Developing a cell-bound detection system for the screening of oxidase activity using the fluorescent peroxide sensor roGFP2-Orp1.

Accurate yet efficient high-throughput screenings have emerged as essential technology for enzyme engineering via directed evolution. Modern high-throughput screening platforms for oxidoreductases are commonly assisted by technologies such as surface display and rely on emulsification techniques to facilitate single-cell analysis via fluorescence-activated cell sorting. Empowered by the dramatically increased throughput, the screening of significantly larger sequence spaces in acceptable time frames is achieved but usually comes at the cost of restricted applicability. In this work, we tackle this problem by utilizing roGFP2-Orp1 as a fluorescent one-component detection system for enzymatic H2O2 formation. We determined the kinetic parameters of the roGFP2-Orp1 reaction with H2O2 and established an efficient immobilization technique for the sensor on Saccharomyces cerevisiae cells employing the lectin Concanavalin A. This allowed to realize a peroxide-sensing shell on enzyme-displaying cells, a system that was successfully employed to screen for H2O2 formation of enzyme variants in a whole-cell setting.

Phenylketonuric diet negatively impacts on butyrate production.

BACKGROUND AND AIMS: Phenylalanine (Phe) restricted diet, combined with Phe-free l-amino acid supplementation, is the mainstay of treatment for phenylketonuria (PKU). Being the diet a key factor modulating gut microbiota composition, the aim of the present paper was to compare dietary intakes, gut microbiota biodiversity and short chain fatty acids (SCFAs) production in children with PKU, on low-Phe diet, and in children with mild hyperphenylalaninemia (MHP), on unrestricted diet. METHODS AND RESULTS: We enrolled 21 PKU and 21 MHP children matched for gender, age and body mass index z-score. Dietary intakes, including glycemic index (GI) and glycemic load (GL), and fecal microbiota analyses, by means of denaturing gradient gel electrophoresis (DGGE) and Real-time PCR were assessed. Fecal SCFAs were quantified by gas chromatographic analysis. RESULTS: We observed an increased carbohydrate (% of total energy), fiber and vegetables intakes (g/day) in PKU compared with MHP children (p = 0.047), as well a higher daily GI and GL (maximum p < 0.001). Compared with MHP, PKU showed a lower degree of microbial diversity and a decrease in fecal butyrate content (p = 0.02). Accordingly, two of the most abundant butyrate-producing genera, Faecalibacterium spp. and Roseburia spp., were found significantly depleted in PKU children (p = 0.02 and p = 0.03, respectively). CONCLUSION: The low-Phe diet, characterized by a higher carbohydrate intake, increases GI and GL, resulting in a different quality of substrates for microbial fermentation. Further analyses, thoroughly evaluating microbial species altered by PKU diet are needed to better investigate gut microbiota in PKU children and to eventually pave the way for pre/probiotic supplementations.

Microbiologic and clinical characteristics of biofilm-forming Candida parapsilosis isolates associated with fungaemia and their impact on mortality.

OBJECTIVES: Biofilm formation (BF) by fungal isolates may dramatically complicate infection. We determined the ability of Candida parapsilosis isolates from single fungaemia episodes to form biofilms and we analysed biofilm subgroups for antifungal susceptibility and pathogenic potential. We then correlated BF with clinical characteristics and outcomes of the episodes. METHODS: BF was measured using the crystal violet biomass assay. Antifungal susceptibility of preformed biofilms was assessed, and virulence was studied using the Galleria mellonella model. A retrospective analysis of patients' clinical records was performed. RESULTS: Of 190 patient-unique isolates, 84, 38 and 68 were identified as having high BF (HBF), moderate BF (MBF) or low BF (LBF), respectively. Among 30 randomly selected isolates, nine (eight HBF and one MBF), six (all HBF) and one (HBF) isolates had elevated sessile minimum inhibitory concentrations to fluconazole, anidulafungin or amphotericin B; all HBF and MBF isolates had elevated voriconazole sessile minimum inhibitory concentrations. G. mellonella killing rates of HBF isolates were significantly greater than MBF (or LBF) isolates (50% vs. 20%, 2 days from infection). By comparing HBF/MBF (106 patients) and LBF (84 patients) groups, we found that HBF/MBF patients had more central venous catheter-related fungaemias (62/106 (58.5%) vs. 29/84 (34.5%), p 0.001) and were more likely to die at 30 days from fungaemia onset (61/106 (57.5%) vs. 28/84 (33.3%), p 0.01). In the HBF/MBF group, azole antifungal therapy and central venous catheter removal were significantly associated with a higher and lower 30-day mortality rate, respectively. CONCLUSIONS: C. parapsilosis BF influences the clinical outcome in patients with fungaemia.

