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- Characterization of all Surgical Specimens Provided by a Portuguese Department of Ophthalmology over a 13 Year PeriodPublication . Ferreira Mendes, J; Ferreira, AM; Freitas, CINTRODUCTION: We intend to evaluate clinically, topographically and morphologically all surgical specimens sent by the Department of Ophthalmology of Hospital de Braga to the Department of Pathology of the same hospital. MATERIAL AND METHODS: Two hundred and fifty eight surgically obtained specimens, from the Department of Ophthalmology of Hospital de Braga, analyzed in the Department of Pathology, from January 2002 to June 2015, were characterized. Data was arranged according to year, age, sex, topography and morphological diagnosis according to the SNOMED® coding system. RESULTS: Mean age at time of diagnosis was 54.6 years old; 52.3% were male subjects. The number of specimens was relatively stable until the year 2010, with a significant increase between 2011 and 2013. Most specimens sent corresponded to eyelid (54.7%), followed by conjunctiva (26.7%); the most common pathological diagnosis was malignant epithelial lesions (22.48%), followed by melanocytic tumours (22.09%) and benign epithelial lesions (17.05%). DISCUSSION: The results are distinct from previous publications presumably because of differences between the populations submitted to analysis. CONCLUSION: This is the first indexed publication characterizing surgical specimens from a Department of Ophthalmology in Portugal; moreover, it also includes an extensive review of global epidemiological data about ophthalmic surgical specimens.
- Characterization and genome sequencing of a Citrobacter freundii phage CfP1 harboring a lysin active against multidrug-resistant isolatesPublication . Oliveira, H; Pinto, G; Oliveira, A; Oliveira, C; Faustino, MA; Briers, Y; Domingues, L; Azeredo, JCitrobacter spp., although frequently ignored, is emerging as an important nosocomial bacterium able to cause various superficial and systemic life-threatening infections. Considered to be hard-to-treat bacterium due to its pattern of high antibiotic resistance, it is important to develop effective measures for early and efficient therapy. In this study, the first myovirus (vB_CfrM_CfP1) lytic for Citrobacter freundii was microbiologically and genomically characterized. Its morphology, activity spectrum, burst size, and biophysical stability spectrum were determined. CfP1 specifically infects C. freundii, has broad host range (>85 %; 21 strains tested), a burst size of 45 PFU/cell, and is very stable under different temperatures (-20 to 50 °C) and pH (3 to 11) values. CfP1 demonstrated to be highly virulent against multidrug-resistant clinical isolates up to 12 antibiotics, including penicillins, cephalosporins, carbapenems, and fluroquinoles. Genomically, CfP1 has a dsDNA molecule with 180,219 bp with average GC content of 43.1 % and codes for 273 CDSs. The genome architecture is organized into function-specific gene clusters typical for tailed phages, sharing 46 to 94 % nucleotide identity to other Citrobacter phages. The lysin gene encoding a predicted D-Ala-D-Ala carboxypeptidase was also cloned and expressed in Escherichia coli and its activity evaluated in terms of pH, ionic strength, and temperature. The lysine optimum activity was reached at 20 mM HEPES, pH 7 at 37 °C, and was able to significantly reduce all C. freundii (>2 logs) as well as Citrobacter koseri (>4 logs) strains tested. Interestingly, the antimicrobial activity of this enzyme was performed without the need of pretreatment with outer membrane-destabilizing agents. These results indicate that CfP1 lysin is a good candidate to control problematic Citrobacter infections, for which current antibiotics are no longer effective.
