José Manuel Correia da Costa

Schistosoma haematobium total antigen induces increased proliferation, migration and invasion, and decreases apoptosis of normal epithelial cells.

Schistosome worms are blood-dwelling flukes that cause chronic infection in more than 200 million people and are thought to be responsible for 500,000 deaths annually. During infection with Schistosoma haematobium, eggs are deposited in the mucosa and submucosa of the bladder and lower ureters. Squamous cell carcinoma (SCC) of the bladder is a long-term sequela of chronic infection. The mechanisms underlying the association between S. haematobium and SCC of the bladder are largely unknown, with all reports to date exclusively demonstrating epidemiological evidence linking S. haematobium infection with SCC of the bladder. We hypothesised that the parasite antigens might induce alterations in epithelial cells towards cancer. For this we used Chinese Hamster Ovary (CHO) cells and treated the cells in culture with S. haematobium total antigen (Sh). Our results showed increased proliferation, increased S-phase and decreased apoptosis, as well as down-regulation of tumour suppressor p27 and up-regulation of anti-apoptotic molecule Bcl-2 in Sh-treated cells compared with controls. We also found increased migration and invasion. To our knowledge, this is the first report demonstrating alterations of normal epithelial cells as a direct effect of S. haematobium antigens.

Salmonella cross-contamination in swine abattoirs in Portugal: Carcasses, meat and meat handlers

In this study the occurrence of Salmonella in swine, pork meat and meat handlers along with their clonal relatedness is evaluated at abattoir level. Samples from the lymph nodes, carcass surface and meat of 100 pigs and 45 meat handlers were collected in eight abattoirs (July 2007-August 2008). Salmonella isolates were serotyped and genotyped by pulsed-field gel electrophoresis (PFGE). From the pigs tested, 42 produced at least one positive sample. A relatively high frequency of Salmonella occurrence was found in the ileoceacal lymph node samples (26.0%), followed by carcass (16.0%) and meat samples (14.0%). However, ileoceacal lymph nodes that test positive for Salmonella are not found to be a predictor of positive test results further on in the process. Besides the slaughterhouse environment, meat handlers were identified as a possible source of subsequent contamination, with 9.3% of the sample testing positive. Diverse Salmonella enterica serotypes were detected, mainly S. Typhimurium and the monophasic variant S. 4,[5],12:i:-, but also S. Derby, S. Rissen, S. Mbandaka, S. London, S. Give, S. Enteritidis and S. Sandiego, in total corresponding to 17 PFGE types. Our results demonstrate that besides a high level of Salmonella swine contamination at pre-harvest level, slaughtering, dressing, cutting and deboning operations are contributing to the occurrence of clinically relevant clones (e.g. S. Typhimurium DT104 and S. 4,[5],12:i:-) in pork products. This study also highlights the possibility of an ongoing Salmonella community being spread by abattoir workers.

The challenge of corporatisation: the experience of Portuguese public hospitals

The inability of traditional state organisations to respond to new economic, technological and social challenges and the associated emerging problems has made it necessary to adopt new methods of health management. As a result, new directions have emerged in the reform of Public Administration together with the introduction of innovative models. The aim is to achieve a type of management that focusses on results as well as on effort and efficiency. We intend to analyse to what extent the adoption of business management models by hospital healthcare units can improve their performance, mainly in terms of standards of efficiency. Data envelopment analysis (DEA) was used to investigate the efficiency of a set of public Portuguese hospitals. The aim was to evaluate the impact of business management in Portuguese public hospitals with regards to efficiency, specifically taking into account the fact that lack of resources and increased health care needs are a present and future reality. From a total of 83 public hospitals, a sample of 59 hospitals was chosen, of which 21 are state-owned hospital enterprises (SA) and 38 are traditional public administration sector hospitals (SPA). This study evaluates hospital performance by calculating two efficiency measures associated with two categories of inputs. The first efficiency measures the costs associated with hospital production lines and the number of beds (representing fixed capacity) as inputs. The annual costs generated by the hospitals in the consumption of capital and work (direct and indirect costs) are used. A second measure of efficiency is calculated separately. This measure includes in the inputs the number of beds as well as the human resources available (number of doctors, number of nurses and other personnel) in each hospital. With regard to output, the variables that best reflect the hospital services rendered were considered: number of inpatient days, patients discharged, outpatient visits, emergencies services, sessions in hospital day care services and the number of surgeries. The results seem to suggest that the introduction of market processes and changes in organisational structure—such as managerial autonomy and corporatisation of public hospitals—have had a positive impact on Portuguese public hospitals. This positive evolution was particularly evident in SA hospitals, but further studies are needed to confirm these preliminary results.

