency ultrasound are promising new imaging techniques supplementing microlaryngoscopy: autofluorescence as well as contact endoscopy are suitable to differentiate dysplasia, carcinoma in situ, microinvasive lesions as well as the evaluation of tumorous margins, while high-frequency ultrasound improves the assessment of tumorous infiltration into the depth of the larynx. These imaging techniques enable the laryngologist to perform a more accurate diagnostic work-up in the assessment of laryngeal cancer and its precursor lesions.

Contact endoscopy of the nasal mucosa.

The possibility of contact endoscopy in the nasal mucosa, using a contact microlaryngoscope is examined. With contact endoscopy it has been possible to visualize in vivo and in situ (60x, 150x) the superficial cell layers of the nasal epithelium, previously stained with methylene blue. Normal mucosa and cases with pathology (chronic rhinitis, nasal polyposis, inverted papilloma, mucormicosis and carcinoma) were assessed with contact endoscopy (50 patients). Squamous epithelium, ciliated epithelium, glandular ostia, mucus secretions, microvascular networks, inflammatory cell infiltrates, tissue inclusions, nuclear abnormalities, and fungal hyphae have been made visible. The potential of contact endoscopy in the nose, the alterations required in the contact endoscope and the intense interdisciplinary work needed are discussed. Evaluation and differentiation of stages of chronic mucosal diseases, nasal polyposis, environmental pathology, allergy, mucocilliary diseases, pharmacotherapy, are some of the entities that will benefit from this technique.

Changing flow pattern of the internal thoracic artery undergoing coronary bypass grafting: continuous-wave Doppler assessment.

Surgeons have limited ability to evaluate intraoperatively the patency of internal thoracic artery graft as a bypass for coronary artery revascularization. We used continuous-wave Doppler ultrasonography to study the velocity of the internal thoracic artery before harvesting and after grafting (scanning probe, 8 MHz). Systolic and diastolic frequency shift (in kilohertz) and systolic frequency/diastolic frequency index were analyzed. Twenty four internal thoracic artery grafts in 15 patients were studied. Fourteen internal thoracic artery grafts were anastomosed to the left anterior descending artery, one to a diagonal artery, and nine to the circumflex artery. The mean systolic frequency before harvesting was 1.19 +/- 0.40 KHz and no significant differences were found between the right and the left internal thoracic artery (right, 1.17 +/- 0.37; left, 1.19 +/- 0.42 KHz). There was a 40% drop in systolic frequency related to the harvesting. Mean systolic frequency decreased after grafting (1.19 +/- 0.40 versus 0.87 +/- 0.32 KHz; p < 0.01) whereas mean diastolic frequency doubled (0.32 +/- 0.12 versus 0.83 +/- 0.4 KHz; p < 0.001) and mean diastolic frequency/systolic frequency index increased from 28% +/- 11% to 101% +/- 39% (p < 0.001), indicating an increased myocardial vascularization during diastole. No significant difference was found between grafted arteries (left anterior descending versus circumflex). All patients had an uneventful postoperative course and no perioperative myocardial infarction was reported. Doppler flow quantification of internal thoracic artery bypasses may give the surgeon an opportunity to evaluate intraoperatively the physiologic features and patency of the internal thoracic artery before and after coronary artery bypasses.

Contact endoscopy during microlaryngeal surgery: a new technique for endoscopic examination of the larynx.

By means of a microcolpohysteroscope during microlaryngoscopy, the cells of superficial layers of the vocal cord epithelium, previously stained with methylene blue, were observed in vivo and in situ. Several parameters were evaluated: regularity of the epithelium, dimensions and color of the nucleus, nucleus-cytoplasm ratio, nucleus and cytoplasm contours, and presence of the nucleolus, mitosis, cytoplasmic inclusions, keratosis, and koilocytes, among others. The normal squamous epithelium of the vocal cord has a homogeneous cell arrangement, with a spheroid and isochromic nucleus, and a uniform nucleus-cytoplasm ratio. Specific cellular epithelial patterns in different disorders were observed, namely, chronic laryngitis, keratosis, dysplasia, papilloma, and malignant tumor. The present study on contact endoscopy during microlaryngeal surgery reports our first results with this original technique, performed in 68 patients.