Does peripheral arterial occlusive disease influence muscle strength and exercise capacity in COPD patients? / A doença arterial periférica obstrutiva influencia a força muscular e a capacidade de exercício nos portadores de doença pulmonar obstrutiva crônica?

Abstract Background The pathophysiology of chronic obstructive pulmonary disease (COPD) is complex and understanding of it has been changing in recent years, with regard to its multisystemic manifestations, especially peripheral dysfunction and its influence on intolerance to exercise. Objectives To evaluate the relationship between peripheral arterial occlusive disease (PAOD) and peripheral muscle strength and exercise capacity in COPD patients. Methods We conducted a cross-sectional study of 35 patients with COPD who were evaluated with the Ankle-Brachial Index, handgrip strength test, 1 repetition maximum (1RM) of knee extensors and flexors, and distance covered in the incremental shuttle walking test (dISWT). Results COPD patients with coexisting PAOD had lower dominant handgrip strength test results (33.00 vs. 26.66 kgf, p = 0.02) and worse performance in the dISWT (297.32 vs. 219.41 m, p = 0.02) when compared to the COPD patients without PAOD. Strong correlations were found between the result of the handgrip strength test and both the dISWT (r = 0.78; p < 0.001) and the 1RM/knee extension (r = 0.71; p = 0.03); and also between the dISWT and both the 1RM/knee extension (r = 0.72; p = 0.02) and the 1RM/knee flexion (r = 0.92; p < 0.001). The linear regression model showed that the dISWT variable alone explains 15.3% of the Ankle-Brachial Index result (p = 0.01). Conclusion COPD patients with PAOD exhibit reduced muscle strength and lower exercise capacity than COPD patients without PAOD.

Resumo Contexto A doença pulmonar obstrutiva crônica (DPOC) apresenta uma complexa fisiopatologia e sua compreensão vem se modificando nos últimos anos, com atenção para as manifestações multissistêmicas, em especial a disfunção periférica e sua influência na intolerância ao exercício físico. Objetivo Avaliar o impacto da doença arterial periférica obstrutiva (DAOP) na força muscular periférica e na capacidade de exercício dos portadores de DPOC. Métodos Estudo transversal realizado com 35 portadores de DPOC, que foram avaliados pelo índice tornozelo-braquial, teste de força de preensão palmar (FPP), uma repetição máxima (1RM) de extensores e flexores de joelho, e a distância no incremental shuttle walking test (dISWT). Resultados Portadores de DPOC com DAP coexistente apresentaram menor FPP da mão dominante (33,00 versus 26,66 kgf, p = 0,02) e pior desempenho no dISWT (297,32 versus 219,41 m, p = 0,02) quando comparados aos portadores de DPOC sem DAP. Fortes correlações foram encontradas entre a medida da FPP e a dISWT (r = 0,78; p < 0,001) e a 1RM/extensão de joelho (r = 0,71; p = 0,03); entre a dISWT e a 1RM/extensão de joelho (r = 0,72; p = 0,02) e a 1RM/flexão de joelho (r = 0,92; p < 0,001). O modelo de regressão linear identificou que a variável dISWT explica isoladamente 15,3% do resultado do índice tornozelo braquial (p = 0,01). Conclusão Portadores de DPOC com DAOP coexistente apresentam maior perda da força muscular periférica e pior desempenho da capacidade de exercício quando comparados aos portadores de DPOC sem DAOP.