Why Does Infection With Some Helminths Cause Cancer

Infections with Opisthorchis viverrini, Clonorchis sinensis, and Schistosoma haematobium are classified as group 1 biological carcinogens: definitive causes of cancer. These worms are metazoan eukaryotes, unlike the other group 1 carcinogens including human papilloma virus, hepatitis C virus, and Helicobacter pylori. By contrast, infections with phylogenetic relatives of these helminths, also trematodes of the phylum Platyhelminthes and major human pathogens, are not carcinogenic. These inconsistencies prompt several questions, including how might these infections cause cancer, and why is infection with only a few helminth species carcinogenic? We present an interpretation of mechanisms contributing to the carcinogenicity of these helminth infections, including roles for metabolites of catechol estrogen and oxysterols of parasite origin as initiators of carcinogenesis.

Mass spectrometry techniques in the survey of steroid metabolites as potential disease biomarkers: A review

Mass spectrometric approaches have been fundamental to the identification of metabolites associated with steroid hormones, yet this topic has not been reviewed in depth in recent years. To this end, and given the increasing relevance of liquid chromatography-mass spectrometry (LC-MS) studies on steroid hormones and their metabolites, the present review addresses this subject. This review provides a timely summary of the use of various mass spectrometry-based analytical techniques during the evaluation of steroidal biomarkers in a range of human disease settings. The sensitivity and specificity of these technologies are clearly providing valuable new insights into breast cancer and cardiovascular disease. We aim to contribute to an enhanced understanding of steroid metabolism and how it can be profiled by LC-MS techniques.

Use of GRA6-Derived Synthetic Polymorphic Peptides in an Immunoenzymatic Assay To Serotype Toxoplasma gondii in Human Serum Samples Collected from Three Continents

ABSTRACT Serotyping is a simple typing method that consists of an immunoenzymatic assay (enzyme-linked immunosorbent assay [ELISA]) using synthetic polymorphic peptides derived from Toxoplasma gondii antigens. We developed a new ELISA based on GRA6 C-terminal polymorphic peptides. Serum samples from 41 human infections due to 23 archetypal (type I, II, or III) and 18 nonarchetypal strains were selected in order to validate this approach. For 20 out of the 23 archetypal infections, there was a clear correlation between microsatellite genotype and GRA6 serotyping. All infections due to nonarchetypal strains were misclassified as archetypal strain infections. The GRA6 C-terminal peptides from these strains were analyzed to explain this misclassification. A second group of 455 patients with acute and chronic toxoplasmosis due to unknown genotypes from different European, African, and Latin American countries were included in this study, and the strain type predicted by this method. The results suggest that serotyping is a promising method for typing strains, although limitations exist for African and South American strains as a consequence of higher peptide polymorphism. Other peptides from different markers must be studied in order to discriminate archetypal from nonarchetypal strains.

Evaluation of a simple sedimentation method (modified McMaster) for diagnosis of bovine fascioliosis.

A coprological sedimentation method is evaluated for quantification of egg shedding in bovine faeces. Through the inclusion of different number of Fasciola hepatica eggs in negative faeces, egg recovery rate and the sensitivity of the method were determined. The mean egg recovery rate of the technique was 76.72+/-15.42%. The sensitivity of the method was 33.3% whenever eggs/g of faeces (EPG) are less than 1.5 and 100% for higher values. To improve the diagnostic accuracy with this technique, it is advisable to increase the sample from 10 to 30g of faeces when manipulating low egg shedding, which allowed for a sensitivity of 83.3%. Regression equations were calculated to quantify the relationship between the number of recovered and incorporated eggs.

