Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

How much cultural events are "worth" for host communities: Case of Faz Música Lisboa

This paper studies the economic and social impact of Faz Música Lisboa on the segment of society that feels from any direct or indirect consequence due to its existence. A qualitative research based on surveys and interviews is made to retract a list of the benefits and costs that each stakeholder perceives. Relying on the quantifiable variables, it is performed a cost-benefit analysis to measure how much the event is “worth” for the community. I conclude this is a viable project, as it brings a positive net benefit to the society, value that could increase with a higher institutional support.

Sweetening abroad

This project aims to plan the internationalization strategy of the Portuguese small firm A Tarte, producer of a gourmet pie, to a European country in order to enlarge its sales. Therefore, a thorough analysis of the company and of the bakery market is performed to understand the complete environment. It is followed by a quantitative analysis of fourteen countries to select the most attractive market, whose results indicated United Kingdom as the best option. After, the direct exporting entry mode was analysed and costs estimated. Lastly, the expansion to London is recommended to be pursued with a UK local agent/distributor support

Competition policy costs to firms

The existence of competition policy forces companies to adjust their behaviour. This is also costly. Using a database from a company on contracts, I will try to estimate if a specific competition policy disposition, supply contracts cannot be longer than 60 months, has costs for the coffee suppliers operating in the Portuguese “on-trade” coffee market. The estimation method used in this paper will be OLS. The results suggest that limiting the duration of exclusivity contracts to 60 months can be harmful to the coffee suppliers and it can even seriously affect the market functioning. Key

Energy efficiency in energy policy: Empirical evidence of direct rebound effect in Portuguese household's demand for electricity

This project the direct rebound effect for the electricity demand in Portugal. While we find evidence of such an effect, the estimations also reflect the institutional arrangement that has characterized the electricity market in the country. Also, issues related to energy efficiency promotion are addressed in general putting into context the case study developed.

Does direct cash flow presentation help in predicting future operating cash flow?

Research literature and regulators are unconditional in pointing the disclosure of operating cash flow through direct method a section of unique information. Besides the intuitive facet, it is also consistent in forecasting future operating cash flows and a cohesive piece to financial statement puzzle. Bearing this in mind, I produce an analysis on the usefulness and predictive ability on the disclosure of gross cash receipts and payments over the disclosure of reconciliation between net income and accruals for two markets with special features, Portugal and Spain. Results validate the usefulness of direct method format in predicting future operating cash flow. Key

Comparison of recombinant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in Northwestern Iran

INTRODUCTION: Various methods are used for the diagnosis of visceral leishmaniasis (VL), such as microscopic examination, culture and inoculation of laboratory animals; however, serological assays are commonly used for the detection of antibodies in serum samples with a wide range of specificity and sensitivity. METHODS: The purpose of this study was to compare three serological methods, including rA2-ELISA, the recombinant KE16 (rKE16) dipstick test and the direct agglutination test (DAT), for the detection of antibodies against VL antigens. The assays utilized 350 statistically based random serum samples from domestic dogs with clinical symptoms as well as samples from asymptomatic and healthy dogs from rural and urban areas of the Meshkinshahr district, northwestern Iran. RESULTS: Samples were assessed, and the following positive rates were obtained: 11.5% by rKE16, 26.9% by DAT and 49.8% by ELISA. The sensitivity among symptomatic dogs was 32.4% with rKE16, 100% with DAT and 52.9% with ELISA. Conversely, rA2-ELISA was less specific for asymptomatic dogs, at 46.5%, compared with DAT, at 88.9%. CONCLUSIONS : This study recommends rA2-ELISA as a parallel assay combined with DAT to detect VL infection among dogs. Further evaluations should be performed to develop an inexpensive and reliable serologic test for the detection of Leishmania infantum among infected dogs.

Performance of direct immunofluorescence assay for the detection of human metapneumovirus under clinical laboratory settings

ABSTRACT INTRODUCTION: Human metapneumovirus (hMPV) is an emergent human respiratory pathogen. This study aimed to evaluate the performance of direct immunofluorescence (DIF) to detect hMPV in a clinical laboratory setting. METHODS: Nasopharyngeal aspirate samples (448) of children and adults with respiratory illness were used to detect hMPV by using DIF and real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays. RESULTS: In all, 36 (8%) samples were positive by DIF and 94 (21%) were positive by qRT-PCR. Direct immunofluorescence specificity was 99% and sensitivity was 38%. CONCLUSIONS: DIF is not very sensitive under clinical laboratory settings.