Does peripheral arterial occlusive disease influence muscle strength and exercise capacity in COPD patients?

BACKGROUND: The pathophysiology of chronic obstructive pulmonary disease (COPD) is complex and understanding of it has been changing in recent years, with regard to its multisystemic manifestations, especially peripheral dysfunction and its influence on intolerance to exercise. OBJECTIVES: To evaluate the relationship between peripheral arterial occlusive disease (PAOD) and peripheral muscle strength and exercise capacity in COPD patients. METHODS: We conducted a cross-sectional study of 35 patients with COPD who were evaluated with the Ankle-Brachial Index, handgrip strength test, 1 repetition maximum (1RM) of knee extensors and flexors, and distance covered in the incremental shuttle walking test (dISWT). RESULTS: COPD patients with coexisting PAOD had lower dominant handgrip strength test results (33.00 vs. 26.66 kgf, p = 0.02) and worse performance in the dISWT (297.32 vs. 219.41 m, p = 0.02) when compared to the COPD patients without PAOD. Strong correlations were found between the result of the handgrip strength test and both the dISWT (r = 0.78; p < 0.001) and the 1RM/knee extension (r = 0.71; p = 0.03); and also between the dISWT and both the 1RM/knee extension (r = 0.72; p = 0.02) and the 1RM/knee flexion (r = 0.92; p < 0.001). The linear regression model showed that the dISWT variable alone explains 15.3% of the Ankle-Brachial Index result (p = 0.01). CONCLUSION: COPD patients with PAOD exhibit reduced muscle strength and lower exercise capacity than COPD patients without PAOD.

CONTEXTO: A doença pulmonar obstrutiva crônica (DPOC) apresenta uma complexa fisiopatologia e sua compreensão vem se modificando nos últimos anos, com atenção para as manifestações multissistêmicas, em especial a disfunção periférica e sua influência na intolerância ao exercício físico. OBJETIVO: Avaliar o impacto da doença arterial periférica obstrutiva (DAOP) na força muscular periférica e na capacidade de exercício dos portadores de DPOC. MÉTODOS: Estudo transversal realizado com 35 portadores de DPOC, que foram avaliados pelo índice tornozelo-braquial, teste de força de preensão palmar (FPP), uma repetição máxima (1RM) de extensores e flexores de joelho, e a distância no incremental shuttle walking test (dISWT). RESULTADOS: Portadores de DPOC com DAP coexistente apresentaram menor FPP da mão dominante (33,00 versus 26,66 kgf, p = 0,02) e pior desempenho no dISWT (297,32 versus 219,41 m, p = 0,02) quando comparados aos portadores de DPOC sem DAP. Fortes correlações foram encontradas entre a medida da FPP e a dISWT (r = 0,78; p < 0,001) e a 1RM/extensão de joelho (r = 0,71; p = 0,03); entre a dISWT e a 1RM/extensão de joelho (r = 0,72; p = 0,02) e a 1RM/flexão de joelho (r = 0,92; p < 0,001). O modelo de regressão linear identificou que a variável dISWT explica isoladamente 15,3% do resultado do índice tornozelo braquial (p = 0,01). CONCLUSÃO: Portadores de DPOC com DAOP coexistente apresentam maior perda da força muscular periférica e pior desempenho da capacidade de exercício quando comparados aos portadores de DPOC sem DAOP.

Efficacy of non-invasive mechanical ventilation in the general ward in patients with chronic obstructive pulmonary disease admitted for hypercapnic acute respiratory failure and pH < 7.35: a feasibility pilot study.