Tumour-like phenotypes in urothelial cells after exposure to antigens from eggs of Schistosoma haematobium: an oestrogen-DNA adducts mediated pathway?

Chronic infection with the blood fluke, Schistosoma haematobium, is associated with squamous cell carcinoma of the bladder. Previously, it has been shown that soluble extracts of mixed sex adult S. haematobium worms (SWAP) are tumourigenic, both in vitro and in vivo. In addition, oestrogen-related molecules in SWAP of S. haematobium down-regulate oestrogen receptors (ERs) alpha and beta in oestrogen responsive cells. Moreover, schistosome oestrogens occur in sera of persons with schistosomiasis haematobia and repress transcription of ERs in urothelial cells. Given that eggs of S. haematobium are the developmental stage directly responsible for urogenital disease during schistosomiasis haematobia, we suspected that soluble antigens from S. haematobium eggs exhibit similar or more potent tumorigenic capacity. Here we investigated the tumorigenic potential of soluble egg antigens (Sh-SEA) of S. haematobium and the endocrine system in favouring parasitism by schistosomes. The findings confirmed that 6.25μg/ml of Sh-SEA was enough to stimulate cell proliferation, reduce apoptosis and increase oxidative stress of Sh-SEA-exposed urothelial cells. In addition, genotoxic effects of Sh-SEA on these cells were determined by using alkaline single-cell gel electrophoresis (Comet). Furthermore, Liquid Chromatography Diode Array Detection Electron Spray Ionisation Mass Spectrometry indicated the presence of catechol-oestrogens in S. haematobium SEA. A prospective oestrogen-DNA adduct mediated pathway in S. haematobium egg induced bladder cancer is also discussed.

Cryptosporidium spp. and Giardia duodenalis in two areas of Galicia (NW Spain).

The aim of the present study was to investigate the environmental dispersal of Cryptosporidium spp. and Giardia duodenalis in two distinct areas (coastal and inland) in Galicia (NW Spain). Faecal samples were collected from healthy asymptomatic domestic (cows and sheep) and wild animals (deer and wild boars) in the selected areas. In each of the selected areas, samples of untreated water (influent) and of treated water (final effluent) were collected from each of the 12 drinking water treatments plants (DWTPs) and 12 wastewater treatment plants (WTPs) under study. Analysis of a single sample from each of the 635 (coastal) and 851 (inland) domestic and wild animals selected at random revealed that the prevalences of cryptosporidiosis and giardiosis in coastal area were 9.2% and 15.9% respectively, and in inland area, 13.7% and 26.7% respectively. In the coastal area, Cryptosporidium spp. oocysts were detected in influent and effluent samples from 2/12 (16.6%) DWTPs and 8/12 (66.6%) WTPs, while G. duodenalis cysts were detected in influent and effluent samples from 3/12 (25.0%) DWTPs and 12/12 (100%) WTPs. The concentrations were notably higher in WTPs; the mean parasite concentrations in the final treated effluent were 10 oocysts per litre and 137.8 cysts per litre for Cryptosporidium and Giardia, respectively. The mean concentration of G. duodenalis cysts per litre was significantly higher (P<0.05) than the mean concentration of Cryptosporidium spp. oocysts per litre in both the influent and the effluent samples from all the treatment plants. In the coastal area, C. parvum, C. hominis and G. duodenalis assemblages A (I and II) and E were most repeatedly detected. In the inland area, C. parvum, C. andersoni and G. duodenalis assemblages A (I and II), B and E were most frequently identified.

Schistosoma haematobium and bladder cancer: what lies beneath?

Schistosoma haematobium is a parasitic flatworm that infects millions of people, mostly in the developing world, and is associated with high incidence of bladder cancer although why is not clear. But our group was able to define the mechanistic relationship for the first time between infection of S. haematobium and cancer. We used in vitro models to demonstrate the presence of informative carcinogenesis-associated phenotypes in CHO cells exposed to Sh total antigen, in which we showed increased cell proliferation, decreased apoptosis, up regulation of the anti-apoptotic molecule Bcl-2, down regulation of the tumor suppressor protein p27, and increased cell migration and invasion. We further discuss the molecular and cellular events that might be responsible for schistosomiasis-related bladder cancer.