Cost of drugs manufactured by the University Hospital - role of the Central Pharmacy

The hospital pharmacy in large and advanced institutions has evolved from a simple storage and distribution unit into a highly specialized manipulation and dispensation center, responsible for the handling of hundreds of clinical requests, many of them unique and not obtainable from commercial companies. It was therefore quite natural that in many environments, a manufacturing service was gradually established, to cater to both conventional and extraordinary demands of the medical staff. That was the case of Hospital das Clinicas, where multiple categories of drugs are routinely produced inside the pharmacy. However, cost-containment imperatives dictate that such activities be reassessed in the light of their efficiency and essentiality. METHODS: In a prospective study, the output of the Manufacturing Service of the Central Pharmacy during a 12-month period was documented and classified into three types. Group I comprised drugs similar to commercially distributed products, Group II included exclusive formulations for routine consumption, and Group III dealt with special demands related to clinical investigations. RESULTS: Findings for the three categories indicated that these groups represented 34.4%, 45.3%, and 20.3% of total manufacture orders, respectively. Costs of production were assessed and compared with market prices for Group 1 preparations, indicating savings of 63.5%. When applied to the other groups, for which direct equivalent in market value did not exist, these results would suggest total yearly savings of over 5 100 000 US dollars. Even considering that these calculations leave out many components of cost, notably those concerning marketing and distribution, it might still be concluded that at least part of the savings achieved were real. CONCLUSIONS: The observed savings, allied with the convenience and reliability with which the Central Pharmacy performed its obligations, support the contention that internal manufacture of pharmaceutical formulations was a cost-effective alternative in the described setting.

Healthcare- Associated infections: An analysis of costs in a Portuguese Hospital

Hospital-acquired infections (HAIs) delay healing, prolong Hospital stay, and increase both Hospital costs and risk of death. This study aims to estimate the extra length of stay and mortality rate attributable to each of the following HAIs: wound infection (WI); bloodstream infection (BSI); urinary infections (UI); and Hospital-acquired pneumonia (HAP). The study population consisted of patients discharged in CHLC in 2014. Data was collected to identify demographic information, surgical operations, development of HAIs and its outputs. The study used regressions and a matched strategy to compare cases (infected) and controls (uninfected). The matching criteria were: age, sex, week and type of admission, number of admissions, major diagnostic category and type of discharge. When compared to matched controls, cases with HAI had a higher mortality rate and greater length of stay. WI related to hip or knee surgery, increased mortality rate by 27.27% and the length of stay by 74.97 days. WI due to colorectal surgery caused an extra mortality rate of 10.69% and an excess length of stay of 20.23 days. BSI increased Hospital stay by 28.80 days and mortality rate by 32.27%. UI caused an average additional length of stay of 19.66 days and risk of death of 12.85%. HAP resulted in an extra Hospital stay of 25.06 days and mortality rate of 24.71%. This study confirms the results of the previous literature that patients experiencing HAIs incur in an excess of mortality rates and Hospital stay, and, overall, it presents worse results comparing with other countries.

Quantitative analysis of collagen and elastic fibers in the transversalis fascia in direct and indirect inguinal hernia

PURPOSE: Our previous studies demonstrated structural and quantitative age-related changes of the elastic fibers in transversalis fascia, which may play a role in inguinal hernia formation. To verify whether there were differences in the extracellular matrix between direct and indirect inguinal hernia, we studied the amount of collagen and elastic fibers in the transversalis fascia of 36 male patients with indirect inguinal hernia and 21 with direct inguinal hernia. MATERIAL AND METHODS: Transversalis fascia fragments were obtained during surgical intervention and underwent histological quantitative analysis of collagen by colorimetry and analysis of elastic fibers by histomorphometry. RESULTS: We demonstrated significantly lower amounts of collagen and higher amounts of elastic fibers in transversalis fascia from patients with direct inguinal hernia compared to indirect inguinal hernia patients. The transversalis fascia from direct inguinal hernia patients showed structural changes of the mature and elaunin elastic fibers, which are responsible for elasticity, and lower density of oxytalan elastic fibers, which are responsible for resistance. These changes promoted loss of resiliency of the transversalis fascia. CONCLUSION: These results improve our understanding of the participation of the extracellular matrix in the genesis of direct inguinal hernia, suggesting a relationship with genetic defects of the elastic fiber and collagen synthesis.