AIM: To date non-invasive (NIV) mechanical ventilation use is not recommended in chronic obstructive pulmonary disease (COPD) patients with acute respiratory failure (ARF) and pH < 7.30 outside a 'protected environment'. We assessed NIV efficacy and feasibility in improving arterial blood gases (ABG) and in-hospital outcome in patients with ARF and severe respiratory acidosis (RA) admitted to an experienced rural medical ward. METHODS: This paper is a prospective pilot cohort study conducted in the General Medicine Ward of Budrio's District Hospital. Two hundred and seventy-two patients with ARF were admitted to our Department, 112, meeting predefined inclusion criteria (pH < 7.35, PaCO2 > 45 mmHg). Patients were divided according to the severity of acidosis into: group A (pH < 7.26), group B (7.26 ≤ pH < 7.30) and group C (7.30 ≤ pH < 7.35). ABG were assessed at admission, at 2-6 h, 24 h, 48 h and at discharge. RESULTS: Group A included 55 patients (24 men, mean age: 80.8 ± 8.3 years), group B 31 (12 men, mean age: 80.3 ± 9.4 years) and group C 26 (15 men, mean age: 78.6 ± 9.9 years). ABG improved within the first hours in 92/112 (82%) patients, who were all successfully discharged. Eighteen percent (20/112) of the patients died during the hospital stay, no significant difference emerged in mortality rate (MR) within the groups (23%, 16% and 8%, for groups A, B and C, respectively) and between patients with or without pneumonia: 8/29 (27%) versus 12/83 (14%). On multivariable analysis, only age and Glasgow Coma Scale had an impact on the clinical outcome. CONCLUSION: In a non-'highly protected' environment such as an experienced medical ward of a rural hospital, NIV is effective not only in patients with mild, but also with severe forms of RA. MR did not vary according to the level of initial pH.

A histological procedure to study fungal infection in the wax moth Galleria mellonella.

The invertebrate model Galleria mellonella is a widely used factitious host to study the microbial pathogenesis in vivo. However, a specific procedure for the recovery and the processing of the infected tissues, important for a better understanding of the host-pathogen interactions, has not been reported to our knowledge. In the present study we describe a new procedure of fixation and processing of larval tissue that allows studying the larval topographic anatomy and assessing the morphological changes due to the fungal infection. Lepidopteran larvae were infected with Candida albicans strains displaying various biofilm-forming abilities. The whole larvae were then examined for tissue changes by histological techniques. We show that comparing cutting planes, serial transversal sections of paraffin-embedded larva result in better accuracy and information recovering. Using this technique, it was possible to preserve the integrity of G. mellonella internal structures allowing the detailed analysis of morphological differences in different experimental groups (i.e., healthy vs infected larvae). We were also able to study strain-related differences in the pathogenesis of C. albicans by observing the immune response elicited and the invasiveness of two isolates within the larval tissues. In general, by processing the whole larva and optimizing routinely histochemical stainings, it is possible to visualize and analyse infected tissues. Various degrees of pathogenicity (strain- or inoculum-related), and the infection time course can be described in details. Moreover, the host immune response events can be followed throughout the infectious process leading to a comprehensive picture of the studied phenomenon.

Candidemia in intensive care unit: a nationwide prospective observational survey (GISIA-3 study) and review of the European literature from 2000 through 2013.

