André Almeida

Purification and in vitro refolding of maize chloroplast transglutaminase over-expressed in Escherichia coli

In contrast to mammalian transglutaminases (TGs), plant members of the superfamily are poorly characterized. In order to produce pure and active TG for its functional and structural studies, variants of maize chloroplast transglutaminase (TGZ, Patent WWO03102128) were sub-cloned into a pET28 vector and overexpressed in Escherichia coli BL21 (DE3) cells. The recombinant proteins were present mainly as insoluble inclusion bodies. The TGZ4p variant with four B-type repeats (Mr∼55xa0kDa), was affinity purified from urea-solubilized inclusion bodies. TGZ4p was refolded by rapid dilution in a Ca2+- and guanidine-containing buffer. Active TGZ4p shows the general catalytic characteristics described for other TGs.

The Rabbit as an Experimental and Production Animal: From Genomics to Proteomics

The rabbit (Oryctolagus cuniculus) is an important animal species widely used for biomedical research purposes, meat production and as a pet animal. There are numerous biomedical and scientific applications that include important areas such as antibody production, muscle, eye and circulatory physiology. The use of proteomics has been limited when considering this species. The aim of this article is to provide a review on applications of proteomics to the rabbit species, including those that are most relevant and where rabbit is a key species: muscle and circulatory system physiology.

Infections on the rise: Raoultella spp., clinical and microbiological findings from a retrospective study, 2010-2014.

We performed a retrospective analysis of clinical and laboratory data over 5 years in a tertiary centre to assess clinical and microbiological characteristics of patients with Raoultella spp. infection. Raoultella spp. were deemed responsible for clinical infections in 57 patients (R. planticola, n = 32 and R. ornithinolytica, n = 25). The most prevalent diagnoses for R. planticola were cystitis (50%; n = 16) followed by bacteraemia and pneumonia (9.4%; n = 3); for R. ornithinolytica, cystitis (36%; n = 9) followed by pneumonia (24%; n = 6). Immunodeficiency was present in 18 patients (56.3%) with R. planticola and in 16 patients (64%) with R. ornithinolytica infection. Of these, 55.6% and 37.5% had diabetes and 27.8% and 18.% were solid organ transplant recipients, respectively. All isolates were sensitive to third-generation cephalosporins, fluoroquinolones and aminoglycosides. Mortality of infections with R. planticola (n = 5; 15.6%) was higher than for R. ornithinolytica (n = 2; 8.0%), but the difference was not statistically significant.

CURB-65 and other markers of illness severity in community-acquired pneumonia among HIV-positive patients:

As the relative burden of community-acquired bacterial pneumonia among HIV-positive patients increases, adequate prediction of case severity on presentation is crucial. We sought to determine what characteristics measurable on presentation are predictive of worse outcomes. We studied all admissions for community-acquired bacterial pneumonia over one year at a tertiary centre. Patient demographics, comorbidities, HIV-specific markers and CURB-65 scores on Emergency Department presentation were reviewed. Outcomes of interest included mortality, bacteraemia, intensive care unit admission and orotracheal intubation. A total of 396 patients were included: 49 HIV-positive and 347 HIV-negative. Mean CURB-65 score was 1.3 for HIV-positive and 2.2 for HIV-negative patients (pu2009<u20090.0001), its predictive value for mortality being maintained in both groups (pu2009=u20090.03 and pu2009<u20090.001, respectively). Adjusting for CURB-65 scores, HIV infection by itself was only associated with bacteraemia (adjusted odds ratio [AOR] 7.1, 95% CI [2.6–19.5]). Patients withu2009<u2009200 CD4 cells/µL presented similar CURB-65 adjusted mortality (aOR 1.7, 95% CI [0.2–15.2]), but higher risk of intensive care unit admission (aOR 5.7, 95% CI [1.5–22.0]) and orotracheal intubation (aOR 9.1, 95% CI [2.2–37.1]), compared to HIV-negative patients. These two associations were not observed in theu2009>u2009200 CD4 cells/µL subgroup (aOR 2.2, 95% CI [0.7–7.6] and aOR 0.8, 95% CI [0.1–6.5], respectively). Antiretroviral therapy and viral load suppression were not associated with different outcomes (pu2009>u20090.05). High CURB-65 scores and CD4 countsu2009<u2009200 cells/µL were both associated with worse outcomes. Severity assessment scales and CD4 counts may both be helpful in predicting severity in HIV-positive patients presenting with community-acquired bacterial pneumonia.