- Acute Alithiasic Cholecystitis and Human Herpes Virus Type-6 Infection: First CasePublication . Gomes, MM; Antunes, H; Lobo, AL; Branca, F; Correia-Pinto, J; Moreira-Pinto, JA three-year-old male child presented with erythematous maculopapular nonpruritic generalized rash, poor feeding, vomiting, and cramping generalized abdominal pain. He was previously healthy and there was no family history of immunologic or other diseases. On examination he was afebrile, hemodynamically stable, with painful palpation of the right upper quadrant and positive Murphy's sign. Laboratory tests revealed elevated inflammatory markers, elevated aminotransferase activity, and features of cholestasis. Abdominal ultrasound showed gallbladder wall thickening of 8 mm with a positive sonographic Murphy's sign, without gallstones or pericholecystic fluid. Acute Alithiasic Cholecystitis (AAC) was diagnosed. Tests for underlying infectious causes were negative except positive blood specimen for Human Herpes Virus Type-6 (HHV-6) by polymerase chain reaction. With supportive therapy the child became progressively less symptomatic with gradual improvement. The child was discharged on the sixth day, asymptomatic and with improved analytic values. Two months later he had IgM negative and IgG positive antibodies (1/160) for HHV-6, which confirmed the diagnosis of previous infection. In a six-month follow-up period he remains asymptomatic. To the best of our knowledge, this represents the first case of AAC associated with HHV-6 infection.
- Carência de Vitamina D numa População Hospitalar: Uma Fotografa pela Perspetiva LaboratorialPublication . Santos, MJ; Fernandes, V; Garcia, FMNTRODUCTION: Although vitamin D deficiency is increasingly recognized around the world, there are few studies on the Portuguese reality. This study aims to analyse vitamin D levels in the assays performed in our hospital and their relationship with age, genre, requesting specialty and moment of sample collection. MATERIAL AND METHODS: Cross-sectional study of measurements of 25(HO)D performed in our Hospital between June 2012 and November 2014. Included variables: gender, age, requesting specialty, month of sample collection. Vitamin D status classified as: 'Deficiency' (≤ 20 ng/mL), 'Insufficiency' (21 - 29 ng/ml) and 'Sufficiency' (≥ 30 ng/mL). RESULTS: We included 5 439 assays; 55.0% from women; the median age was 64.0 years. Sixty per cent had 'Deficiency', 20.7% 'Insufficiency' and 18.9% 'Sufficiency'. We found a negative correlation between age and vitamin D level (p < 0.001). We didn't find differences in vitamin D levels between genres. Nine specialties requested 98% of the assays, namely Nephrology (56.2%). We found differences between specialties based on age and vitamin D level (p < 0.001). Vitamin D levels changed throughout the year, with higher levels in the summer, followed by autumn, spring and winter (p < 0.001). Despite this seasonal fluctuation, vitamin D sufficiency was only present in a minority of assays (27.8% in summer and 9.2% in winter). DISCUSSION: Vitamin D deficiency is prevalent in this population, affects individuals of all ages and is not offset by the seasonal variation of sunlight. CONCLUSION: Vitamin D deficiency is a real and prevalent problem in our population that needs further attention and action, given its clinical implications.
- Acute pancreatitis in children : a tertiary hospital reportPublication . Antunes, H; Nascimento, J; Mesquita, A; Correia-Pinto, JINTRODUCTION: The incidence of acute pancreatitis (AP) in children has increased significantly in the past two decades. OBJECTIVE: All cases of AP, acute recurrent pancreatitis (ARP), and chronic pancreatitis examined between May 2002 and May 2012 at Hospital de Braga, Portugal, were reviewed. MATERIAL AND METHODS: Patients were identified by searching the hospital's electronic discharge records for the International Classification of Disease, Ninth Revision (ICD-9) code 577.0 (acute pancreatitis). ARP was considered as two or more episodes of AP per year or more than three episodes over a lifetime with intervening return to baseline. The following data were analyzed: demographic information, clinical, laboratory and imaging test results, etiology of pancreatitis, medical and surgical management, length of hospitalization, and outcome. The clinical and laboratory factors used in the pediatric acute pancreatitis severity score system and computed tomography severity index (CTSI) score were compared between patients with mild and severe disease. RESULTS: A total of 37 patients, 31 episodes of AP and 6 patients with ARP, were documented. The most prevalent etiologies were biliary stones/sludge (24.3%) and trauma (16.2%). Admission elevated white blood cell count (p=0.011), 48-h trough calcium (p=0.007), and 48-h rise in blood urea nitrogen (p=0.025) correlated significantly with disease severity. CTSI on admission had a score below 4 in three patients with severe disease. CONCLUSION: This Portuguese pediatric pancreatitis report highlights the multiple and complex etiology of this disease. Better pediatric scoring systems and management algorithms are needed.