BACKGROUND: Candida bloodstream infections (BSI) represent an important problem in Intensive Care Units (ICUs). The epidemiology of candidemia is changing with an increase in the proportion of Candida (C.) non-albicans. OBJECTIVES: An Italian 2-year observational survey on ICU was conducted to evaluate the species distribution and possible differences between BSI caused by C. albicans and C. non-albicans. For comparative purposes, we performed a European literature-based review to evaluate distribution and frequency of Candida spp. causing ICU candidemia, during the period 2000-2013. MATERIALS AND METHODS: This laboratory-based survey involved 15 microbiology centers (GISIA-3 study). All candidemia episodes in adult patients were considered. Data were prospectively collected from 2007 to 2008. PubMed was searched for peer-reviewed articles. RESULTS: In total, 462 candidemia episodes were collected. C. albicans accounted for 49.4% of the isolates, followed by C. parapsilosis (26.2%) and C. glabrata (10.4%). Mortality was higher in patients with C. non-albicans than C. albicans (47.3% vs. 32.4 %, p > 0.05). Among risk factors, parenteral nutrition was more common (p = 0.02) in non-albicans candidemia, while surgery was more frequent (p = 0.02) in C. albicans candidemia. Twenty-four relevant articles were identified. C. albicans was the predominant species in almost all studies (range 37.9% -76.3%). C. glabrata was commonly isolated in the German-speaking countries, France, UK and North Europe; C. parapsilosis in Turkey, Greece and Spain. CONCLUSIONS: Although C. non-albicans BSI is increasing, our study shows that C. albicans is still the predominant species in ICU candidemia. There are differences in the epidemiology of Candida BSI among European countries, with a prevalence of C. glabrata and C. parapsilosis in Northern and Southern countries, respectively.

Reliability and validity of the Italian version of the Consensus Auditory-Perceptual Evaluation of Voice (CAPE-V).

OBJECTIVES: To evaluate the reliability and validity of the Italian version of the Consensus Auditory-Perceptual Evaluation of Voice (CAPE-V). PATIENTS AND METHODS: Eighty dysphonic patients and 120 asymptomatic subjects were enrolled. The voice signal of each participant was recorded, listened to and rated by 3 licensed speech-language pathologists using the GRBAS scale and the Italian version of the CAPE-V. The intra- and interrater reliability of the CAPE-V was assessed as well as the degree of association between the CAPE-V and GRBAS judgments. The CAPE-V values were also compared between the patients with dysphonia and the asymptomatic subjects. RESULTS: The intra- and interrater reliability appeared to be good for all the parameters except for the strain parameter. The attributes 'consistent' and 'intermittent' demonstrated optimal intra- and interrater reliability. The difference between pathological and control groups was significant for six perceptual parameters. The highest average correlation between GRBAS and CAPE-V judgments was found between overall severity and grade while the lowest was found between the two strain scales. CAPE-V profiles differed significantly between different pathological groups. CONCLUSION: The Italian version of CAPE-V appears to be a reliable and valid tool for the perceptual analysis of the voice signal.

Segurança da intervenção fisioterápica precoce após o infarto agudo do miocárdio / Safety of early physical therapy intervention after acute myocardial infarction

INTRODUÇÃO: A fisioterapia na fase I da reabilitação cardiovascular (FTCV) pode ser iniciada de 12 a 24 horas após o infarto agudo do miocárdio (IAM), no entanto, é comum o repouso prolongado no leito em razão do receio de instabilização do paciente. OBJETIVOS: Avaliar as respostas autonômicas e hemodinâmicas de pacientes pós-IAM submetidos ao primeiro dia de protocolo de FTCV fase I, bem como sua segurança. MATERIAIS E MÉTODOS: Foram estudados 51 pacientes com primeiro IAM não complicado, 55 ± 11 anos, 76% homens. Foram submetidos ao primeiro dia do protocolo de FTCV fase I, em média 24 horas pós-IAM. A frequência cardíaca (FC) instantânea e os intervalos R-R do ECG foram captados pelo monitor de FC (Polar®S810i) e a pressão arterial (PA) aferida pelo método auscultatório. A variabilidade da FC foi analisada nos domínios do tempo (RMSSD e RMSM dos iR-R em ms) e da frequência. A densidade espectral de potência foi expressa em unidades absolutas (ms²/Hz) e normalizada (un) para as bandas de baixa (BF) e alta frequência (AF) e pela razão BF/AF. RESULTADOS: O índice RMSSD, a AF e a AFun apresentaram redução na execução dos exercícios em relação ao repouso pré e pós-exercício (p < 0,05), a BFun e a razão BF/AF aumentaram (p < 0,05). A FC e a PA sistólica apresentaram aumento durante a execução dos exercícios em relação ao repouso (p < 0,05). Não foi observado qualquer sinal e/ou sintoma de intolerância ao esforço. CONCLUSÕES: O exercício realizado foi eficaz, pois promoveu alterações hemodinâmicas e na modulação autonômica nesses pacientes, sem ocasionar qualquer intercorrência clínica.