Custos indirectos associados à obesidade em Portugal

RESUMO - A obesidade constitui um importante problema de saúde pública com consequências económicas de grande dimensão. Os obesos têm um risco acrescido de contrair doenças e de sofrer morte prematura devido a problemas como a diabetes, hipertensão arterial, AVC, insuficiência cardíaca e algumas neoplasias malignas. O presente estudo tem como objectivo estimar o custo económico indirecto (valor da produção perdida) associado à obesidade em Portugal no ano de 2002. O estudo adopta uma abordagem tipo custos da doença baseada na prevalência. Os dados são retirados do Inquérito Nacional de Saúde e estatísticas de rotina publicadas pelo INE e por outros organismos oficiais. Consideram-se como obesas pessoas com índice de massa corporal (IMC) ≥ 30 kg/m2 e estabelecem-se como limites etários para participação em actividades económicas produtivas as idades compreendidas entre os 15 e os 64 anos. A estratégia de imputação de custos ao factor de risco obesidade caracteriza- se por estimar, para a população portuguesa, as proporções de doença e morte prematura atribuíveis à obesidade e em multiplicar as estimativas populacionais encontradas pelo valor da produtividade económica potencial das pessoas afectadas. O custo indirecto total da obesidade em Portugal no ano de 2002 foi estimado em 199,8 milhões de euros. A mortalidade contribuiu com 58,4% deste valor (117 milhões de euros) e a morbilidade com 41,6% (83 milhões de euros). Os custos da morbilidade advêm de mais de 1,6 milhões de dias de incapacidade anuais, principalmente por faltas ao trabalho associadas a doenças do sistema circulatório e diabetes tipo II. Os custos da mortalidade são o resultado de 18 733 potenciais anos de vida activa perdidos, numa razão de 3 mortes masculinas por cada morte feminina. Os resultados indicam que a obesidade acarreta consideráveis perdas económicas para o país. Comparando os resultados com um estudo complementar que calculou os custos directos (em cuidados de saúde) da obesidade, verifica-se que a componente indirecta representa 40,2% do total dos custos da obesidade. A implementação de estratégias que prevenissem ou reduzissem a incidência e prevalência de obesidade em Portugal poderia gerar ganhos de produtividade elevados. Para conhecer a dimensão destes ganhos é necessária mais investigação sobre os benefícios clínicos e relação custo-efectividade de estratégias para a redução da obesidade.

Polyhydroxyalkanoates production by photosynthetic mixed cultures

Polyhydroxyalkanoates (PHAs) are natural biologically synthesized polymers that have been the subject of much interest in the last decades due to their biodegradability. Thus far, its microbial production is associated with high operational costs, which increases PHA prices and limits its marketability. To address this situation, this thesis’ work proposes the utilization of photosynthetic mixed cultures (PMC) as a new PHA production system that may lead to a reduction in operational costs. In fact, the operational strategies developed in this work led to the selection of PHA accumulating PMCs that, unlike the traditional mixed microbial cultures, do not require aeration, thus permitting savings in this significant operational cost. In particular, the first PHA accumulating PMC tested in this work was selected under non-aerated illuminated conditions in a feast and famine regime, being obtained a consortium of bacteria and algae, where photosynthetic bacteria accumulated PHA during the feast phase and consumed it for growth during the famine phase, using the oxygen produced by algae. In this symbiotic system, a maximum PHA content of 20% cell dry weight (cdw) was reached, proving for the first time, the capacity of a PMC to accumulate PHA. During adaptation to dark/light alternating conditions, the culture decreased its algae content but maintained its viability, achieving a PHA content of 30% cdw. Also, the PMC was found to be able to utilize different volatile fatty acids for PHA production, accumulating up to 20% cdw of a PHA co-polymer composed of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (HV) monomers. Finally, a new selective approach for the enrichment of PMCs in PHA accumulating bacteria was tested. Instead of imposing a feast and famine regime, a permanent feast regime was used, thus selecting a PMC that was capable of simultaneously growing and accumulating PHA, being attained a maximum PHA content of 60% cdw, the highest value reported for a PMC thus far. The results presented in this thesis prospect the utilization of cheap, VFA-rich fermented wastes as substrates for PHA production, which combined with this new photosynthetic technology opens up the possibility for direct sunlight illumination, leading to a more cost-effective and environmentally sustainable PHA production process.

Marketing plan for online aqctivity of Rota Vicentina boosting direct website sales

The nature tourism experienced a great expansion of its market with the appearance of different lifestyles. In this Work Project a study regarding the website direct sales of Rota Vicentina was developed. Its website shows the idea of being solely an information structure and not a purchase one, leading to a current absence of online sales. Hence, it is suggested the modification of its business model, using different instruments and channels. Some digital marketing recommendations were developed in order to boost website sales, such as a platform for online reviews, remarketing campaigns and social media activity.

Firing and training costs and labour market segmentation

This dissertation consists of three essays on the labour market impact of firing and training costs. The modelling framework resorts to the search and matching literature. The first chapter introduces firing costs, both liner and non-linear, in a new Keynesian model, analysing business cycle effects for different wage rigidity degrees. The second chapter adds training costs in a model of a segmented labour market, accessing the interaction between these two features and the skill composition of the labour force. Finally, the third chapter analyses empirically some of the issues raised in the second chapter.