- Use of newly isolated phages for control of Pseudomonas aeruginosa PAO1 and ATCC 10145 biofilmsPublication . Pires, D; Sillankorva, S; Faustino, A; Azeredo, JPseudomonas aeruginosa is a relevant opportunistic pathogen involved in nosocomial infections that frequently shows low antibiotic susceptibility. One of its virulence factors is associated with the ability to adhere to surfaces and form virulent biofilms. This work describes the isolation and characterization of lytic phages capable of infecting antibiotic-resistant P. aeruginosa strains. In addition, characterization of P. aeruginosa biofilms and the potential of newly isolated phages for planktonic and biofilm control was accessed. According to the results, the isolated phages showed different spectra of activity and efficiency of lysis. Four broad lytic phages were selected for infection of planktonic cells; however, despite their broad range of activity, two of the selected phages failed to efficiently control planktonic cultures. Therefore, only two phages (phiIBB-PAA2 and phiIBB-PAP21), highly capable of causing strong biomass reduction of planktonic cells, were tested against 24 h biofilms using a m.o.i. of 1. Both phages reduced approximately 1-2 log the biofilm population after 2 h of infection and reduction was further enhanced after 6 h of biofilm infection. However, biofilm cells of P. aeruginosa PAO1 acquired resistance to phiIBB-PAP21; consequently, an increase in the number of cells after 24 h of treatment was observed. Conversely, phage phiIB-PAA2 for P. aeruginosa ATCC10145 continued to destroy biofilm cells, even after 24 h of infection. In these biofilms, phages caused a 3 log reduction in the number of viable counts of biofilm cells.
- mTOR pathway overactivation in BRAF mutated papillary thyroid carcinomaPublication . Faustino, A; Couto, JP; Pópulo, H; Rocha, AS; Pardal, F; Cameselle-Teijeiro, JM; Lopes, JM; Sobrinho-Simões, M; Soares, PCONTEXT: There are several genetic and molecular evidences suggesting dysregulation of the mammalian target of rapamycin (mTOR) pathway in thyroid neoplasia. Activation of the phosphatidylinositol-3-kinase/AKT pathway by RET/PTC and mutant RAS has already been demonstrated, but no data have been reported for the BRAF(V600E) mutation. OBJECTIVE: The aim of this study was to evaluate the activation pattern of the mTOR pathway in malignant thyroid lesions and whether it may be correlated with known genetic alterations, as well as to explore the mechanisms underlying mTOR pathway activation in these neoplasias. RESULTS: We observed, by immunohistochemical evaluation, an up-regulation/activation of the mTOR pathway proteins in thyroid cancer, particularly in conventional papillary thyroid carcinoma (cPTC). Overactivation of the mTOR signaling was particularly evident in cPTC samples harboring the BRAF(V600E) mutation. Transfection assays with BRAF expression vectors as well as BRAF knockdown by small interfering RNA revealed a positive association between BRAF expression and mTOR pathway activation, which appears to be mediated by pLKB1 Ser428, and emerged as a possible mechanism contributing to the association between BRAF mutation and mTOR pathway up-regulation. When we evaluated the rapamycin in the growth of thyroid cancer cell lines, we detected that cell lines with activating mutations in the MAPK pathway show a higher sensitivity to this drug. CONCLUSIONS: We determined that the AKT/mTOR pathway is particularly overactivated in human cPTC harboring the BRAF(V600E) mutation. Moreover, our results suggest that the mTOR pathway could be a good target to enhance therapy effects in certain types of thyroid carcinoma, namely in those harboring the BRAF(V600E) mutation.