INTRODUCTION: Physical therapy during phase I of cardiac rehabilitation (CPT) can be started 12 to 24 hours after acute myocardial infarction (AMI), however, it is common to extend the bed rest due to fear of patient’s instability. OBJECTIVES: To assess the hemodynamic and autonomic responses to post-AMI patients when subjected to first day of phase I protocol of CPT, as well as their safety. MATERIALS AND METHODS: We studied 51 patients with first uncomplicated AMI, 55 ± 11 years, 76% men. The patients were subjected to first day protocol phase I CPT, on average, 24 hours after AMI. The Instantaneous heart rate (HR) and RR interval were acquired by HR monitor (Polar™S810i) and blood pressure (BP) checked by auscultation. HR variability was analyzed in the time (RMSSD and RMSM-Ri in ms) and frequency domains. Power spectral density was expressed in absolute (ms²/Hz) and normalized (nu) units for the bands of low (LF) and high frequencies (HF) and as LF/HF ratio. RESULTS: The RMSSD, HF and HFnu have reduced performance of the exercises in relation to rest and post-exercise (p < 0.05), LFnu and LF/HF ratio increased (p < 0.05). HR and systolic BP showed an increase during the execution of the exercises in relation to rest (p < 0.05). There were no any signs and/or symptoms of exercise intolerance. CONCLUSION: The exercise was effective, because it caused changes hemodynamic and autonomic modulation in these patients, without causing any medical complications.

Tinea corporis due to Trichophyton rubrum in a woman and Tinea capitis in her 15-day-old baby: molecular evidence of vertical transmission.

We report a case of a 40-year-old Caucasian woman who came under our observation with a 7-year history of a chronic erythematous scaly dermatitis, diagnosed as psoriasis, involving gluteal area and thighs, and treated with topical steroids without benefit. During pregnancy, a progressive worsening of her condition and an extension of cutaneous lesions were observed. Her newborn, a 15-day-old girl, presented a similar scaly and squamous lesion on her scalp. Mycological examination was positive for Trichophyton rubrum in both cases, and random amplified polymorphic DNA analysis confirmed the isogenicity of the two isolates. We performed a diagnosis of T. rubrum tinea corporis and tinea capitis. The case we describe illustrates an unusual clinical presentation of tinea corporis with remarkable extension of cutaneous lesions due to the diagnostic delay and the continuous use of local steroids, together with a rare tinea capitis in the newborn. Our experience highlights the possibility of mother-child transmission and the importance of an early diagnosis.

Characterization of Candida parapsilosis complex strains isolated from invasive fungal infections.

In the present work, we studied the distribution of Candida parapsilosis complex species and the antifungal susceptibility of clinical isolates collected during an Italian surveillance study of yeast invasive fungal infections (IFIs) in intensive care units (ICUs). Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution method. BanI digestion patterns of the secondary alcohol dehydrogenase polymerase chain reaction (PCR) products were used to identify C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis. A total of 138 C. parapsilosis isolates were stored (January 2007-December 2008). The overall frequency of C. parapsilosis complex in IFIs was 22%. Of the 138 tested isolates, 95% were C. parapsilosis sensu stricto, 3.6% were C. orthopsilosis, and 1.4% were C. metapsilosis. The MIC(50) values (expressed as µg/ml) for anidulafungin, caspofungin, and micafungin for C. parapsilosis complex were 2, 1, and 2, respectively, and the MIC(90) values were 4, 2, and 4, respectively. The MIC(50) and MIC(90) values for itraconazole and posaconazole were 0.12 and 0.25, respectively, and for fluconazole, they were 1 and 4, respectively. This study, the most comprehensive study conducted to date to evaluate the frequency and antifungal susceptibility profiles of C. parapsilosis complex isolates from critically ill patients in Italy, highlights the low prevalence of C. orthopsilosis and C. metapsilosis in IFIs.