Prevalence of and factors mediating HIV infection among sex workers in Lisbon, Portugal: the 5-year experience of a community organisation

Studies from Western European countries report that HIV prevalence varies widely among sex workers (SW), with figures generally indicating a higher burden of infection among male and transgender workers.1–3 However, large variations in sample sizes and sampling methods are reported, so caution should be taken when making direct comparisons between countries. Portuguese data are scarce and sourced mostly from behavioural surveys. A recent report in 2012 described a country prevalence of 8.9% among SW (7.9% and 13.5% for female and male workers, respectively), showing that Portugal has the highest prevalence in Western Europe and the sixth highest in Europe. …

Chronic Q Fever with No Elevation of Inflammatory Markers: A Case Report

We describe the case of a 55-year-old man with a biological prosthetic aortic valve who suffered from epigastrium and right hypochondrium pain associated with intermittent night sweats. Liver biopsy showed infectious hepatitis pattern without pathognomonic features. Coxiella burnetii serology was suggestive of chronic Q fever, and modified Dukes criteria for endocarditis were also fulfilled. The authors present a brief literature review concerning chronic Q fever, emphasizing absent previous reports of chronic Q fever with hepatitis and endocarditis and no increase in inflammatory markers.

Community-Acquired Pneumonia in HIV-Positive Patients: an Update on Etiologies, Epidemiology and Management

Purpose of ReviewCommunity-acquired pneumonia (CAP) is the cause of a considerable burden of morbidity and mortality among HIV-infected patients. The purpose of this study was to review recent literature for epidemiological trends and management of CAP in HIV patients.Recent FindingsWhile the specific infectious etiologies are very diverse, bacterial infection and tuberculosis are the main causes of CAP in this population. Bacterial CAP mostly affects high-income countries, while tuberculosis is mostly in low-/middle-income countries. Early antiretroviral therapy is of benefit for all patients regardless of etiology and immune status.SummarySpecific etiologies require individualized approaches, both therapeutically and prophylatically. Antiretroviral therapy is recommended for all patients.

Nursing home-acquired pneumonia presenting at the emergency department

Nursing home-acquired pneumonia (NHAP) is one of the most common infections arising amongst nursing home residents, and its incidence is expected to increase as population ages. The NHAP recommendation for empiric broad-spectrum antibiotic therapy, arising from the concept of healthcare-associated pneumonia, has been challenged by recent studies reporting low rates of multidrug-resistant (MDR) bacteria. This single center study analyzes the results of NHAP patients admitted through the Emergency Department (ED) at a tertiary center during the year 2010. There were 116 cases, male gender corresponded to 34.5xa0% of patients and median age was 84xa0years old (IQR 77–90). Comorbidities were present in 69.8xa0% of cases and 48.3xa0% of patients had used healthcare services during the previous 90xa0days. In-hospital mortality rate was 46.6xa0% and median length-of-stay was 9xa0days. Severity assessment at the Emergency Department provided CURB65 index score and respective mortality (%) results: zero: nxa0=xa00; one: nxa0=xa07 (0xa0%); two: nxa0=xa018 (38.9xa0%); three: nxa0=xa026 (38.5xa0%); four: nxa0=xa030 (53.3xa0%); and five; nxa0=xa022 (68.2xa0%); and sepsis nxa0=xa050 (34.0xa0%), severe sepsis nxa0=xa043 (48.8xa0%) and septic shock nxa0=xa022 (72.7xa0%). Significant risk factors for in-hospital mortality in multivariate analysis were polypnea (pxa0=xa00.001), agexa0≥xa075xa0years (pxa0=xa00.02), and severe sepsis or shock (pxa0=xa00.03) at the ED. Microbiological testing in 78.4xa0% of cases was positive in 15.4xa0% (nxa0=xa015): methicillin-resistant Staphylococcus aureus (26.7xa0%), Pseudomonas aeruginosa (20.0xa0%), S. pneumoniae (13.3xa0%), Escherichia coli (13.3xa0%), others (26.7xa0%); the rate of MDR bacteria was 53.3xa0%. This study reveals high rates of mortality and MDR bacteria among NHAP hospital admissions supporting the use of empirical broad-spectrum antibiotic therapy in these patients.

Erratum to: Nursing home-acquired pneumonia presenting at the emergency department

The authors would like to thank for the contribution of Ana Catarina Patrı́cio, Filipa Silva, Joana Ferreira, Pedro la Féria, Pedro Silva, João Oliveira, Lara câmara, Rui Malheiro, Rodrigo Leão, Mário Silva, Pedro Russo, Ana Lladó and Paulo Barreto. Also the authors would like to thank the Epidemiology and Statistics Office of the Research Unit, Centro Hospitalar de Lisboa Central for their contribution.

Streptococcus gallolyticus: a single bacteria, two different conditions

The authors describe the case of a 48-year-old woman presenting with fever, joint pain and migratory skin lesions. She had no other symptoms or medical history. After an extensive and inconclusive work up, she was admitted to the hospital for further study. This patient was ultimately found to have Streptococcus gallolyticus subsp gallolyticus bacteraemia. This finding led to the diagnosis of mitral valve infective endocarditis related to an underlying rectum adenocarcinoma. This article points out diagnostic difficulties related to an unusual presentation of the underlying disease. Furthermore, the authors reinforce the need of keeping a high level of suspicion and a systematic approach in every case of fever of unknown origin. This case highlights the importance of performing a colonoscopy in the event of S. gallolyticus subsp gallolyticus bacteraemia, as it may provide an opportunity for detecting colonic lesions at an earlier stage.