- Determinantes da colonização materna e da infecção neonatal por Streptococcus do grupo BPublication . Areal, A; Moreira, M; Nunes, S; Faustino, MA; Cardoso, L; Sá, CAim and Objective: During the past three decades, group B Streptococcus (GBS) neonatal infection has been the subject of little research. The aim of this study was to evaluate the association between maternal risk factors, as established by the Center for Disease Control and Prevention (CDC), and maternal colonization. We also analysed the association between risk factors present in newborns and early-onset GBS disease. Study design: Cross-sectional study. Population: All pregnant women admitted for delivery in our institution and their newborns, between 1st February and 31st July 2005. Methods: Maternal and neonatal characteristics were collected from hospital clinical data, including information on risk factors established by the CDC. Descriptive statistics was used to characterize the study sample. Qui-square and Mantel-haenszel tests were applied to compare proportions and to measure the strength of associations, respectively, setting significance at p < 0,05. Results: In this sample only 47% of women were screened for GBS colonization in suitable time and 34,9% of these women were colonized. The incidence of early neonatal infection by SGB was 9/1000 neonates. Significant associations between GBS maternal colonization ant the following parameters were observed: maternal age [p=0,012; OR=1,659 (IC a 95%, 1,218-2,260)], gestational age at labour [p=0,001; OR= 2,621 (IC a 95%, 1,641- 4,188)], and urinary GBS infection during pregnancy (p<0,001). Maternal colonization occurred in women without CDC defined risk factors. Early neonatal infection by SGB was strongly associated with unscreened women (p=0,014). Conclusion: In this study, maternal GBS colonization occurred in the absence of CDC defined risk factors and varied according to maternal age and gestational week. Neonatal GBS infection was more frequent in unscreened women.
- A infecção peri-natal por Streptococcus agalactiae pode ser evitada: prevalência da colonização em parturientes no Hospital de S. Marcos, factores de risco e sua relação com a infecção peri-natalPublication . Areal, A; Nunes, S; Moreira, M; Faustino, MA; Cardoso, L; Sá, Cntrodução: O Streptococcus agalactiae (SGB) é o agente mais frequente de infecção neonatal precoce, sendo possível a sua prevenção. Em Portugal é desconhecida a prevalência de mulheres colonizadas por SGB. O estudo da Unidade de Vigilância Pediátrica refere uma prevalência nacional de infecção neonatal por SGB de 0,5:1000 nados-vivos. Objectivo: Determinar a prevalência da colonização materna e da infecção perinatal por SGB no Hospital de S. Marcos, Braga (HSM), de modo a avaliar a importância da implementação do rastreio universal e o uso de medidas profiláticas. Método: De 1 de fevereiro a 31 de julho de 2005 foi realizado um estudo transversal com análise de coortes anichada, avaliando todas as grávidas assistidas para trabalho de parto no HSM e respectivos recém-nascidos. Os dados foram submetidos a análise estatística bi-variada pelo teste do qui-quadrado, com um nível de significância de 5% (p<0,05). Resultados: A prevalência da colonização nas grávidas rastreadas na região de Braga foi de 34,9 % [intervalo de confiança a 95% (IC95%), 31,5-38,3%]. No HSM o diagnóstico de infecção neonatal precoce por SGB ocorreu em 9:1000 recém-nascidos. O risco relativo de ocorrência de infecção perinatal entre os recém-nascidos dos grupos de mães rastreadas e não rastreadas foi de 0,3 [IC95% 0,2,7-0,32]. Conclusão: O rastreio bacteriológico positivo para colonização materna por SGB associado à adequada profilaxia intra-parto reduziu significativamente a infecção neonatal precoce, em relação ao grupo de gestantes não rastreadas (p=0,014). Consideramos que é recomendável a instituição do rastreio universal das grávidas e a profilaxia adequada quando indicada.
- Group B streptococcal colonisation in pregnant women: turnaround time of three culture methodsPublication . Areal, A; Faustino, MA
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