Fungal infections in ICU patients: epidemiology and the role of diagnostics.

Invasive fungal infections (IFIs) are on the increase not only among oncology and transplant patients but also among patients admitted to intensive care units (ICU). The rise in ICU IFIs can be attributed to the growing use of complex surgical procedures, invasive medical devices, and long-term, broad-spectrum antibiotic therapy. The majority of these life-threatening infections are caused by the well-known opportunistic pathogens Candida albicans and Aspergillus fumigatus, but new opportunistic pathogens, including yeast-like and other filamentous fungi, have emerged as additional causes. Invasive Candida infections, particularly candidemia, represent the most common IFI in critically ill patients. The species that cause candidemia markedly differ in their responses to antifungal drugs; for this reason, therapy must be tailored to the susceptibility characteristics of the infectious agent. Candidemia caused by non-albicans Candida species is increasing worldwide, and these infections are generally associated with high mortality rates, particularly bloodstream infections caused by C. krusei, which is innately resistant to fluconazole, or C. glabrata, which easily develops azole resistance. Although invasive yeast infections can be considered the most important causes of morbidity and mortality in ICU patients, pulmonary aspergillosis has recently emerged as an additional complication. Diagnosis of IFIs can be achieved using conventional approaches (microscopy, culture, and serology) and newer methods, including antigen detection and polymerase chain reaction (PCR) assays. Because most of the conventional approaches lack sensitivity, antigen detection and PCR assays could represent a valid alternative; however, these procedures need to be standardized and evaluated in a large number of patients.

Adaptation and validation of the Italian MD Anderson Dysphagia Inventory (MDADI).

INTRODUCTION: Oropharyngeal dysphagia is a common symptom in patients with head and neck tumours. The MD Anderson Dysphagia Inventory (MDADI) is a questionnaire currently used in North America for the assessment of dysphagia-related disability in patients with head and neck cancer. The aim of the study is to analyze reliability and clinical validity of the Italian MDADI. MATERIAL AND METHOD: 48 persons with no history of dysphagia and 50 head and neck cancer patients with a chronic and stable dysphagia have been included in the study. Each subject completed alone the Italian MDADI twice with a week interval between the two questionnaire completion. Intra-subject reliability was analyzed through Pearson test in both groups of subjects. Clinical validity was calculated through the non parametric Mann Whitney test of the first MDADI assessment in both groups. RESULTS: Internal consistency and test-retest reliability were high for each MDADI subscale in subjects without dysphagia as well as in those with dysphagia. The difference between MDADI values in subjects with and without dysphagia was significant for each subscale. DISCUSSION: The Italian MDADI is reliable and clinically valid. The application of the MDADI is recommended in clinical practice as well as in descriptive, outcome and efficacy research.

Oocyte euploidy, pronuclear zygote morphology and embryo chromosomal complement.

BACKGROUND: Pronuclear morphology has been proposed as an indicator of embryo development and chromosomal complement. In this study, the morphology of pronuclear zygotes generated from euploid oocytes [diagnosed by first polar body (PB1) analysis] was evaluated and compared with the configurations observed in chromosomally normal embryos (diagnosed by blastomere analysis). MATERIALS AND METHODS: Group 1--238 patients underwent 273 assisted conception cycles in combination with the screening of aneuploidy on PB1 for the chromosomes 13, 15, 16, 18, 21 and 22. Only normal oocytes were inseminated. Group 2--218 patients underwent 318 assisted conception cycles with aneuploidy screening on day 3 embryos. In both groups, oocytes were checked for fertilization and pronuclear morphology at 16 h after insemination. RESULTS: Seventy-three percent of zygotes from Group 1 had the configurations with centralized and juxtaposed pronuclei, large-size aligned or scattered nucleoli and PB located in the longitudinal or perpendicular axis of pronuclei. In Group 2, these configurations corresponded to those with the highest proportion of chromosomally normal embryos. Accordingly, in both groups, these configurations had a higher implantation rate than all the others. CONCLUSIONS: These observations confirm that some patterns of pronuclear morphology are associated with a higher proportion of euploidy and implantation reaffirming the relevance of this scoring system for the prediction of zygote viability.

Influence of noninvasive ventilation by BiPAP on exercise tolerance and respiratory muscle strength in chronic obstructive pulmonary disease patients (COPD).

This study aimed to assess the effect of BiPAP, by nasal mask, on exercise tolerance and respiratory muscle strength in patients with a clinical and spirometric diagnosis of moderate/severe COPD (FEV1 < 60% of predicted). Ten patients of 59.4+/-8.9 years old, with FEV1/FVC <70% of predicted level, were treated with 30 minutes of BiPAP (IPAP:10 and 15 cmH2O; EPAP:4 cmH2O), three days per week, during two months. Before and after the treatment, spirometry, inspiratory (MIP) and expiratory (MEP) muscle strength and the distance walked in six minutes (6MWT) were measured. We observed a significant increase (Wilcoxon, p<0.05) in the mean values of MIP (from -55+/-17 to -77+/-19, respectively), MEP (from 75+/-20 to 109+/-36, respectively) and walking distance (from 349+/-67 to 448+/-75). Based on these results, we concluded that BiPAP improves respiratory muscle strength and exercise tolerance in these COPD patients.

Influence of noninvasive ventilation by BiPAP® on exercise tolerance and respiratory muscle strength in chronic obstructive pulmonary disease patients (COPD)

O objetivo deste estudo foi avaliar o efeito do BiPAP®, através de máscara nasal, na tolerância ao exercício físico e no desempenho muscular respiratório em pacientes com diagnóstico clínico e espirométrico de DPOC, moderado/grave (VEF1 < 60% do previsto). Com VEF1/CVF <70% do previsto e idade média de 59,4±8,9 anos, dez pacientes com doença pulmonar obstrutiva crônica (DPOC) foram tratados com 30 minutos de BiPAP® (IPAP=10-15 e EPAP=4 cmH2O), em três sessões semanais, durante dois meses. Antes e após o tratamento mediu-se a espirometria, a força muscular inspiratória (PImax) e expiratória (PEmax) e a distância percorrida em seis minutos (TC6). Foram constatados aumentos significativos (Wilcoxon, p<0,05) na média da PImax (de -55±17 para -77±19 cmH2O), da PEmax (de 75±20 para 109±36 cmH2O) e da distância percorrida (de 349±67 para 448±75 metros). Com base nesses resultados conclui-se que o BiPAP® melhorou o desempenho muscular respiratório e a tolerância ao exercício físico nesses pacientes com DPOC.

Construction of the World Health Organization child growth standards: selection of methods for attained growth curves.

The World Health Organization (WHO), in collaboration with a number of research institutions worldwide, is developing new child growth standards. As part of a broad consultative process for selecting the best statistical methods, WHO convened a group of statisticians and child growth experts to review available methods, develop a strategy for assessing their strengths and weaknesses, and discuss methodological issues likely to be faced in the process of constructing the new growth curves. To select the method(s) to be used, the group proposed a two-stage decision-making process. First, to select a few relevant methods based on a list of set criteria and, second, to compare the methods using available tests or other established procedures. The group reviewed 30 methods for attained growth curves. Using the pre-defined criteria, a few were selected combining five distributions and two smoothing techniques. Because the number of selected methods was considered too large to be fully tested, a preliminary study was recommended to evaluate goodness of fit of the five distributions. Methods based on distributions with poor performance will be eliminated and the remaining methods fully tested and compared.